Hot water extract of Pleurotus pulmonarius stalk waste enhances innate immune response and immune-related gene expression in red hybrid tilapia Oreochromis sp. following challenge with pathogen-associated molecular patterns.

In aquaculture, commercial fish such as red hybrid tilapia are usually raised at high density to boost the production within a short period of time. This overcrowded environment, however, may cause stress to the cultured fish and increase susceptibility to infectious diseases. Antibiotics and chemotherapeutics are used by fish farmers to overcome these challenges, but this may increase the production cost. Studies have reported on the potential of mushroom polysaccharides that can act as immunostimulants to enhance the immune response and disease resistance in fish. In the current study, hot water extract (HWE) from mushroom stalk waste (MSW) was used to formulate fish feed and hence administered to red hybrid tilapia to observe the activation of immune system. Upon 30 days of feeding, the fish were challenged with pathogen-associated molecular patterns (PAMPs) such as lipopolysaccharides (LPS) and polyinosinic:polycytidylic acid (poly (I:C)) to mimic bacterial and viral infection, respectively. HWE supplementation promoted better feed utilisation in red hybrid tilapia although it did not increase the body weight gain and specific growth rate compared to the control diet. The innate immunological parameters such as phagocytic activity and respiratory burst activity were significantly higher in HWE-supplemented group than that of the control group following PAMPs challenges. HWE-supplemented diet also resulted in higher mRNA transcription of il1b and tnfa in midgut, spleen and head kidney at 1-day post PAMPs injection. Tlr3 exhibited the highest upregulation in the HWE fed fish injected with poly (I:C). At 3-days post PAMPs injection, both ighm and tcrb expression were upregulated significantly in the spleen and head kidney. Results showed that HWE supplementation enhances the immune responses of red hybrid tilapia and induced a higher serum bactericidal activity against S. agalactiae.

Chromosome behavior during meiosis in pollen mother cells from Saccharum officinarum × Erianthus arundinaceus F1 hybrids.

BACKGROUND: In recent years, sugarcane has attracted increasing attention as an energy crop. Wild resources are widely used to improve the narrow genetic base of sugarcane. However, the infertility of F1 hybrids between Saccharum officinarum (S. officinarum) and Erianthus arundinaceus (E. arundinaceus) has hindered sugarcane breeding efforts. To discover the cause of this infertility, we studied the hybridization process from a cytological perspective. RESULTS: We examined the meiotic process of pollen mother cells (PMCs) in three F1 hybrids between S. officinarum and E. arundinaceus. Cytological analysis showed that the male parents, Hainan 92-77 and Hainan 92-105, had normal meiosis. However, the meiosis process in F1 hybrids showed various abnormal phenomena, including lagging chromosomes, micronuclei, uneven segregation, chromosome bridges, and inability to form cell plates. Genomic in situ hybridization (GISH) showed unequal chromatin distribution during cell division. Interestingly, 96.70% of lagging chromosomes were from E. arundinaceus. Furthermore, fluorescence in situ hybridization (FISH) was performed using 45S rDNA and 5S rDNA as probes. Either 45S rDNA or 5S rDNA sites were lost during abnormal meiosis, and results of unequal chromosomal separation were also clearly observed in tetrads. CONCLUSIONS: Using cytogenetic analysis, a large number of meiotic abnormalities were observed in F1. GISH further confirmed that 96.70% of the lagging chromosomes were from E. arundinaceus. Chromosome loss was found by further investigation of repeat sequences. Our findings provide insight into sugarcane chromosome inheritance to aid innovation and utilization in sugarcane germplasm resources.

Transcriptional and proteomic insights into phytotoxic activity of interspecific potato hybrids with low glycoalkaloid contents.

