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Background CBA/J mice are standard experimental animals in auditory studies, and age-related changes in auditory pathways are well documented. However, changes in locomotion-related brain regions have not been systematically explored. Results We showed an increase in immunoreactivity for glial fibrillary acidic protein (GFAP) in the cerebellar molecular layer associated with Purkinje cells in mice at 24 weeks of age but not in the younger mice. Increased GFAP immunoreactivity appeared in the form of clusters and distributed multifocally consistent with hyperplasia of astrocytes that were occasionally associated with Purkinje cell degeneration. Three out of 12 animals at 16 and 24 weeks of age exhibited pre-convulsive clinical signs. Two of these 3 animals also showed increased GFAP immunoreactivity in the cerebellum. Rotarod behavioral assessments indicated decreased performance at 24 weeks of age. Conclusions These results suggest minimal to mild reactive astrocytosis likely associated with Purkinje cell degeneration in the cerebellum at 24 weeks of age in CBA/J mice. These findings should be taken into consideration prior to using this mouse strain for studying neuroinflammation or aging.

In recent years, the excellent curative effect of CD19‐specific chimeric antigen receptor (CAR) T‐cell therapy has brought hope to patients with relapsing or refractory B‐cell hematological malignancies, however relapse after CAR T‐cell infusion has hindered the widespread clinical application of this immunotherapy and targeted antigen‐negative relapse has caused widespread concern. Consequently, strategies for increasing targeted antigens have been created. In addition to the most widely applied target, namely CD19, researchers have further explored the possibility of other targets, such as CD20, CD22, CD33, and CD123, and have tested a series of combination antigen CAR T‐cell therapies. Here, we summarize the current preclinical and clinical studies of dual‐target CAR T cells., CD19‐specific chimeric antigen receptor (CAR) T‐cell therapy has excellent efficacy in patients with relapsed or refractory B‐cell hematological malignancies, however antigen loss after single‐target CAR T‐cell infusion hinders the clinical application of this immunotherapy. Researchers have proposed a dual‐target chimeric antigen receptor‐armed T cell (CART) strategy for more comprehensive tumor coverage. Here we summarize current preclinical and clinical studies on dual‐target CAR T cells.

Soil microbes have a direct impact on plant metabolism and health. The current study investigates the comparative rhizobiome between sugarcane monoculture and sugarcane-soybean intercropping. A greenhouse experiment was performed with two treatments: 1) sugarcane monoculture and 2) Sugarcane-Soybean intercropped. We performed a high throughput sequencing (HTS) platform to analyze the microbial community. We used the 16S rRNA gene and the internal transcribed spacer (ITS) region primers to identify the microbial diversity. HTS results revealed that a total of 2979 and 124 bacterial and fungal operational taxonomic units (OTUs) were observed, respectively. Microbial diversity results concluded that the intercropping system has a beneficial impact on soil microbes. The highest numbers of bacterial and fungal OTUs were found in the intercropping system, and these results also collaborated with quantitative PCR results. Additionally, intercropped sugarcane plants were showed a higher weight of above and below ground parts than the monoculture. Soil chemical analysis results also complemented that the intercropping system nourished the quantity of organic carbon, total nitrogen, and soil enzyme activities. Correlation analysis of diversity index and abundance concluded that soil nutrient content positively influenced the microbial abundance that improves plant growth. The present study framed out the profound insights of microbial community interaction under the sugarcane soybean intercropping system, and this information could help to. This information could help improve or increase the sugarcane crop production without causing any negative impact on the sugarcane plant growth and development.

Macrophage activation participates in the pathogenesis of pulmonary inflammation. As a coenzyme, vitamin B6 (VitB6) is mainly involved in the metabolism of amino acids, nucleic acids, glycogen and lipids. We have previously reported that activation of AMP‐activated protein kinase (AMPK) produces anti‐inflammatory effects both in vitro and in vivo. Whether VitB6 via AMPK activation prevents pulmonary inflammation remains unknown. The model of acute pneumonia was induced by injecting mice with lipopolysaccharide (LPS). The inflammation was determined by measuring the levels of interleukin‐1 beta (IL‐1β), IL‐6 and tumour necrosis factor alpha (TNF‐α) using real time PCR, ELISA and immunohistochemistry. Exposure of cultured primary macrophages to VitB6 increased AMP‐activated protein kinase (AMPK) Thr172 phosphorylation in a time/dose‐dependent manner, which was inhibited by compound C. VitB6 downregulated the inflammatory gene expressions including IL‐1β, IL‐6 and TNF‐α in macrophages challenged with LPS. These effects of VitB6 were mirrored by AMPK activator 5‐aminoimidazole‐4‐carboxamide ribonucleoside (AICAR). However, VitB6 was unable to inhibit LPS‐induced macrophage activation if AMPK was in deficient through siRNA‐mediated approaches. Further, the anti‐inflammatory effects produced by VitB6 or AICAR in LPS‐treated macrophages were abolished in DOK3 gene knockout (DOK3 −/−) macrophages, but were enhanced in macrophages if DOK3 was overexpressed. In vivo studies indicated that administration of VitB6 remarkably inhibited LPS‐induced both systemic inflammation and acute pneumonia in wild‐type mice, but not in DOK3 −/− mice. VitB6 prevents LPS‐induced acute pulmonary inflammation in mice via the inhibition of macrophage activation.

