Changes in gene expression in Arabidopsis shoots during phosphate starvation and the potential for developing smart plants

Our aim was to generate and prove the concept of "smart" plants to monitor plant phosphorus (P) status in Arabidopsis. Smart plants can be genetically engineered by transformation with a construct containing the promoter of a gene up-regulated specifically by P starvation in an accessible tissue upstream of a marker gene such as beta-glucuronidase (GUS). First, using microarrays, we identified genes whose expression changed more than 2.5-fold in shoots of plants growing hydroponically when P, but not N or K, was withheld from the nutrient solution. The transient changes in gene expression occurring immediately (4 h) after P withdrawal were highly variable, and many nonspecific, shock-induced genes were up-regulated during this period. However, two common putative cis-regulatory elements (a PHO-like element and a TATA box-like element) were present significantly more often in the promoters of genes whose expression increased 4 h after the withdrawal of P compared with their general occurrence in the promoters of all genes represented on the microarray. Surprisingly, the expression of only four genes differed between shoots of P-starved and -replete plants 28 h after P was withdrawn. This lull in differential gene expression preceded the differential expression of a new group of 61 genes 100 h after withdrawing P. A literature survey indicated that the expression of many of these "late" genes responded specifically to P starvation. Shoots had reduced P after 100 h, but growth was unaffected. The expression of SQD1, a gene involved in the synthesis of sulfolipids, responded specifically to P starvation and was increased 100 h after withdrawing P. Leaves of Arabidopsis bearing a SQD1::GUS construct showed increased GUS activity after P withdrawal, which was detectable before P starvation limited growth. Hence, smart plants can monitor plant P status. Transferring this technology to crops would allow precision management of P fertilization, thereby maintaining yields while reducing costs, conserving natural resources, and preventing pollution.     

高寒草甸黄帚橐吾种群根际/非根际土壤可培养微生物群落特征

黄帚橐吾（Ligularia virgaurea）是高寒草甸常见的毒杂草,被认为是指示一个地区草地植被退化的重要物种,研究其根际/非根际土壤微生物在不同海拔梯度上的群落特征具有重要意义。以甘南州高寒草甸不同海拔梯度黄帚橐吾根际/非根际土壤可培养微生物为研究对象,采用稀释涂布平板法和最大可能数法（MPN）测定了土壤微生物的数量及土壤理化因子的变化。结果表明：细菌在微生物总数中占比最大,根际微生物数量随海拔升高呈先增加后减小的变化,非根际则表现为递增的趋势,微生物功能群在根际和非根际土壤中均逐渐增加;根际土壤的微生物和功能群数量均高于非根际土壤。RDA分析发现,土壤温度、有机碳、电导率、pH、全氮、全磷、速效氮及脲酶对根际/非根际土壤微生物数量及功能群变化影响较大。通径分析可知：根际土壤中,细菌和真菌受速效氮和有机碳影响较大,放线菌主要受土壤温度和电导率的影响;根际土壤固氮菌和氨化细菌决策系数速效氮 > 有机碳 > 全氮;根际和非根际土壤中硝化细菌的影响因子各不相同,根际土壤决策系数最大和最小分别为全磷和全氮,非根际则是pH和脲酶。     

Hypoxia‐induced miR‐3677‐3p promotes the proliferation,migration and invasion of hepatocellular carcinoma cells by suppressing SIRT5

Hepatocellular carcinoma (HCC), with life‐threatening malignant behaviours, often develops distant metastases and is the fourth most common primary cancer in the world, having taken millions of lives in Asian countries such as China. The novel miR‐3677‐3p is involved in a high‐expression‐related poor prognosis in HCC tissues and cell lines, indicating oncogenesis functions in vitro and in vivo. Initially, we confirmed the inhibition of proliferation, migration and invasion in miR‐3677‐3p knock‐down MHCC‐97H and SMMC‐7721 cell lines, which are well known for their high degree of invasiveness. Then, we reversed the functional experiments in the low‐miR‐3677‐3p‐expression Hep3B cell line via overexpressing miR‐3677‐3p. In nude mice xenograft and lung metastasis assays, we found suppressor behaviours, smaller nodules and low density of organ spread, after injection of cells transfected with shRNA‐miR‐3677‐3p. A combination of databases (Starbase, TargetScan and MiRgator) illustrated miR‐3677‐3p targets, and it was shown to suppress the expression of SIRT5 in a dual‐luciferase reporter system. To clarify the conclusions of previous ambiguous research, we up‐regulated SIRT5 in Hep3B cells, and rescue tests were established for confirmation that miR‐3677‐3p suppresses SIRT5 to enhance the migration and invasion of HCC. Interestingly, we discovered hypoxia‐induced miR‐3677‐3p up‐regulation benefited HCC malignancy and invasiveness. In conclusion, the overexpression of miR‐3677‐3p mediated SIRT5 inhibition, which could increase proliferation, migration and invasion of HCC in hypoxic microenvironments.     