BACKGROUND: Glycoalkaloids are bioactive compounds that contribute to the defence response of plants against herbivore attack and during pathogenesis. Solanaceous plants, including cultivated and wild potato species, are sources of steroidal glycoalkaloids. Solanum plants differ in the content and composition of glycoalkaloids in organs. In wild and cultivated potato species, more than 50 steroidal glycoalkaloids were recognized. Steroidal glycoalkaloids are recognized as potential allelopathic/phytotoxic compounds that may modify the growth of target plants. There are limited data on the impact of the composition of glycoalkaloids on their phytotoxic potential. RESULTS: The presence of α-solasonine and α-solamargine in potato leaf extracts corresponded to the high phytotoxic potential of the extracts. Among the differentially expressed genes between potato leaf bulks with high and low phytotoxic potential, the most upregulated transcripts in sample of high phytotoxic potential were anthocyanin 5-aromatic acyltransferase-like and subtilisin-like protease SBT1.7-transcript variant X2. The most downregulated genes were carbonic anhydrase chloroplastic-like and miraculin-like. An analysis of differentially expressed proteins revealed that the most abundant group of proteins were those related to stress and defence, including glucan endo-1,3-beta-glucosidase acidic isoform, whose expression level was 47.96× higher in potato leaf extract with low phytotoxic. CONCLUSIONS: The phytotoxic potential of potato leaf extract possessing low glycoalkaloid content is determined by the specific composition of these compounds in leaf extract, where α-solasonine and α-solamargine may play significant roles. Differentially expressed gene and protein profiles did not correspond to the glycoalkaloid biosynthesis pathway in the expression of phytotoxic potential. We cannot exclude the possibility that the phytotoxic potential is influenced by other compounds that act antagonistically or may diminish the glycoalkaloids effect.

Flavonoid Profiles of Two New Approved Romanian Ocimum Hybrids.

Basil (Ocimum spp.) is a traditional herbal medicine abundant in antioxidants such as phenolic compounds. As part of a diet, this herb is proved to have some roles in decreasing the risk of cancer, and in the treatment of inflammation and neurodegenerative diseases. This study aims to explore the total phenolic and flavonoid content of two new basil hybrids growing in Romania, namely "Aromat de Buzau" (AB) and "Macedon" (MB). The antioxidant capacity of those two species was also analyzed by DPPH and cyclic voltammetry. Six different flavonoids, such as catechin (+), rutin, hyperoside, naringin, naringenin, and genistein, were separated, identified, and quantified by HPLC-DAD chromatography, for the first time, from romanian basil hybrids. The main flavonoid of the extracts was found to be naringin which is present in the highest amount (26.18 mg/kg) in "Aromat de Buzau" (O. basilicum) methanolic extract. These results suggest that dietary intake of these new hybrids can be a source of antioxidant compounds.

Comprehensive physiological and transcriptomic analysis revealing the responses of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) to the replacement of fish meal with soy protein concentrate.

Plant proteins are suitable and alternative to fish meals (FMs), with less cost compared with that of all other types of fish feeds. In recent years, soy protein concentrate (SPC) has emerged as a cost-effective alternative to FM; however, little is known regarding the effects of dietary SPC on general fish physiology and well-being. This study aimed to perform comprehensive physiological and transcriptomic analysis for testing the applicability of SPC as fish feeds in hybrid grouper (Epinephelus fuscoguttatus♀ נE. lanceolatus♂) [SPC replaced 0% (CK), 30% (SPC30), and 75% (SPC75) of FM protein]. Generally, SPC30 promoted fish survival and had less effects on the phenotype, while SPC75 reduced fish survival, promoted inflammation, and regulated multiple physiological responses. Thousands of differentially expressed genes (DEGs) by SPC were identified in the intestine, liver, and muscle, which were enriched in biological regulation, cellular process, metabolic process, single-organism process, cell, cell part, membrane, binding, and catalytic activity based on RNA-seq. Notably, some DEGs involved in amino acid and lipid metabolism in the digestive system highlighted the modulatory effect of SPC on these metabolic processes, consistent with the physiological responses including enzyme activities. The enriched aspects of these predominant DEGs might be directly related to the different effects of SPC30 and SPC75 on fish growth, digestibility, and underlying enzyme activities and histology. In conclusion, the comprehensive physiological and transcriptomic comparative analysis of CK, SPC30, and SPC75 was also effective in testing the applicability of SPC as fish feeds and in designing a proper diet with the best impact on the growth performance and health of fish in hybrid grouper.

