Serotonin and dopamine regulate the aggressiveness of swimming crabs (Portunus trituberculatus) in different ways.

Bioamines act as a pivotal part in the regulation of aggressive behavior in animals as a type of neuroendocrine, but the patterns of how they regulate aggressiveness in crustaceans are still unclear due to species-specific responses. To determine the effects of serotonin (5-HT) and dopamine (DA) on the aggressiveness of swimming crabs (Portunus trituberculatus), we quantified their behavioral and physiological characteristics. The results showed that an injection of 5-HT at 0.5 mmol L-1 and 5 mmol L-1 could significantly enhance the aggressiveness of swimming crabs, as well as an injection of DA at 5 mmol L-1. The regulation of 5-HT and DA on aggressiveness is dose-dependent, and these two bioamines have different concentration thresholds that can trigger aggressiveness changes. 5-HT could up-regulate the 5-HTR1 gene expression and increase lactate content at the thoracic ganglion as the aggressiveness enhances, suggesting that 5-HT may activate related receptors and neuronal excitability to regulate aggressiveness. As a result of DA injection at 5 mmol L-1, lactate content in the chela muscle and hemolymph increased, glucose content in the hemolymph increased, and the CHH gene was significantly up-regulated. Pyruvate kinase and hexokinase enzyme activities in the hemolymph increased, which accelerated the glycolysis process. These results demonstrate that DA regulates the lactate cycle, which provides substantial short-term energy for aggressive behavior. Both 5-HT and DA can mediate aggressive behavior in the crab by activating calcium regulation in muscle tissue. We conclude that the enhancement of aggressiveness is a process of energy consumption, in which 5-HT acts on the central nervous system to induce aggressive behavior, and DA affects muscle and hepatopancreas tissue to provide a large amount of energy. This study expands upon the knowledge of regulatory mechanisms of aggressiveness in crustaceans and offers a theoretical foundation for enhancing crab culture management.

The Effect of Intruder Density on Territoriality and Dominance in Male Swimming Crab (Portunus trituberculatus).

Territorial behavior of animals is affected by numerous factors, one being the number of intruders. The swimming crab (Portunus trituberculatus), an important commercial and ecological species on the continental shelf of Asia, usually needs to defend its territory from intrusion by other crabs, especially in habitats with high densities of conspecifics. To clarify the underlying patterns of how P. trituberculatus protects its territory, we assessed the territorial behavior of occupant crabs (territory holders) when presented with different numbers of intruders using an indoor observation system. We calculated the territory size of the occupants and quantified their behavioral responses to intruders. With an increased number of intruders, the territory size the occupants owned significantly decreased, and their behavior adjusted accordingly. Besides, the territorial behavior score, reflecting the territoriality of crab, decreased significantly. Furthermore, in a high density group that had seven intruders, the occupants showed a higher dominance hierarchy than the intruders, indicating the ascendancy of occupants in territorial competition with intruders. These results revealed that as the number of intruders increased, the territory size of P. trituberculatus shrunk because the fight for territory became more intense.

Epigenetic inactivation of hydroxymethylglutaryl CoA synthase reduces ketogenesis and facilitates tumor cell motility in clear cell renal carcinoma.

Previously, we have reported that the dysregulation of ketogenesis plays an important role in the carcinogenesis of clear cell renal cell carcinoma (ccRCC). Here, we demonstrate decreased expression of the HMGCS2 gene in ccRCC, a critical enzyme for the synthesis of the ketone body ß-hydroxybutyrate (ß-OHB). We found that the reduced transcription of the HMGCS2 gene in ccRCC cells was significantly correlated to a higher relative methylation rate in its promotor region. The higher methylation rate in the region of the transcription start site and 1st exon of the HMGCS2 gene was, in turn, correlated with a worse clinical outcome for patients. The transcription of HMGCS2 was possible to restore by treatment with 5-aza-2'-deoxycytidine and with the histone deacetylase inhibitor ß-OHB. Therefore, the low levels of the HMGCS2 enzyme in ccRCC may be the consequence of hypermethylation of the HMGCS2 promotor. The ensuing reduction in the ketone body levels further suppresses the transcription of HMGCS2 via a feedback loop. Ectopic expression of HMGCS2 attenuates the migration and invasion of ccRCC but does not affect the proliferative capacity of ccRCC cells in vitro. In addition, we showed that ectopic expression of HMGCS2 boosts the intracellular levels of ß-OHB and that exogenously applied ß-OHB suppresses the motility and invasion of ccRCC. Our study reveals crosstalk between genes that regulate metabolism and their metabolites, thus providing a better understanding of the epigenetic mechanism involved in ccRCC carcinogenesis and suggesting opportunities for metabolic therapy of tumors. Initially, we suggest that the mRNA level of HMGCS2 could serve as a potentially valuable diagnostic (AUC = 0.918, p < 0.001) and prognostic biomarker.

