Wogonoside impedes the progression of acute myeloid leukemia through inhibiting bone marrow angiogenesis

Decreasing bone marrow (BM) microvessel density and circulating angiogenic cytokine levels are promising strategies for the treatment of relapsed and resistant acute myeloid leukemia (AML). Previous studies have reported that wogonoside could inhibit the progression of AML and suppress angiogenesis in a solid tumor, but the correlation of these two effects was ignored. In this research, we determined whether wogonoside could inhibit angiogenesis in this hematologic malignancy. We found that wogonoside could inhibit tumor growth and progression, and prolong the survival of nude mice inoculated with U937/MDR. Besides, reducing BM angiogenesis might cause therapeutic effect against resistant AML. Therefore, coculture between AML cells and BM stromal cells was established to imitate their crosstalk. Then, the effect of wogonoside on BM angiogenesis was tested in vitro and in vivo. We found that wogonoside could suppress microvessel formation in the chicken chorioallantoic membrane assay model and matrigel plug assay. The mechanism research revealed that wogonoside could block the JAK2-STAT3 pathway in AML cells and stromal cells to break their positive feedback. We detected several cytokines related to AML or angiogenesis and found that secreted interleukin-8 was a significant angiogenic cytokine to induce BM angiogenesis. These findings supported that new diagnostics and promising treatment strategies could be developed in relapsed and resistant AML patients.     

A polysaccharide from the dried rhizome of Drynaria fortunei (Kunze) J. Sm. prevents ovariectomized (OVX)-induced osteoporosis in rats

In the present study, a homogenous polysaccharide (DFPW) was isolated and purified from the dried rhizome of Drynaria fortunei, and its protective effect against osteoporosis was investigated in ovariectomized (OVX) rats. Histological analysis indicated that oral administration of DFPW (100 and 400 mg/kg) for 12 weeks significantly improved trabecular bone mass, as demonstrated by the increase in trabecular area, trabecular thickness and its number in OVX rats. Furthermore, the decline of bone mineral density and bone mineral content including Ca, P and Mg induced by OVX was reversed by the DFPW administration. This function was achieved by the decreased levels of the bone turnover markers, such as serum ALP, urinary deoxypyridinoline (DPD), Ca and P excretions. Besides, DFPW improved biomechanical parameters (maximum load, energy, Young's, modulus and maximum stress) to strengthen the hardness and strength femoral diaphysis in OVX rats. These results strongly suggested that DFPW might be a hopeful alternative therapeutics to treat postmenopausal osteoporosis.     

Molecular Detection of Novel Anammox Bacterial Clusters in the Sediments of the Shallow Freshwater Lake Taihu

Previous studies have demonstrated the wide occurrence of anaerobic ammonium oxidizing (anammox) bacteria; however, there is very limited information on the distribution of these bacteria in freshwater habitats. In this study, the anammox bacterial communities were detected by molecular analysis targeting the 16S rRNA genes in the sediments of Lake Taihu, a large and shallow eutrophic freshwater lake in China. The recovery of specific 16S rRNA sequences with two stable monophyletic clusters indicated that anammox bacteria were present in Lake Taihu. A phylogenetic analysis indicated that these two groups represent two novel lineages within the first subgroup of anammox bacteria, independent of the treeing methods. High intra-lake variability in anammox bacterial diversity and community composition was observed, in particular, based on a 1% cut-off of 16S rRNA sequence variation. The spatial variability was largely related to the substrate availability, which was denoted by the correlations between the relative abundance of the two Taihu anammox bacterial groups and the concentrations of ammonium and nitrite. This indicates that the niche differentiation of anammox bacteria is linked to the environmental heterogeneity. These findings suggest that the freshwater lakes may accommodate different anammox bacterial communities and, thus, expand our knowledge on the diversity and distribution of anammox bacteria. Supplemental materials are available for this article. Go to the publisher's online edition of Geomicrobiology Journal to view the free supplemental files.     

An efficient method for protein function annotation based on multilayer protein networks

Background

Accurate annotation of protein functions is still a big challenge for understanding life in the post-genomic era. Many computational methods based on protein-protein interaction (PPI) networks have been proposed to predict the function of proteins. However, the precision of these predictions still needs to be improved, due to the incompletion and noise in PPI networks. Integrating network topology and biological information could improve the accuracy of protein function prediction and may also lead to the discovery of multiple interaction types between proteins. Current algorithms generate a single network, which is archived using a weighted sum of all types of protein interactions.

Method

The influences of different types of interactions on the prediction of protein functions are not the same. To address this, we construct multilayer protein networks (MPN) by integrating PPI networks, the domain of proteins, and information on protein complexes. In the MPN, there is more than one type of connections between pairwise proteins. Different types of connections reflect different roles and importance in protein function prediction. Based on the MPN, we propose a new protein function prediction method, named function prediction based on multilayer protein networks (FP-MPN). Given an un-annotated protein, the FP-MPN method visits each layer of the MPN in turn and generates a set of candidate neighbors with known functions. A set of predicted functions for the testing protein is then formed and all of these functions are scored and sorted. Each layer plays different importance on the prediction of protein functions. A number of top-ranking functions are selected to annotate the unknown protein.

