The lack of PPARα exacerbated the progression of non-alcoholic steatohepatitis in mice with spleen deficiency syndrome by triggering an inflammatory response.

Background: In addition to abnormal liver inflammation, the main symptoms of non-alcoholic steatohepatitis (NASH) are often accompanied by gastrointestinal digestive dysfunction, consistent with the concept of spleen deficiency (SD) in traditional Chinese medicine. As an important metabolic sensor, whether peroxisome proliferator-activated receptor alpha (PPARα) participates in regulating the occurrence and development of NASH with SD (NASH-SD) remains to be explored. Methods: Clinical liver samples were collected for RNA-seq analysis. C57BL/6J mice induced by folium sennae (SE) were used as an SD model. qPCR analysis was conducted to evaluate the inflammation and metabolic levels of mice. PPARα knockout mice (PPARαko) were subjected to SE and methionine-choline-deficient (MCD) diet to establish the NASH-SD model. The phenotype of NASH and the inflammatory indicators were measured using histopathologic analysis and qPCR as well. Results: The abnormal expression of PPARα signaling, coupled with metabolism and inflammation, was found in the results of RNA-seq analysis from clinical samples. SD mice showed a more severe inflammatory response in the liver evidenced by the increases in macrophage biomarkers, inflammatory factors, and fibrotic indicators in the liver. qPCR results also showed differences in PPARα between SD mice and control mice. In PPARαko mice, further evidence was found that the lack of PPARα exacerbated the inflammatory response phenotype as well as the lipid metabolism disorder in NASH-SD mice. Conclusion: The abnormal NR signaling accelerated the vicious cycle between lipotoxicity and inflammatory response in NAFLD with SD. Our results provide new evidence for nuclear receptors as potential therapeutic targets for NAFLD with spleen deficiency.

Physiological and Transcriptome Analyses Revealed the Mechanism by Which Deferoxamine Promotes Iron Absorption in Cinnamomum camphora.

Iron deficiency causes chlorosis and growth inhibition in Cinnamomum camphora, an important landscaping tree species. Siderophores produced by plant growth-promoting rhizobacteria have been widely reported to play an indispensable role in plant iron nutrition. However, little to date has been determined about how microbial siderophores promote plant iron absorption. In this study, multidisciplinary approaches, including physiological, biochemical and transcriptome methods, were used to investigate the role of deferoxamine (DFO) in regulating Fe availability in C. camphora seedlings. Our results showed that DFO supplementation significantly increased the Fe2+ content, SPAD value and ferric-chelate reductase (FCR) activity in plants, suggesting its beneficial effect under Fe deficiency. This DFO-driven amelioration of Fe deficiency was further supported by the improvement of photosynthesis. Intriguingly, DFO treatment activated the metabolic pathway of glutathione (GSH) synthesis, and exogenous spraying reduced glutathione and also alleviated chlorosis in C. camphora. In addition, the expression of some Fe acquisition and transport-related genes, including CcbHLH, CcFRO6, CcIRT2, CcNramp5, CcOPT3 and CcVIT4, was significantly upregulated by DFO treatment. Collectively, our data demonstrated an effective, economical and feasible organic iron-complexing agent for iron-deficient camphor trees and provided new insights into the mechanism by which siderophores promote iron absorption in plants.

Chronic toxicity of PFAS-free AFFF alternatives in terrestrial plant Brassica rapa.

