Ligand-Receptor Interaction-Induced Intracellular Phase Separation: A Global Disruption Strategy for Resistance-Free Lethality of Pathogenic Bacteria.

Addressing the global challenge of bacterial resistance demands innovative approaches, among which multitargeting is a widely used strategy. Current strategies of multitargeting, typically achieved through drug combinations or single agents inherently aiming at multiple targets, face challenges such as stringent pharmacokinetic and pharmacodynamic requirements and cytotoxicity concerns. In this report, we propose a bacterial-specific global disruption approach as a vastly expanded multitargeting strategy that effectively disrupts bacterial subcellular organization. This effect is achieved through a pioneering chemical design of ligand-receptor interaction-induced aggregation of small molecules, i.e., DNA-induced aggregation of a diarginine peptidomimetic within bacterial cells. These intracellular aggregates display affinity toward various proteins and thus substantially interfere with essential bacterial functions and rupture bacterial cell membranes in an "inside-out" manner, leading to robust antibacterial activities and suppression of drug resistance. Additionally, biochemical analysis of macromolecule binding affinity, cytoplasmic localization patterns, and bacterial stress responses suggests that this bacterial-specific intracellular aggregation mechanism is fundamentally different from nonselective classic DNA or membrane binding mechanisms. These mechanistic distinctions, along with the peptidomimetic's selective permeation of bacterial membranes, contribute to its favorable biocompatibility and pharmacokinetic properties, enabling its in vivo antimicrobial efficacy in several animal models, including mice-based superficial wound models, subcutaneous abscess models, and septicemia infection models. These results highlight the great promise of ligand-receptor interaction-induced intracellular aggregation in achieving a globally disruptive multitargeting effect, thereby offering potential applications in the treatment of malignant cells, including pathogens, tumor cells, and infected tissues.

Enhanced Insecticidal Activity of Chlorfenapyr against Spodoptera frugiperda by Reshaping the Intestinal Microbial Community and Interfering with the Metabolism of Iron-Based Metal-Organic Frameworks.

Spodoptera frugiperda (S. frugiperda) is an invasive pest that threatens global crop production and food security and poses a serious threat to maize production worldwide. Metal-organic framework (MOF) nanocarriers have great potential for agricultural pest control applications. The present study successfully prepared the chemical cross-linking of iron-based metal-organic framework nanoparticles (MIL-101(Fe)-NH2 NPs) with sodium lignosulfonate (SL) as a pH/laccase double stimuli-responsive pesticide release system. The average particle size of the prepared chlorfenapyr (CF)-loaded nanoparticles (CF@MIL-101-SL NPs) was 161.54 nm, and the loading efficiency was 44.52%. Bioactivity assays showed that CF@MIL-101-SL NPs increased the toxicity of CF to S. frugiperda and caused the rupture of the peritrophic membrane and enlargement of the midgut. Data from 16S rRNA gene sequencing showed that CF@MIL-101-SL treatment reduced the resistance of S. frugiperda to pesticides and pathogens and affected nutrient and energy availability by remodeling the intestinal microbiota of S. frugiperda. The dysregulated microbial community interacted with the broken peritrophic membrane, which exacerbated damage to the host. Nontargeted metabolomic results showed that ABC transporters may be a potential mechanism for the enhanced toxicity of CF@MIL-101-SL to S. frugiperda. In summary, the present study provides effective strategies for toxicological studies of nanopesticides against insects.

Strengthening a Medium-Carbon Low-Alloy Steel by Nanosized Grains: The Role of Asymmetrical Rolling.

A medium-carbon low-alloy steel was prepared via the asymmetric rolling process with different ratios of upper and down roll velocities. Subsequently, the microstructure and mechanical properties were explored by using SEM, EBSD, TEM, tensile tests and nanoindentation. The results show that asymmetrical rolling (ASR) can significantly improve strength while retaining good ductility compared with conventional symmetrical rolling. The yield strength and tensile strength of the ASR-steel are 1292 ± 10 MPa and 1357 ± 10 MPa, respectively, which are higher than the values of 1113 ± 10 MPa and 1185 ± 10 MPa for the SR-steel. The ASR-steel retains good ductility of 16.5 ± 0.5%. The significant increase in strength is related to the joint actions of the ultrafine grains, dense dislocations and a large number of nanosized precipitates. This is mainly because of the introduction of extra shear stress on the edge under asymmetric rolling, which induces gradient structural changes hence increasing the density of geometrically necessary dislocations.

