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Umbilical hernia (UH) and inguinal hernia (IH) are among the most common defects in pigs, affecting their welfare and resulting in economic losses. In this study, we aimed to verify the association of previously reported differences in transcript levels of the ACAN , COL6A5 , MMP13 , and VIT genes with the occurrence of UH and IH. We examined mRNA levels in muscle and connective tissue from 68 animals-34 affected by UH and 34 controls. In a second cohort, we examined inguinal channel samples from 46 pigs (in four groups). We determined DNA methylation levels in muscle tissue for the UH and control animals. The transcript level of MMP13 changed in the UH cases, being upregulated and downregulated in muscle and connective tissue, respectively, and the VIT gene also showed an increased muscular mRNA level. The transcript of the ACAN gene significantly decreased in old pigs with IH. We further observed an increased DNA methylation level for one CpG site within the MMP13 gene in UH individuals. We conclude that these alterations in gene mRNA levels in the UH animals depend on the tissue and can sometimes be a consequence of, not a cause of, the affected phenotype.

Catch-and-release (C&R) angling is a conservation-oriented practice intended to reduce the impact recreational angling has on fish populations. Even though most recreationally angled fish are released, little is known about how C&R angling impacts fish at the cellular or tissue level. As the first to explore the impacts of C&R angling on mRNA abundances, our study aimed to identify how the stress of angling influenced metabolism, acid-base regulation and cellular stress in the gills of lake trout ( Salvelinus namaycush) . Because gills are responsible for metabolic gas exchange, are crucial sites of acid-base homeostasis and respond to stressors quickly, we hypothesized that the relative mRNA abundance of genes related to these three physiological processes would be altered after angling. We took gill samples of live lake trout at 0, 2 or 48 h after fish were angled by rod and reel, and then used quantitative PCR (qPCR) to measure the relative abundance of nine candidate mRNA transcripts. Heat shock protein 70 ( hsp70 ) mRNA levels significantly increased over 5-fold 2 h after angling, indicating a potential activation of a cytoprotective response. However, contrary to our hypothesis, we observed no change in the relative mRNA abundance of genes related to metabolism or acid-base regulation in response to C&R angling within a 48-h period. As C&R angling can negatively impact fish populations, further use of transcript-level studies will allow us to understand the impact C&R has on specific tissues and improve our knowledge of how C&R influences overall fish health., (© The Author(s) 2023. Published by Oxford University Press and the Society for Experimental Biology.)

Smoking has been linked to male infertility by affecting the sperm epigenome and genome. In this study, we aimed to determine possible changes in the transcript levels of PGAM5 (the phosphoglycerate mutase family member 5), PTPRN2 (protein tyrosine phosphatase, N2-type receptor), and TYRO3 (tyrosine protein kinase receptor) in heavy smokers compared to non-smokers, and to investigate their association with the fundamental sperm parameters. In total, 118 sperm samples (63 heavy-smokers (G1) and 55 non-smokers (G2)) were included in this study. A semen analysis was performed according to the WHO guidelines. After a total RNA extraction, RT-PCR was used to quantify the transcript levels of the studied genes. In G1, a significant decrease in the standard semen parameters in comparison to the non-smokers was shown ( p < 0.05). Moreover, PGAM5 and PTPRN2 were differentially expressed ( p ≤ 0.03 and p ≤ 0.01, respectively) and downregulated in the spermatozoa of G1 compared to G2. In contrast, no difference was observed for TYRO3 ( p ≤ 0.3). In G1, the mRNA expression level of the studied genes was correlated negatively with motility, sperm count, normal form, vitality, and sperm membrane integrity ( p < 0.05). Therefore, smoking may affect gene expression and male fertility by altering the DNA methylation patterns in the genes associated with fertility and sperm quality, including PGAM5 , PTPRN2 , and TYRO3 .

This study looked at genetic polymorphisms and transcript levels of immune, antioxidant, and erythritol-related markers for postparturient endometritis prediction and tracking in Holstein dairy cows. One hundred and thirty female dairy cows (65 endometritis affected and 65 apparently healthy) were used. Nucleotide sequence variations between healthy and endometritis-affected cows were revealed using PCR-DNA sequencing for immune ( TLR4 , TLR7 , TNF-α , IL10 , NCF4 , and LITAF ), antioxidant ( ATOX1 , GST , and OXSR1 ), and erythritol-related ( TKT , RPIA , and AMPD1 ) genes. Chi-square investigation exposed a noteworthy variance amongst cow groups with and without endometritis in likelihood of dispersal of all distinguished nucleotide variants ( p < 0.05). The IL10 , ATOX1 , and GST genes were expressed at substantially lower levels in endometritis-affected cows. Gene expression levels were considerably higher in endometritis-affected cows than in resistant ones for the genes TLR4 , TLR7 , TNF-α , NCF4 , LITAF , OXSR1 , TKT , RPIA , and AMPD1 . The sort of marker and vulnerability or resistance to endometritis had a significant impact on the transcript levels of the studied indicators. The outcomes might confirm the importance of nucleotide variants along with gene expression patterns as markers of postparturient endometritis susceptibility/resistance and provide a workable control plan for Holstein dairy cows.

Leptographium qinlingensis is one of the major fungal associates of the Chinese white pine beetle (Dendroctonus armandi) and is an active participant in the large-scale death of Pinus armandi. Beetles and associated fungi have evolved efficient systems for overcoming the toxicity of host defense chemicals, which consist of a multitude of monoterpenes and diterpenes. As fungal cytochromes (CYPs) can detoxify and degrade various xenobiotic compounds, we identified 11 CYPs from L. qinlingensis to analyze their potential function in detoxifying or degrading host chemical defense. These 11 CYPs of L. qinlingensis belong to 6 clans and cluster into 3 clades, clade 2, clade 8 and clade 10, based on their phylogenetic relationships. Moreover, we also analyzed the transcript levels of CYPs following treatment with terpenes or expression in mycelia grown on terpenoids as a carbon source. The results in this paper showed that several CYPs were upregulated after treatment with terpenes or growth on terpenoids as the only carbon source. Our research provides some insight into the function of CYPs from bark beetle symbiotic fungi in the detoxification of pine defense compounds or their relationships with the utilization of terpenoids., Competing Interests: Declaration of competing interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

The flavonoid metabolites were compared between red 'Summer Black' (SB) and white 'Shine Muscat' (SM) table grapes during fruit development based on widely targeted metabolome. A total of 134 flavonoids were identified in two cultivars, including 37 flavones, 33 flavonols, and 11 anthocyanidins, and so on. From young to veraison, the composition and the content of most flavonoids were decreasing in both cultivars but increased at maturation in SB. In general, SB has higher flavonoid compositions and content than SM during the whole fruit development, especially the content of anthocyanin after veraison. While the SM had higher content of flavonols such as quercetin, kaempferol and their derivatives. The expression of anthocyanin-related genes such as UFGT, OMT, GST, MATE, MYBA1, and MYBA2 was remarkably higher in SB than those in SM, which may attribute to higher anthocyanin content, while the higher expression of F3H and FLS resulted higher level of flavonols in SM. These results improve our understanding of flavonoid profiles and molecular mechanism in table grape cultivars., (© 2022 Institute of Food Technologists®.)

