Conductive Polymer-Based Interlayers in Restraining the Polysulfide Shuttle of Lithium-Sulfur Batteries.

Lithium-sulfur batteries (LSBs) are considered a promising candidate for next-generation energy storage devices due to the advantages of high theoretical specific capacity, abundant resources and being environmentally friendly. However, the severe shuttle effect of polysulfides causes the low utilization of active substances and rapid capacity fading, thus seriously limiting their practical application. The introduction of conductive polymer-based interlayers between cathodes and separators is considered to be an effective method to solve this problem because they can largely confine, anchor and convert the soluble polysulfides. In this review, the recent progress of conductive polymer-based interlayers used in LSBs is summarized, including free-standing conductive polymer-based interlayers, conductive polymer-based interlayer modified separators and conductive polymer-based interlayer modified sulfur electrodes. Furthermore, some suggestions on rational design and preparation of conductive polymer-based interlayers are put forward to highlight the future development of LSBs.

Genome-Wide Analysis of Long Non-Coding RNAs Related to UV-B Radiation in the Antarctic Moss Pohlia nutans.

Antarctic organisms are consistently suffering from multiple environmental pressures, especially the strong UV radiation caused by the loss of the ozone layer. The mosses and lichens dominate the vegetation of the Antarctic continent, which grow and propagate in these harsh environments. However, the molecular mechanisms and related regulatory networks of these Antarctic plants against UV-B radiation are largely unknown. Here, we used an integrated multi-omics approach to study the regulatory mechanism of long non-coding RNAs (lncRNAs) of an Antarctic moss (Pohlia nutans) in response to UV-B radiation. We identified a total of 5729 lncRNA sequences by transcriptome sequencing, including 1459 differentially expressed lncRNAs (DELs). Through functional annotation, we found that the target genes of DELs were significantly enriched in plant-pathogen interaction and the flavonoid synthesis pathway. In addition, a total of 451 metabolites were detected by metabonomic analysis, and 97 differentially change metabolites (DCMs) were found. Flavonoids account for 20% of the total significantly up-regulated metabolites. In addition, the comprehensive transcriptome and metabolome analyses revealed the co-expression pattern of DELs and DCMs of flavonoids. Our results provide insights into the regulatory network of lncRNA under UV-B radiation and the adaptation of Antarctic moss to the polar environments.

Taxonomic Identification of the Arctic Strain Nocardioides Arcticus Sp. Nov. and Global Transcriptomic Analysis in Response to Hydrogen Peroxide Stress.

Microorganisms living in polar regions rely on specialized mechanisms to adapt to extreme environments. The study of their stress adaptation mechanisms is a hot topic in international microbiology research. In this study, a bacterial strain (Arc9.136) isolated from Arctic marine sediments was selected to implement polyphasic taxonomic identification based on factors such as genetic characteristics, physiological and biochemical properties, and chemical composition. The results showed that strain Arc9.136 is classified to the genus Nocardioides, for which the name Nocardioides arcticus sp. nov. is proposed. The ozone hole over the Arctic leads to increased ultraviolet (UV-B) radiation, and low temperatures lead to increased dissolved content in seawater. These extreme environmental conditions result in oxidative stress, inducing a strong response in microorganisms. Based on the functional classification of significantly differentially expressed genes under 1 mM H2O2 stress, we suspect that Arc9.136 may respond to oxidative stress through the following strategies: (1) efficient utilization of various carbon sources to improve carbohydrate transport and metabolism; (2) altering ion transport and metabolism by decreasing the uptake of divalent iron (to avoid the Fenton reaction) and increasing the utilization of trivalent iron (to maintain intracellular iron homeostasis); (3) increasing the level of cell replication, DNA repair, and defense functions, repairing DNA damage caused by H2O2; (4) and changing the composition of lipids in the cell membrane and reducing the sensitivity of lipid peroxidation. This study provides insights into the stress resistance mechanisms of microorganisms in extreme environments and highlights the potential for developing low-temperature active microbial resources.