Epigenetic Inactivation of Acetyl-CoA Acetyltransferase 1 Promotes the Proliferation and Metastasis in Nasopharyngeal Carcinoma by Blocking Ketogenesis.

The dysregulation of epigenetic modification and energy metabolism cooperatively contribute to the tumorigenesis of nasopharyngeal carcinoma (NPC). However, the detailed mechanisms underlying their joint contribution to NPC development and progression remain unclear. Here, we investigate the role of Acy1 Coenzyme A Acyltransferases1 (ACAT1), a key enzyme in the metabolic pathway of ketone bodies, in the proliferation and metastasis of NPC and to elucidate the underlying molecular mechanisms. Ketogenesis, plays a critical role in tumorigenesis. Previously, we reported two enzymes involved in ketone body metabolism mediate epigenetic silencing and act as tumor suppressor genes in NPC. Here, we identify another key enzyme, Acetyl-CoA acetyltransferase 1 (ACAT1), and show that its transcriptional inactivation in NPC is due to promoter hypermethylation. Ectopic overexpression of ACAT1 significantly suppressed the proliferation and colony formation of NPC cells in vitro. The migratory and invasive capacity of NPC cells was inhibited by ACAT1. The tumorigenesis of NPC cells overexpressing ACAT1 was decreased in vivo. Elevated ACAT1 in NPC cells was accompanied by an elevated expression of CDH1 and a reduced expression of vimentin and SPARC, strongly indicating that ACAT1 is involved in regulating epithelial-mesenchymal transition (EMT). We also found that ACAT1 contributes to increased intracellular levels of ß-hydroxybutyrate (ß-HB). Exogenously supplied ß-HB significantly inhibits the growth of NPC cells in a dose-dependent manner. In summary, ACAT1 may function as a tumor suppressor via modulation of ketogenesis and could thus serve as a potential therapeutic target in NPC. In summary, our data suggest that regulation of ketogenesis may serve as adjuvant therapy in NPC.

Inverse correlation of miR-27a-3p and CDH5 expression serves as a diagnostic biomarker of proliferation and metastasis of clear cell renal carcinoma.

BACKGROUND: Cadherin-5 (CDH5) is aberrantly expressed in a variety of human cancers and plays an important role in angiogenesis. The present study provides further insight into the role of miR-27a-3p in the regulation of CDH5 expression in renal clear cell carcinoma (ccRCC). METHODS: Thedysregulation of CDH5 expression in ccRCC and its association with clinicopathological characteristics were analyzed using the TCGA database. A meta-analysis was performed to verify the alteration of CDH5 expression in ccRCC using the GEO database. Quantitative RT-PCR and immunohistochemical staining were applied to assess the transcriptional and protein levels of CDH5. TargetScan and Tarbase were employed to predict the miRNAs with the potential to target mRNA of CDH5. RESULTS: The mRNA level of CDH5 in ccRCCwas significantly higher than in normal tissue. CDH5 mRNA expression could therefore serve as a potential diagnostic biomarker for ccRCC (AUC = 0.844). However, the reduced CDH5 transcription levels were significantly correlated with patients in the T3-4 stage, lymph node, and distant metastasis, as well as with a worse clinical outcome. We further observed that CDH5, at the protein level, was almost absent in ccRCC samples. In addition, a few databases screen showed that mir-27a-3p is a highly conserved miRNA targeting CDH5. The expression of mir-27a-3p was significantly elevated in ccRCC tissues in contrast to normal tissues. Importantly, it was positively associated with the T3-4 stage and M stage, respectively, suggesting that the expression level of mir-27a-3p could serve as a diagnostic biomarker for ccRCC (AUC = 0.775). CONCLUSION: Our data suggest that thereduced translational level of CDH5 in ccRCC was related to the overexpression of mir-27a-3p. The higher mir-27a-3p and lower CDH5 expression significantly correlated with advanced clinical stages for ccRCC patients.

Insight into AMPK regulation mechanism in vivo and in vitro: Responses to low temperatures in the olive flounder Paralichthys olivaceus.