Rodent studies often must be conducted during an animal's active phase (that is, in darkness) yet also during a typical day shift for staff. Low-pressure sodium lighting (LPSL), to which human retinas are more sensitive than rodents' at low intensity, has been used to facilitate study conduct in dark phase. The assumption was that LPSL would be equivalent to total darkness due to low rodent retinal sensitivity but provide enough lighting for safe technical manipulations due to higher human retinal sensitivity. Unlike other light sources, LPSL has been tested for effects on circadian rhythm specific to locomotive activities in albino mice. Whether LPSL affects circadian rhythms in rats is unknown. In this study, circadian endpoints were derived from body temperature and locomotor activity via telemeters in 8 adult male Wistar rats. When moved from a 12:12-h white-light (that is, cold white fluorescent light):dark (LD) cycle to a 12:12-h white-light:sodium-light cycle, rats demonstrated free-running and disrupted circadian rhythms (that is, lengthened circadian period and reduced circadian robustness and amplitude). Body temperature and locomotor activity were significantly lower in the LPSL phase as compared with dark phase under the baseline condition. When exposed to a 12:12 h sodium-light:dark (SD) cycle, rats entrained with a circadian period similar to 12:12-h white-light:dark (LD), but significantly different from the period under constant darkness (DD). Circadian onset and acrophase were delayed under SD compared with LD. When illuminated with a LPSL pulse under DD, rats showed phase shifts similar to white-light pulse effects, consistent with the phase response curve. To determine whether the image-forming photoreceptors are involved in this process, we used electroretinography. Compared with white light, 589-nm light generated during electroretinography elicited rod photoreceptors responses with longer latency and cone photoreceptor responses with lower amplitude. These results indicate that LPSL is a weaker zeitgeber than white light and may alter the circadian system in rats. Furthermore, because LPSL appeared to be visible to rats, it may not be an appropriate substitute for actual darkness.

Fusarium verticillioides is the pathogen associated with pokkah boeng disease (PBD), the most significant airborne disease of sugarcane. The molecular mechanisms that regulate the defense responses of sugarcane towards this fungus are not yet fully known. Samples of 'YT 94/128' (resistant, R) and 'GT 37' (susceptible, S) inoculated with F. verticillioides on the 14 days post-inoculation were used to analyze the transcriptome to screen R genes. In total, 80.93 Gb of data and 76,175 Unigenes were obtained after assembling the sequencing data, and comparisons of Unigenes with NR, Swiss-prot, KOG, and KEGG databases confirmed 42,451 Unigenes. The analysis of differentially expression genes (DEGs) in each sample revealed 9092 DEGs in 'YT 94/128,' including 8131 up-regulated DEGs and 961 down-regulated DEGs; there were 9829 DEGs in 'GT 37,' including 7552 up-regulated DEGs and 2277 down-regulated DEGs. The identified DEGs were mainly involved in catalytic enzyme activity, cell protease, hydrolytic enzymes, peptide enzyme, protein metabolism process of negative regulation, phenylpropanoid metabolism, extracellular region, aldehyde dehydrogenase, endopeptidase, REDOX enzyme, protein kinases, and phosphoric acid transferase categories. KEGG pathway clustering analysis showed that the DEGs involved in resistance were significantly related to metabolic pathways of phenylpropanoid biosynthesis, cutin, suberine and wax biosynthesis, nitrogenous metabolism, biosynthesis of secondary metabolites, and plant-pathogen interactions. This application of transcriptomic data clarifies the mechanism of interactions between sugarcane and F. verticillioides, which can help to reveal disease-related metabolic pathways, molecular regulatory networks, and key genes involved in sugarcane responses to F. verticillioides.

Microbial agents are the best alternative to the chemical fertilizers that promote the plant growth in an eco-friendly manner. This study was intended to characterize Burkholderia anthina strain MYSP113 and to evaluate its mechanisms linked to sugarcane growth promotion. The morphological, biochemical, and molecular characterization (gyrB and recA) results identified the bacteria as B. anthina. To determine the plant bacterial association, MYSP113 strain was tagged with a plasmid-bear green fluorescent protein (gfp) gene, and the tagged strain was inoculated to the sugarcane (variety GXB-9), and bacterial colonization analyzed by confocal laser fluorescence microscopy. The results revealed that strain MYSP113 had root-colonizing abilities, and colonization was detected at the root and leaves maturation zones. Bacteria entered the roots via cracks at the surfacing site of the roots and disrupted epidermis. Bacterial cells were observed in the substomatal chambers of leaves. Furthermore, to evaluate the mechanisms related to sugarcane growth promotion and bacterial competence in soil and plant, greenhouse experiment performed, and bacterial competence, plant growth parameters, and phytoenzymes, and hormones were assessed with or without MYSP113. We examined bacterial competence in soil and plant via qPCR and direct plant count at 45 and 90 DAI, and results recommended that strain MYSP113 has the ability to colonize in sugarcane rhizosphere and in the aerial part of the plant. Furthermore, strain MYSP113 significantly induced the plant fresh weight, dry weight, and chlorophyll content of sugarcane seedlings at 45 and 90 days after inoculation. Additionally, strain MYSP113 triggered chitinase, cellulose, indole acetic acid, and abscisic acid production in the plant roots and aerial parts, and these results signifying that bacterial inoculation modulates the plant metabolism and boost the health. The present study concluded that strain MYSP113 has beneficial linkages with the sugarcane plant, and it would be utilized as a bioinoculant in future.

In the era of climate change, due to increased incidences of a wide range of various environmental stresses, especially biotic and abiotic stresses around the globe, the performance of plants can be affected by these stresses. After oxygen, silicon (Si) is the second most abundant element in the earth’s crust. It is not considered as an important element, but can be thought of as a multi-beneficial quasi-essential element for plants. This review on silicon presents an overview of the versatile role of this element in a variety of plants. Plants absorb silicon through roots from the rhizospheric soil in the form of silicic or monosilicic acid. Silicon plays a key metabolic function in living organisms due to its relative abundance in the atmosphere. Plants with higher content of silicon in shoot or root are very few prone to attack by pests, and exhibit increased stress resistance. However, the more remarkable impact of silicon is the decrease in the number of seed intensities/soil-borne and foliar diseases of major plant varieties that are infected by biotrophic, hemi-biotrophic and necrotrophic pathogens. The amelioration in disease symptoms are due to the effect of silicon on a some factors involved in providing host resistance namely, duration of incubation, size, shape and number of lesions. The formation of a mechanical barrier beneath the cuticle and in the cell walls by the polymerization of silicon was first proposed as to how this element decreases plant disease severity. The current understanding of how this element enhances resistance in plants subjected to biotic stress, the exact functions and mechanisms by which it modulates plant biology by potentiating the host defence mechanism needs to be studied using genomics, metabolomics and proteomics. The role of silicon in helping the plants in adaption to biotic stress has been discussed which will help to plan in a systematic way the development of more sustainable agriculture for food security and safety in the future.