Complete genome sequencing and comparison of two nitrogen-metabolizing bacteria isolated from Antarctic deep-sea sediment.

BACKGROUND: Bacteria are an essential component of the earth`s biota and affect circulation of matters through their metabolic activity. They also play an important role in the carbon and nitrogen cycle in the deep-sea environment. In this paper, two strains from deep-sea sediments were investigated in order to understand nitrogen cycling involved in the deep-sea environment. RESULTS: In this paper, the basic genomic information of two strains was obtained by whole genome sequencing. The Cobetia amphilecti N-80 and Halomonas profundus 13 genome sizes are 4,160,095 bp with a GC content of 62.5% and 5,251,450 bp with a GC content of 54.84%. Through a comparison of functional analyses, we predicted the possible C and N metabolic pathways of the two strains and determined that Halomonas profundus 13 could use more carbon sources than Cobetia amphilecti N-80. The main genes associated with N metabolism in Halomonas profundus 13 are narG, narY, narI, nirS, norB, norC, nosZ, and nirD. On the contrast, nirD, using NH4+ for energy, plays a main role in Cobetia amphilecti N-80. Both of them have the same genes for fixing inorganic carbon: icd, ppc, fdhA, accC, accB, accD, and accA. CONCLUSION: In this study, the whole genomes of two strains were sequenced to clarify the basic characteristics of their genomes, laying the foundation for further studying nitrogen-metabolizing bacteria. Halomonas profundus 13 can utilize more carbon sources than Cobetia amphilecti N-80, as indicated by API as well as COG and KEGG prediction results. Finally, through the analysis of the nitrification and denitrification abilities as well as the inorganic carbon fixation ability of the two strains, the related genes were identified, and the possible metabolic pathways were predicted. Together, these results provide molecular markers and theoretical support for the mechanisms of inorganic carbon fixation by deep-sea microorganisms.

Source and Distribution of Emerging and Legacy Persistent Organic Pollutants in the Basins of the Eastern Indian Ocean.

Persistent organic pollutants (POPs) have received significant and ongoing attention. To establish favorable regulatory policies, it is vital to investigate the occurrence, source, and budgets of POPs worldwide. POPs including phthalic acid esters (PAEs), organophosphate esters (OPEs), brominated flame retardants (BFRs), and highly chlorinated flame retardants (HFRs) have not yet been examined in the Eastern Indian Ocean (EIO). In this study, the distribution of POPs has been investigated from surface sediments with the depth of 4369-5742 m in the Central Indian Ocean Basin (CIOB) and Wharton Basin (WB) of EIO. The average (±SD) concentrations of ∑11PAEs, ∑11OPEs, ∑4 BFRs, and ∑5HFRs were 1202.0 ± 274.36 ng g-1 dw, 15.3 ± 7.23 ng g-1 dw, 327.6 ± 211.74 pg g-1 dw, and 7.9 ± 7.45 pg g-1 dw, respectively. The high abundance of low-molecular-weight (LMW) PAEs, chlorinated OPEs, LMW BDEs, and anti-Dechlorane Plus indicated the pollution characteristics in the EIO. Correlation analysis demonstrated that LMW compounds may be derived from the high-molecular-weight compounds. The monsoon circulation, currents, and Antarctic Bottom Water may be the main drivers. POP accumulation rate, depositional flux, and mass inventory in the Indian Ocean were also estimated.

The flavonoid 3'-hydroxylase gene from the Antarctic moss Pohlia nutans is involved in regulating oxidative and salt stress tolerance.