ß-glucan alleviates the immunosuppressive effects of oxytetracycline on the non-specific immune responses and resistance against Vibrio alginolyticus infection in Epinephelus fuscoguttatus × Epinephelus lanceolatus hybrids.

This study was conducted to examine the combinatory effects of ß-glucan and oxytetracycline (OTC) on hybrid giant tiger groupers (Epinephelus fuscoguttatus × Epinephelus lanceolatus). In vitro tests, OTC significantly reduced superoxide anion production and phagocytic activity in primary head kidney leukocytes. However, this suppressive effect was alleviated by co-treatment with ß-glucan. Subsequently, feeding trials were performed to investigate the potential immunomodulatory effects of dietary ß-glucan alone or in combination with OTC on groupers. A total of 210 healthy groupers (368.00 ± 51.03 g) were divided into six groups. Group 1 was the control group, group 2 (BG) received 5 g ß-glucan per kg feed weight, groups 3-5 received 5 g/kg ß-glucan in combination with 10, 30, or 50 mg OTC/kg fish weight/day (groups M1, M2, and M3, respectively), and group 6 (O) received 50 mg OTC/kg fish weight/day. Fish were sampled to determine the innate immunity parameters and residual OTC levels in the muscle tissue during a 28-day feeding regimen. Residual OTC levels were considerably higher in groups M3 and O compared with the other groups, and peaked on day 14. This was followed by a slight decrease on day 28, despite a continuous supply of OTC. Notably, fish fed with OTC alone had significantly decreased phagocytic rates and superoxide anion production observed in head kidney leukocytes, as well as poorer protection against Vibrio alginolyticus infection. These immunosuppressive effects were not observed in the fish fed with ß-glucan in combination with a lower dose of OTC (group M2). Thus, these data suggest that the combination of dietary ß-glucan and OTC exerts synergistic immunostimulating effects that protect groupers from bacterial infection.

Antioxidant defenses and non-specific immunity at enzymatic and transcriptional levels in response to dietary carbohydrate in a typical carnivorous fish, hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂).

The present study was conducted to explore the influence of dietary carbohydrate on antioxidant capacity and non-specific immunity of hybrid grouper, which would contribute to determine the tolerable dietary carbohydrate content. Seven diets with grade levels of carbohydrate (5.27, 8.95, 11.49, 14.37, 17.78, 20.82 and 23.65%) were fed to triplicate groups of fish for 10 weeks. Results showed that the inclusion of carbohydrate above 11.49% produced significant increased content of hydrogen peroxide (H2O2) in liver and malondialdehyde (MDA) in both serum and liver. The specific activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (Gpx) and total antioxidative capacity (T-AOC) were significantly elevated with the increase of dietary carbohydrate from 8.95 to 23.65%, which may be associated with the reduced hepatic soluble protein content. However, opposite variation was observed in the expression of antioxidant related genes (SOD1 and Gpx), which was partly caused by the activation of NF-E2-related factor 2 (Nrf2) and inhibition of Kelch-like-ECH-associated protein 1 (Keap1) at the transcriptional level. The immunoglobulin M (lgM) content and activity of lysozyme and CCP in serum significantly depressed when dietary carbohydrate was above 11.49%. The expression of pro-inflammatory cytokines (TNF-α, IL-1ß and IL-8) was significantly increased with the increase of dietary carbohydrate from 5.27 to 8.95% and thereafter significantly reduced, which was consistent with the changed expression of toll-like receptor 2 (TLR2) and nuclear factor κΒ (NF-κΒ). In above, high dietary carbohydrate significantly impaired the antioxidant capacity and reduced the non-specific immunity of hybrid grouper, and the tolerable dietary carbohydrate content should not exceed 11.49%.