Plants are associated with the combination of microbiomes, which can improve plant growth, stress tolerance, and control plant pathogens. The incorporation of a beneficial microbiome represents a promising sustainable key to enhance agricultural production. Sugarcane is an essential crop with many agricultural and industrial uses including animal feed, biofuels and bagasse. There is a need to strengthen sugarcane crop production sustainably and to locate solutions to control pathogens, abiotic stress, and pests. The utilization of beneficial microbes as a biofertilizer has become essential in sugarcane agriculture for sustainable crop production. In this perspective, the sugarcane microbiome becomes a promising approach for sustainable production. The chapter describes the application of sugarcane-associated microbiome.

As the polyploidy progenitor of modern sugarcane, Saccharum spontaneum is considered to be a valuable resistance source to various biotic and abiotic stresses. However, little has been reported on the mechanism of drought tolerance in S. spontaneum. Herein, the physiological changes of S. spontaneum GXS87-16 at three water-deficit levels (mild, moderate, and severe) and after re-watering during the elongation stage were investigated. RNA sequencing was utilized for global transcriptome profiling of GXS87-16 under severe drought and re-watered conditions. There were significant alterations in the physiological parameters of GXS87-16 in response to drought stress and then recovered differently after re-watering. A total of 1569 differentially expressed genes (DEGs) associated with water stress and re-watering were identified. Notably, the majority of the DEGs were induced by stress. GO functional annotations and KEGG pathway analysis assigned the DEGs to 47 GO categories and 93 pathway categories. The pathway categories were involved in various processes, such as RNA transport, mRNA surveillance, plant hormone signal transduction, and plant-pathogen interaction. The reliability of the RNA-seq results was confirmed by qRT-PCR. This study shed light on the regulatory processes of drought tolerance in S. spontaneum and identifies useful genes for genetic improvement of drought tolerance in sugarcane.

Several factors influenced the sugarcane production, and among them, higher use of nitrogen and depletion of soil nutrient constitutes a significant concern in China. Sugarcane-legume intercropping may help to regulate the microbial structure and functions. In the present study, sugarcane rhizosphere soils of three cropping systems: Sugarcane only (S-only), sugarcane with peanut (S + P), and sugarcane + soybean (S + S) were sampled in tillering, elongation, and maturation stages from two different experimental fields. High-throughput sequencing technologies applied to assess the effects of different cropping systems on the structure of nitrogenase (nifH) gene communities. A total of 3818 OTUs (operational taxonomic units) were acquired from all soil samples. Intercropping systems noticeably increased the relative abundance of Proteobacteria in the tillering stage. The increased microbial diversity in the rhizosphere was mainly due to soil organic carbon and total soil N. In contrast, intercropping has no significant negative impact on microbial abundance, but sugarcane growth stages influence it significantly, and two bacteria (Bradyrhizobium and Pseudacidovorax) showed significant shift during plant growth. The results provide insight into the microbial structure of Proteobacteria in the sugarcane legume-intercropping field, and how microbial community behaves in different growth stages. It can boost the microbial activity of the soil, and that could be a new strategy to stimulate soil fertility without causing any negative impact on crop production.

Sugarcane-legume intercrops have gained attention because of their ability to improve crop productivity in an ecofriendly manner. We investigated the roles of rhizospheric and endospheric diazotrophs in soil nutrient transformation. We conducted two field experiments with three treatments: sugarcane monoculture, sugarcane + peanut intercrop, and sugarcane + soybean intercrop. Soil and plant samples were collected and the nutrient content, enzymatic activity, and diazotroph population were assessed. The nitrogenase (nifH) gene and 16S rRNA gene were used for molecular characterization. Correlation analysis was performed to discover the relations among the soil variables, plant variables, and microbial communities. The multiple variance analysis results indicated that the intercropping systems had a significant (p

In plants, both abscisic acid (ABA) dependent and independent pathways form the basis for the response to environmental stresses. Sucrose non-fermenting 1-related protein kinase 2 (SnRK2) plays a central role in plant stress signal transduction. However, complete annotation and specific expression patterns of SnRK2s in sugarcane remain unclear. For the present study, we performed a full-length cDNA library survey of sugarcane, thus identifying ten SoSnRK2 genes via phylogenetic, local BLAST methods, and various bioinformatics analyses. Phylogenetic analysis indicated division of SoSnRK2 genes into three subgroups, similar to other plant species. Gene structure comparison with Arabidopsis suggested a unique evolutionary imprint of the SnRK2 gene family in sugarcane. Both sequence alignment and structural annotation provided an overview of the conserved N-terminal and variations of the C-terminal, suggesting functional divergence. Transcript and transient expression assays revealed SoSnRK2s to be involved in the responses to diverse stress signals, and strong ABA induction of SoSnRK2s in subgroup III. Co-expression network analyses indicated the existence of both conserved and variable biological functions among different SoSnRK2s members. In summary, this comprehensive analysis will facilitate further studies of the SoSnRK2 family and provide useful information for the functional validation of SoSnRK2s.

Background Stress evokes lipolytic release of free fatty acid (FFA) and low-grade inflammation in visceral adipose tissue, mediated by increased adipokine secretion, and contributes to glucose metabolism disorder and prothrombotic state. We tested the hypothesis that alogliptin, a dipeptidyl peptidase-4 inhibitor, can ameliorate the biological effects of chronic stress in mice. Method and results C57BL/6J mice were subjected to 2-week intermittent restraint stress and orally treated with vehicle or alogliptin (dose: 15 or 45 mg/kg/day). Plasma levels of lipids, proinflammatory cytokines (monocyte chemoattractant protein-1, tumor necrosis factor-α, and interleukin-6), and 8-hydroxydeoxyguanosine were measured with enzyme-linked immunosorbent assay. Monocyte/macrophage accumulation in inguinal white adipose tissue (WAT) was examined by CD11b-positive cell count and mRNA expression of CD68 and F4/80 was examined by immunohistochemistry and RT-PCR, respectively. The mRNA levels of the above-mentioned proinflammatory cytokines, NADPH oxidase 4, adiponectin, and coagulation factors (plasminogen activation inhibitor-1 and tissue factor) in WAT were also assessed with RT-PCR. Glucose metabolism was assessed by glucose and insulin tolerance tests, plasma levels of DPP-4 activity, glucagon-like peptide-1, expression of DPP-4, insulin receptor substrate-1 and glucose transporter 4 in WAT and skeletal muscle. Alogliptin administration suppressed stress-induced FFA release, oxidative stress, adipose tissue inflammation, DPP-4 activation, and prothrombotic state in a dose-dependent manner, and improved insulin sensitivity in stressed mice. Conclusions The results indicate that alogliptin improves stress-induced prothrombotic state and insulin resistance; suggesting that alogliptin could have beneficial therapeutic effects against cardiovascular complications in diabetic patients under stress.