The olive flounder, Paralichthys olivaceus, is a commercially important maricultured fish in China, Japan, and Korea. Low winter temperatures influence its survival and growth and affect the output of the aquaculture industry. Energy metabolism is essential for fish survival, and the central energy-regulating factor - 5'-AMP-activated protein kinase (AMPK) - plays an important role in responses to cold stress. However, the mechanism of AMPK pathway regulation in fish coping with cold stress remains poorly understood. In the present study, the expression of AMPK and its upstream (LKB1 and CaMKKß) and downstream genes (SITR1, FOXO1A, and TFAM) in the brain, muscle, and heart was analyzed while the flounder was under cold stress (0.2 ± 0.2 °C). The results showed that low temperatures activated LKB1, CaMKKß, and AMPK genes in the brain, and the activated AMPK induced expression of SITR1, FOXO1A, and TFAM. In the muscle tissue, the expression patterns of these genes presented a trend of initially decreasing and then increasing, and there was a delay in the response to low temperatures. At the cellular level, comparative analysis of the effects of the activator 5-aminoimidazole-4-carboxamide1-ß-D-ribofuranoside (AICAR) and inhibitor compound C of the AMPK pathway demonstrated that cold stress was similar to AICAR, which activated the AMPK pathway with hysteresis. Thus, the regulation mechanism of AMPK under cold stress was preliminarily analyzed. In general, AMPK was involved not only in responses to low temperatures but also in energy regulation under cold stress.

Network of microRNA-transcriptional factor-mRNA in cold response of turbot Scophthalmus maximus.

The aim of this study is to understand fish cold-tolerant mechanism. We analyzed the transcriptional reactions to the cold condition in turbot Scophthalmus maximus by using RNA-seq and microRNA (miRNA)-seq. Meio-gynogenetic diploid turbots were treated at 0 °C to distinguish the cold-tolerant (CT) and cold-sensitive (CS) groups. The results showed that there were quite different responses at both mRNA and miRNA levels, with more up-regulated mRNAs (1069 vs. 194) and less down-regulated miRNAs (4 vs. 1) in CT versus CS relative to the control group. The network of miRNA-transcription factor-mRNA, regulating turbot different response to cold stress, was constructed, which involved in cell cycle, component of cell membrane, signal transduction, and circadian rhythm pathways. The above information demonstrates mechanisms by which cold tolerance is increased in fish.

Dysregulation of Ketone Body Metabolism Is Associated With Poor Prognosis for Clear Cell Renal Cell Carcinoma Patients.

Kidney is an important organ for ketone body metabolism. However, the role of abnormal ketone metabolism and its possible function in tumorigenesis of clear cell renal cell carcinoma (ccRCC) have not yet been elucidated. Three differentially expressed key enzymes involved in ketone body metabolism, ACAT1, BDH2, and HMGCL, were screened out between ccRCC and normal kidney tissues using the GEO and TCGA databases.We confirmed that the transcription and protein expression of ACAT1, BDH2, and HMGCL were significantly lower in ccRCC by real-time RT-PCR and IHC assays. Those patients with lower expression of these three genes have a worse outcome. In addition, we demonstrated that ectopic expression of each of these genes inhibited the proliferation of ccRCC cells. The overexpressed ACAT1 and BDH2 genes remarkably impeded the migratory and invasive capacity of ccRCC cells. Furthermore, exogenous ß-hydroxybutyrate suppressed the growth of ccRCC cells in vitro in a dose-dependent manner. Our findings suggest that ACAT1, BDH2, and HMGCL are potential tumor suppressor genes, and constitute effective prognostic biomarkers for ccRCC. Ketone body metabolism might thus be a promising target in a process for developing novel therapeutic approaches to treat ccRCC.

Insight into the heat resistance of fish via blood: Effects of heat stress on metabolism, oxidative stress and antioxidant response of olive flounder Paralichthys olivaceus and turbot Scophthalmus maximus.