Flavonoids are the important secondary metabolites. They are thought to play an important role in plant adaptation to terrestrial environment. However, the downstream branching pathway of flavonoids in bryophytes, which are the most ancient of terrestrial plants, remains unclear. Here, we cloned a flavonoid 3'-hydroxylase gene (PnF3'H) from the Antarctic moss Pohlia nutans and studied its function in plant stress tolerance. The Arabidopsis with overexpressing PnF3'H (AtOE) were constructed. The AtOE plants had more lateral roots and higher activities of antioxidant enzymes than the wild-type plants under oxidative stress. Meanwhile, the gene expression levels of reactive oxygen species (ROS) scavengers (i.e., AtCAT3, AtFeSOD1, and AtCu-ZnSOD3) were upregulated in the AtOE plants, and the transcription levels of ROS producing enzyme genes were significantly downregulated. The AtOE plans showed increased sensitivity to NaCl stress or abscisic acid (ABA) treatment during seed germination and early root development. Furthermore, several stress-resistant genes in the ABA signaling pathway were also downregulated in the AtOE plants when compared with the wild-type plants. These results suggested that PnF3'H participates in regulating the oxidative tolerance and ABA sensitivity to enable P. nutans to adapt to polar environments.

A Novel Ag@AgCl Nanoparticle Synthesized by Arctic Marine Bacterium: Characterization, Activity and Mechanism.

An additive- and pollution-free method for the preparation of biogenic silver and silver chloride nanoparticles (Ag@AgCl NPs) was developed from the bacteria Shewanella sp. Arc9-LZ, which was isolated from the deep sea of the Arctic Ocean. The optimal synthesizing conditions were explored, including light, pH, Ag+ concentration and time. The nanoparticles were studied by means of ultraviolet-visible (UV-Vis) spectrophotometry, energy dispersive spectrometry (EDS), X-ray diffraction (XRD) and inductively coupled plasma optical emission spectrometers (ICP-OES). The transmission electron microscope (TEM) showed that the nanoparticles were spherical and well dispersed, with particle sizes less than 20.00 nm. With Ag@AgCl nanoparticles, the kinetic rate constants for congo red (CR) and rhodamine B (RhB) dye degradation were 2.74 × 10-1 min-1 and 7.78 × 10-1 min-1, respectively. The maximum decolourization efficiencies of CR and RhB were 93.36% and 99.52%, respectively. Ag@AgCl nanoparticles also showed high antibacterial activities against the Gram-positive and Gram-negative bacteria. The Fourier transform infrared spectroscopy (FTIR) spectrum indicated that the O-H, N-H and -COO- groups in the supernatant of Arc9-LZ might participate in the reduction, stabilization and capping of nanoparticles. We mapped the schematic diagram on possible mechanisms for synthesizing Ag@AgCl NPs.

Insights into the Jasmonate Signaling in Basal Land Plant Revealed by the Multi-Omics Analysis of an Antarctic Moss Pohlia nutans Treated with OPDA.

12-oxo-phytodienoic acid (OPDA) is a biosynthetic precursor of jasmonic acid and triggers multiple biological processes from plant development to stress responses. However, the OPDA signaling and relevant regulatory networks were largely unknown in basal land plants. Using an integrated multi-omics technique, we investigated the global features in metabolites and transcriptional profiles of an Antarctic moss (Pohlia nutans) in response to OPDA treatment. We detected 676 metabolites based on the widely targeted metabolomics approach. A total of 82 significantly changed metabolites were observed, including fatty acids, flavonoids, phenolic acids, amino acids and derivatives, and alkaloids. In addition, the transcriptome sequencing was conducted to uncover the global transcriptional profiles. The representative differentially expressed genes were summarized into functions including Ca2+ signaling, abscisic acid signaling, jasmonate signaling, lipid and fatty acid biosynthesis, transcription factors, antioxidant enzymes, and detoxification proteins. The integrated multi-omics analysis revealed that the pathways of jasmonate and ABA signaling, lipid and fatty acid biosynthesis, and flavonoid biosynthesis might dominate the molecular responses to OPDA. Taken together, these observations provide insights into the molecular evolution of jasmonate signaling and the adaptation mechanisms of Antarctic moss to terrestrial habitats.