Effect of dietary threonine on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂.

The experiment was conducted to investigate the effects of dietary threonine (Thr) on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. A total of 1200 fish (14·19 (se 0·13) g) were randomly distributed into six groups with four replicates each, fed six diets with graded level of Thr (9·5, 11·5, 13·5, 15·4, 17·4 and 19·3 g/kg diets) for 56 d. Results showed (P < 0·05) that dietary Thr (1) increased percentage weight gain, specific growth rate, feed efficiency and protein efficiency ratio; (2) up-regulated growth hormone (GH), insulin-like growth factor 1 (IGF-1), proliferating cell nuclear antigen, myogenic regulation factors (MyoD, Myf5, MyoG and Mrf4) and myosin heavy chain (MyHC) mRNA levels; (3) increased muscle protein content via regulating the protein kinase B/target of rapamycin signalling pathway and (4) decreased malondialdehyde and protein carbonyl contents, increased catalase, glutathione-S-transferase, glutathione reductase and GSH activities, up-regulated mRNA levels of antioxidant enzymes related to NFE2-related factor 2 and γ-glutamylcysteine ligase catalytic subunit. These results suggest that Thr has a potential role to improve muscle growth and protein synthesis, which might be due to the regulation of GH-IGF system, muscle growth-related gene, antioxidative capacity and protein synthesis-related signalling pathways. Based on the quadratic regression analysis of specific growth rate, the Thr requirement of hybrid catfish (14·19-25·77 g) was estimated to be 13·77 g/kg of the diet (33·40 g/kg of dietary protein).

Standard Deviations: The Biological Bases of Transmission Ratio Distortion.

The rule of Mendelian inheritance is remarkably robust, but deviations from the equal transmission of alternative alleles at a locus [a.k.a. transmission ratio distortion (TRD)] are also commonly observed in genetic mapping populations. Such TRD reveals locus-specific selection acting at some point between the diploid heterozygous parents and progeny genotyping and therefore can provide novel insight into otherwise-hidden genetic and evolutionary processes. Most of the classic selfish genetic elements were discovered through their biasing of transmission, but many unselfish evolutionary and developmental processes can also generate TRD. In this review, we describe methodologies for detecting TRD in mapping populations, detail the arenas and genetic interactions that shape TRD during plant and animal reproduction, and summarize patterns of TRD from across the genetic mapping literature. Finally, we point to new experimental approaches that can accelerate both detection of TRD and characterization of the underlying genetic mechanisms.

Embryonic Chimeras with Human Pluripotent Stem Cells.

Human pluripotent stem (PS) cells can be isolated from preimplantation embryos or by reprogramming of somatic cells or germline progenitors. Human PS cells are considered the "holy grail" of regenerative medicine because they have the potential to form all cell types of the adult body. Because of their similarity to humans, nonhuman primate (NHP) PS cells are also important models for studying human biology and disease, as well as for developing therapeutic strategies and test bed for cell replacement therapy. This chapter describes adjusted methods for cultivation of PS cells from different primate species, including African green monkey, rhesus monkey, chimpanzee, and human. Supplementation of E8 medium and inhibitors of the Tankyrase and GSK3 kinases to various primate PS cell media reduce line-dependent predisposition for spontaneous differentiation in conventional PS cell cultures. We provide methods for basic characterization of primate PS cell lines, which include immunostaining for pluripotency markers such as OCT4 and TRA-1-60, as well as in vivo teratoma formation assay. We provide methods for generating alternative PS cells including region-selective primed PS cells, two different versions of naïve-like cells, and recently reported extended pluripotent stem (EPS) cells. These derivations are achieved by acclimation of conventional PS cells to target media, episomal reprogramming of somatic cells, or resetting conventional PS cells to a naïve-like state by overexpression of KLF2 and NANOG. We also provide methods for isolation of PS cells from human blastocysts. We describe how to generate interspecies primate-mouse chimeras at the blastocyst and postimplantation embryo stages. Systematic evaluation of the chimeric competency of human and primate PS cells will aid in efforts to overcome species barriers and achieve higher grade chimerism in postimplantation conceptuses that could enable organ-specific enrichment of human xenogeneic PS cell derivatives in large animals such as pigs and sheep.