Fluorine (F)-free firefighting foams will be replacing per- and polyfluoroalkyl substances (PFAS)-containing aqueous film-forming foams (AFFFs) at U.S. military installations imminently, yet the environmental impacts of F-free foams are largely unknown. Ecotoxicity assessment of F-free foams is urgently needed to avoid replacement regret. In this study, we comparatively assessed phytotoxicity of six F-free formulations and one current short-chain fluorinated AFFF in terrestrial plant Brassica rapa. Five of six F-free formulations exerted higher toxicity than the short-chain AFFF to the growth and reproduction of B. rapa, with 8-51 times and > 10 times lower EC50 values, respectively. Nontargeted analysis indicated the occurrence of transformation products of the test formulations in the above-ground plant tissues. In agreement with their phytotoxicity, the five highly toxic F-free formulations generated more transformation products with higher peak intensities in plant tissues than the two weakly toxic formulations. The most abundant transformation products detected in plant extracts were suspect transformation products derived from diethylene glycol monobutyl ether, a common ingredient of the five toxic formulations. This study provides ecotoxicological data that, combined with data from all related ongoing research, should be used in decision making regarding recommendations for manufacturing and use of candidate F-free foams.

Comparison of the effects of Tai Chi and general aerobic exercise on weight, blood pressure and glycemic control among older persons with depressive symptoms: a randomized trial.

BACKGROUND: Blood pressure and glycemic control are associated with the management of depressive symptoms in patients with depression. Previous studies have demonstrated that both Tai Chi and aerobic exercise have positive effects on blood pressure and glycemic control. Few studies have compared the physiological effects of Tai Chi versus aerobic exercise in older adults with depressive symptoms. The objective of this study was to compare the effects of Tai Chi and aerobic exercise on weight, body mass index, blood pressure and glycosylated hemoglobin (HbA1c) level in older persons with mild to moderate-severe depressive symptoms. METHODS: A randomized controlled trial was performed. The older persons (age ≥ 60 years old) with depressive symptoms were recruited. Then, participants were randomly allocated to the Tai Chi group and the aerobic exercise group received a 12-week 24-movement Yang's Tai Chi intervention and aerobic exercise, respectively. Data collection occurred at baseline and after completion of the interventions (week 12). RESULTS: A total of 238 participants with mild to moderate-to-severe depressive symptoms were included in the final analysis, including 120 in the Tai Chi group and 118 in the aerobic exercise group. The difference in weight and body mass index in the Tai Chi group was 2.0 kg (Z = -4.930, P < 0.001) and 0.77 kg/m2 (Z = -5.046, P < 0.001) higher than that in the aerobic exercise group, respectively. After the 12-week intervention, the systolic pressure and diastolic pressure in the Tai Chi group were 5.50 mmHg (Z = -2.282, P = 0.022) and 8.0 mmHg (Z = -3.360, P = 0.001) lower than that in the aerobic exercise group, respectively. The difference in HbA1c level in the Tai Chi group was 0.50% higher than that in the aerobic exercise group (Z = -4.446, P < 0.001). CONCLUSION: This study showed that Tai Chi exercise was more effective in improving blood pressure and HbA1c level than general aerobic exercise. It suggested that Tai Chi might be an effective approach for the management of blood pressure and long-term glucose control in older persons with depressive symptoms. TRIAL REGISTRATION: Trial registration: ChiCTR, ChiCTR2100042534 . Registration date: 23/01/2021, http://www.chictr.org.cn/showproj.aspx?proj=120602 .

Characterizing Natural Organic Matter Transformations by Microbial Communities in Terrestrial Subsurface Ecosystems: A Critical Review of Analytical Techniques and Challenges.

Determining the mechanisms, traits, and pathways that regulate microbial transformation of natural organic matter (NOM) is critical to informing our understanding of the microbial impacts on the global carbon cycle. The capillary fringe of subsurface soils is a highly dynamic environment that remains poorly understood. Characterization of organo-mineral chemistry combined with a nuanced understanding of microbial community composition and function is necessary to understand microbial impacts on NOM speciation in the capillary fringe. We present a critical review of the popular analytical and omics techniques used for characterizing complex carbon transformation by microbial communities and focus on how complementary information obtained from the different techniques enable us to connect chemical signatures with microbial genes and pathways. This holistic approach offers a way forward for the comprehensive characterization of the formation, transformation, and mineralization of terrestrial NOM as influenced by microbial communities.