Otus scops adenovirus: the complete genome sequence of a novel aviadenovirus discovered in a wild owl.

We present the complete genome sequence of an aviadenovirus obtained by metagenomics from cloacal swabs taken from a free-living Eurasian scops owl (Otus scops, a small raptor distributed in Europe and several parts of Asia) in China. Thirty protein coding genes were predicted in this 40,239-bp-long genome, which encodes the largest fiber protein among all reported aviadenoviruses. The viral genome sequence is highly divergent, and the encoded proteins have an average of only 55% amino acid sequence identity to those of other adenoviruses. In phylogenetic analysis, the new owl virus grouped with members of the genus Aviadenovirus and formed a common clade with another owl adenovirus reported previously in Japan. This is the second complete genome sequence of an aviadenovirus discovered in owls, and its proteins have an average of 62% amino acid sequence identity to those of the previously reported owl adenovirus. Combining this result with comparative genomic analysis of all aviadenoviruses, we propose that this owl virus and the previously described Japanese owl adenovirus can be assigned to two new species in the genus Aviadenovirus. This study provides new data on the diversity of aviadenoviruses in wild birds.

Whole genome analysis of a novel adenovirus discovered from Oriolus chinesis.

We present the first complete genome sequence of an aviadenovirus Oriolus adenovirus (OrAdV) sequenced from the cloaca of a Oriolus chinensis (a passerine bird widely distributed in Asia), which was collected from an island off the east coast of China. Thirty-one protein coding genes were predicted in this 40425-bp-long genome. OrAdV genome is highly divergent and has only 57% average protein identity compared with other aviadenovirus genomes. Comparative genomic analysis indicates that this passerine virus is a new species of aviadenovirus. One unique thymidylate kinase gene was discovered in OrAdV genome. This gene is absent in other adenovirus genomes and usually reported to occur in herpesvirus. Protein sequence alignment against all known proteins indicates that this gene may be originated from ancient horizontal gene transfer event between virus and parasitic eukaryote like protozoan. This new aviadenovirus genome enriches the genomic information of adenovirus and suggests that there is a large unknown space of adenovirus world.

Analyses of open-access multi-omics data sets reveal genetic and expression characteristics of maize ZmCCT family genes.

Flowering in maize (Zea mays) is influenced by photoperiod. The CO, CO-like/COL and TOC1 (CCT) domain protein-encoding genes in maize, ZmCCTs, are particularly important for photoperiod sensitivity. However, little is known about CCT protein-encoding gene number across plant species or among maize inbred lines. Therefore, we analysed CCT protein-encoding gene number across plant species, and characterized ZmCCTs in different inbred lines, including structural variations (SVs), copy number variations (CNVs), expression under stresses, dark-dark (DD) and dark-light (DL) cycles, interaction network and associations with maize quantitative trait loci (QTLs) by referring to the latest v4 genome data of B73. Gene number varied greatly across plant species, more in polyploids than in diploids. The numbers of ZmCCTs identified were 58 in B73, 59 in W22, 48 in Mo17, and 57 in Huangzao4 for temperate maize inbred lines, and 68 in tropical maize inbred line SK. Some ZmCCTs underwent duplications and presented chromosome collinearity. Structural variations and CNVs were found but they had no germplasm specificity. Forty-two ZmCCTs responded to stresses. Expression of 37 ZmCCTs in embryonic leaves during seed germination of maize under DD and DL cycles was roughly divided into five patterns of uphill pattern, downhill-pattern, zigzag-pattern, └-pattern and ⅃-pattern, indicating some of them have a potential to perceive dark and/or dark-light transition. Thirty-three ZmCCTs were co-expressed with 218 other maize genes; and 24 ZmCCTs were associated with known QTLs. The data presented in this study will help inform further functions of ZmCCTs.