Since 1980s, the Chinese sugar industry has been developing at a fast pace. In the milling year 2007/2008, the total sugar production in China reached 14.83 MT, which was 24.04 % higher than that in previous milling year, and cane sugar production reached 13.67 MT, which was 92.18 % of the total. Guangxi is the largest sugarcane and sugar producer in China, which produced 9.41 MT sugar in 2007/2008 and 8.56 MT sugar in 2013/2014. China has developed a series of unique farming technologies in commercial sugarcane production and has developed an advanced technology system since 1990s. These technologies include deep ploughing and fine preparation of soil, economical fertilization system; trash addition to field, water-saving irrigation, use of pathogen-free healthy seedcane, rational application of vinasse in sugarcane field, chemical control, mechanization for sugarcane management, and comprehensive control of diseases, pests and weeds. The exploitation and comprehensive application of the new sugarcane farming technologies have promoted the Chinese sugar industry to a new level in about every 5 years, and made China become the third biggest sugar-producing country in the world. However, the sugar industry has been experiencing a very difficult time in the last two milling years because of the worldwide low sugar price and the high production cost at the domestic level, which led to a substantial reduction in sugarcane-growing areas and sugar production. Mechanization and the related sugarcane variety selection and farming technology development have become the bottleneck for sustaining the development of the sugar industry in China.

Soil survey was conducted at four different locations of Guangxi, China to investigate the effect of peanut and soybean inter-cropping in sugarcane cultivation on the soil properties and diazotrophic bacterial diversity. Principal component analysis result shown (PCA) showed difference impact of intercropping and monoculture on the basis of biological and chemical properties of soil. Two-way analysis of variance of soil properties showed a significant interactive effect with locations and cultivation systems. Microbial enumeration results demonstrated that intercropping system enhanced the diazotrophic population. After isolation of diazotrophs, 21 selected bacteria were characterized by siderophore, phosphate, indole acetic acid, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and nitrogenase assay. Among the identified bacteria, isolate MYSP104 showed maximum nitrogenase activity and isolate MYSS78 showed maximum ACC deaminase activity. Partial 16S rRNA gene sequencing results classified these diazotrophs in six phyla (Firmicutes, α, β, Υ-proteobacteria, actinobacteria and bacteroidetes). Diazotrophs such as Bacillus tequilensis, Variovorax paradoxus, Acidovorax facilis, Leucobacter aridicollis, Streptomyces fimicarius and Pseudomonas nitroreducens were reported for the first time from the sugarcane rhizosphere of Guangxi, China. Venn diagram explained that seven bacterial species (Brevibacterium, Burkholderia, Delftia, Leucobacter, Pseudomonas, Sinorhizobium and Variovorax) were recognized with soybean and sugarcane intercropping cultivation systems. This study concluded that intercropping system could enhance the population of N-fixers in soil. Soybean intercropping influenced the soil chemical and biological properties better than peanut. Moreover, the isolated diazotrophs need further characterization and they might be utilized as bio-inoculums for commercial sugarcane production in the future.

The diazotrophic Burkholderia anthina MYSP113 is a vital plant growth-promoting bacteria and sugarcane root association. The present study based on a detailed analysis of sugarcane root transcriptome by using the HiSeq-Illumina platform in response to the strain MYSP113. The bacterium was initially isolated from the rhizosphere of sugarcane. To better understand biological, cellular, and molecular mechanisms, a de novo transcriptomic assembly of sugarcane root was performed. HiSeq-Illumina platformwas employed for the sequencing of an overall of 16 libraries at a 2×100 bp configuration. Differentially expressed genes (DEGs) analysis identified altered gene expression in 370 genes (total of 199 up-regulated genes and 171 down-regulated genes). Deciphering the huge datasets, concerning the functioning and production of biological systems, a high throughput genome sequencing analysis was attempted with Gene ontology functional analyses and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The report revealed a total of 148930 unigenes. 70414 (47.28%) of them were annotated successfully to Gene Ontology (GO) terms. 774 at 45 days, 4985 of 30 days and 15 days of 6846 terms were significantly regulated. GO analysis revealed that many genes involved in the metabolic, oxidation-reduction process and biological regulatory processes in response to strain MYSP113 and significantly enriched as compare to the control. Moreover, KEGG enriched results show that differentially expressed genes were classified into different pathway categories involved in various processes, such as nitrogen metabolism, plant hormone signal transduction, etc. The sample correlation analyses could help examine the similarity at the gene expression level. The reliability of the observed differential gene expression patterns was validated with quantitative real-time PCR (qRT-PCR). Additionally, plant enzymes activities such as peroxidase and superoxide dismutase were significantly increased in plant roots after the inoculation of strain MYSP113. The results of the research may help in understanding the plant growth-promoting rhizobacteria and plant interaction.