Advances in Molecular Mechanisms for Traditional Chinese Medicine Actions in Regulating Tumor Immune Responses.

Traditional Chinese medicine (TCM) has been developed for thousands of years with its various biological activities. The interest in TCM in tumor prevention and treatment is rising with its synergistic effect on tumor cells and tumor immunosuppressive microenvironment (TIM). Characteristic of TCM fits well within the whole system and multi-target cancer treatment. Herein we discuss the underlying mechanisms of TCM actions in TIM via regulating immunosuppressive cells, including restoring the antigen presentation function of dendritic cells, enhancing NK cells-mediated killing activity, restraining the functions of myeloid cell-derived suppressor cells, and inhibiting cancer-associated fibroblasts. TCM also regulates tumor progression through enhancing immune response, preventing immune escape and inducing cell death of tumor cells, which triggers immune response in nearby cells. In addition, we discuss TCM in clinical applications and the advantages and disadvantages of TCM in cancer prevention and treatment, as well as current therapeutic challenges and strategies. It might be helpful for understanding the therapeutic potential of TCM for cancer in clinic.

Characterization of subsurface media from locations up- and down-gradient of a uranium-contaminated aquifer.

The processing of sediment to accurately characterize the spatially-resolved depth profiles of geophysical and geochemical properties along with signatures of microbial density and activity remains a challenge especially in complex contaminated areas. This study processed cores from two sediment boreholes from background and contaminated core sediments and surrounding groundwater. Fresh core sediments were compared by depth to capture the changes in sediment structure, sediment minerals, biomass, and pore water geochemistry in terms of major and trace elements including pollutants, cations, anions, and organic acids. Soil porewater samples were matched to groundwater level, flow rate, and preferential flows and compared to homogenized groundwater-only samples from neighboring monitoring wells. Groundwater analysis of nearby wells only revealed high sulfate and nitrate concentrations while the same analysis using sediment pore water samples with depth was able to suggest areas high in sulfate- and nitrate-reducing bacteria based on their decreased concentration and production of reduced by-products that could not be seen in the groundwater samples. Positive correlations among porewater content, total organic carbon, trace metals and clay minerals revealed a more complicated relationship among contaminant, sediment texture, groundwater table, and biomass. The fluctuating capillary interface had high concentrations of Fe and Mn-oxides combined with trace elements including U, Th, Sr, Ba, Cu, and Co. This suggests the mobility of potentially hazardous elements, sediment structure, and biogeochemical factors are all linked together to impact microbial communities, emphasizing that solid interfaces play an important role in determining the abundance of bacteria in the sediments.

In silico Identification and Mechanism Exploration of Hepatotoxic Ingredients in Traditional Chinese Medicine.

BACKGROUNDS AND AIMS: Recently, a growing number of hepatotoxicity cases aroused by Traditional Chinese Medicine (TCM) have been reported, causing increasing concern. To date, the reported predictive models for drug induced liver injury show low prediction accuracy and there are still no related reports for hepatotoxicity evaluation of TCM systematically. Additionally, the mechanism of herb induced liver injury (HILI) still remains unknown. The aim of the study was to identify potential hepatotoxic ingredients in TCM and explore the molecular mechanism of TCM against HILI. MATERIALS AND METHODS: In this study, we developed consensus models for HILI prediction by integrating the best single classifiers. The consensus model with best performance was applied to identify the potential hepatotoxic ingredients from the Traditional Chinese Medicine Systems Pharmacology database (TCMSP). Systems pharmacology analyses, including multiple network construction and KEGG pathway enrichment, were performed to further explore the hepatotoxicity mechanism of TCM. RESULTS: 16 single classifiers were built by combining four machine learning methods with four different sets of fingerprints. After systematic evaluation, the best four single classifiers were selected, which achieved a Matthews correlation coefficient (MCC) value of 0.702, 0.691, 0.659, and 0.717, respectively. To improve the predictive capacity of single models, consensus prediction method was used to integrate the best four single classifiers. Results showed that the consensus model C-3 (MCC = 0.78) outperformed the four single classifiers and other consensus models. Subsequently, 5,666 potential hepatotoxic compounds were identified by C-3 model. We integrated the top 10 hepatotoxic herbs and discussed the hepatotoxicity mechanism of TCM via systems pharmacology approach. Finally, Chaihu was selected as the case study for exploring the molecular mechanism of hepatotoxicity. CONCLUSION: Overall, this study provides a high accurate approach to predict HILI and an in silico perspective into understanding the hepatotoxicity mechanism of TCM, which might facilitate the discovery and development of new drugs.