New Enantiomers of a Nor-Bisabolane Derivative and Two New Phthalides Produced by the Marine-Derived Fungus Penicillium chrysogenum LD-201810.

Marine-derived fungi are a treasure house for the discovery of structurally novel secondary metabolites with potential pharmaceutical value. In this study, a pair of new nor-bisabolane derivative enantiomers (±)-1 and two new phthalides (4 and 5), as well as four known metabolites, were isolated from the culture filtrate of the marine algal-derived endophytic fungus Penicillium chrysogenum LD-201810. Their structures were established by detailed interpretation of spectroscopic data (1D/2D NMR and ESI-MS). The optical resolution of compound (±)-1 by chiral HPLC successfully afforded individual enantiomers (+)-1 and (-)-1, and their absolute configurations were determined by TDDFT-ECD calculations. Compound (±)-1 represents the first example of bisabolane analogs with a methylsulfinyl substituent group, which is rare in natural products. All of the isolated compounds 1-7 were evaluated for their cytotoxic activity against A549, BT-549, HeLa, HepG2, MCF-7, and THP-1 cell lines, as well as for antifungal activity against four plant pathogenetic fungi (Alternaria solani, Botrytis cinerea, Fusarium oxysporum, and Valsa mali). Compound 2, a bisabolane-type sesquiterpenoid, was shown to possess excellent activity for control of B. cinerea with half-maximal inhibitory concentration (IC50) of 13.6 µg/mL, whereas the remaining investigated compounds showed either weak or no cytotoxic/antifungal activity in this study.

PM2.5 in poultry houses synergizes with Pseudomonas aeruginosa to aggravate lung inflammation in mice through the NF-κB pathway.

BACKGROUND: High concentrations of particulate matter less than 2.5 µm in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. OBJECTIVES: In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. METHODS: High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. RESULTS: Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. CONCLUSIONS: The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.

Cytotoxic Secondary Metabolites Isolated from the Marine Alga-Associated Fungus Penicillium chrysogenum LD-201810.

A new pentaketide derivative, penilactonol A (1), and two new hydroxyphenylacetic acid derivatives, (2'R)-stachyline B (2) and (2'R)-westerdijkin A (3), together with five known metabolites, bisabolane-type sesquiterpenoids 4-6 and meroterpenoids 7 and 8, were isolated from the solid culture of a marine alga-associated fungus Penicillium chrysogenum LD-201810. Their structures were elucidated based on extensive spectroscopic analyses, including 1D/2D NMR and high resolution electrospray ionization mass spectra (HRESIMS). The absolute configurations of the stereogenic carbons in 1 were determined by the (Mo2(OAc)4)-induced circular dichroism (CD) and comparison of the calculated and experimental electronic circular dichroism (ECD) spectra, while the absolute configuration of the stereogenic carbon in 2 was established using single-crystal X-ray diffraction analysis. Compounds 2 and 3 adapt the 2'R-configuration as compared to known hydroxyphenylacetic acid-derived and O-prenylated natural products. The cytotoxicity of 1-8 against human carcinoma cell lines (A549, BT-549, HeLa, HepG2, MCF-7, and THP-1) was evaluated. Compound 3 exhibited cytotoxicity to the HepG2 cell line with an IC50 value of 22.0 µM. Furthermore, 5 showed considerable activities against A549 and THP-1 cell lines with IC50 values of 21.2 and 18.2 µM, respectively.

New Cyclic Diarylheptanoids from Platycarya strobilacea.