High temperature has direct confinement on fish survival and growth, especially under the background of global warming. Selection of fish line with heat resistance is an important means to address this problem. In the present study, we analyzed the difference in heat resistance between families of olive flounder Paralichthys olivaceus and turbot Scophthalmus maximus, two flatfish species occupying slightly different thermal niches. Then the chosen families were tested to determine their differential response to heat stress (ΔT = +8 °C and +12 °C) in blood, including anaerobic metabolism (lactate), oxidative stress (lipid peroxidation and protein carbonylation) and antioxidant enzymes. Results showed a difference in heat resistance between families of the two species. Among the chosen parameters, growth traits had a significant effect on contents of lactate and malondialdehyd (MDA), and activities of catalase (CAT) and glutathione S-transferase in flounder (P < 0.05), and on MDA content and CAT activity in turbot (P < 0.05). In comparison with heat-sensitive family of each species, levels of all studied parameters were lower and more stable in heat-resistant families after heat stress. What's more, heat resistance of fish significantly influenced contents of lactate and MDA and activity of CAT in flounder (P < 0.05), as well as contents of lactate, MDA and carbonyl and activity of superoxide dismutate (SOD) in turbot (P < 0.05). These results demonstrated that such physiological phenotypes as anaerobic metabolism, oxidative stress and antioxidant enzymes are good biomarkers of fish heat-resistance, being potentially valuable in fish breeding. However, these markers should be applied with more caution when there is a growth discrepancy between fish families.

Hypothermal effects on survival, energy homeostasis and expression of energy-related genes of swimming crabs Portunus trituberculatus during air exposure.

Previously, dry or semi-dry approach under the hypothermal condition is proved to be an alternative method in transport of live swimming crabs Portunus trituberculatus. However, we wondered whether this method can improve crab survival when temperature is kept as cool as possible. In this study, we hypothesized that there is a thermal threshold below which dry or semi-dry approach (air exposure) could cause crab physiological disruption and therefore aggravate their mortality. To test the above hypothesis, crabs (23°C) were exposed to air at temperatures ranging from 4 to 16°C. Results showed that crabs had a worse survival and vigor at temperatures below 12°C. Then we tested crab energy metabolism to explore the possible reason. It was shown that total adenine nucleotide and adenylate energy charge in gills were remarkably reduced by air exposure of below 12°C. This increased the need for crabs to re-balance energy metabolism, which was indicated by the upregulation of AMPKα and HIF-1α. Meanwhile, there was a significant increase of the expression of Na(+)/K(+)-ATPase, V-type ATPase and HSP90 at temperatures below 12°C, while all treatments shared a similar level of hemocyanin, urate and lactate in hemolymph and expression of cytochrome c oxidase and NADH-ubiquinone reductase in gills. These results implied that dry or semi-dry approach below 12°C could exert detrimental effects on P. trituberculatus, and perturbation of energy homeostasis, which is more related with changes of energy-demanding physiological pathways, is a possible reason of crab death and poor vigor.

De novo assembly and transcriptome analysis of osmoregulation in Litopenaeus vannamei under three cultivated conditions with different salinities.

Litopenaeus vannamei, one of the most important euryhaline crustaceans, is cultured in seawater, brackish water, and freshwater worldwide. We performed Illumina RNA sequencing of L. vannamei gills, generating 124,914,870; 119,250,450; and 105,487,350 raw reads from the shrimps cultured in seawater, brackish water, and freshwater, respectively. From these reads, 466,293 transcripts were de novo assembled and annotated. Comparative genomic analysis showed that 1752 genes were significantly differentially expressed in the freshwater group compared with the seawater group, including 1242 upregulated and 510 downregulated genes. In addition, 1246 genes were differentially expressed in the brackish group vs. the seawater water group, including 659 upregulated and 587 downregulated genes. These differentially expressed genes were mainly involved in energy metabolism, substance metabolism, ion transport and signal transduction, and genetic process. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyze the functional significance of the differentially expressed genes, included those responding to salinity through diverse biological functions and processes and numerous potential genes associated with the osmotic response. L. vannamei responses to the three cultivated salinities were analyzed using next-generation sequencing. The transcriptional database established from the current research adds to the information available on L. vannamei and the findings expand our knowledge of the molecular basis of osmoregulation mechanisms in this species.

Three-dimensional face recognition in the presence of facial expressions: an annotated deformable model approach.

In this paper, we present the computational tools and a hardware prototype for 3D face recognition. Full automation is provided through the use of advanced multistage alignment algorithms, resilience to facial expressions by employing a deformable model framework, and invariance to 3D capture devices through suitable preprocessing steps. In addition, scalability in both time and space is achieved by converting 3D facial scans into compact metadata. We present our results on the largest known, and now publicly available, Face Recognition Grand Challenge 3D facial database consisting of several thousand scans. To the best of our knowledge, this is the highest performance reported on the FRGC v2 database for the 3D modality.