High-speed silicon Michelson interferometer modulator and streamlined IMDD PAM-4 transmission of Mach-Zehnder modulators for the 2 µm wavelength band.

We demonstrate high-speed silicon modulators optimized for operating at the wavelength of 2 µm. The Mach-Zehnder interferometer (MZI) carrier-depletion modulator with 2 mm phase shifter has a single-arm modulation efficiency (Vπ ·Lπ) of 2.89 V·cm at 4 V reverse bias. Using a push-pull configuration it operates at a data rate of 25 Gbit/s OOK with an extinction ratio of 6.25 dB. We also proposed a mathematically-analysed streamlined IMDD PAM-4 scheme and successfully demonstrated a 25 Gbit/s datarate PAM-4 with the same 2 mm modulator. A Michelson interferometer carrier-depletion modulator with 0.5 mm phase shift length has also been shown with modulation efficiency (Vπ ·Lπ) of 1.36 V·cm at 4 V reverse bias and data rate of 20 Gbit/s OOK. The Michelson interferometer modulator performs similarly to a Mach-Zehnder modulator with twice the phase shifter length.

Co-design of a differential transimpedance amplifier and balanced photodetector for a sub-pJ/bit silicon photonics receiver.

This paper presents the design and implementation of a fully differential optical receiver, which is aimed for short reach intensity modulation and direct detection (IMDD) transceiver links. A Si-Ge balanced photodetector (PD) has been co-designed and packaged with a novel differential transimpedance amplifier (TIA). The TIA design is realized with a standard 28 nm CMOS process and operates with a standard digital supply (1V). Without using any equalization or DSP techniques, the proposed receiver can operate up to 54 Gb/s with a BER less than the KP4 limit (2.2×10-4) under an optical modulation amplitude (OMA) of -8.6 dBm, while the power efficiency has been optimized to 0.55 pJ/bit (0.98 pJ/bit if output buffer is included).

Isolation, characterization and transcriptome analysis of a novel Antarctic Aspergillus sydowii strain MS-19 as a potential lignocellulosic enzyme source.

BACKGROUND: With the growing demand for fossil fuels and the severe energy crisis, lignocellulose is widely regarded as a promising cost-effective renewable resource for ethanol production, and the use of lignocellulose residues as raw material is remarkable. Polar organisms have important value in scientific research and development for their novelty, uniqueness and diversity. RESULTS: In this study, a fungus Aspergillus sydowii MS-19, with the potential for lignocellulose degradation was screened out and isolated from an Antarctic region. The growth profile of Aspergillus sydowii MS-19 was measured, revealing that Aspergillus sydowii MS-19 could utilize lignin as a sole carbon source. Its ability to synthesize low-temperature lignin peroxidase (Lip) and manganese peroxidase (Mnp) enzymes was verified, and the properties of these enzymes were also investigated. High-throughput sequencing was employed to identify and characterize the transcriptome of Aspergillus sydowii MS-19. Carbohydrate-Active Enzymes (CAZyme)-annotated genes in Aspergillus sydowii MS-19 were compared with those in the brown-rot fungus representative species, Postia placenta and Penicillium decumbens. There were 701CAZymes annotated in Aspergillus sydowii MS-19, including 17 cellulases and 19 feruloyl esterases related to lignocellulose-degradation. Remarkably, one sequence annotated as laccase was obtained, which can degrade lignin. Three peroxidase sequences sharing a similar structure with typical lignin peroxidase and manganese peroxidase were also found and annotated as haem-binding peroxidase, glutathione peroxidase and catalase-peroxidase. CONCLUSIONS: In this study, the fungus Aspergillus sydowii MS-19 was isolated and shown to synthesize low-temperature lignin-degrading enzymes: lignin peroxidase (Lip) and manganese peroxidase (Mnp). These findings provide useful information to improve our understanding of low-temperature lignocellulosic enzyme production by polar microorganisms and to facilitate research and applications of the novel Antarctic Aspergillus sydowii strain MS-19 as a potential lignocellulosic enzyme source.