Conclusions

The method proposed in this paper was a better predictor when used on Saccharomyces cerevisiae protein data than other function prediction methods previously used. The proposed FP-MPN method takes different roles of connections in protein function prediction into account to reduce the artificial noise by introducing biological information.

     

Antifungal property of dihydrodehydrodiconiferyl alcohol 9'-O-β-d-glucoside and its pore-forming action in plasma membrane of Candida albicans

The aims of this study were to investigate the antifungal activity as a bioactive property of dihydrodehydrodiconiferyl alcohol 9'-O-β-d-glucoside (DDDC9G) and the mode of action(s) involved in its effect. Antifungal susceptibility testing showed that DDDC9G possessed potent antifungal activities toward various fungal strains with almost no hemolytic effect. To understand the antifungal mechanism(s) of DDDC9G, we conducted the following experiments in this study using Candida albicans. Fluorescence experiments using the probes, 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) and propidium iodide suggested that DDDC9G perturbed the fungal plasma membrane. Consecutively, the analysis of the transmembrane electrical potential (ΔΨ) with 3, 3'-dipropylthiadicarbocyanine iodide [DiSC(3)(5)] and bis-(1, 3-dibutylbarbituric acid) trimethine oxonol [DiBAC(4)(3)] indicated that DDDC9G induced membrane-depolarization. Furthermore, model membrane studies were performed with rhodamine-labeled giant unilamellar vesicles (GUVs), calcein encapsulating large unilamellar vesicles (LUVs), and FITC-dextran (FD) loaded LUVs. These results demonstrated that the antifungal effects of DDDC9G upon the fungal plasma membrane were through the formation of pores with the radii between 0.74nm and 1.4nm. Finally, in three dimensional (3D) flow cytometric contour plots, a reduced cell size was observed as a result of osmolarity changes from DDDC9G-induced structural and functional membrane damages. Therefore, the present study suggests that DDDC9G exerts its antifungal effect by damaging the membrane through pore formation in the fungal plasma membrane.     

Low Taxon Richness of Bacterioplankton in High-Altitude Lakes of the Eastern Tibetan Plateau,with a Predominance of Bacteroidetes and Synechococcus spp.

Plankton samples were collected from six remote freshwater and saline lakes located at altitudes of 3,204 to 4,718 m and 1,000 km apart within an area of ca. 1 million km² on the eastern Tibetan Plateau to comparatively assess how environmental factors influence the diversity of bacterial communities in high-altitude lakes. The composition of the bacterioplankton was investigated by analysis of large clone libraries of 16S rRNA genes. Comparison of bacterioplankton diversities estimated for the six Tibetan lakes with reference data previously published for lakes located at lower altitudes indicated relatively low taxon richness in the Tibetan lakes. The estimated average taxon richness in the four Tibetan freshwater lakes was only one-fifth of the average taxon richness estimated for seven low-altitude reference lakes. This cannot be explained by low coverage of communities in the Tibetan lakes by the established libraries or by differences in habitat size. Furthermore, a comparison of the taxonomic compositions of bacterioplankton across the six Tibetan lakes revealed low overlap between their community compositions. About 70.9% of the operational taxonomic units (99% similarity) were specific to single lakes, and a relatively high percentage (11%) of sequences were <95% similar to publicly deposited sequences of cultured or uncultured bacteria. This beta diversity was explained by differences in salinity between lakes rather than by distance effects. Another characteristic of the investigated lakes was the predominance of Cyanobacteria (Synechococcus) and Bacteroidetes. These features of bacterioplankton diversity may reflect specific adaptation of various lineages to the environmental conditions in these high-altitude lakes.Bacterioplankton is a major component of aquatic ecosystems (13). The advent of molecular techniques during the past 2 decades has advanced our knowledge of the diversity and ecology of bacterioplankton in various aquatic habitats (7, 14, 19, 20, 39, 63, 64, 67). Based on comparative analyses of bacterial 16S rRNA sequences obtained from diverse freshwater habitats, typical freshwater bacterial clusters have been proposed (67), and this list of clusters was subsequently expanded (e.g., 2, 15, 23, 57). Many of these freshwater bacterial clusters appeared to be widely distributed in lakes with different ecological characteristics and in different geographic regions (34, 38, 44, 57, 67). However, most of the knowledge about freshwater bacterioplankton diversity originated from investigations of low-altitude lake systems. Differences in elevation result in pronounced differences in environmental conditions in lakes (59). Lakes at high altitudes are often characterized by oligotrophy, low temperature, low productivity, simple food web structure, and strong UV radiation in surface water layers. All these factors have been shown to strongly influence bacterial community composition and bacterial taxon richness (e.g., 25, 32, 33, 35, 48, 58, 60). These in situ environmental conditions, which change with the elevation, may result in a change in the bacterioplankton community and taxon richness at high elevations. Since lake ecosystems at high altitude are quite sensitive to the impacts of climate change, i.e., global warming, and these rarely explored lakes may harbor new microbial species, our knowledge of the bacterial diversity in high-altitude lakes is also crucial to species protection and ecosystem conservation.In this study, sequence analysis of large 16S rRNA gene clone libraries was applied in order to obtain deep insights into the phylogenetic diversity of bacterioplankton in six permanent high altitude lakes located ca. 1,000 km apart within an area of ca. 1 million km² on the eastern Tibetan Plateau (Fig. ​(Fig.1).1). We selected these lakes because (i) they are permanent lakes that have existed for thousands of years and lack direct anthropogenic influences (66); (ii) they posses typical characteristics of high-altitude lakes, like low nutrient concentration, low temperature, low primary productivity, and transparent water; (iii) they are located distant from each other in three different regions of the plateau; and (iv) they cover gradients in salinity and chemical composition that are typical of the ecologically broad spectrum of Tibetan lakes (62). By performing interlake comparisons across these Tibetan lakes and comparisons with the available information on bacterioplankton diversity in low-altitude lakes, we aimed to (i) evaluate the taxon richness of bacterioplankton in lakes in relation to the indirect influence of elevation via the environmental conditions, (ii) test the importance of spatial and local environmental factors for bacterioplankton community composition (BCC) in these remote high-altitude lakes, and (iii) characterize the phylogenetic diversity of bacterioplankton and assess the community features in these remote ecosystems.Open in a separate windowFIG. 1.Map of central Tibet indicating the locations of the investigated lakes. 1, Lake Qinghai; 2, Lake Zhaling; 3, Lake Tuosuhai; 4, Lake E''ling; 5, Lake Xinxinhai; 6, Lake Namucuo. (Adapted from reference 61a, with kind permission from Springer Science+Business Media.)     