Macronutrients influence yield and oil quality of hybrid maize (Zea mays L.).

In the present two-year study, an attempt was made to estimate the grain yield, grain nutrient uptake, and oil quality of three commonly grown maize (Zea mays L.) hybrids fertilized with varied levels of nitrogen (N), phosphorus (P) and potassium (K). Results obtained from both the experimental years indicated that application of 125% of recommended dose of fertilizer (RDF) recorded maximum grain yield (10.37 t ha-1; 124% higher than control). When compared with 100% RDF, grain yield reduction with nutrient omission was 44% for N omission, 17% for P omission, and 27% for K omission. Nitrogen uptake was increased with increasing NPK levels up to 150% RDF that was statistically at par (p ≥ 0.01) with 125% RDF. Increasing trend in P and K uptake was observed with successive increase in NPK levels up to 125% RDF, above which it declined. The protein content was significantly higher in grains of var. P 3396 with 125% RDF. Nutrient management has significant (p ≤ 0.01) role in the grain oil content. Saturated fatty acids (palmitic, stearic and arachidic acid) content decreased, and unsaturated fatty acid (oleic, linoleic and linolenic acid) increased with increasing NPK levels. The average oleic acid desaturation and linoleic acid desaturation ratios were increased with increasing NPK levels up to 100 and 125% RDF, respectively. However, average monounsaturated fatty acids (MUFA): poly-unsaturated fatty acids (PUFA), saturated: unsaturated as well as linoleic: linolenic acid ratios were increased on receiving 75% RDF, and beyond that it showed decreasing trend. The omission of K had the highest inhibitory effect on corn oil quality followed by N and P omission.

Generation of ERK-Independent Human and Non-Human Primate Pluripotent Stem Cells.

The production of human organs inside human-animal interspecies chimeras might one day comprise a viable strategy for generating patient-specific organs, but such experiments will require human chimera-competent pluripotent stem (PS) cells. The stabilization of PS cell self-renewal in serum-free medium and ERK blockade might be critical for capturing primate chimera-competent pluripotency. It has recently been shown that shielding primate cells from the activation of ERK, WNT, and PKC signaling is crucial for deriving African green monkey ERK-independent PS cells. Here, I show that this principle is generalizable to human cells. In this chapter, methods are provided to reset conventional human PS cells to ERK-independence using histone deacetylase inhibitors and PGCX media comprised of N2B27 medium supplemented with LIF, PD0325901, Go6983, CHIR99021, and XAV939. The novel stem cells exhibit higher levels of KLF4 and manifest increased mitochondrial membrane depolarization. However, the author observed that not all PS cell lines are amenable to small molecule-mediated resetting. The ERK-independent PS cells described herein will provide a useful resource for testing interspecies organogenesis strategies. © 2019 by John Wiley & Sons, Inc.

Heterotic patterns of primary and secondary metabolites in the oilseed crop Brassica juncea.