Molecular characterization and functional analysis of four teleostean K+ channels in macrophages of sea perch (Lateolabrax japonicas).

Potassium ion channels are one of the most diversely and widely distributed channels, which are involved in all kinds of physiological functions in both excitable and non-excitable cells. The expression of voltage-gated potassium ion (Kv) channels is highly variable according to the state of macrophages activation. Macrophages have an important function in innate immunity against intruding pathogens. They produce a variety of inflammatory and immunoactive molecules that modulate imflammatory responses. Here we show that blockade of K+ channels by non-selective Kv channel inhibitor tetraethylammonium chloride (TEA), and 4-aminopyridine (4-AP) inhibited proinflammatory cytokines expression, cell proliferation, and reactive oxygen species (ROS) production in LPS-stimulated macrophages of Sea perch (Lateolabrax japonicas). Then we isolated four Kv channels genes (spKv1.1, spKv1.2, spKv1.5 and spKv3.1) in LPS-activated fish macrophages. These channels genes were up-regulated after LPS stimulation except spKv3.1, which remained unchanged during the test. The results of this study indicate that Kv channels could be required for modulating the immune function of fish macrophages.

Enhanced dipicolinic acid production during the stationary phase in Bacillus subtilis by blocking acetoin synthesis.

Bacterial bio-production during the stationary phase is expected to lead to a high target yield because the cells do not consume the substrate for growth. Bacillus subtilis is widely used for bio-production, but little is known about the metabolism during the stationary phase. In this study, we focused on the dipicolinic acid (DPA) production by B. subtilis and investigated the metabolism. We found that DPA production competes with acetoin synthesis and that acetoin synthesis genes (alsSD) deletion increases DPA productivity by 1.4-fold. The mutant showed interesting features where the glucose uptake was inhibited, whereas the cell density increased by approximately 50%, resulting in similar volumetric glucose consumption to that of the parental strain. The metabolic profiles revealed accumulation of pyruvate, acetyl-CoA, and the TCA cycle intermediates in the alsSD mutant. Our results indicate that alsSD-deleted B. subtilis has potential as an effective host for stationary-phase production of compounds synthesized from these intermediates.

Mechanisms of Gills Response to Cadmium Exposure in Greenfin Horse-Faced Filefish (Thamnaconus septentrionalis): Oxidative Stress, Immune Response, and Energy Metabolism.

Cadmium (Cd) pollution has become a global issue due to industrial and agricultural developments. However, the molecular mechanism of Cd-induced detrimental effects and relevant signal transduction/metabolic networks are largely unknown in marine fishes. Here, greenfin horse-faced filefish (Thamnaconus septentrionalis) were exposed to 5.0 mg/L Cd up to 7 days. We applied both biochemical methods and multi-omics techniques to investigate how the gills respond to Cd exposure. Our findings revealed that Cd exposure caused the formation of reactive oxygen species (ROS), which in turn activated the MAPK and apoptotic pathways to alleviate oxidative stress and cell damage. Glycolysis, protein degradation, as well as fatty acid metabolism might assist to meet the requirements of nutrition and energy under Cd stress. We also found that long-term (7 days, "long-term" means compared to 12 and 48 h) Cd exposure caused the accumulation of succinate, which would in turn trigger an inflammatory response and start an immunological process. Moreover, ferroptosis might induce inflammation. Overall, Cd exposure caused oxidative stress, energy metabolism disturbance, and immune response in greenfin horse-faced filefish. Our conclusions can be used as references for safety risk assessment of Cd to marine economic fishes.

Next-generation sequencing-based transcriptome profiling analysis of Pohlia nutans reveals insight into the stress-relevant genes in Antarctic moss.