Intercropping significantly improves land use efficiency and soil fertility. This study examines the impact of three cultivation systems (monoculture sugarcane, peanut-sugarcane and soybean-sugarcane intercropping) on soil properties and diazotrophs. Sugarcane rhizosphere soil was sampled from the farmers’ field. Soil properties and nifH gene abundance were analyzed by high throughput sequencing. Moreover, a total of 436,458 nifH gene sequences were obtained and classified into the 3201 unique operational taxonomic units (OTUs). Maximum unique OTUs resulted with soybean-sugarcane intercropping (

The apelin/APJ system has been implicated in obesity-related hypertension. We investigated the mechanism responsible for the pathogenesis of obesity-related hypertension with a special focus on the crosstalk between AngII/its type 1 receptor (AT1R) signaling and apelin/APJ expression. Sprague-Dawley rats fed a high-fat (obesity-related hypertension, OH) or normal-fat diet (NF) for 15 weeks were randomly assigned to one of two groups and administered vehicle or perindopril for 4 weeks. Compared to the NF rats, the OH rats showed lower levels of plasma apelin and apelin/APJ mRNAs of perirenal adipose tissues, and these changes were restored by perindopril. Administration of the AT1R antagonist olmesartan resulted in the restoration of the reduction of apelin and APJ expressions induced by AngII for 48 h in 3T3-L1 adipocytes. Among several inhibitors for extracellular signal-regulated kinases 1/2 (ERK1/2) PD98059, p38 mitogen-activated protein kinase (p38MAPK) SB203580 and phosphatidylinositol 3-kinase (PI3K) LY294002, the latter showed an additive effect on AngII-mediated inhibitory effects. In addition, the levels of p-Akt, p-ERK and p38MAPK proteins were decreased by long-term treatment with AngII (120 min), and these changes were restored by Olmesartan. Apelin/APJ appears to be impaired in obesity-related hypertension. The AngII inhibition-mediated beneficial effects are likely attributable, at least in part, to restoration of p38/ERK-dependent apelin/APJ expression in diet-induced obesity-related hypertension.

Abscisic acid (ABA) plays important roles in adaptive responses to various environmental stresses. The rate-limiting step in ABA biosynthesis is the oxidative cleavage of cis-epoxycarotenoids, which is catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED). In this experiment, a full-length cDNA encoding NCED gene was cloned by RT-PCR and RACE from sugarcane (Saccharum officinarum L.). The full-length of SoNCED is 2,521 bp with 1,827 bp open reading frame, encoding a peptide of 608 amino acids. The calculated molecular weight of protein was 65.9 kDa with isoelectric point of 6.04. Conserved domains prediction indicated a chloroplast-targeting peptide located at N-terminus of SoNCED. Phylogenetic tree, constructed by Neighbor-Joining method indicated that SoNCED shared high identity with the NCEDs reported from other plant species. Sequence alignment revealed that the basic secondary structure including α-helices, β-strands, β-propeller and His residues coordinating catalytic sites of SoNCED were highly conserved as in the NCEDs from other plants. Tissue specific expression analysis using quantitative real-time PCR showed a significant increase in SoNCED mRNA level and its correlation with O2 – production rate and ABA accumulation in leaves and roots of sugarcane variety GT21 when exposed to water stress. Further, the stimulation of SoNCED mRNA level, O2 – production rate and ABA content after exogenous application of ABA (100 μMol l−1) proved its involvement in pathways providing tolerance to drought stress.

Water stress causes considerable yield losses in sugarcane. To investigate differentially expressed genes under water stress, a pot experiment was performed with the sugarcane variety GT21 at three water-deficit levels (mild, moderate and severe) during the elongation stage and gene expression was analyzed using microarray technology. Physiological parameters of sugarcane showed significant alterations in response to drought stress. Based on the expression profile of 15,593 sugarcane genes, 1,501 (9.6%) genes were differentially expressed under different water-level treatments; 821 genes were upregulated and 680 genes were downregulated. A gene similarity analysis showed that approximately 62.6% of the differentially expressed genes shared homology with functional proteins. In a Gene Ontology (GO) analysis, 901 differentially expressed genes were assigned to 36 GO categories. Moreover, 325 differentially expressed genes were classified into 101 pathway categories involved in various processes, such as the biosynthesis of secondary metabolites, ribosomes, carbon metabolism, etc. In addition, some unannotated genes were detected; these may provide a basis for studies of water-deficit tolerance. The reliability of the observed expression patterns was confirmed by RT-PCR. The results of this study may help identify useful genes for improving drought tolerance in sugarcane.

The acetylcholinesterase inhibitor (AChEI)-based therapeutic strategies have been shown to have vasculoprotective properties in the animal model of hindlimb ischemia due to its activation of the endothelial cholinergic system. However, little is know about whether other cell types (myocytes, immunocytes) are involved in the AChEI-related therapeutic benefits in peripheral artery disease. Therefore, we review the multiple cell-targeted effects of AChEI on the animal model of hindlimb ischemia and explore its clinical application in angiomyogenesis.

Vascular disease is the leading cause of morbidity and mortality. Oxidative stress can cause endothelial cell apoptosis. Low insulin like growth factor-1 (IGF-1) has been linked to adverse risk profile and increased vascular disease incidence. Since IGF-1 acts as an important survival factor for multiple cell types, we undertook this study to investigate whether IGF-1 favorably affects oxidative-stress mediated apoptosis of vascular endothelial cells. Exposure to hydrogen peroxide induced apoptotic changes (e.g. DNA fragmentation, altered mitochondrial membrane potential and caspase-3 activity) in human umbilical vein endothelial cells (HUVECs) in a time dependent manner. Addition of IGF-1 blocked the oxidative-stress effect parallel to IGF-1 receptor (IGF-1R) expression, and silencing the IGF-1R with small interference RNA attenuated the IGF-1 influence. Our findings show that enhanced IGF-1 signaling inhibits oxidative-stress induced apoptosis in HUVECs by reducing mitochondrial dysfunction. Specifically the protective mechanism of IGF-1 involves preserving the mitochondrial membrane potential, maintaining the mitochondrial retention of cytochrome-c, and reducing caspase-3 activity. These results may have therapeutic implications in preventing/reducing vascular disease associated endothelial dysfunction.