Isolation and characterization of two phosphate-solubilizing fungi from rhizosphere soil of moso bamboo and their functional capacities when exposed to different phosphorus sources and pH environments.

Phosphate-solubilizing fungi (PSF) generally enhance available phosphorus (P) released from soil, which contributes to plants' P requirement, especially in P-limiting regions. In this study, two PSF, TalA-JX04 and AspN-JX16, were isolated from the rhizosphere soil of moso bamboo (Phyllostachys edulis) widely distributed in P-deficient areas in China and identified as Talaromyces aurantiacus and Aspergillus neoniger, respectively. The two PSF were cultured in potato dextrose liquid medium with six types of initial pH values ranging from 6.5 to 1.5 to assess acid resistance. Both PSF were incubated in Pikovskaya's liquid media with different pH values containing five recalcitrant P sources, including Ca3(PO4)2, FePO4, CaHPO4, AlPO4, and C6H6Ca6O24P6, to estimate their P-solubilizing capacity. No significant differences were found in the biomass of both fungi grown in media with different initial pH, indicating that these fungi could grow well under acid stress. The P-solubilizing capacity of TalA-JX04 was highest in medium containing CaHPO4, followed by Ca3(PO4)2, FePO4, C6H6Ca6O24P6, and AlPO4 in six types of initial pH treatments, while the recalcitrant P-solubilizing capacity of AspN-JX16 varied with initial pH. Meanwhile, the P-solubilizing capacity of AspN-JX16 was much higher than TalA-JX04. The pH of fermentation broth was negatively correlated with P-solubilizing capacity (p<0.01), suggesting that the fungi promote the dissolution of P sources by secreting organic acids. Our results showed that TalA-JX04 and AspN-JX16 could survive in acidic environments and both fungi had a considerable ability to release soluble P by decomposing recalcitrant P-bearing compounds. The two fungi had potential for application as environment-friendly biofertilizers in subtropical bamboo ecosystem.

1-Methylcyclopropene Treatment on Phenolics and the Antioxidant System in Postharvest Peach Combined with the Liquid Chromatography/Mass Spectrometry Technique.

In the present study, the potential effect of 1-methylcyclopropene (1-MCP) treatment on phenolics and antioxidant capacity in postharvest peach was assessed. Peach fruit (cv. Xiahui-8) treated with 1-MCP or without treatment was stored in 25 °C for 2, 4, 6, and 8 days. The phenolic composition and change trend were evaluated by liquid chromatography/mass spectrometry. The reactive oxygen species production and scavenging capacity against DPPH, O2• -, and HO• were determined. Gene expression of enzymes in the flavonoid biosynthetic pathway was assayed by quantitative reverse transcription polymerase chain reaction analysis. 1-MCP application inhibited the ethylene and CO2 production and stimulated the total phenol and total flavonoid contents. Total anthocyanin formation may be influenced directly or indirectly by the level of ethylene. The scavenging capacities of DPPH, HO•, and O2• - after 1-MCP treatment were enhanced. 1-MCP treatment affected the tissue color change, stimulated gene expression of PpaPAL, PpaCHS, PpaF3H, and PpaUFGT, and promoted the biosynthesis of flavonoids and stability of anthocyanin. PpaDFR and PpaUFGT played crucial roles in rapid color change stages. Kaempferol and kaempferol 3- O-galactoside increased distinctively during storage time.