The F-box domain is a conserved structural protein motif that most frequently interacts with the SKP1 protein, the core of the SCFs (SKP1-CULLIN-F-box protein ligase) E3 ubiquitin protein ligases. As part of the SCF complexes, the various F-box proteins recruit substrates for degradation through ubiquitination. In this study, we functionally characterized an F-box gene (MtF-box) identified earlier in a population of Tnt1 retrotransposon-tagged mutants of Medicago truncatula and its Arabidopsis thaliana homolog (AtF-box) using gain- and loss-of-function plants. We highlighted the importance of MtF-box in leaf development of M. truncatula. Protein-protein interaction analyses revealed the 2-isopropylmalate synthase (IPMS) protein as a common interactor partner of MtF-box and AtF-box, being a key enzyme in the biosynthesis pathway of the branched-chain amino acid leucine. For further detailed analysis, we focused on AtF-box and its role during the cell division cycle. Based on this work, we suggest a mechanism for the role of the studied F-box gene in regulation of leucine homeostasis, which is important for growth., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Objectives: Ras-related dexamethasone-induced 1 ( RASD1 ) is abnormally expressed in many solid cancers. However, its potential role in adults with B-cell acute lymphoblastic leukemia (B-ALL) is unclear. Therefore, we aim to clarify the abnormal expression of the tumor-associated biomarker, RASD1 , as a potential target for diagnosis and prognosis in adult Philadelphia-negative B-ALL., Methods: The expression of RASD1 was detected with RT-qPCR in 92 adults with de novo Ph-negative B-ALL and 40 healthy controls. The correlation between RASD1 transcript levels and relapse was assessed., Results: RASD1 transcript levels in patients with Ph-negative B-ALL (median 81.76%, range 0.22%-1824.52%) were significantly higher than those in healthy controls (7.59%, 0.46%-38.66%; P <0.0001). Patients with low RASD1 transcript levels had a lower 5-year relapse-free survival (RFS, 47.5% [32.9%, 62.1%] vs. 63.1% [49.0%, 77.2%]; P  = 0.012) and a higher 5-year cumulative incidence of relapse (CIR, 52.0% [37.4%, 66.6%] vs. 36.2% [22.2%, 50.2%]; P  = 0.013) especially in patients receiving chemotherapy only. Multivariate analysis showed that a low RASD1 transcript level was an independent risk factor for RFS (HR = 2.938 [1.427, 6.047], P  = 0.003) and CIR (HR = 3.367 [1.668, 6.796], P  = 0.001) in patients with Ph-negative B-ALL., Conclusions: RASD1 transcript levels were significantly higher in patients with Ph-negative B-ALL and a low RASD1 transcript level was independently correlated with increased relapse risk.

The tolerance mechanism of chemical pesticide is necessary to combat the pest infestation challenges. This study intended to analyze the responses of enzymatic activity and expression level of an antioxidant gene to organophosphate pesticide stress. The alteration of anti-oxidative correlated with pesticide treatment in eggplant ( S. melongena L.cv. Longai) using varying concentrations (0, 50, 100, 150 and 200 ppm) of malathion (PM) and tatafen (PTF) each. The enzyme activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were observed to be elevated with pesticide treatment in eggplant seedling. FeSOD (iron SOD) , CAT and APX genes associated in defense mechanisms were significantly expressed under PM and PTF stress which contributed to stress tolerance to the plant. The different concentration of both pesticide stresses altered the expression level of mRNA, FeSOD , CAT and APX genes in comparison to the non-treated plant. While mRNA level of three antioxidant genes were evaluated and found to be APX gene expression was more potent than the CAT and FeSOD gene subjected to different concentrations of PM and PTF in eggplant. The current experiment highlights the presence of minimum level of pesticide concentration impacted positively towards the plant growth and metabolism, while high level of pesticide concentration impacted negatively. In summary, antioxidant enzymes activity responded to both pesticide stresses at an early stage of exposure and their gene expression profiles provided more details about their complex interaction and effectively scavenge reactive oxygen species. This allows the plant to maintain growth under pesticide stress., Competing Interests: Conflict of interestOn behalf of all authors, the corresponding author states that there is no conflict of interest., (© King Abdulaziz City for Science and Technology 2021.)

Seagrass ecosystems are among the most productive marine ecosystems, and diazotrophic communities play a crucial role in sustaining the productivity and stability of such ecosystems by introducing fixed nitrogen. However, information concerning both total and active diazotrophic groups existing in different compartments of seagrass is lacking. This study comprehensively investigated the diversity, structure, and abundance of diazotrophic communities in different parts of the seagrass Halophila ovalis at the DNA and RNA level from clone libraries and real-time quantitative PCR. Our results indicated that nearly one-third of existing nitrogen-fixing bacteria were active, and their abundance might be controlled by nitrogen to phosphorus ratio (N:P). Deltaproteobacteria and Gammaproteobacteria were dominant groups among the total and active diazotrophic communities in all samples. These two groups accounted for 82.21% and 70.96% at the DNA and RNA levels, respectively. The genus Pseudomonas and sulfate-reducing bacteria (genera: Desulfosarcina, Desulfobulbus, Desulfocapsa, and Desulfopila) constituted the significant fraction of nitrogen-fixing bacteria in the seagrass ecosystem, playing an additional role in denitrification and sulfate reduction, respectively. Moreover, the abundance of the nitrogenase gene, nifH, was highest in seawater and lowest in rhizosphere sediments from all samples. This study highlighted the role of diazotropic communities in the subtropical seagrass ecosystem., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Efficiency is the basis for the application of RNA interference (RNAi) technology. Actually, RNAi efficiency varies greatly among insect species, tissues and genes. Previous efforts have revealed the mechanisms for variation among insect species and tissues. Here, we investigated the reason for variable efficiency among the target genes in the same insect. First, we tested the genes sampled randomly from Tribolium castaneum, Locusta migratoria and Drosophila S2 cells for both their expression levels and sensitivity to RNAi. The results indicated that the genes with higher expression levels were more sensitive to RNAi. Statistical analysis showed that the correlation coefficients between transcript levels and knockdown efficiencies were 0.8036 (n = 90), 0.7255 (n = 18) and 0.9505 (n = 13), respectively in T. castaneum, L. migratoria and Drosophila S2 cells. Subsequently, ten genes with varied expression level in different tissues (midgut and carcass without midgut) of T. castaneum were tested. The results indicated that the higher knockdown efficiency was always obtained in the tissue where the target gene expressed higher. In addition, three genes were tested in different developmental stages, larvae and pupae of T. castaneum. The results found that when the expression level increased after insect pupation, these genes became more sensitive to RNAi. Thus, all the proofs support unanimously that transcript level is a key factor affecting RNAi sensitivity. This finding allows for a better understanding of the RNAi efficiency variation and lead to effective or efficient use of RNAi technology., (Copyright © 2021. Published by Elsevier Inc.)