Heterosis refers to the superior performance of F1 hybrids over their respective parental inbred lines. Although the genetic and expression basis of heterosis have been previously investigated, the metabolic basis for this phenomenon is poorly understood. In a preliminary morphological study in Brassica juncea, we observed significant heterosis at the 50% flowering stage, wherein both the growth and reproduction of F1 reciprocal hybrids were greater than that of their parents. To identify the possible metabolic causes or consequences of this heterosis, we carried out targeted LC-MS analysis of 48 primary (amino acids and sugars) and secondary metabolites (phytohormones, glucosinolates, flavonoids, and phenolic esters) in five developmental tissues at 50% flowering in hybrids and inbred parents. Principal component analysis (PCA) of metabolites clearly separated inbred lines from their hybrids, particularly in the bud tissues. In general, secondary metabolites displayed more negative heterosis values in comparison to primary metabolites. The tested primary and secondary metabolites displayed both additive and non-additive modes of inheritance in F1 hybrids, wherein the number of metabolites showing an additive mode of inheritance were higher in buds and siliques (52.77-97.14%) compared to leaf tissues (47.37-80%). Partial least regression (PLS) analysis further showed that primary metabolites, in general, displayed higher association with morphological parameters in F1 hybrids. Overall, our results are consistent with a resource-cost model for heterosis in B. juncea, where metabolite allocation in hybrids appears to favor growth, at the expense of secondary metabolism.

Generation of a new thermo-sensitive genic male sterile rice line by targeted mutagenesis of TMS5 gene through CRISPR/Cas9 system.

BACKGROUND: Two-line hybrid rice with high yield potential is increasingly popular and the photo- and temperature-sensitive male sterile line is one of the basic components for two-line hybrid rice breeding. The development of male sterile lines through conventional breeding is a lengthy and laborious process, whereas developing thermo-sensitive genic male sterile (TGMS) lines for two-line hybrid breeding by editing a temperature-sensitivity gene by CRISPR/Cas9 is efficient and convenient. RESULTS: Here, thermo-sensitive genic male sterility (TGMS) was induced by employing the CRISPR/Cas9 gene editing technology to modify the gene TMS5. Two TGMS mutants, tms5-1 and tms5-2, both lacking any residual T-DNA, were generated in the indica rice cultivar Zhongjiazao17 (cv. YK17) background. When grown at a sub-optimal temperature (22 °C), both mutants produced viable pollen and successfully produced grain through self-fertilization, but at temperatures 24 and 26 °C, their pollen was sterile and no grain was set. F1 hybrids derived from the crosses between YK17S (tms5-1) and three different restorer lines outperformed both parental lines with respect to grain yield and related traits. CONCLUSION: The YK17S generated by CRISPR/Cas9 system was proved to be a new TGMS line with superior yield potential and can be widely utilized in two-line hybrid breeding of indica rice.

The response of maize protoplasts to cadmium stress mitigated by silicon.

The aim of this article was to evaluate the viability of maize protoplasts, cell wall regeneration, Cd uptake by protoplasts, and the impact of silicon under cadmium cations (Cd) stress in two maize hybrids with contrasting tolerances to Cd toxicity. The differences in protoplast viability between the sensitive (Novania) and tolerant (Almansa) hybrids were noticeable even at the beginning of culture. The percentage of living protoplasts in the presence of Cd was higher in the tolerant hybrid. In both hybrids, Si supplementation significantly increased the viability of protoplasts exposed to Cd. The percentage of protoplasts with regenerated cell walls gradually increased in both hybrids and by the end of the culture it had reached almost identical values. Differences were observed during the first four days, when a lag phase occurred in the protoplasts of the sensitive hybrid accompanied by a rapid decrease in protoplast viability in all the variants tested. The addition of Si increased the cell wall regeneration compared with the Cd variant in both hybrids. The Cd content was higher in the tolerant hybrid than in the sensitive one during the first four days and declined on the seventh day. This may be connected with the increasing intensity of cell wall formation from the fourth up to the seventh day. The addition of Si decreased the Cd uptake into protoplasts of both hybrids. Despite the higher content of Cd, the protoplasts of the tolerant hybrid showed higher viability, obviously indicating unequal mechanisms of Cd processing in studied hybrids. CAPSULE: Protoplasts of two maize hybrids were tested for their viability, regeneration, Cd-uptake and the mitigation of cadmium stress by silicon.