Spectacular modification of Gambogic acid on microwave irradiation in methanol: Isolation and structure identification of two products with potent anti-tumor activity

Treatment of gambogic acid with methanol in acidic condition under microwave irradiation led to the formation of two new products bearing spectacular A ring systems which were different from reported Garcinia natural products. The chemical structures of these two compounds were elucidated by extensive NMR and MS spectroscopic analysis as well as crystallographic study. Both of the two products could significantly inhibit the growth of various tumor cell lines in vitro with IC₅₀ values at μM level through apoptotic pathway.     

Overexpression of ZEB1 associated with metastasis and invasion in patients with gastric carcinoma

The aim of this study was to investigate the expression of ZEB1 in gastric carcinoma, its correlation with the clinicopathology of gastric carcinoma, and the role of ZEB1 in invasion and metastasis in gastric carcinoma. ZEB1 expression was analyzed by immunohistochemistry and Western blot in 45 gastric carcinoma tissue samples that contained the adjacent gastric mucosa. The correlation between ZEB1 expression, the occurrence and development of gastric cancer, and clinical pathology was investigated. ZEB1 expression in the human gastric carcinoma cell line AGS was downregulated by RNA interference, and changes in ZEB1 expression corresponded with changes in the invasive and metastatic ability of AGS cells. Immunohistochemistry revealed that ZEB1 protein expression in gastric carcinoma tissues was significantly higher than in normal gastric mucosa tissues (p < 0.001). A lower degree of differentiation of gastric cancer (p = 0.009), a higher TNM (tumor, node, and metastasis) stage (p = 0.010), and a larger scope of invasion were correlated with higher expression of ZEB1 (p = 0.041, 0.002). However, the expression of ZEB1 in gastric carcinoma tissue was independent of gender, age, and tumor size (p > 0.05). Western blot results also showed that ZEB1 protein expression was significantly higher in gastric carcinoma tissue than in the adjacent normal gastric mucosa tissue (p = 0.008). A lower degree of differentiation of the gastric carcinoma correlated with a higher TNM stage, and a larger scope of invasion correlated with increased ZEB1 expression (p = 0.023). Transfection of ZEB1 siRNA in AGS cells significantly decreased the expression level of ZEB1 protein (p = 0.035). Furthermore, the number of cells that could pass through the Transwell chamber was significantly lower in the transfected group than in the non-transfected control group (p = 0.039), indicating that the suppression of ZEB1 expression could significantly reduce the invasive and metastatic ability of AGS cells (p = 0.005). Concluding, in gastric carcinoma tissue, overexpression of ZEB1 may be related to the occurrence and development as well as invasion and metastasis of gastric carcinoma.