Fall armyworm (FAW), Spodoptera frugiperda a recent invasive pest in India is reported to cause significant damage by feeding voraciously on maize and other economic crops from tropical to temperate provinces. It is becoming an arduous challenge to control the pest as it can survive in a wide range of temperature conditions and is already said to develop resistance towards certain insecticides. The small Heat shock proteins (hereafter, sHsps) are known to play an important role in adaptation of insects under such stress conditions. Our present study involved characterization of the three sHsps genes (sHsp19.74, sHsp20.7 and sHsp19.07) which encoded proteins of about 175, 176 and 165 amino acids with a conserved α-crystalline domain. Phylogenetic analysis of deduced amino acid sequences of the three genes showed strong similarity with the other lepidopteran sHsps. The effect of different growth stages on the expression profile of these stress proteins has also been studied and the Quantitative real time PCR (qRT-PCR) analysis revealed that the transcript level of sHsp19.07 and sHsp20.7 were significantly upregulated under extreme heat (44 °C) and cold (5 °C) stress. However, sHsp19.74 responded only to heat treatment but not to the cold treatment. In addition, the expression profile of all three sHsps was significantly lower in the larval stage (5 th instar). Chlorantraniliprole treatment resulted in maximum expression of sHsp19.07 and sHsp20.7 after 12hr of exposure to the insecticide. Meanwhile, the same expression was observed after 8hr of exposure in case of sHsp19.74. These results proved that the sHsp genes of S. frugiperda were induced and modulated in response to abiotic stress, thus influencing the physiological function leading to survival of FAW in diversified climate in India., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Published by Elsevier Ltd.)

The aim of this current study was to investigate the influence of tobacco smoke on sperm quality determined by standard parameters, on sperm DNA maturity tested by chromomycin A3 (CMA3) staining, on sperm DNA fragmentation tested by TUNEL assay and on the transcript level of sperm nuclear proteins H2BFWT, PRM1, PRM2, TNP1 and TNP2 genes quantified by RT-PCR. One hundred forty-one (141) sperm samples (43 nonsmokers (G.1) and 98 heavy smokers (G.2)) of couples undergoing ICSI were enrolled in this study. In G2, a significant decrease in standard semen parameters in comparison with nonsmokers was shown (p < .01). In contrast, protamine deficiency (CMA3 positivity) and sperm DNA fragmentation (sDF) were significantly higher in G2 than in G1 (p < .01). Furthermore, the studied genes were differentially expressed (p < .01), down-regulated in the spermatozoa of G.2 compared to that of G.1 (fold change <0.5) and were significantly correlated between each other (p < .01). Moreover, in comparison with G1, the protamine mRNA ratio in G2 was significantly higher (p < .01). It can therefore be concluded that smoking alters mRNA expression levels of H2BFWT, TNP1, TNP2, PRM1 and PRM2 genes and the protamine mRNA ratio and consequently alters normal sperm function., (© 2021 The Authors. Andrologia published by Wiley-VCH GmbH.)

Testosterone can induce impulsivity, a behavioral impairment associated with various psychiatric illnesses. The molecular mechanisms associated with testosterone-induced impulsivity are unclear. Our earlier studies showed that supraphysiological doses of testosterone to rats induced impulsive behavior, impacted hypothalamic-pituitary-adrenal axis (HPA) and hypothalamic-pituitary-gonadal axis interactions, and altered α 2A adrenergic receptors in prefrontal cortex (PFC). Owing to the importance of GABAergic system in impulsivity and memory, the present study examines whether testosterone-mediated impulsivity is associated with changes in the expression of Gamma-Aminobutyric Acid (GABA) A and B receptor subunit transcripts ( Gabra1, Gabra2, Gabra2 transcript variant 2, Gabra3, Gabra4, Gabra5, Gabra6, Gabrb1, Gabrb2, Gabrb3, Gabrg1, Gabrg2, Gabrg3, Gabbr1, Gabbr2 ) in rat PFC, and whether testosterone influences GABA A receptor subunit organization. We studied GABA receptor functions by examining GABA receptor-mediated calcium/calmodulin-dependent kinase signaling genes ( Calm1, Calm2, Calm3, Camk2a, Camk2b, Camk2g, Camk2d, Camk4 ) in the testosterone-induced impulsivity model. Rats were left untreated as controls (C), gonadectomized (GDX), or GDX and injected with supraphysiological doses of testosterone (T). Impulsive behavior was examined using the go/no-go paradigm. Gene expression was studied using qRT-PCR and GABA A subunit reorganization using cross correlation. Our findings show that expressions of select GABA A receptor subunits ( Gabra3, Gabra5, Gabra6 ) were significantly upregulated in PFC of T group compared to GDX or C groups. GABA A receptor subunit organization was different in C, T, and GDX groups. Additionally, Camk4 expression was significantly downregulated in T compared to C group. Our findings suggest that specific GABA A receptor subunit expression, their reorganization, and Camk4 -mediated functions may be associated with testosterone-mediated impulsivity., (Copyright © 2020 Agrawal and Dwivedi.)