The pollen morphology in 15 species representing five genera in the tribe Lithospermeae of Boraginoideae (Boraginaceae) has been investigated and illustrated using light microscopy and scanning electron microscopy. The tribe Lithospermeae is very diverse in pollen morphology. The pollen grains are 10.4–41.8 × 7–33.1 μm in size with subspheroidal, prolate, cocoon, dumbbell, and ovoid shapes. The pollen apertures are of five types: 3-colporate, 3-syncolporate, 4-8-colporate, 4-6-syncolpate, and 6-7-colpate types. The exine ornamentations are generally smooth or rugulose, sparsely echinulate, and rarely rugulate. Based on pollen morphology, we developed a key to identify the genera of the tribe Lithospermeae, compared the pollen apertures among tribes of Boraginoideae, explained the evolutionary trends of the pollen grains, and discussed the taxonomic position of the tribe Lithospermeae. The palynological data suggest pollen of the eurypalynous type and support the proposal that the tribe Lithospermeae is in the primitive position of the subfamily Boraginoideae, and Echium Linn. is in the tribe Lithospermeae rather than in a novel tribe. Our observations have application potential for identification of pollen fossils of the tribe Lithospermeae.

We read with great interest the important article by Sayed and coworkers1 on the role of innate immunity in somatic reprogramming. They discovered that activation of innate immune signaling by small molecules and an agonist of Toll-like receptor 3, combined with endothelial growth factors, is sufficient to transdifferentiate human fibroblasts to endothelial cells. The same research group reported previously that stimulation of Toll-like receptor 3 causes global changes in the expression of epigenetic modifiers to enhance nuclear reprogramming.2 …

Background/Aims: Odontogenic keratocysts (OKC) are aggressive lesions in the jaws, which can occur as isolated cases or in association with nevoid basal cell carcinoma syndrome (NBCCS). Mutations on PTCH gene have been identified in patients with NBCCS. It was hypothesized that PTCH mutations may be causative in isolated OKC. This study aims to investigate germline mutations of PTCH in families with OKC and NBCCS. Methods: Three Chinese families with OKC and NBCCS were enrolled in the study. The diagnosis was based on examination and medical history. Mutation analysis was performed by amplifying all exons of PTCH and sequencing the products. Results: One family with isolated OKC (family 1) and the other two families with NBCCS were diagnosed. Three novel germline mutations in PTCH were identified, including a missense mutation (p.S1089>P) in family 1, a nonsense mutation (p.Q160X) in family 2 and a de novo mutation (c.768_777delGACAAACTTC) in family 3. Conclusions: It is proposed that isolated OKC can be inherited in an autosomal dominant mode. The results suggest that germline mutations on PTCH can cause isolated OKC, and that the PTCH gene responsible for NBCCS plays an important role in the formation of OKCs even when they are not syndrome-related.

Background: The interaction between the renin-angiotensin system and toll-like receptors (TLRs) in the pathogenesis of advanced atherosclerotic plaques is not well understood. We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient (ApoE –/– ) mice with a special focus on plaque neovessel formation. Methods and Results: Four-wk-old ApoE –/– mice were fed a high-fat diet for 8 wks, and the mice were randomly assigned to one of three groups and administered a vehicle, hydralazine, or aliskiren for an additional 12 wks. Aliskiren reduced the atherosclerotic plaque area and plaque neovessel density. It increased the plaque collagen and elastin contents, and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S (CatS) protein. Aliskiren also decreased the levels of AT1R, gp91phox, TLR2, monocyte chemotactic protein-1, and CatS mRNAs in the aortic roots. Hydralazine had no beneficial vascular effects, although its administration resulted in the same degree of blood pressure reduction as aliskiren. CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE –/– mice, but aliskiren showed no further benefits in ApoE –/– CatS –/– mice. In vitro , CatS mRNA expressions were induced by angiotensin II, and TLR2 silencing reduced CatS mRNA expression. Moreover, the genetic inhibition of CatS impaired the endothelial cell angiogenic action in vitro and ex vivo . Conclusion: Renin inhibition appears to inhibit advanced plaque neovessel formation in ApoE –/– mice and to decrease the vascular inflammatory action and extracellular matrix degradation, partly by reducing AT1R/TLR2-mediated CatS activation and activity, thus regressing advanced atherosclerotic plaques.

Crizotinib (Xalkori) is a tyrosine kinase inhibitor of both anaplastic lymphoma kinase (ALK) and mesenchymal-epithelial transition factor (c-Met). Though not predicted from standard nonclinical toxicological evaluation, visual disturbance became a frequently observed adverse event in humans. To understand the possible mechanism of this vision effect, an in vivo electroretinogram (ERG) study was conducted to assess retinal functional changes following oral administration of crizotinib. Immunohistochemical (IHC) staining of ALK and c-Met in the neural retinas of human, non-human primate, dog, rat, and mouse was used to aid in the animal model selection. ALK IHC staining was identified predominantly in the ganglion cell and inner nuclear layers of most species evaluated, in the inner plexiform layer in human and rodent, and in the nerve fiber layer in human and rat only. There was no apparent staining of any layer of the neural retina for c-Met in any of the species evaluated. ERG measurements identified a significant reduction in b-wave amplitude during the initial phase of dark adaptation in the crizotinib-treated rats. ERGs were also taken following oral administration of PF-06463922 (an ALK-selective inhibitor), for an understanding of potential kinase involvement. ERG effects were not observed in PF-06463922-treated animals when comparable exposures in the vitreous humor were achieved. Collectively, our results suggest that the ERG b-wave amplitude decreases during dark adaption following crizotinib administration may be related to signaling changes within the retina in rats, likely independent of ALK inhibition.

The interaction between the renin-angiotensin system and toll-like receptors (TLRs) in the pathogenesis of advanced atherosclerotic plaques is not well understood. We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient (ApoE(-/-)) mice with a special focus on plaque neovessel formation.Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks, and the mice were randomly assigned to one of three groups and administered a vehicle, hydralazine, or aliskiren for an additional 12 wks. Aliskiren reduced the atherosclerotic plaque area and plaque neovessel density. It increased the plaque collagen and elastin contents, and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S (CatS) protein. Aliskiren also decreased the levels of AT1R, gp91phox, TLR2, monocyte chemotactic protein-1, and CatS mRNAs in the aortic roots. Hydralazine had no beneficial vascular effects, although its administration resulted in the same degree of blood pressure reduction as aliskiren. CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice, but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice. In vitro, TLR2 silencing reduced CatS expression induced by angiotensin II. Moreover, aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo.Renin inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation, partly by reducing AT1R/TLR2-mediated CatS activation and activity, thus regressing advanced atherosclerosis.

Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing. To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene. The HTR gene consists of Exon I, which contains the 5'-regulatory region and 85 nucleotides of coding sequence; a approximately 15-kb intron; and Exon II, which contains the remainder of the coding sequence and the entire 3'-untranslated region. Multiple transcription initiation sites were identified by S1 mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence of two SP-1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and approximately3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms that regulate its expression within the blood vessel wall.

Although sodium iodate (NaIO3)-induced retinal injury model has been widely used in rodents, its application in large animal species has encountered variation in retinal toxicity. NaIO3 induced retinal degeneration and functional changes in sheep, but not in swine. In monkeys, administration of NaIO3 via a carotid artery affected only the cell function of ipsilateral retinal pigment epithelium. The aim of the present study was to identify the dosage and route of NaIO3 administration resulting in morphologic and functional retinal changes in cynomolgus monkeys. Separate groups of animals received NaIO3 intravenously in three different dosing paradigms. Vehicle control animals received phosphate-buffered saline. At selected time points following dosing, flash electroretinograms (ERGs) were recorded followed by necropsy. The eyes were examined microscopically post-necropsy and the levels of circulating microRNA-183 cluster were evaluated in the blood samples collected on days 1, 4, and 5 postdose. A statistically significant reduction in both scotopic a-wave and scotopic and photopic b-wave signals ( p < 0.05) were observed between the ERG signals acquired from NaIO3-treated and vehicle control animals, coupled with time-dependent elevations in plasma miR-183 cluster. Mild to moderate retinal degeneration was observed in the outer layer of the retina, which correlated well with the functional and clinical observations. There were no statistically significant differences in scotopic oscillatory potentials. These findings suggest that intravenous injection of sublethal NaIO3 markedly damaged the cone and rod photoreceptors both functionally and morphologically, and plasma miR-183 reflected the retinal toxicity in those animals with moderate retinal damage.

The mechanism by which angiogenesis declines with aging is not fully understood. Soluble vascular endothelial growth factor receptor 1 (VEGFR1) form (sFlt1) contributes to endothelial dysfunction in pathological conditions. However, the roles of sFlt1 in ischemia-induced neovascularizationof aged animals have not been investigated. To study aging-related sFlt1 change and its impact on ischemia-induced neovascularization, a hindlimb ischemia model was applied to young and aged mice. Blood flow imaging assay revealed that the blood flow recovery remained impaired throughout the follow-up period. At day 14, immunostaining showed lesser capillary formation in the aged mice. An ELISA showed that the aged mice had increased plasma sFlt-1 levels at indicated time points after surgery. On operative day 4, the aged ischemic muscles had decreased levels of p-VEGFR2 and p-Akt and increased levels of sFlt-1, Wnt5a, and SC35 genes or/and protein as well as increased numbers of inflammatory cells (macrophages and leucocytes) and matrix metalloproteinase-9 activity. Immnunofluorescence showed that Flt-1 was co-localized with CD11b+ macrophages of aged ischemic muscles. Hypoxia stimulated sFlt1 expression in CD11b+ cells of aged bone-marrow (BM), and this effect was diminished by siWnt5a. The cultured medium of aged mice BM-derived CD11b+ cells suppressed human endothelial cell (EC) and endothelial progenitor cell (EPC) angiogenic actions induced by VEGF, and these decreases were improved by treatment with siWnt5a-conditioned medium. Thus, aging appears to decline neovascularization in response to ischemic stress via the VEGFR2/Akt signaling inactivation in ECs and ECPs that is mediated by Wnt5a/SC35 axis activated macrophages-derived sFlt1 production in advanced age.

Telithromycin, a ketolide antibiotic, is reported to exacerbate myasthenia gravis, potentially leading to respiratory failure and death. However, telithromycin is not associated with neuromuscular effects in animal toxicity studies. The objective of this study was to examine the effect of telithromycin on the neuromuscular junction in the isolated rat phrenic nerve-diaphragm preparation and to investigate its postsynaptic effects on the muscle-like nicotinic acetylcholine (ACh) receptors expressed on human TE671 cells. Telithromycin decreased the twitch contraction force of the rat diaphragm muscle in response to phrenic nerve stimulation in a concentration-dependent manner with an IC(50) of 22.3 microM and a maximal inhibition of approximately 70%. The trans-membrane current from the ACh receptors expressed in the TE671 neuromedulloblastoma cells was recorded in the whole-cell patch-clamp configuration. When applied to the TE671 cells, telithromycin caused a dose-dependent inhibition of the nicotinic ACh current with an IC(50) of 3.5 microM and maximal inhibition of nearly 100%. These results indicate that telithromycin inhibits postsynaptic nicotinic ACh receptors in vitro and partially blocks neuromuscular transmission in the isolated rat phrenic nerve-diaphragm preparation. Based on these findings, we propose that exacerbation of myasthenia gravis reported in some patients taking telithromycin results in part from postsynaptic neuromuscular transmission block.

Background Therapeutic ultrasound (TUS) has been proposed as a potential strategy to accelerate proliferation and regeneration to the damaged tissue. However, the underline mechanism remains modest clarified. The purpose of this study was to determine the mechanism of TUS on ischemia-induced angiogenesis using mice model of hindlimb ischemia. Methods and results Hindlimb ischemia was induced by unilateral femoral artery resection using 8–12 week-old male C57BL6 mice. External TUS (1.0 MHz, 0.3W/cm 2 ) was then performed for 9 min/day. At postoperative day 14, decreased prevalence of gangrene, increased capillary density and skin temperature ratio between ischaemic/normal extremities were observed in TUS-treated mice. Moreover, the levels of nitric oxide (NO) expression, endothelial nitric oxide synthase (eNOS) and Akt phosphorylation contained in the ischaemic muscles were increased in response to TUS. In vitro , TUS interrupted the process of hypoxia-induced apoptosis and contributed to the tube-formative and proliferative capacities of human umbilical vein endothelial cells. TUS increased the level of NO in the conditioned media, as well as the eNOS and Akt phosphorylation in the cell lysate. Finally, administration of PI3K inhibitor LY294002 and NG-Nitro-L-arginine Methyl Ester (L-NAME) could block these TUS-mediated angiogenic benefits. Conclusions In conclusion, the present study indicates that external ultrasound exposure contributes to ischemia-induced neovascularization through promoting endothelial functions via Akt-eNOS signal pathway.