Characteristics of Organic Acid Secretion Associated with the Interaction between Burkholderia multivorans WS-FJ9 and Poplar Root System.

The objective of this study was to investigate whether plant-bacteria interaction affects the secretion of organic acids by both organisms and to assess whether the production of IAA by the bacterium increases the secretion of organic acids by root exudates, and if the stress produced by low available phosphorus (P) affects the production of organic acids by bacteria, by roots, or by root exudates in presence of bacterial cultures. With this purpose, we used as a biological model poplar plants and one strain of Burkholderia multivorans able to solubilize P. High performance liquid chromatography was utilized to measure organic acids. The tests, the inductive effects of exogenous indole-3-acetic acid (IAA) on secretion of organic acids, the 2 × 4 × 2 factorial design experiment, and the ability of organic acids to solubilize tricalcium phosphate were performed to investigate the interactive effects. The results showed that, after B. multivorans WS-FJ9 interacted with the poplar root system, the key phosphate-solubilizing driving force was gluconic acid (GA) which was produced in three ways: (1) secreted by the root system in the presence of IAA produced by B. multivorans WS-FJ9; (2) secreted by B. multivorans WS-FJ9; and (3) secreted by the poplar root system in the presence of phosphorus stress. When phosphorus stress was absent, the GA was produced as outlined in (1) and (2) above. These results demonstrated that inoculating B. multivorans WS-FJ9 into the poplar root system could increase the amount of GA secretion and implied that the interaction between B. multivorans WS-FJ9 and the poplar root system could contribute to the increase of P available fraction for poplar plants.

The antitumor effect of TIG3 in liver cancer cells is involved in ERK1/2 inhibition.

Tazarotene-induced gene 3 (TIG3) was first characterized in tazarotene-treated human keratinocytes and identified as a retinoic acid responder gene, an important mediator of antitumor effects by retinoids. In this study, we aim to investigate the inhibitory effect of TIG3 on the growth of liver cancer and explore its underlying mechanism. Human hepatocellular carcinoma (HCC) Hep3B cells were transfected with plasmid GV141 carrying full-length TIG3 complementary DNA (cDNA). The effects of TIG3 on cell proliferation, apoptosis, and migration were determined in vitro. The suppressor effect of TIG3 on tumor growth was evaluated in vivo in a nude mouse HCC model. We observed that TIG3 expression is decreased in the Hep3B cell line as well as primary HCC tumors, and TIG3 expression inversely correlates with Ki-67 expression. Overexpression of TIG3 suppresses tumor growth in HCC both in vitro and in vivo via ERK1/2 inhibition by promoting apoptosis and inhibiting proliferation and migration. These findings identify TIG3 as an attractive therapeutic target for HCC.

Biosafety and colonization of Burkholderia multivorans WS-FJ9 and its growth-promoting effects on poplars.

