Abundance and Diversity of Ammonia-Oxidizing Bacteria and Archaea in Cold Springs on the Qinghai-Tibet Plateau

The cold springs underlain by gas hydrates on the Qinghai-Tibet Plateau (QTP) are similar to deep-sea cold seeps with respect to methane biogeochemistry. Previous studies have shown that ammonia oxidizing bacteria (AOB) and archaea (AOA) are actively present and play important roles in the carbon/nitrogen cycles in cold seeps. Studying AOA and AOB communities in the QTP cold springs will be of great importance to our understanding of carbon and nitrogen cycling dynamics related to the underlying gas hydrates on the QTP. Thus, the abundance and diversity of AOB and AOA in sediments of four cold springs underlain by gas hydrates on the QTP were determined by using quantitative polymerase chain reaction and amoA gene (encoding ammonia monooxygenase involved in ammonia oxidation) phylogenetic analysis. The results showed that the AOB and AOA amoA gene abundances were at 10³–10⁴ copies per gram of the sediments in the investigated cold springs. The AOB population consisted of Nitrosospira and Nitrosomonas in contrast with the mere presence of Nitrosospira in marine cold seeps. The AOB diversity was higher in cold springs than in cold seeps. The AOA population was mainly composed of Nitrososphaera, in contrast with the dominance of Nitrosopumilus in cold seeps. The terrestrial origin and high level of dissolved oxygen of the cold springs may be the main factors accounting for the observed differences in AOB and AOA populations between the QTP cold springs and marine cold seeps.     

S‐nitrosylated protein disulfide isomerase contributes to mutant SOD1 aggregates in amyotrophic lateral sclerosis

A major hallmark of mutant superoxide dismutase (SOD1)‐linked familial amyotrophic lateral sclerosis is SOD1‐immunopositive inclusions found within motor neurons. The mechanism by which SOD1 becomes aggregated, however, remains unclear. In this study, we aimed to investigate the role of nitrosative stress and S‐nitrosylation of protein disulfide isomerase (PDI) in the formation of SOD1 aggregates. Our data show that with disease progression inducible nitric oxide synthase (iNOS) was up‐regulated, which generated high levels of nitric oxide (NO) and subsequently induced S‐nitrosylation of PDI in the spinal cord of mutant SOD1 transgenic mice. This was further confirmed by in vitro observation that treating SH‐SY5Y cells with NO donor S‐nitrosocysteine triggered a dose‐dependent formation of S‐nitrosylated PDI. When mutant SOD1 was over‐expressed in SH‐SY5Y cells, the iNOS expression was up‐regulated, and NO generation was consequently increased. Furthermore, both S‐nitrosylation of PDI and the formation of mutant SOD1 aggregates were detected in the cells expressing mutant SOD1^G93A. Blocking NO generation with the NOS inhibitor N‐nitro‐l ‐arginine attenuated the S‐nitrosylation of PDI and inhibited the formation of mutant SOD1 aggregates. We conclude that NO‐mediated S‐nitrosylation of PDI is a contributing factor to the accumulation of mutant SOD1 aggregates in amyotrophic lateral sclerosis.     

Cadmium removal and 2,4-dichlorophenol degradation by immobilized Phanerochaete chrysosporium loaded with nitrogen-doped TiO2 nanoparticles

Phanerochaete chrysosporium has been identified as an effective bioremediation agent for its biosorption and degradation ability. However, the applications of P. chrysosporium are limited owing to its long degradation time and low resistance to pollutants. In this research, nitrogen-doped TiO₂ nanoparticles were loaded on P. chrysosporium to improve the remediation capacity for pollutants. The removal efficiencies were maintained at a high level: 84.2 % for Cd(II) and 78.9 % for 2,4-dichlorophenol (2,4-DCP) in the wide pH range of 4.0 to 7.0 in 60 h. The removal capacity of immobilized P. chrysosporium loaded with nitrogen-doped TiO₂ nanoparticles (PTNs) was strongly affected by the initial Cd(II) and 2,4-DCP concentrations. The hyphae of PTNs became tight, and a large amount of crystals adhered to them after the reaction. Fourier transform infrared spectroscopy showed that carboxyl, amino, and hydroxyl groups on the surface of PTNs were responsible for the biosorption. In the degradation process, 2,4-DCP was broken down into o-chlorotoluene and 4-hexene-1-ol. These results showed that PTNs is promising for simultaneous removal of Cd(II) and 2,4-DCP from wastewater.     

Seed rain and its relationship with above-ground vegetation of degraded Kobresia meadows

Seed rain is a crucial element in vegetation regeneration, but has been rarely studied in high altitude regions, particularly degraded Kobresia meadow. Weed infestation is a distinctive feature of pasture degradation in Kobresia meadows on the Tibetan plateau, the ecological mechanism of which is closely related with vegetation’s seed rain. In this paper we assess the effect of vegetation degradation on seed rain and consider its implication for restoration of degraded Kobresia meadows in the headwater area of Yellow river, through analysis of seed species composition, number of seeds landing per m² of soil surface, and their relationship with above ground vegetation. Vegetation degradation had an impact on the species composition and numbers of seeds in seed rain and their relationship with above-ground vegetation. Within the un-degraded meadow, which provided a closed vegetation cover, 35 % of the seed rain was of sedge and gramineae species. However, within the degraded meadows, as the extent of degradation increased, so the total number of seeds m^?2 increased, with those derived from sedge and gramineae species forming a declining proportion of the total. Degradation of Kobresia meadow on the Tibetan plateau is exacerbated by the seed input of weed species (such as Oxytropis ochrocephala, Carum carvi, Aconitum pendulum, Pedicularis kansuensis in this study). Therefore, a major priority for the restoration of such degraded meadows should be the elimination of these weeds from the above ground vegetation by human intervention.     