Aerobic CH 4 oxidation coupled to denitrification (AME-D) can not only mitigate the emission of greenhouse gas (e.g., CH 4 ) to the atmosphere, but also reduce NO 3 - and/or NO 2 - and alleviate nitrogen pollution. The effects of O 2 tension on the community and functional gene expression of methanotrophs and denitrifiers were investigated in this study. Although higher CH 4 oxidation occurred in the AME-D system with an initial O 2 concentration of 21% (i.e., the O 2 -sufficient condition), more NO 3 - -N was removed at the initial O 2 concentration of 10% (i.e., the O 2 -limited environment). Type I methanotrophs, including Methylocaldum, Methylobacter, Methylococcus, Methylomonas, and Methylomicrobium, and type II methanotrophs, including Methylocystis and Methylosinus, dominated in the AME-D systems. Compared with type II methanotrophs, type I methanotrophs were more abundant in the AME-D systems. Proteobacteria and Actinobacteria were the main denitrifiers in the AME-D systems, and their compositions varied with the O 2 tension. Quantitative PCR of the pmoA, nirS, and 16S rRNA genes showed that methanotrophs and denitrifiers were the main microorganisms in the AME-D systems, accounting for 46.4% and 24.1% in the O 2 -limited environment, respectively. However, the relative transcripts of the functional genes including pmoA, mmoX, nirK, nirS, and norZ were all less than 1%, especially the functional genes involved in denitrification under the O 2 -sufficient condition, likely due to the majority of the denitrifiers being dormant or even nonviable. These findings indicated that an optimal O 2 concentration should be used to optimize the activity and functional gene expression of aerobic methanotrophs and denitrifiers in AME-D systems.

Background: Studies have indicated that graphene oxide (GO) could regulated Brassica napus L. root growth via abscisic acid (ABA) and indole-3-acetic acid (IAA). To study the mechanism and interaction between GO and IAA further, B. napus L (Zhongshuang No. 9) seedlings were treated with GO and IAA accordance with a two factor completely randomized design., Results: GO and IAA cotreatment significantly regulated the root length, number of adventitious roots, and contents of IAA, cytokinin (CTK) and ABA. Treatment with 25 mg/L GO alone or IAA (> 0.5 mg/L) inhibited root development. IAA cotreatment enhanced the inhibitory role of GO, and the inhibition was strengthened with increased in IAA concentration. GO treatments caused oxidative stress in the plants. The ABA and CTK contents decreased; however, the IAA and gibberellin (GA) contents first increased but then decreased with increasing IAA concentration when IAA was combined with GO compared with GO alone. The 9-cis-epoxycarotenoid dioxygenase (NCED) transcript level strongly increased when the plants were treated with GO. However, the NCED transcript level and ABA concentration gradually decreased with increasing IAA concentration under GO and IAA cotreatment. GO treatments decreased the transcript abundance of steroid 5-alpha-reductase (DET2) and isochorismate synthase 1 (ICS), which are associated with brassinolide (BR) and salicylic acid (SA) biosynthesis, but increased the transcript abundance of brassinosteroid insensitive 1-associated receptor kinase 1 (BAK1), cam-binding protein 60-like G (CBP60) and calmodulin binding protein-like protein 1, which are associated with BR and SA biosynthesis. Last, GO treatment increased the transcript abundance of 1-aminocyclopropane-1-carboxylic acid synthase 2 (ACS2), which is associated with the ethylene (ETH) pathway., Conclusions: Treatment with 25 mg/L GO or IAA (> 0.5 mg/L) inhibited root development. However, IAA and GO cotreatment enhanced the inhibitory role of GO, and this inhibition was strengthened with increased IAA concentration. IAA is a key factor in the response of B. napus L to GO and the responses of B. napus to GO and IAA cotreatment involved in multiple pathways, including those involving ABA, IAA, GA, CTK, BR, SA. Specifically, GO and IAA cotreatment affected the GA content in the modulation of B. napus root growth.

The aim of this study was to determine the influence of two types of UV-A LEDs on the growth and accumulation of phytochemicals in kale ( Brassica oleracea var. acephala ). Fourteen-day-old kale seedlings were transferred to a growth chamber and cultivated for 3 weeks. The kale plants were subsequently subjected to two types of UV-A LEDs (370 and 385 nm) of 30 W/m 2 for 5 days. Growth characteristics were all significantly increased in plants exposed to UV-A LEDs, especially at the 385 nm level, for which dry weight of shoots and roots were significantly increased by 2.22 and 2.5 times, respectively, at 5 days of treatment. Maximum quantum efficiency of photosystem II photochemistry (Fv/Fm ratio) began to decrease after 3 h of treatment compared to the control. The total phenolic content of plants exposed to the two types of UV-A LEDs increased by 25% at 370 nm and 42% at 385 nm at 5 days of treatment, and antioxidant capacity also increased. The two types of UV-A LEDs also induced increasing contents of caffeic acid, ferulic acid, and kaempferol. The reactive oxygen species (ROS) temporarily increased in plants exposed to the two types of UV-A LEDs after 3 h of treatment. Moreover, transcript levels of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), and flavanone 3-hydroxylase (F3H) genes and PAL enzyme activity were higher in plants treated with UV-A LEDs. Our results suggested that short-term UV-A LEDs were effective in increasing growth and improving antioxidant phenolic compounds in kale, thereby representing a potentially effective strategy for enhancing the production of phytochemicals.

Senna obtusifolia is a widely used medicinal herb in Asian countries. To select elite cultivars, S. obtusifolia seeds were carried by "ShenZhou Ⅷ" recoverable satellite to space. Three spaceflight-subjected lines (SP-lines), namely QC10, QC29, QC46, and their ground control line (GC-line) were cultivated on the ground. Previous studies demonstrated that biological traits and secondary metabolites are different between SP-lines and GC-line. Here, we combined physiological, transcriptional, and metabolic studies to compare the differences between SP-lines and GC-line. The results showed that activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and monodehydroascorbate reductase (MDHAR) were dramatically increased in SP-lines as compared to that of GC-line. Transcript levels of SOD, POD, CAT, APX, and MDHAR were significantly up-regulated in SP-lines. Malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ) contents decreased in SP-lines. Seed yields of QC29 and QC46 were considerably higher than that of GC-line. Besides, QC29 had significantly higher aurantio-obtusin content. Pearson correlation coefficient analysis revealed positive relationships between POD and aurantio-obtusin, as well as APX and aurantio-obtusin. In conclusion, SP-lines have higher antioxidant gene expression level and antioxidant enzyme activity as compared to that of GC-line. With higher seed yield and aurantio-obtusin content, QC29 can be used to breed elite S. obtusifolia cultivars. This study provides a new insight in SP-lines and paves the way to breed elite S. obtusifolia cultivars in the future., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

The aim of this study is to determine the key laccase-encoding gene in the life cycle of Morchella importuna SCYDJ1-A1, and to characterize the biochemical properties of the laccase. Two laccase-like multicopper oxidase (LMCO) genes were identified in the genome of M. importuna SCYDJ1-A1 as putative laccase-encoding genes. The two genes, belonging to Auxiliary Activity family 1 subfamily 3, were named as MiLacA and MiLacB. Phylogenetic analysis of deduced amino acid sequences showed that MiLacA is closest to a LMCO of M. importuna 22J1, while MiLacB had low similarity with known Morchella LMCOs. Real-time quantitative PCR results showed that MiLacA was expressed at much higher levels than MiLacB throughout the entire course of artificial cultivation. MiLacA was overexpressed in Pichia pastoris as a recombinant protein. Biochemical characterization of the purified enzyme showed that MiLacA simultaneously possessed laccase and polyphenol-oxidase activities. MiLacA could be strongly inhibited by Fe 2+ , which is unusual. The optimum pH was four and optimum temperature was 60 °C. The enzyme retained over 74% of the laccase activity after 16-h incubation at 60 °C, which means that its thermostability is at the forefront among the currently known laccases. Our findings may help to elucidate how the laccase of M. importuna is involved in decaying lignin in plant litter, and could also provide a candidate thermostable laccase for potential industrial application., Competing Interests: Compliance with ethical standardsThe authors declare no conflict of interest.