Genome-wide characterization of the Pohlia nutans transcriptome is essential for clarifying the role of stress-relevant genes in Antarctic moss adapting to the extreme polar environment. High-throughput Illumina sequencing was used to analyze the gene expression profile of P. nutans after cold treatment. A total of 93,488 unigenes, with an average length of 405 bp, were obtained. Gene annotation showed that 16,781 unigenes had significant similarity to known functional protein-coding genes, most of which were annotated using the GO, KOG and KEGG pathway databases. Global profiling of the differentially expressed genes revealed that 3,796 unigenes were significantly upregulated after cold treatment, while 1,405 unigenes were significantly downregulated. In addition, 816 receptor-like kinases and 1,309 transcription factors were identified from P. nutans. This overall survey of transcripts and stress-relevant genes can contribute to understanding the stress-resistance mechanism of Antarctic moss and will accelerate the practical exploitation of the genetic resources for this organism.

Taxonomic identification and temperature stress tolerance mechanisms of Aequorivita marisscotiae sp. nov.

The deep sea harbours microorganisms with unique life characteristics and activities due to adaptation to particular environmental conditions, but the limited sample collection and pure culture techniques available constrain the study of deep-sea microorganisms. In this study, strain Ant34-E75 was isolated from Antarctic deep-sea sediment samples and showed the highest 16 S rRNA gene sequence similarity (97.18%) with the strain Aequorivita viscosa 8-1bT. Strain Ant34-E75 is psychrotrophic and can effectively increase the cold tolerance of Chlamydomonas reinhardtii (a model organism). Subsequent transcriptome analysis revealed multiple mechanisms involved in the Ant34-E75 response to temperature stress, and weighted gene co-expression network analysis (WGCNA) showed that the peptidoglycan synthesis pathway was the key component. Overall, this study provides insights into the characteristics of a deep-sea microorganism and elucidates mechanisms of temperature adaptation at the molecular level.

Spatial distribution, source identification and flux estimation of polycyclic aromatic hydrocarbons and organochlorine pesticides in basins of the Eastern Indian Ocean.

Although polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCPs) have been recorded worldwide, information on their presence in the Eastern Indian Ocean (EIO), especially south of 10°S, remains limited. We investigated the distribution and depositional fluxes of PAHs and OCPs, and the major sources and ecological risks of PAHs in EIO surface sediments from the Central Indian Ocean (CIOB) and Wharton Basin (WB). The concentration of Σ18 PAHs and ∑10 OCPs had an average value (± SD) of 138.4 ± 52.34 and 0.8 ± 0.20 ng g-1, respectively. PAHs may mainly affected by traffic emission and biomass and wood combustion. Persistent organic pollutant accumulation rate (PAR) and depositional flux (DF) values showed that abundant PAHs might lost during top-down transport. The low trans- chordane (CHL)/cis-CHL ratio and PAR of OCPs may indicated few OCPs were inputted into the EIO recently. The results of binary isotope mixing modeling indicate the predominance of marine organic matter (MOM) in total organic carbon (TOC) of sediments. Fluoranthene (Flour) and pyrene (Py) might have potential biological effects in the EIO. The study provided background values for PAHs and OCPs in the Indian Ocean, and preliminarily revealed the fate of POPs in the open oceans.

Estimating the Spatial Distribution and Future Conservation Requirements of the Spotted Seal in the North Pacific.

Local adaptation has been increasingly involved in the designation of species conservation strategies to response to climate change. Marine mammals, as apex predators, are climatechange sensitive, and their spatial distribution and conservation requirements are critically significant for designing protection strategies. In this study, we focused on an ice-breeding marine mammal, the spotted seal (Phoca largha), which exhibits distinct morphological and genetic variations across its range. Our objectives were to quantify the ecological niches of three spotted seal populations, construct the species-level model and population-level models that represent different regions in the Bering population (BDPS), Okhotsk population (ODPS) and southern population (SDPS), and conduct a conservation gap analysis. Our findings unequivocally demonstrated a clear niche divergence among the three populations. We predicted habitat contraction for the BDPS and ODPS driven by climate change; in particular, the spotted seals inhabiting Liaodong Bay may face breeding habitat loss. However, most spotted seal habitats are not represented in existing marine protected areas. Drawing upon these outcomes, we propose appropriate conservation policies to effectively protect the habitat of the different geographical populations of spotted seals. Our research addresses the importance of incorporating local adaptation into species distribution modeling to inform conservation and management strategies.