Background Therapeutic ultrasound (TUS) has been proposed as a potential strategy for therapeutic revascularisation. However, the specific mechanism remains to be explored in detail. The purpose of this study was to determine the mechanism of TUS on ischaemic angiogenesis in mice with acute ischaemic hind limb and proliferation, migration, tube formation and potential signalling pathways of human umbilical vein endothelial cells (HUVECs). Methods TUS was set by a combination of various ultrasound-related variables: frequency of 1.0 MHz; intensity of 0.3 W/cm2; exposure time of 3, 6, or 9 min. We stimulated the HUVECs in culture daily with TUS for 3 days. The effects of therapeutic ultrasound on HUVECs proliferation were evaluated using the CCK-8 assay. Cell migration was assayed by scratch test. Angiogenesis was observed by tube formation assay. The potential involvement of PI3K-Akt-eNOS signalling pathways was explored using selective chemical inhibitor or Western-blot analysis. Left femoral artery ligation was used to establish the model of acute hind limb ischaemic in mice (C57BL/6J male mice, 18–22 g, 2–3 months old), TUS was exposed subcutaneously once everyday for 14 days. The mice were randomly divided into four experimental groups: sham operated group; TUS 3 min group; TUS 6 min group and TUS 9 min group. The experiment was carried out on the 1st and14th after ligation. To determine whether TUS affected the angiogenesis activity of mice with ischaemic hind limb, we measured temperature of hind limb by using infrared thermometer, and microvessel density (MVD) was determined by CD31 staining. Results The results show that proliferation, migration and tube formation in HUVECs could be promoted by TUS at exposure time of 9 min, and inhibited by NG-Nitro-L-arginine Methyl Ester (L-NAME). eNOS and p-eNOS expression increased significantly when TUS 9 min was added, but could be attenuated by L-NAME. p-Akt expression augmented by TUS at exposure time of 9 min, but could be inhibited by LY294002. Compared to the temperature of hind limb, the group of TUS 9 min at the 14th was increased, while TUS 6 min group increased, but only TUS 9 min was there a significant difference. Except that, the MVD was promoted in TUS 9 min group significantly. Conclusions In conclusion, the present study indicates that 9 min of exposure to TUS promote angiogenesis among ischaemic hind limb in mice and that this process is mediated through the PI3K-Akt-eNOS signal pathway in ECs.

Abstract —The mechanisms by which reactive species (RS) participate in the development of atherosclerosis remain incompletely understood. The present study was designed to test the hypothesis that RS produced in the vascular environment cause mitochondrial damage and dysfunction in vitro and, thus, may contribute to the initiating events of atherogenesis. DNA damage was assessed in vascular cells exposed to superoxide, hydrogen peroxide, nitric oxide, and peroxynitrite. In both vascular endothelial and smooth muscle cells, the mitochondrial DNA (mtDNA) was preferentially damaged relative to the transcriptionally inactive nuclear β -globin gene. Similarly, a dose-dependent decrease in mtDNA-encoded mRNA transcripts was associated with RS treatment. Mitochondrial protein synthesis was also inhibited in a dose-dependent manner by ONOO − , resulting in decreased cellular ATP levels and mitochondrial redox function. Overall, endothelial cells were more sensitive to RS-mediated damage than were smooth muscle cells. Together, these data link RS-mediated mtDNA damage, altered gene expression, and mitochondrial dysfunction in cell culture and reveal how RS may mediate vascular cell dysfunction in the setting of atherogenesis.

Spike activity in dorsal root ganglion (DRG) neurons depolarizes passive neighbors that share the same ganglion. We asked whether age or prior nerve injury affect this 'cross-depolarization' signal. Intracellular recordings made from excised DRGs in vitro revealed that the prevalence and duration of cross-depolarization were no greater in adult than in young rats, and that its amplitude was significantly smaller in adults. The amplitude of cross-depolarization was not affected by nerve injury. The decrease in membrane input resistance (R(in)) observed during cross-depolarization was less than that expected from equivalent depolarization alone. This affirms prior evidence that the neural process underlying cross-depolarization causes a net increase in R(in).

The capacity of antisense sequences to the thrombin receptor to selectively inhibit thrombin receptor expression and limit mitogenic responses in vascular wall cells was investigated in vitro. Eight phosphorothioate oligodeoxynucleotides based on the sequences of the rat thrombin receptor (including sense, antisense, scrambled, and missense controls) were synthesized, characterized, and purified by high performance liquid chromatography. The antisense oligodeoxynucleotide (ODN 4) inhibitory effect was sequence-specific and both time-and concentration-dependent. A reduction in serum or alpha-thrombin-induced smooth muscle cell (SMC) proliferation was noted as early as 3 days at 30 microM (82%; 6.17 +/- 1.01 versus 34.08 +/- 3.89 x 10(4) cells/well; p0.05) and at a dose as low as 15 microM after 4 days in culture (19%; p0.05). Nonspecific effects were enhanced after prolonged exposure of SMC to the antisense oligodeoxynucleotide (or = 6 days). A reduction of inositol phosphate generation greater than 50% (p0.05) was detected after exposure of SMC to antisense but not to sense or scrambled nucleotide sequences. This was observed after stimulation with both thrombin and SFFLRN (thrombin receptor peptide agonist). Northern blot analysis and enzyme-linked immunosorbent assays revealed 50 and 22% decreases, respectively, in thrombin receptor mRNA and protein (cell surface) levels in antisense oligonucleotide-treated (72 h) SMC as compared to untreated cells, suggesting that thrombin receptor down-regulation occurred at the pretranslational level. Thus, thrombin receptor-specific antisense sequences inhibit growth-related effects both of serum and thrombin on smooth muscle cells, potentially providing a new strategy for selective inhibition of receptor-mediated arterial injury responses.