Five new cyclic diarylheptanoids (platycary A-E, compounds 1-5) and three previously identified analogues (i.e., phttyearynol (compound 6), myricatomentogenin (compound 7), and juglanin D (compound 8)) were isolated from the stem bark of Platycarya strobilacea. The structures of these compounds were determined using NMR, HRESIMS, and electronic circular dichroism (ECD) data. The cytotoxicity of compounds 1-5 and their ability to inhibit nitric oxide (NO) production, as well as protect against the corticosterone-induced apoptosis of Pheochromocytoma (PC12) cells, were evaluated in vitro using the appropriate bioassays. Compounds 1 and 2 significantly inhibited the corticosterone-induced apoptosis of PC12 cells at a concentration of 20 µΜ.

Trends and Health Risks of Dissolved Heavy Metal Pollution in Global River and Lake Water from 1970 to 2017.

Heavy metal pollution in surface water is a global environmental problem. This study analyzed the trends, health risks, and sources of eight dissolved heavy metal species in river and lake water across five continents (Africa, Asia, Europe, North America, and South America; Oceania was excluded owing to a lack of data) for the period 1970-2017. We wanted to assess the effects of various implemented countermeasures to pollution and to determine those that could be adopted worldwide. Collectively, the water system showed increasing trends for Cd, Cr, Cu, Ni, Mn, and Fe and decreasing trends for Pb and Zn. The mean dissolved concentrations of most heavy metals were highest in Asia and lowest in Europe. Most heavy metals had low non-carcinogenic risks over this period. The cancer risks associated with Pb were lower than the hazardous level on all five continents over the five decades, whereas the cancer risks related to Cr exceeded the hazardous level in the 1970s, 2000s, and 2010s, and in Africa, Asia, and North America over the entire period. Mining and manufacturing were consistently found to be critical sources of metal pollution from 1970 to 2017. However, the heavy metal sources differed significantly by continent, with waste discharge and rock weathering dominant in Africa; mining and manufacturing, along with rock weathering, are dominant in Asia and South America; fertilizer and pesticide use, along with rock weathering, are dominant in North America; and mining and manufacturing, waste discharge, and rock weathering are dominant in Europe. Global trends in the metal loadings in water and in relevant pollution-control measures suggest that countermeasures in Europe have successfully controlled heavy metal pollution. The successful measures include implementing rigorous standards for metal emissions, limiting the metal concentrations in products, and rigorously treating metal-contaminated waste. Therefore, the measures implemented in Europe should be extended worldwide to treat heavy metal pollution in water.

Malignant catarrhal fever: An emerging yet neglected disease in captive sika deer (Cervus nippon) herds in China.

Malignant catarrhal fever (MCF) is a fatal lymphoproliferative disease that represents a serious problem in the deer-rearing industry. To better understand an MCF-like disease that has emerged in northern China since 2015, we investigated ten cases by documenting clinical and epidemiological data and analysing causative agents and histopathological changes. In addition, a retrospective screen for Macavirus DNA and a questionnaire-based survey were conducted. Epizootic MCF in Chinese sika deer herds has emerged with a low morbidity of 3.8% (95% CI: 2.5%-5.1%) and a high mortality of 93.2% (95% CI: 86.6%-99.9%). The disease course varied from 3 to 12 days. Aetiologically, OvHV-2 was predominant in the MCFV, accounting for most MCF cases (21/23). In contrast, only two CpHV-2 isolates were phylogenetically closely related to CpHV-2. Diarrhoea and nasal discharges were the most frequent manifestations, although clinical signs varied in some cases. Pathologically typical lesions of haemorrhage, necrosis and lymphoid cell infiltration were readily observed in a variety of organs. Vasculitis caused by vascular and perivascular lymphoid cell infiltration was common. The retrospective survey suggested a low positive rate (3/275) of MCFV DNA in peripheral blood lymphocytes (PBL). The questionnaire-based survey suggested the disease was neglected by local veterinarians, who did not acknowledge the risk of co-rearing deer with reservoir species. Collectively, the emerging epizootic MCF in Chinese sika deer herds remains neglected, emphasizing the urgency of initiating full-field diagnoses and control strategies.

Cassava AGPase genes and their encoded proteins are different from those of other plants.