The underlying pathway involved in inter-subspecific hybrid male sterility in rice.

f5 locus in rice (Oryza sativa L.) confers significant effects on hybrid male sterility and segregation distortion. BC14F2 plants with f5-i/i, f5-j/j and f5-i/j genotypes were used to dissect the underlying pathway of f5-caused hybrid male sterility via comparative transcriptome analysis. A total of 350, 421, and 480 differentially expressed genes (DEGs) were identified from f5-i/j vs f5-j/j, f5-j/j vs f5-i/i, and f5-i/j vs f5-i/i, respectively. 145 DEGs were identified simultaneously in f5-i/j vs f5-j/j and f5-i/j vs f5-i/i. Enrichment analysis indicated that stress and cell control related processes were enriched. The expression of ascorbate peroxidase (APX) and most of the heat shock proteins (HSPs) were decreased, which might result in higher sensitivity to various stresses in pollen cells. A model was proposed to summarize the underlying process for f5-caused hybrid male sterility. These results would provide significant clues to further dissecting the molecular mechanism of f5-caused inter-subspecific reproductive isolation.

Mechanisms involved in the death of steatotic WIF-B9 hepatocytes co-exposed to benzo[a]pyrene and ethanol: a possible key role for xenobiotic metabolism and nitric oxide.

We previously demonstrated that co-exposing pre-steatotic hepatocytes to benzo[a]pyrene (B[a]P), a carcinogenic environmental pollutant, and ethanol, favored cell death. Here, the intracellular mechanisms underlying this toxicity were studied. Steatotic WIF-B9 hepatocytes, obtained by a 48h-supplementation with fatty acids, were then exposed to B[a]P/ethanol (10 nM/5 mM, respectively) for 5 days. Nitric oxide (NO) was demonstrated to be a pivotal player in the cell death caused by the co-exposure in steatotic hepatocytes. Indeed, by scavenging NO, CPTIO treatment of co-exposed steatotic cells prevented not only the increase in DNA damage and cell death, but also the decrease in the activity of CYP1, major cytochrome P450s of B[a]P metabolism. This would then lead to an elevation of B[a]P levels, thus possibly suggesting a long-lasting stimulation of the transcription factor AhR. Besides, as NO can react with superoxide anion to produce peroxynitrite, a highly oxidative compound, the use of FeTPPS to inhibit its formation indicated its participation in DNA damage and cell death, further highlighting the important role of NO. Finally, a possible key role for AhR was pointed out by using its antagonist, CH-223191. Indeed it prevented the elevation of ADH activity, known to participate to the ethanol production of ROS, notably superoxide anion. The transcription factor, NFκB, known to be activated by ROS, was shown to be involved in the increase in iNOS expression. Altogether, these data strongly suggested cooperative mechanistic interactions between B[a]P via AhR and ethanol via ROS production, to favor cell death in the context of prior steatosis.

Acylsugar amount and fatty acid profile differentially suppress oviposition by western flower thrips, Frankliniella occidentalis, on tomato and interspecific hybrid flowers.

Tomatoes (Solanum lycopersicum L.) have been bred to exude higher amounts or different types of the specialized plant metabolites, acylsugars, from type IV trichomes. Acylsugars are known to deter several herbivorous insect pests, including the western flower thrips (WFT), Frankliniella occidentalis (Pergande); however, all previous studies investigated the effect of acylsugars on leaves, or acylsugar extracts obtained from leaves. In spite of the WFT predilection for flowers, there is a gap in knowledge about flower defenses against thrips damage. This is especially important in light of their capacity to acquire and inoculate viruses in the genus Orthotospovirus, such as Tomato spotted wilt orthotospovirus (TSWV), in flowers. Therefore, we turned our attention to assessing thrips oviposition differences on flowers of 14 entries, including 8 interspecific hybrids, 5 tomato lines bred for specific acylsugar-related characteristics (type IV trichome densities, acylsugar amount, sugar moiety and fatty acid profile), and a fresh market tomato hybrid, Mt. Spring, which only produces trace amounts of acylsugars. Our results show that the density of the acylsugar droplet bearing type IV trichomes is greatest on sepals, relative to other flower structures, and accordingly, WFT avoids oviposition on sepals in favor of trichome-sparse petals. In concordance with past studies, acylsugar amount was the most important acylsugar-related characteristic suppressing WFT oviposition. Certain acylsugar fatty acids, specifically i-C5, i-C9 and i-C11, were also significantly associated with changes in WFT oviposition. These results support continued breeding efforts to increase acylsugar amounts and explore modifications of fatty acid profile and their roles in deterring thrips oviposition. The finding that acylsugar production occurs and reduces thrips oviposition in tomato flowers will be important in efforts to use acylsugar-mediated resistance to reduce incidence of orthotospoviruses such as TSWV in tomato by deterring virus transmission and development of thrips vector populations in the crop.