Burkholderia cepacia complex (Bcc) is a group of bacteria with conflicting biological characteristics, which make them simultaneously beneficial and harmful to humans. They have been exploited for biocontrol, bioremediation, and plant growth promotion. However, their capacity as opportunistic bacteria that infect humans restricts their biotechnological applications. Therefore, the risks of using these bacteria should be assessed. In this study, Burkholderia multivorans WS-FJ9 originally isolated from pine rhizosphere, which was shown to be efficient in solubilizing phosphate, was evaluated with respect to its biosafety, colonization in poplar rhizosphere, and growth-promoting effects on poplar seedlings. Pathogenicity of B. multivorans WS-FJ9 on plants was determined experimentally using onion and tobacco as model plants. Onion bulb inoculated with B. multivorans WS-FJ9 showed slight hypersensitive responses around the inoculation points, but effects were not detectable based on the inner color and odor of the onion. Tobacco leaves inoculated with B. multivorans WS-FJ9 exhibited slightly water-soaked spots around the inoculation points, which did not expand or develop into lesions even with repeated incubation. Pathogenicity of the strain in alfalfa, which has been suggested as an alternative Bcc model for mice, was not detectable. Results from gene-specific polymerase chain reactions showed that the tested B. multivorans WS-FJ9 strain did not possess the BCESM and cblA virulence genes. Scanning electron microscopy revealed that the colonization of the WS-FJ9 strain reached 1.4 × 10(4) colony forming units (cfu) g(-1) rhizosphere soil on day 77 post-inoculation. The B. multivorans WS-FJ9 strain could colonize the rhizosphere as well as the root tissues and cells of poplars. Greenhouse evaluations in both sterilized and non-sterilized soils indicated that B. multivorans WS-FJ9 significantly promoted growth in height, root collar diameter, and plant biomass of inoculated poplar seedlings compared with controls. Phosphorus contents of roots and stems of treated seedlings were 0.57 and 0.55 mg g(-1) higher than those of the controls, respectively. Phosphorus content was lower in the rhizosphere soils by an average of 1.03 mg g(-1) compared with controls. The results demonstrated that B. multivorans WS-FJ9 is a nonpathogenic strain that could colonize the roots and significantly promote the growth of poplar seedlings.

Relationship between total antioxidant capacities of cereals measured before and after in vitro digestion.

Total antioxidant capacity (TAC), which is the cumulative capacity of different antioxidants to reduce oxidative stress, proved to be very useful to correlate the antioxidant capacity of the diet with the incidence of some disease or with modification of disease biomarkers. Therefore, it is important to estimate the real antioxidant potential of cereals. Most methods are based mainly on extractive methods which may largely underestimate the TAC of cereals. Several recent articles have reported a direct approach, the Quencher procedure, which produces a higher TAC compared to extractive methods. However, both the extractive methods and the Quencher procedure are performed prior to digestion. In this article, an in vitro approach using enzymatic digestion, designed to mimic digestion in the gastrointestinal tract (physiological conditions), had been used to release antioxidants from cereals. Seven whole cereals were employed for the determination of DPPH as well as ABTS radical scavenging activity before and after simulated digestion. The objective was to compare the in vitro procedure of antioxidant extraction with the Quencher procedure and water extraction. The values of the TAC's from the cereal grains obtained from the in vitro procedure were 1.8-10.3 times higher than the Quencher procedure and 3.5-10.5 times higher than water extraction. Correlation between the results of the TAC obtained using the three different procedures was also investigated. The in vitro gastrointestinal digestion procedure is more useful in the screening of grains, assessing their beneficial health effects compared to the Quencher procedure or water extraction.

Study on mitigation of acrylamide formation in cookies by 5 antioxidants.

UNLABELLED: This study investigated the capacity of various antioxidants in reducing the formation of acrylamide during cookie processing. Five antioxidants, antioxidants of bamboo leaves (AOB), sodium erythorbate (SE), tea polyphenols (TP), vitamin E (VE), and tert-butyl hydroquinone (TBHQ), were individually added into cookie formulas, and acrylamide content was determined by liquid chromatography tandem mass spectrometry (LC-MS/MS). Cookie quality indexes, including flavor, brittleness, and water activity, were also evaluated. Results showed that the maximum inhibitory rate of acrylamide by AOB was achieved with addition of 0.2 g/kg AOB. Addition of AOB (0.2 g/kg), TP (0.1 g/kg), VE (0.1 g/kg), SE (0.1 g/kg), and TBHQ (0.2 g/kg) mitigated the formation of acrylamide by 63.9%, 43.0%, 71.2%, 49.6%, and 54.1%, respectively. Sensory evaluation showed that the color, texture, and flavor of cookies processed with either AOB (0.2 g/kg) or VE (0.1 g/kg) had no significant difference compared to control cookies (P > 0.05). The present study indicated that AOB (0.2 g/kg) and VE (0.1 g/kg) could not only effectively mitigate the formation of acrylamide, but also retain acceptable sensory attributes of cookies. This work shows the potential effectiveness of antioxidants in food processing to decrease acrylamide formation. PRACTICAL APPLICATION: There is an urgent need for reducing the level of acrylamide produced during food processing. This study found that certain antioxidants (antioxidant of bamboo leaves and vitamin E) could effectively inhibit acrylamide formation in cookie processing without affecting sensory properties. The results suggested that the application of antioxidants could be an effective method to decrease acrylamide formation.