Nowadays, modern plant physiology focuses on complex behavior of metal co-contaminants in agrosystems. Keeping this in view, the current study was conducted to investigate the response of two Brassica napus cultivars (Zheda 622 and ZS 758) under co-contamination of copper (Cu 2+ ) and chromium (Cr 6+ ) to observe their effects on plant growth, photosynthetic parameters, and subcellular distribution of these metals in leaves and roots. The results showed that exposure to Cu and Cr causes decline in plant growth, including biomass and plant height. Significant decrease in pigment concentration and the photosynthetic activity [photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (E), maximal quantum yield of photosystem II (Fv/Fm)] in leaves was also observed. Results of subcellular distribution of metals showed that Cu and Cr were predominantly distributed in cell wall and soluble fraction of roots and leaves. Moreover, Cu and Cr in cellular fractions showed a synergistic accumulation pattern under combined metal stress treatment. Both cultivars showed increased levels of reactive oxygen species (ROS), i.e., hydrogen peroxide (H 2 O 2 ) and superoxide radical (O 2 •- ), and significant modulation in the activities of antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX)] under Cu/Cr alone or their combined treatments. Similarly, expression levels of defense-related genes, such as BnCat, BnApx, BnPrx, and BnSod, were also generally up-regulated compared with control. Electron micrographs (TEM) of the mesophyll and root tip cells indicated prominent alterations both in cellular and organelle levels. Additionally, Cr was found to be more toxic than Cu but less than their combined effect, as revealed by enhanced production of oxidative stress and a reduction in biomass production and photosynthetic activity. The present results also suggest that cultivar ZS 758 is more resistant to Cu/Cr than Zheda 622, due to better adapted metabolism and maintenance of structural integrity under metal stress.

Cantharidin (CTD), a terpenoid defensive toxin mainly produced by blister beetles, is widely known by its toxicity to both cancer cells and pests. However, little information is known about its biosynthesis in blister beetles. In this study, first we determined the CTD content in various tissues of adult blister beetles on different days after mating, and then detected the temporal and spatial expression patterns of genes related to CTD biosynthesis in Epicauta chinensis. Results revealed that the accessory gland is the source of the highest CTD production. The second highest level was in the fat body in male blister beetles after mating. In females, the highest CTD content was in the reproductive system except the ovary after mating. As revealed by messenger RNA expression level analysis, the highest levels of 3-hydroxy-3-methylglutary-CoA reductase (HMGR) and juvenile hormone epoxide hydrolase (JHEH) transcripts of E. chinensis were observed in the fat body in males after mating. However, the highest transcript level of EcHMGR was in the ovary and EcJHEH was maintained at a nearly similar level in females. The transcript level of methyl-farnesoate epoxide was significantly higher in the head and that of CYP4BM1 in the midgut in both male and female E. chinensis. We speculate that the fat body may play a more important role than other tissues on the CTD biosynthesis in male E. chinensis after mating. There may be multiple tissues involved in the process of CTD biosynthesis. These four genes probably play regulatory roles in different tissues in males., (© 2017 Institute of Zoology, Chinese Academy of Sciences.)

Background: Bark beetles rely on detoxifying enzymes to resist the defensive terpenoids of their host trees. Research on carboxylesterases (CarEs) has focused on their multiple functions in the metabolic detoxification of pesticides and plant allelochemicals, drug resistance, and juvenile hormone and pheromone degradation., Result: We identified eight new CarE genes in the Chinese white pine beetle (Dendroctonus armandi) and carried out bioinformatics analysis on the deduced full-length amino acid sequences. Differential transcript levels of CarE genes were observed between sexes; within these levels, significant differences were found among the different development stages, and between insects fed on the phloem of Pinus armandi and exposed to five stimuli [(-)-α-pinene, (-)-β-pinene, (+)-3-carene, limonene and turpentine] at 8 and 24 h., Conclusion: Transcription levels of CarE genes suggest some relationship with the detoxification of terpenoids released by host trees. The functions of bark beetle esterase are mainly in hydrolyzing the host chemical defense and degrading odorant molecules during host selection and colonization. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Key Message: Tomato pollen grains have the capacity for ethylene production, possessing specific components of the ethylene-biosynthesis and -signaling pathways, being affected/responsive to high-temperature conditions. Exposure of plants to heat stress (HS) conditions reduces crop yield and quality, mainly due to sensitivity of pollen grains. Recently, it was demonstrated that ethylene, a gaseous plant hormone, plays a significant role in tomato pollen heat-tolerance. It is not clear, however, whether, or to what extent, pollen grains are dependent on the capacity of the surrounding anther tissues for ethylene synthesis and signaling, or can synthesize this hormone and possess an active signaling pathway. The aim of this work was (1) to investigate if isolated, maturing and mature, tomato pollen grains have the capacity for ethylene production, (2) to find out whether pollen grains possess an active ethylene-biosynthesis and -signaling pathway and characterize the respective tomato pollen components at the transcript level, (3) to look into the effect of short-term HS conditions. Results from accumulation studies showed that pollen, anthers, and flowers produced ethylene and HS affected differentially ethylene production by (rehydrated) mature pollen, compared to anthers and flowers, causing elevated ethylene levels. Furthermore, several ethylene synthesis genes were expressed, with SlACS3 and SlACS11 standing out as highly HS-induced genes of the pollen ethylene biosynthesis pathway. Specific components of the ethylene-signaling pathway as well as several ethylene-responsive factors were expressed in pollen, with SlETR3 (ethylene receptor; named also NR, for never ripe) and SlCTR2 (constitutive triple response2) being HS responsive. This work shows that tomato pollen grains have the capacity for ethylene production, possessing active ethylene-biosynthesis and -signaling pathways, highlighting specific pollen components that serve as a valuable resource for future research on the role of ethylene in pollen thermotolerance.