The hepatoprotective effects of Radix Bupleuri extracts against D-galactosamine/lipopolysaccharide induced liver injury in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

The present study is aiming at evaluating the hepatoprotective of Radix Bupleuri extracts (RBE) on the d-galactosamine/lipopolysaccharide (D-GalN/LPS) induced liver injury of hybrid grouper in vitro and in vivo. In vitro, RBE (0, 200, 400 and 800 µg/ml) was added to the hybrid grouper primary hepatocytes before (pretreatment) the incubation of the hepatocytes with D-GalN (20 mM) plus LPS (1 µg/ml) in the culture medium. RBE at concentrations of 200, 400 and 800 µg/ml significantly improved cell viability and inhibited the elevation of TNF-α, IL-1ß and IL-6 and significantly down-regulated the caspase-3, caspase-9 and P53 mRNA levels. In vivo administration of RBE at the doses of 0, 200, 400, 800 and 1600 mg/kg in the diet for 8 weeks prior to D-GalN (500 mg/kg) and LPS (20 µg/kg) intoxication. The study indicated that the RBE not only ameliorated liver injury, as evidenced by well-preserved liver architecture, but also significantly increased hepatic antioxidant enzymes activities in the D-GalN/LPS-induced liver injury animal model. Further demonstrating the protective effects of the RBE, we found that pretreatment with the RBE up-regulated the expression of antioxidant genes (GPx and MnSOD), while down-regulated apoptosis-related genes (caspase-3, caspase-9 and P53), immune related genes (MHC2 and TLR3) and pro-inflammatory cytokines (TOR and IKKα) mRNA expression in the liver of hybrid grouper. In brief, the present study showed that RBE can protect hepatocyte injury induced by D-GalN/LPS through elevating antioxidant enzyme activity and suppressing apoptosis and immune inflammatory responses. The results support the use of RBE as a hepatoprotective in fish.

Productivity and Antioxidant Activity of Wild, Reconstituted, and Hybrid Strains of the Pink Oyster Mushroom, Pleurotus djamor (Agaricomycetes), from Mexico.

The genus Pleurotus is the third most commonly produced edible fungi in the world. In addition, species of genus Pleurotus have functional properties such as anticancer, antiviral, antimicrobial, anti-inflammatory, and antioxidant activities, which are mainly attributed to phenolic compounds. For these reasons, this study evaluated the productivity and antioxidant activity (AA) of 2 wild strains (white and pink), 2 reconstituted strains (called "BB" and "RR"), and 4 hybrid strains (H1, H2, H3, and H4) of P. djamor from monokaryotic components (neohaplonts). The results showed that the white wild-type strain and the reconstituted strains exhibited the best production potential, expressed as biological efficiency and mycelial growth rate. The carpophores of hybrid strains H1 and H3 had the greatest AA, as evaluated with DPPH radical scavenging and reducing power assays, respectively. The H3 strain had the highest total phenol (TP) content. Pearson correlations led us to conclude that the mycelial growth rate has a regular inverse correlation with TP and a regular direct correlation with AA of methanolic extracts from carpophores and myce-lia. This is, to our knowledge, the first report in the literature about the effect of Pleurotus strain hybridization through a chemical de-dikaryotization process on TP content.