Hypolipidemic effect of bamboo shoot oil (P. pubescens) in Sprague-Dawley rats.

Atherosclerosis and its related complications are the leading causes of death in the West and in many developed countries. This study aims to investigate the hypolipidemic effect of bamboo shoot oil (BSO) in Sprague-Dawley rats. A group of rats had induced hyperlipidemia, hypercholesterolemia, and fatty liver by being fed with a high-fat, high-cholesterol diet for 4 wk. The control group was administered 10 mL distilled water per kg body weight, while the other groups were, respectively, administered 250 mg beta-sitosterol, 250 mg BSO, 500 mg BSO, and 1000 mg BSO per kg body weight by oral gavage. The results demonstrated that BSO could significantly decrease the levels of total cholesterol, triacylglycerol, low-density lipoprotein-cholesterol, phytosterol, lipoprotein lipase, hepatic lipase, and atherogenic index in serum, and increase the levels of cholesterol in feces. It could also significantly decrease the level of relative liver weight and liver lipids. The pronounced hypolipidemic effects of BSO might be attributed to its ability to inhibit cholesterol absorption and increase cholesterol excretion. These results suggest that consuming BSO may provide benefits in managing hypercholesterolemia. Therefore, BSO may be a good candidate for development as a functional food and nutraceutical.

Mutagenicity and safety evaluation of ethanolic extract of Prunus mume.

The ethanolic extract of Prunus mume (EPM) is a novel polyphenol preparation derived from branches (with leaves) of Prunus mume, which could be used as a functional ingredient for antioxidant and antiobesity therapy. The purpose of this study was to evaluate the safety of EPM. An EPM was prepared and evaluated for oral acute and subacute toxicity in Sprague-Dawley rats, while its mutagenic potential was assessed by a reverse mutation test using Salmonella typhimurium, by a bone marrow cell micronucleus test using ICR mice, and by a sperm abnormality test using ICR mice. The results showed no acute lethal effects at the maximal tested EPM dose of 20 g/kg bw in either rats or mice, suggesting that EPM can be regarded as virtually nontoxic. Administration at levels of 0.84, 1.67, and 3.33 g/kg bw to rats for 30 d did not induce any significant hematological, clinical, chemical, or histopathological changes. No mutagenicity evidence was detected in any of the 3 mutagenic tests. The level of "no observed adverse effect" (NOAEL) for EPM was above 3.33 g/kg bw for the subacute toxicity study.

Combined proteomic and cytological analysis of Ca2+-calmodulin regulation in Picea meyeri pollen tube growth.