Main Conclusion: The new model orange callus line, similar to carrot root, was rich in carotenoids due to altered expression of some carotenogenesis-associated genes and possessed unique diversity of chromoplast ultrastructure. Callus induced from carrot root segments cultured in vitro is usually pale yellow (p-y) and poor in carotenoids. A unique, non-engineered callus line of dark orange (d-o) colour was developed in this work. The content of carotenoid pigments in d-o callus was at the same level as in an orange carrot storage root and nine-fold higher than in p-y callus. Carotenoids accumulated mainly in abundant crystalline chromoplasts that are also common in carrot root but not in p-y callus. Using transmission electron microscopy, other types of chromoplasts were also found in d-o callus, including membranous chromoplasts rarely identified in plants and not observed in carrot root until now. At the transcriptional level, most carotenogenesis-associated genes were upregulated in d-o callus in comparison to p-y callus, but their expression was downregulated or unchanged when compared to root tissue. Two pathway steps were critical and could explain the massive carotenoid accumulation in this tissue. The geranylgeranyl diphosphate synthase gene involved in the biosynthesis of carotenoid precursors was highly expressed, while the β-carotene hydroxylase gene involved in β-carotene conversion to downstream xanthophylls was highly repressed. Additionally, paralogues of these genes and phytoene synthase were differentially expressed, indicating their tissue-specific roles in carotenoid biosynthesis and metabolism. The established system may serve as a novel model for elucidating plastid biogenesis that coincides with carotenogenesis.

DNA methylation is a heritable epigenetic mechanism that participates in gene regulation under abiotic stresses in plants. Sesame (Sesamum indicum) is typically considered a drought-tolerant crop but highly susceptible to waterlogging, probably because of its origin in Africa or India. Understanding DNA methylation patterns under drought and waterlogging conditions can provide insights into the regulatory mechanisms underlying sesame contrasting responses to these abiotic stresses. We combined Methylation-Sensitive Amplified Polymorphism and transcriptome analyses to profile cytosine methylation patterns, transcript accumulation, and their interplay in drought-tolerant and waterlogging-tolerant sesame genotypes. Drought stress strongly induced de novo methylation (DNM) whereas most of the loci were demethylated (DM) during the recovery phase. In contrast, waterlogging stress decreased the level of methylation but during the recovery phase, both DM and DNM were concomitantly deployed. In both stresses, the levels of the differentially accumulated transcripts (DATs) highly correlated with the methylation patterns. We observed that DM was associated with an increase of DAT levels while DNM was correlated with a decrease of DAT levels. Altogether, sesame has divergent epigenetic programs that respond to drought and waterlogging stresses and an interplay among DNA methylation and transcript accumulation may partly modulate the contrasting responses to these stresses., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Glyphosate (N-phosphonomethyl-glycine) (GLY) is the active ingredient of the most used herbicides in the world. GLY is applied in formulated products known as glyphosate-based herbicides (GBH), which could induce effects that are not predicted by toxicity assays with pure GLY. This herbicide is classified as organophosphorus compound, which is known to induce neurotoxic effects. Although this compound is classified as non-neurotoxic by regulatory agencies, acute exposure to GBH causes neurological symptoms in humans. However, there is no consensus in relation to neurotoxic effects of GBH. Thus, the aim of this study was to investigate the neurotoxic effects of the GBH in the zebrafish Danio rerio, focusing on acute toxicity, the activity and transcript levels of mitochondrial respiratory chain complexes, mitochondrial membrane potential, reactive species (RS) formation, and behavioral repertoire. Adult zebrafish were exposed in vivo to three concentrations of GBH Scout ® , which contained GLY in formulation (fGLY) (0.065, 1.0 and 10.0 mg L -1 fGLY) for 7 d, and an in vitro assay was performed using also pure GLY. Our results show that GBH induced in zebrafish brain a decrease in cell viability, inhibited mitochondrial complex enzymatic activity, modulated gene expression related to mitochondrial complexes, induced an increase in RS production, promoted hyperpolarization of mitochondrial membrane, and induced behavioral impairments. Together, our data contributes to the knowledge of the neurotoxic effects of GBH. Mitochondrial dysfunction has been recognized as a relevant cellular response that should not be disregarded. Moreover, this study pointed to the mitochondria as an important target of GBH., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The pathogenesis of diabetes mellitus is a complicated process involving much gene regulation. The molecular mechanism of mulberry ( Morus alba L.) leaf in the treatment of diabetes is not fully understood. In this study, we used the Illumina HiSeq™ 2,500 platform to explore the liver transcriptome of normal mice, STZ-induced diabetic mice, and mulberry leaf-treated diabetic mice, and we obtained 52,542,956, 52,626,414, and 52,780,196 clean reads, respectively. We identified differentially expressed genes (DEGs) during the pathogenesis of diabetes in mice. The functional properties of DEGs were characterized by comparison with the GO and KEGG databases, and the results show that DEGs are mainly involved in the metabolic pathway. qRT-PCR was used to analyse 27 differential genes involved in liver expression in different groups of diabetic mice. Among the DEGs, the expression of Scube1, Spns3, Ly6a, Igf2 , and other genes between the control (C) and diabetic control (DC) groups was significantly upregulated; the expression of Grb10, Mup2 , and Fasn was significantly downregulated; the expression of the Sqle, Lss , and Irs2 genes between the C group and diabetic group treated with mulberry (DD) was significantly upregulated; the expression of Fabp2, Ly6a , and Grb10 was significantly downregulated; and the expression of Sqle and Lss was significantly upregulated in the DC and DD groups, but Tap1, Igf2 , and Spns3 were significantly downregulated. The results of Western blot validation showed that dynamic changes in proteins, such as IGF2, Ly6a, Grb10, and UBD, occurred to regulate the incidence of diabetes by influencing the insulin receptor substrate (IRS) signaling pathway.