Insulin detemir enhances proglucagon gene expression in the intestinal L cells via stimulating ß-catenin and CREB activities.

Insulin therapy using insulin detemir (d-INS) has demonstrated weight-sparing effects compared with other insulin formulations. Mechanisms underlying these effects, however, remain largely unknown. Here we postulate that the intestinal tissues' selective preference allows d-INS to exert enhanced action on proglucagon (Gcg) expression and the production of glucagon-like peptide (GLP)-1, an incretin hormone possessing both glycemia-lowering and weight loss effects. To test this hypothesis, we used obese type 2 diabetic db/db mice and conducted a 14-day intervention with daily injection of a therapeutic dose of d-INS or human insulin (h-INS) in these mice. The body weight of the mice after 14-day daily injection of d-INS (5 IU/kg) was decreased significantly compared with those injected with the same dose of h-INS or saline. The weight-sparing effect of d-INS was associated with significantly elevated circulating levels of total GLP-1 and reduced food intake. Histochemistry analysis demonstrated that d-INS induced rapid phosphorylation of protein kinase B (Akt) in the gut L cells of normal mice. Western blotting showed that d-INS stimulated Akt activation in a more rapid and enhanced fashion in the mouse distal ileum compared with those by h-INS. In vitro investigation in primary fetal rat intestinal cell (FRIC) cultures showed that d-INS increased Gcg mRNA expression as determined by Northern blotting and real-time RT-PCR. Consistent with these in vivo investigations, d-INS significantly increased GLP-1 secretion in FRIC cultures. Consistently, d-INS was also shown to induce rapid phosphorylation of Akt in the clonal gut cell line GLUTag. Furthermore, d-INS increased ß-catenin phosphorylation, its nuclear translocation, and enhanced cAMP response element-binding protein (CREB) phosphorylation in a phosphatidylinositol 3-kinase and/or mitogen-activated protein kinase kinase/extracellular signal-regulated kinase-sensitive manner. We suggest that the weight-sparing benefit of d-INS in mice is related to its intestinal tissues preference that leads to profound stimulation of Gcg expression and enhanced GLP-1 secretion in intestinal L cells, potentially involving the activation of insulin/ß-catenin/CREB signaling pathways.

Validation of housekeeping genes for gene expression studies in an ice alga Chlamydomonas during freezing acclimation.

Antarctic ice alga Chlamydomonas sp. ICE-L can endure extreme low temperature and high salinity stress under freezing conditions. To elucidate the molecular acclimation mechanisms using gene expression analysis, the expression stabilities of ten housekeeping genes of Chlamydomonas sp. ICE-L during freezing stress were analyzed. Some discrepancies were detected in the ranking of the candidate reference genes between geNorm and NormFinder programs, but there was substantial agreement between the groups of genes with the most and the least stable expression. RPL19 was ranked as the best candidate reference genes. Pairwise variation (V) analysis indicated the combination of two reference genes was sufficient for qRT-PCR data normalization under the experimental conditions. Considering the co-regulation between RPL19 and RPL32 (the most stable gene pairs given by geNorm program), we propose that the mean data rendered by RPL19 and GAPDH (the most stable gene pairs given by NormFinder program) be used to normalize gene expression values in Chlamydomonas sp. ICE-L more accurately. The example of FAD3 gene expression calculation demonstrated the importance of selecting an appropriate category and number of reference genes to achieve an accurate and reliable normalization of gene expression during freeze acclimation in Chlamydomonas sp. ICE-L.