MAIN CONCLUSION: Cassava AGPase and AGPase genes have some unique characteristics. ADP-glucose pyrophosphorylase (AGPase) is a rate-limiting enzyme for starch synthesis. In this study, cassava AGPase genes (MeAGP) were analyzed based on six cultivars and one wild species. A total of seven MeAGPs was identified, including four encoding AGPase large subunits (MeAGPLs 1, 2, 3 and 4) and three encoding AGPase small subunits (MeAGPSs 1, 2 and 3). The copy number of MeAGPs varied in cassava germplasm materials. There were 14 introns for MeAGPLs 1, 2 and 3, 13 introns for MeAGPL4, and 8 introns for other three MeAGPSs. Multiple conservative amino acid sequence motifs were found in the MeAGPs. There were differences in amino acids at binding sites of substrates and regulators among different MeAGP subunits and between MeAGPs and a potato AGPase small subunit (1YP2:B). MeAGPs were all located in chloroplasts. MeAGP expression was not only associated with gene copy number and types/combinations, regions and levels of the DNA methylation but was also affected by environmental factors with the involvement of various transcription factors in multiple regulation networks and in various cis-elements in the gene promoter regions. The MeAGP activity also changed with environmental conditions and had potential differences among the subunits. Taken together, MeAGPs differ in number from those of Arabidopsis, potato, maize, banana, sweet potato, and tomato.

Bioaccumulation in Miscanthus sacchariflorus grown on cadmium-contaminated sediments: a comparative study between submerged and non-submerged environments.

Bioaccumulation of heavy metals in aquatic plants is significantly affected by hydrological regime and therefore the accumulation and translocation of cadmium in five organs-panicle, leaf, stem, root, and bud-of an emergent plant (Miscanthus sacchariflorus) were compared between the submerged environment and non-submerged environment. In the submerged condition, the cadmium concentration was higher in the panicle and leaf than in the stem, root, and bud. Cadmium concentration in the root exhibited a positive regression with cadmium concentration in the sediment. However, cadmium concentration in the panicle, leaf, stem, and bud exhibited no significant regression with cadmium concentration in the sediment. In the non-submerged environment, the cadmium concentration was higher in the below-ground organs than in the aboveground organs. The mean bioaccumulation coefficient in the 24 investigated plots in the submerged environment was higher than that in the 20 and 40 mg kg-1 cadmium treatments in the non-submerged environment. The mean translocation factor in the submerged environment was nine times higher than that in non-submerged environment. These results indicate that submergence enhanced cadmium bioaccumulation in the aboveground organs and that this plant can be used to remove heavy metals from polluted rivers and lakes.

MePHD1 as a PHD-Finger Protein Negatively Regulates ADP-Glucose Pyrophosphorylase Small Subunit1a Gene in Cassava.

ADP-glucose pyrophosphorylase (AGPase) is an important enzyme in the starch synthesis pathway. Its enzyme activity can determine the efficiency of starch biosynthesis. Cassava (Manihot esculenta Crantz) is the main staple crop worldwide and has a high starch content in its storage root. However, the inner regulatory mechanism of AGPase gene family is unclear. MePHD1; a plant homeodomain transcription factor; was isolated through a yeast one-hybrid screening using the promoter of ADP-glucose pyrophosphorylase small subunit1a (MeAGPS1a) as bait, and cassava storage root cDNA library as prey. This factor could bind to the MeAGPS1a promoter in vitro and in vivo, and its predicted binding region ranged from -400 bp to -201 bp, at the translation initiation site. The transcript level of MePHD1 could be induced by five plant hormones, and a temperature of 42 °C. This was down-regulated during the maturation process of the storage root. MePHD1 protein could repress the promoter activity of MeAGPS1a gene by a dual-luciferase assay; which indicated that MePHD1 is a negative regulator for the transcript level of MeAGPS1a gene.

Evidence of two genetically different lymphotropic herpesviruses present among red deer, sambar, and milu herds in China.