3,5-Diiodo-l-thyronine ameliorates diabetic nephropathy in streptozotocin-induced diabetic rats

3, 5-Diiodothyronine (T2), a natural metabolite of triiodothyronine (T3) from deiodination pathway, can mimic biologic effects of T3 without inducing thyrotoxic effects. Recent studies revealed T3 acted as a protective factor against diabetic nephropathy (DN). Nevertheless, little is known about the effect of T2 on DN. This study was designed to investigate whether and how T2 affects experimental models of DN in vivo and in vitro. Administration of T2 was found to prevent significant decrease in SIRT1 protein expression and activity as well as increases in blood glucose, urine albumin excretion, matrix expansion, transforming growth factor-β1 expression, fibronectin and type IV collagen deposition in the diabetic kidney. Concordantly, similar effects of T2 were exhibited in the cultured rat mesangial cells (RMC) exposed to high glucose and that could be abolished by a known SIRT1 inhibitor, sirtinol. Moreover, enhanced NF-κB acetylation and JNK phosphorylation present in both diabetic rats and high glucose-treated RMC were distinctly dampened by T2. Collectively, these results suggested that T2 was a protective agent against renal damage in diabetic nephropathy, whose action involved regulation of SIRT1.     

Cancer-initiating cells derived from human rectal adenocarcinoma tissues carry mesenchymal phenotypes and resist drug therapies

Accumulating evidence indicates that cancer-initiating cells (CICs) are responsible for cancer initiation, relapse, and metastasis. Colorectal carcinoma (CRC) is typically classified into proximal colon, distal colon, and rectal cancer. The gradual changes in CRC molecular features within the bowel may have considerable implications in colon and rectal CICs. Unfortunately, limited information is available on CICs derived from rectal cancer, although colon CICs have been described. Here we identified rectal CICs (R-CICs) that possess differentiation potential in tumors derived from patients with rectal adenocarcinoma. The R-CICs carried both CD44 and CD54 surface markers, while R-CICs and their immediate progenies carried potential epithelial–mesenchymal transition characteristics. These R-CICs generated tumors similar to their tumor of origin when injected into immunodeficient mice, differentiated into rectal epithelial cells in vitro, and were capable of self-renewal both in vitro and in vivo. More importantly, subpopulations of R-CICs resisted both 5-fluorouracil/calcium folinate/oxaliplatin (FolFox) and cetuximab treatment, which are the most common therapeutic regimens used for patients with advanced or metastatic rectal cancer. Thus, the identification, expansion, and properties of R-CICs provide an ideal cellular model to further investigate tumor progression and determine therapeutic resistance in these patients.     

Three‐dimensional neural differentiation of embryonic stem cells with ACM induction in microfibrous matrices in bioreactors

The clinical use of pluripotent stem cell (PSC)‐derived neural cells requires an efficient differentiation process for mass production in a bioreactor. Toward this goal, neural differentiation of murine embryonic stem cells (ESCs) in three‐dimensional (3D) polyethylene terephthalate microfibrous matrices was investigated in this study. To streamline the process and provide a platform for process integration, the neural differentiation of ESCs was induced with astrocyte‐conditioned medium without the formation of embryoid bodies, starting from undifferentiated ESC aggregates expanded in a suspension bioreactor. The 3D neural differentiation was able to generate a complex neural network in the matrices. When compared to 2D differentiation, 3D differentiation in microfibrous matrices resulted in a higher percentage of nestin‐positive cells (68% vs. 54%) and upregulated gene expressions of nestin, Nurr1, and tyrosine hydroxylase. High purity of neural differentiation in 3D microfibrous matrix was also demonstrated in a spinner bioreactor with 74% nestin + cells. This study demonstrated the feasibility of a scalable process based on 3D differentiation in microfibrous matrices for the production of ESC‐derived neural cells. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1013–1022, 2013     

Hela细胞HGPRT缺陷型细胞系构建

建立稳定的次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)缺陷的Hela细胞系,为细胞融合相关研究和人源化单克隆抗体制备提供有利于筛选的亲本细胞。通过诱变剂N-甲基-N′-硝基-N-亚硝基胍(MNNG)对Hela细胞进行诱变,逐步提高培养基中6-巯基鸟嘌呤(6-TG)的浓度,筛选出对6-TG稳定耐受的细胞,在次黄嘌呤-氨基喋呤-胸腺嘧啶(hypoxanthine-aminopterin-thymidine,HAT)培养基中鉴定其敏感性,最后对筛选得到的Hela-HGPRT-进行生物学鉴定。在此基础上,将Hela-HGPRT-细胞系与人淋巴细胞融合,在HAT培养基中筛选杂交细胞。筛选得到了能够长期在含20μg/mL 6-TG培养基中生长的Hela-HGPRT-细胞,并且在HAT培养基中不能存活。Hela-HGPRT-细胞与人淋巴细胞成功融合,获得能够连续传代培养的杂交瘤细胞。经MNNG诱导和6-TG筛选,得到了稳定传代的Hela-HGPRT-细胞系,该细胞系可用于细胞融合相关研究。