Vitamin A excess alters membrane flow in rat liver

Pulse-chase methodology with [35S]methionine as label was employed to determine flow kinetics through the endoplasmic reticulum-Golgi apparatus-(lysosome-) secretory vesicle-plasma membrane export route in livers of animals receiving vitamin A excess by gavage. Overall fraction composition determined by morphometry and by analyses of marker enzymes was unchanged by vitamin administration. The vitamin modified the pattern of flow of proteins through the Golgi apparatus to the cell surface and to lysosomes. Altered flux was evidenced by a markedly reduced rate of labeling of lysosomes and a slightly increased rate of labeling of both total membrane proteins of the plasma membrane and of a specific membrane glycoprotein GP80. Also reduced was overall labeling of the Golgi apparatus. Differences in the rate or routes of trafficking of glycoproteins through the Golgi apparatus together with altered opportunities for processing might account for some of the alterations in glycoconjugate glycosylation associated with excess vitamin A administration.     

Rat Sertoli cells in culture undergo metabolic co-operation with each other and with fibroblasts

Metabolic co-operation between Sertoli cells from adult rats was detected by adding one group of cells, which were the recipients, to a second group of cells, which had been labelled for 3 h with [3H]uridine and were the donors. Metabolic co-operation also was studied by co-culturing Sertoli cells, which were the donors, with human or Chinese hamster HGPRT- fibroblasts (recipients) in the presence of [3H]hypoxanthine. With both techniques the recipients in contact with donors had significantly more radioactive grains than did the recipients alone. In all cases the proportion of interactions that were positive for metabolic co-operation was greater than 80%.     

The fine structure of the planarianPolycelis tenuis ijima

Summary The fine structure of the pharynx is presented and demonstrates that the pharyngeal epithelial system is a continuous one. The epithelial lining of the pharyngeal cavity with its characteristic fibrous secretory bodies merges with the outer pharyngeal epithelium at the point of anchorage of the pharynx. A few of these cells are insunk, the nuclei occurring beneath the underlying muscular layers. The nature of the outer epithelium changes towards the free end of the pharynx; the cells become ciliated and in contents come to resemble the inner epithelium which it joins at the tip.The gut cells merge at a transitional zone with the inner pharyngeal epithelium and at this point both bear microvilli and contain rod-shaped apical bodies. Some of these cells are also insunk. Towards the mouth the epithelium shows a greater degree of insinking and exhibits microapocrine secretion. Both inner and outer epithelia bear sense receptors which are concentrated at the lip.At the point of pharyngeal insertion, the sub-epithelial tissue resembles planarian parenchyma, but is rich in gland cells. These glands open on to the outer epithelium especially towards the free end of the pharynx.This research was supported by the Scientific Research Council. Grant No. B/RG/086.     

A case of Ebola virus infection.

In November 1976 an investigator at the Microbiological Research Establishment accidentally inoculated himself while processing material from patients in Africa who had been suffering from a haemorrhagic fever of unknown cause. He developed an illness closely resembling Marburg disease, and a virus was isolated from his blood that resembled Marburg virus but was distinct serologically. The course of the illness was mild and may have been modified by treatment with human interferon and convalescent serum. Convalescence was protracted; there was evidence of bone-marrow depression and virus was excreted in low titre for some weeks. Recovery was complete. Infection was contained by barrier-nursing techniques using a negative-pressure plastic isolator and infection did not spread to attendant staff or to the community.     

Engineering a therapeutic IgG molecule to address cysteinylation,aggregation and enhance thermal stability and expression

Antibodies can undergo a variety of covalent and non-covalent degradation reactions that have adverse effects on efficacy, safety, manufacture and storage. We had identified an antibody to Angiopoietin 2 (Ang2 mAb) that neutralizes Ang2 binding to its receptor in vitro and inhibits tumor growth in vivo. Despite favorable pharmacological activity, the Ang2 mAb preparations were heterogeneous, aggregated rapidly and were poorly expressed. Here, we report the engineering of the antibody variable and constant domains to generate an antibody with reduced propensity to aggregate, enhanced homogeneity, 11°C elevated T_m, 26-fold improved level of expression and retained activity. The engineered molecule, MEDI-3617, is now compatible with the large scale material supply required for clinical trials and is currently being evaluated in Phase 1 in cancer patients. This is the first report to describe the stability engineering of a therapeutic antibody addressing non canonical cysteine residues and the design strategy reported here is generally applicable to other therapeutic antibodies and proteins.