Background: One of evolution's most important achievements is the development and radiation of multicellular organisms with different types of cells. Complex multicellularity has evolved several times in eukaryotes; yet, in most lineages, an investigation of its molecular background is considerably challenging since the transition occurred too far in the past and, in addition, these lineages evolved a large number of cell types. However, for volvocine green algae, such as Volvox carteri, multicellularity is a relatively recent innovation. Furthermore, V. carteri shows a complete division of labor between only two cell types - small, flagellated somatic cells and large, immotile reproductive cells. Thus, V. carteri provides a unique opportunity to study multicellularity and cellular differentiation at the molecular level., Results: This study provides a whole transcriptome RNA-Seq analysis of separated cell types of the multicellular green alga V. carteri f. nagariensis to reveal cell type-specific components and functions. To this end, 246 million quality filtered reads were mapped to the genome and valid expression data were obtained for 93% of the 14,247 gene loci. In the subsequent search for protein domains with assigned molecular function, we identified 9435 previously classified domains in 44% of all gene loci. Furthermore, in 43% of all gene loci we identified 15,254 domains that are involved in biological processes. All identified domains were investigated regarding cell type-specific expression. Moreover, we provide further insight into the expression pattern of previously described gene families (e.g., pherophorin, extracellular matrix metalloprotease, and VARL families). Our results demonstrate an extensive compartmentalization of the transcriptome between cell types: More than half of all genes show a clear difference in expression between somatic and reproductive cells., Conclusions: This study constitutes the first transcriptome-wide RNA-Seq analysis of separated cell types of V. carteri focusing on gene expression. The high degree of differential expression indicates a strong differentiation of cell types despite the fact that V. carteri diverged relatively recently from its unicellular relatives. Our expression dataset and the bioinformatic analyses provide the opportunity to further investigate and understand the mechanisms of cell type-specific expression and its transcriptional regulation.

Ixeris dentata var. albiflora is considered as a potential therapeutic agent against mithridatism, calculous, indigestion, pneumonia, hepatitis, and tumors as well as good seasoned vegetable in Far East countries. Phytoene synthase (PSY), phytoene desaturase (PDS) ξ-carotene desaturase (ZDS), lycopene β-cyclase (LCYB), lycopene ε-cyclase (LCYE), ε-ring carotene hydroxylase (CHXB), and zeaxanthin epoxidase (ZDS) are vital enzymes in the carotenoid biosynthesis pathway. We have examined these seven genes from I. dentat a that are participated in carotenoid biosynthesis utilizing an Illumina/Solexa HiSeq 2000 platform. In silico analysis of the seven deduced amino acid sequences were revealed its closest homology with other Asteracea plants. Further, we explored transcript levels and carotenoid accumulation in various organs of I. dentata using quantitative real time PCR and high-performance liquid chromatography, respectively. The highest transcript levels were noticed in the leaf for all the genes while minimal levels were noticed in the root. The maximal carotenoid accumulation was also detected in the leaf. We proposed that these genes expressions are associated with the accumulation of carotenoids. Our findings may suggest the fundamental clues to unravel the molecular insights of carotenoid biosynthesis in various organs of I. dentata ., Competing Interests: The authors declare no conflict of interest.

Microcystins (MCs) in freshwater and marine waters released by toxin-producing cyanobacteria have negative impacts to the aquatic environment. This study aimed to investigate the effect of pure microcystin-LR on activity and transcript level of immune-related enzymes in the white shrimp Litopenaeus vannamei. After exposed to varying concentrations of pure microcystin-LR (MC-LR) for 30 days, the activity of superoxide dismutase (SOD), lysozyme (LZM), glutathione peroxidase (GPx), peroxidase (POD), acid phosphatase (ACP), alkaline phosphatase (AKP) and transcript level of cMn-sod, lzm, gpx were investigated in the hepatopancreas of white shrimp (L. vannamei). Immune-related enzyme activities responded differently to MC-LR exposure. SOD, GPx, and POD activity in the hepatopancreas were activated in a concentration-dependent manner while LZM activity was significantly inhibited in the treatment groups. ACP and AKP activity showed an increase, followed by a decrease. The transcript levels of cMn-sod, lzm, and gpx were consistent with changes in their encoding enzyme activity. These results demonstrated that sub-chronical exposure to MC-LR induced the alteration of immune-related enzymes and corresponding genes in the hepatopancreas, which may help explain the presence of detoxification mechanisms in crustaceans and how they were protected from MC-LR stress for a long period of time.

Members of the genus Ixeris have long been used in traditional medicines as stomachics, sedatives, and diuretics. Phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate: coenzyme-A (CoA) ligase (4CL), chalcone synthase (CHS), and dihydroflavonol 4-reductase (DFR) are important enzymes in the phenylpropanoid pathway. In this study, we analyzed seven genes from Ixeris dentata var. albiflora that are involved in phenylpropanoid biosynthesis, using an Illumina/Solexa HiSeq 2000 platform. The amino acid sequence alignments for IdPAL s, IdC4H, Id4CL s, IdCHS , and IdDFR showed high identity to sequences from other plants. We also investigated transcript levels using quantitative real-time PCR, and analyzed the accumulation of phenylpropanoids in different organs of I. dentata var. albiflora using high-performance liquid chromatography. The transcript levels of IdC4H, Id4CL1 , IdCHS , and IdDFR were highest in the leaf. The catechin, chlorogenic acid, ferulic acid, and quercetin contents were also highest in the leaf. We suggest that expression of IdC4H, Id4CL1 , IdCHS , and IdDFR is associated with the accumulation of phenylpropanoids. Our results may provide baseline information for elucidating the mechanism of phenylpropanoid biosynthesis in different organs of I. dentata var. albiflora ., Competing Interests: The authors declare no conflict of interest.