Ca2+-calmodulin (Ca2+-CaM) is a critical molecule that mediates cellular functions by interacting with various metabolic and signaling pathways. However, the protein expression patterns and accompanying serial cytological responses in Ca2+-CaM signaling deficiency remain enigmatic. Here, we provide a global analysis of the cytological responses and significant alterations in protein expression profiles after trifluoperazine treatment in Picea meyeri, which abrogates Ca2+-CaM signaling. Ninety-three differentially displayed proteins were identified by comparative proteomics at different development stages and were assigned to different functional categories closely related to tip growth machinery. The inhibition of Ca2+-CaM signaling rapidly induced an increase in extracellular Ca2+ influx, resulting in dramatically increased cytosolic Ca2+ concentrations and ultrastructural abnormalities in organelles as the primary responses. Secondary and tertiary alterations included actin filament depolymerization, disrupted patterns of endocytosis and exocytosis, and cell wall remodeling, ultimately resulting in perturbed pollen tube extension. In parallel with these cytological events, time-course experiments revealed that most differentially expressed proteins showed time-dependent quantitative changes (i.e. some signaling proteins and proteins involved in organelle functions and energy production changed first, followed by alterations in proteins related to cytoskeletal organization, secretory pathways, and polysaccharide synthesis). Taken together, Ca2+-CaM dysfunction induced serial cytological responses and temporal changes in protein expression profiles, indicating the pivotal role of Ca2+-CaM in the regulation of tip growth machinery.

Integrative proteomic and cytological analysis of the effects of extracellular Ca(2+) influx on Pinus bungeana pollen tube development.

Ca (2+) is an essential ion in the control of pollen germination and tube growth. However, the control of pollen tube development by Ca (2+) signaling and its interactions with cytoskeletal components, energy-providing pathways, and cell-expansion machinery remain elusive. Here, we used nifedipine (Nif) to study Ca (2+) functions in differential protein expression and other cellular processes in Pinus bungeana pollen tube growth. Proteomics analysis indicated that 50 proteins showed differential expression with varying doses of Nif. Thirty-four of these were homologous to previously reported proteins and were classified into different functional categories closely related to tip-growth machinery. Blocking the L-type Ca (2+) channel with Nif in the pollen tube membrane induced several early alterations within a short time, including a reduction of extracellular Ca (2+) influx and a subsequently dramatic decrease in cytosolic free Ca (2+) concentration ([Ca (2+)] c), concomitant with ultrastructural abnormalities and changes in the abundance of proteins involved in energy production and signaling. Secondary alterations included actin filament depolymerization, disrupted patterns of endocytosis/exocytosis, and cell wall remodeling, along with changes in the proteins involved in these processes. These results suggested that extracellular Ca (2+) influx was necessary for the maintenance of the typical tip-focused [Ca (2+)] c gradient in the P. bungeana pollen tube, and that reduced adenosine triphosphate production (ATP), depolymerization of the cytoskeleton, and abnormal endocytosis/exocytosis, together with enhanced rigidity of cell walls, were responsible for the growth arrest observed in pollen tubes treated with Nif.

Antihyperlipidemic and antihypertensive effect of a triterpenoid-rich extract from bamboo shavings and vasodilator effect of friedelin on phenylephrine-induced vasoconstriction in thoracic aortas of rats.

Triterpenoids, which are widely distributed throughout the plant kingdom, were applied to the development of functional foods as physiological and pharmacological active agents. Some physiological activities of a triterpenoid-rich extract from bamboo shavings (EBS), i.e. antihyperlipidemic and antihypertensive effects, were systematically evaluated in the present study. The results demonstrated that EBS could reduce the serum total cholesterol (TC) and total triglyceride (TG) levels, which are the main factors directly leading to pathological changes in cardiovascular diseases, in hyperlipidemic rats (p < 0.05). Furthermore, EBS could significantly reduce the systolic pressure of spontaneously hypertensive rats (SHR) but its maximum effect time was less than 24 h. Meanwhile, EBS did not influence the heart rate of SHR. On the other hand, the vasodilator effects of friedelin, a main triterpenoid compound separated from EBS, on phenylephrine-induced vasoconstriction in the thoracic aortas of rats were also discussed. The results showed that friedelin probably served a function of vasodilator responses in the thoracic aortas of rats in accordance with the antihypertensive effect of EBS. The results also suggested that EBS might prevent some cardiovascular diseases by its impact on lipid metabolism.