Herpesvirus infections in Cervidae are a serious threat affecting some deer species worldwide. In our attempt to identify malignant catarrhal fever-associated herpesviruses in deer herds, ten gammaherpesviral DNA fragments were identified in five species of deer in herds in China by using a pan-herpesvirus polymerase chain reaction assay targeting viral DNA polymerase. Notably, in sambar (Rusa unicolor), a novel gamma-2 herpesvirus was identified that showed a close relationship with fallow deer lymphotropic herpesvirus (LHV), while the other fragments were phylogenetically grouped together with Elk-LHV. Determination of whether these viruses have any clinical implication in these deer species should be undertaken urgently.

Napabucasin versus placebo in refractory advanced colorectal cancer: a randomised phase 3 trial.

BACKGROUND: Napabucasin is a first-in-class cancer stemness inhibitor that targets STAT3, which is a poor prognostic factor in colorectal cancer. This study aimed to test napabucasin in advanced colorectal cancer. METHODS: This study was a double-blind randomised phase 3 trial done at 68 centres in Canada, Australia, New Zealand, and Japan. Patients with advanced colorectal cancer with a good Eastern Cooperative Oncology Group (ECOG) performance status (0-1) for whom all available standard therapies had failed were eligible for the study. Patients were randomly assigned (1:1) to receive placebo or napabucasin through a web-based system with a permuted block method, after stratification by ECOG performance status, KRAS status, previous VEGF inhibitor treatment, and time from diagnosis of metastatic disease. Napabucasin 480 mg or matching placebo was taken orally every 12 h. All patients received best supportive care. The primary endpoint was overall survival assessed in an intention-to-treat analysis. This is the final analysis of this trial, which is registered at ClinicalTrials.gov, number NCT01830621. FINDINGS: Accrual began on April 15, 2013, and was stopped for futility on May 23, 2014, at which point 282 patients had undergone randomisation (138 assigned to the napabucasin group and 144 to the placebo group). Overall survival did not differ significantly between groups: median overall survival was 4·4 months (95% CI 3·7-4·9) in the napabucasin group and 4·8 months (4·0-5·3) in the placebo group (adjusted hazard ratio [HR] 1·13, 95% CI 0·88-1·46, p=0·34). The safety population included 136 patients in the napabucasin group and 144 patients in the placebo group. More patients who received napabucasin had any grade of treatment-related diarrhoea (108 [79%] of 136 patients), nausea (69 [51%]), and anorexia (52 [38%]) than did patients who received placebo (28 [19%] of 144 patients, 35 [24%], and 23 [16%], respectively). The most common severe (grade 3 or worse) treatment-related adverse events were abdominal pain (five [4%] patients receiving napabucasin vs five [3%] receiving placebo), diarrhoea (21 [15%] vs one [1%]), fatigue (14 [10%] vs eight [6%]), and dehydration (six [4%] vs one [1%]). 251 (89%) patients had data on pSTAT3 expression, of whom 55 (22%) had pSTAT3-positive tumours (29 in the napabucasin group, 26 in the placebo group). In a prespecified biomarker analysis of pSTAT3-positive patients, overall survival was longer in the napabucasin group than in the placebo group (median 5·1 months [95% CI 4·0-7·5] vs 3·0 months [1·7-4·1]; HR 0·41, 0·23-0·73, p=0·0025). INTERPRETATION: Although there was no difference in overall survival between groups in the overall unselected population, STAT3 might be an important target for the treatment of colorectal cancer with elevated pSTAT3 expression. Nevertheless, these results require validation. FUNDING: Canadian Cancer Society Research Institute and Boston Biomedical.

Effect of water cadmium concentration and water level on the growth performance of Salix triandroides cuttings.