The fat mass and obesity associated (Fto) and iroquois homeobox 3 (Irx3) genes have been recognised as important obesity-related genes. Studies on the expression of these genes in the fat tissue of human and mouse have produced inconsistent results, while similar data on rat are limited. Environmental factors such as diet, should be considered as potential modulators of gene transcript levels through epigenetic mechanisms including DNA methylation. The aim of this study was to evaluate transcription levels and DNA methylation profiles of rat Fto and Irx3 genes in two white adipose tissue depots in response to high-fat and high-protein diets. The relative transcript levels of Fto and Irx3 were shown to be tissue-specific with higher levels detected in subcutaneous fat tissue than in abdominal fat tissue. Moreover, negative correlations between the transcripts of both genes were observed for subcutaneous fat tissue. The identified interactions (e.g. diet×duration of diet regimen) indicated that the diet had an impact on the transcript level; however, this effect was dependent on the duration of the diet regimen. The high-fat diet led to upregulation of Fto and Irx3 linearly with time across the two tissues. DNA methylation of the regulatory regions of the studied genes was very low and not related with the tissue, diet, or duration of diet regimen. Our study revealed that diet was an important factor modulating transcription of Fto and Irx3, but its effect is time-dependent. In contrast, the DNA methylation profiles of Fto and Irx3 were not altered by nutrition, which may indicate that the feeding type, when applied postnatally, did not affect DNA methylation of these genes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Cyperus esculentus is unique in that it can accumulate rich oil in its tubers. However, the underlying mechanism of tuber oil biosynthesis is still unclear. Our transcriptional analyses of the pathways from pyruvate production up to triacylglycerol (TAG) accumulation in tubers revealed many distinct species-specific lipid expression patterns from oil seeds and fruits, indicating that in C. esculentus tuber: (i) carbon flux from sucrose toward plastid pyruvate could be produced mostly through the cytosolic glycolytic pathway; (ii) acetyl-CoA synthetase might be an important contributor to acetyl-CoA formation for plastid fatty acid biosynthesis; (iii) the expression pattern for stearoyl-ACP desaturase was associated with high oleic acid composition; (iv) it was most likely that endoplasmic reticulum (ER)-associated acyl-CoA synthetase played a significant role in the export of fatty acids between the plastid and ER; (v) lipid phosphate phosphatase (LPP)-δ was most probably related to the formation of the diacylglycerol (DAG) pool in the Kennedy pathway; and (vi) diacylglyceroltransacylase 2 (DGAT2) and phospholipid:diacylglycerolacyltransferase 1 (PDAT1) might play crucial roles in tuber oil biosynthesis. In contrast to oil-rich fruits, there existed many oleosins, caleosins and steroleosins with very high transcripts in tubers. Surprisingly, only a single ortholog of WRINKLED1 (WRI1)-like transcription factor was identified and it was poorly expressed during tuber development. Our study not only provides insights into lipid metabolism in tuber tissues, but also broadens our understanding of TAG synthesis in oil plants. Such knowledge is of significance in exploiting this oil-rich species and manipulating other non-seed tissues to enhance storage oil production., (© The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Interleukin-6 (IL-6) is one of the most important multifunctional cytokines, playing essential roles in mediating the innate and adaptive immune responses. In this study, il-6 gene and its promoter from blunt snout bream, Megalobrama amblycephala, were characterized, and its expression at the transcript level in healthy fish and after bacterial infection was determined by quantitative real-time PCR. The results showed that the M. amblycephala il-6 (Mamil-6) cDNA had an ORF of 699 bp, encoding 232 amino acids, and contained 9 instable motifs in the 3' UTR. The deduced MamIL-6 possessed a 24-amino acid signal peptide and was located in the cytoplasm. Although sequence alignment and phylogenetic analysis revealed that IL-6 is poorly conserved in vertebrates, the protein and genomic structure of il-6 gene was well conserved. Analysis of the Mamil-6 promoter revealed the presence of a conserved TATA box and six major cis-regulatory elements, including C/EBPβ (NF-IL6), AP-1, CRE, GRE, GATA and NF-κB binding sites. In healthy fish, Mamil-6 was the most abundant in the spleen. After Aeromonas hydrophila infection, Mamil-6 was significantly up-regulated in all 6 immune-related tissues examined, suggesting that Mamil-6 plays an important role in the blunt snout bream immune system.

Background: Bark beetles rely on their detoxifying enzymes to resist the defensive terpenoids of host trees. Glutathione S-transferases (GSTs) conjugate xenobiotic compounds with a glutathione moiety (GSH) and often work in tandem with cytochromes P450 or other enzymes that aid in the detoxification, sequestration or excretion of toxic compounds., Result: We identified nine new GST genes in the Chinese white pine beetle (Dendroctonus armandi) and carried out a bioinformatics analysis on the deduced full-length amino acid sequences. These genes belong to four different classes (epsilon, sigma, omega and theta). Differential transcript levels of each class of GST genes were observed between sexes, and, within these levels, significant differences were found among the different adult substages that were fed phloem of Pinus armandi and exposed to six stimuli [(±)-α-pinene, (-)-α-pinene, (-)-β-pinene, (+)-3-carene, (±)-limonene and turpentine] at 8 and 24 h., Conclusion: The increased transcription levels of GST genes suggested that they have some relationship with the detoxification of terpenoids that are released by host trees. The mediating oxidative stress that is caused by monoterpene might be the main role of the bark beetle GSTs., (© 2015 Society of Chemical Industry.)

Though iron- and sulfate-reducing bacteria are well known for mediating uranium(VI) reduction in contaminated subsurface environments, quantifying the in situ activity of the microbial groups responsible remains a challenge. The objective of this study was to demonstrate the use of quantitative molecular tools that target mRNA transcripts of key genes related to Fe(III) and sulfate reduction pathways in order to monitor these processes during in situ U(VI) remediation in the subsurface. Expression of the Geobacteraceae-specific citrate synthase gene (gltA) and the dissimilatory (bi)sulfite reductase gene (dsrA), were correlated with the activity of iron- or sulfate-reducing microorganisms, respectively, under stimulated bioremediation conditions in microcosms of sediments sampled from the U.S. Department of Energy's Oak Ridge Integrated Field Research Challenge (OR-IFRC) site at Oak Ridge, TN, USA. In addition, Geobacteraceae-specific gltA and dsrA transcript levels were determined in parallel with the predominant electron acceptors present in moderately and highly contaminated subsurface sediments from the OR-IFRC. Phylogenetic analysis of the cDNA generated from dsrA mRNA, sulfate-reducing bacteria-specific 16S rRNA, and gltA mRNA identified activity of specific microbial groups. Active sulfate reducers were members of the Desulfovibrio, Desulfobacterium, and Desulfotomaculum genera. Members of the subsurface Geobacter clade, closely related to uranium-reducing Geobacter uraniireducens and Geobacter daltonii, were the metabolically active iron-reducers in biostimulated microcosms and in situ core samples. Direct correlation of transcripts and process rates demonstrated evidence of competition between the functional guilds in subsurface sediments. We further showed that active populations of Fe(III)-reducing bacteria and sulfate-reducing bacteria are present in OR-IFRC sediments and are good potential targets for in situ bioremediation.

Identification of prognostic factors and risk-based post-remission therapy was proposed to improve the outcomes of acute myeloid leukemia (AML) and a mutation of FLT3 has been reported to be a risk factor, especially for pediatric patients. Recently, FLT3 expression level was implicated to have prognostic significance in adults, but little is known for childhood AML. To define the prognostic significance, transcript level of FLT3 was analyzed in 52 pediatric AML patients. The median copy number of FLT3 was 4.6x10(3) (40-5.9x10(7) copies)/1.0x10(6) GAPDH copy, and the relapse free survival of patients with high transcript level of FLT3 (>10(6) copy number) (0%) was significantly lower than that of the others (53.2%). High transcript level of FLT3 was associated with a markedly high risk of relapse. The development of new therapeutic scheme such as a frontline allogeneic stem cell transplantation or administration of FLT3 inhibitor is needed to improve outcomes.