The growth performance of Salix triandroides cuttings at three water cadmium (Cd) concentrations (0, 20, and 40 mg L-1) and three water levels (- 40 cm, water level 40 cm below the soil surface; 0 cm, water level even with the soil surface; and 40 cm, water level 40 cm above soil surface) was investigated to evaluate its potential in phytoextraction strategies. Compared to cuttings in the - 40 or 0 cm water levels, cuttings in the 40 cm water level showed significantly lower biomass, height, and adventitious root length and significantly fewer leaves and adventitious roots. However, these growth and morphological parameters were not different among the three water Cd concentrations. Water level decreased stomatal conduction and transpiration rate but showed no significant effects on chlorophyll concentration or photosynthetic rate. Chlorophyll concentration and stomatal conductance were higher at 40 mg L-1 Cd treatment than at 0 or 20 mg L-1 Cd treatment; yet, photosynthetic rate and transpiration rate were not different. Cd concentration in the leaves and stems increased as the water level increased, but the highest Cd concentration in the roots occurred in the 0 cm water level. As water Cd concentration increased, Cd concentration in the leaves, stems, and roots increased in all three water levels, except in stems in the - 40 cm water level. Under Cd stress, cuttings in the - 40 or 0 cm water levels were characterized by a higher bioaccumulation coefficient, but a lower translocation factor, than those in the 40 cm water level. However, the bioaccumulation coefficient increased with increasing water Cd concentration in all three water levels, as did the translocation factor in the 40 cm water level. The tolerance index for the cuttings was the same among all water levels and water Cd concentrations. The results clearly indicated that the low water level increased plant growth and Cd accumulation in underground parts, while the high water level decreased plant growth but increased Cd accumulation in aboveground parts.

A proteomic-based investigation of potential copper-responsive biomarkers: Proteins, conceptual networks, and metabolic pathways featuring Penicillium janthinellum from a heavy metal-polluted ecological niche.

Filamentous fungi-copper (Cu) interactions are very important in the formation of natural ecosystems and the bioremediation of heavy metal pollution. However, important issues at the proteome level remain unclear. We compared six proteomes from Cu-resistant wild-type (WT) Penicillium janthinellum strain GXCR and a Cu-sensitive mutant (EC-6) under 0, 0.5, and 3 mmol/L Cu treatments using iTRAQ. A total of 495 known proteins were identified, and the following conclusions were drawn from the results: Cu tolerance depends on ATP generation and supply, which is relevant to glycolysis pathway activity; oxidative phosphorylation, the TCA cycle, gluconeogenesis, fatty acid synthesis, and metabolism are also affected by Cu; high Cu sensitivity is primarily due to an ATP energy deficit; among ATP generation pathways, Cu-sensitive and Cu-insensitive metabolic steps exist; gluconeogenesis pathway is crucial to the survival of fungi in Cu-containing and sugar-scarce environments; fungi change their proteomes via two routes (from ATP, ATP-dependent RNA helicases (ADRHs), and ribosome biogenesis to proteasomes and from ATP, ADRHs to spliceosomes and/or stress-adapted RNA degradosomes) to cope with changes in Cu concentrations; and unique routes exist through which fungi respond to high environmental Cu. Further, a general diagram of Cu-responsive paths and a model theory of high Cu are proposed at the proteome level. Our work not only provides the potential protein biomarkers that indicate Cu pollution and targets metabolic steps for engineering Cu-tolerant fungi during bioremediation but also presents clues for further insight into the heavy metal tolerance mechanisms of other eukaryotes.

New 30-Noroleanane Triterpenoid Saponins from Holboellia coriacea Diels.

Three new 30-noroleanane triterpenoid saponins, akebonoic acid 28-O-ß-d-glucopyranosyl-(1''→6')-ß-d-glucopyranosyl ester (1), akebonoic acid 28-O-(6''-O-caffeoyl)-ß-d-glucopyranosyl-(1''→6')-ß-d-glucopyranosyl ester (Holboelliside A, 2) and 3ß,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 3-O-(6'-O-caffeoyl)-ß-d-glucopyranoside (Holboelliside B, 3) were isolated from the stems of Holboellia coriacea Diels, together with five known compounds, eupteleasaponin VIII (4), 3α-akebonoic acid (5), quinatic acid (6), 3ß-hydroxy-30-norhederagenin (7) and quinatoside A (8). The structures of these compounds were determined on the basis of spectral and chemical evidence. Compounds 1-5 were evaluated for their inhibitory activity against three human tumors HepG2, HCT116 and SGC-7901 cell lines in vitro.