[Microbial Community Structure Shift During Bioremediation of Petroleum Contaminated Soil Using High Throughput Sequencing].

The shift in microbial community structure during the bioremediation of oil-polluted soil was analyzed by high-throughput sequencing. The results demonstrated obvious changes in the soil microbial community structure and diversity during bioremediation. The species richness and evenness of the microbial community decreased substantially due to the bioaugmentation treatment. Proteobacteria became the predominant phylum, with a relative increase in abundance from 37.44% to 87.44%. Pseudomonas was the most dominant genus, which increased in abundance from 2.99% to 76.37%. In the biostimulation treated soil, the relative abundance of Proteobacteria decreased from 37.44% to 10.90%, while the phylum Firmicutes increased from 9.16% to 35.32%. At the genus level, the relative abundances of Exiguobacterium and Promicromonospora decreased from 8.49% and 18.96% to 2.19% and 14.97%, respectively. Nocardioides and Bacillus became the dominant genera and increased from 5.56% and 0.29% to 28.95% and 22.70%, respectively. The results indicated that bioaugmentation substantially influenced the soil microbial diversity and community structure. Additionally, the biostimulation treatment maintained the balance in the soil microbial community structure. The stabilization of bacteria community structure is beneficial to petroleum biodegradation in the soil.

[Isolation, Identification and Degradation Characteristics of a 17ß-estradiol Degrading Strain Fusarium sp. KY123915].

A 17ß-estradiol (E2) degrading strain (designated as Wu-SP1) was isolated from the activated sludge collected from a wastewater treatment plant (WWTP) in Xi'an. The strain was identified as Fusarium sp. according to 18S rDNA sequence and phylogenetic analysis. The optimal pH and temperature for E2 degradation were 6 and 30℃, respectively. Under these conditions, the E2 biodegradation rate of 2 mg·L-1 E2 amounted to 92.5% within 48 h by this strain. The kinetics of E2 degradation by the strain KY123915 were in good accord with the first-order equation, with the concentration ranged from 10 to 500 mg·L-1. UV spectrum analysis showed the strength of maximum absorption of metabolites became weak compared to E2, indicating that E2 may be degraded via estrone (E1) by Fusarium sp. KY123915.

[Expression and preliminary applications of hepatitis B surface antigen mutants].

AIM: To clone and express HBsAg mutant in the Pichia pastoris. METHODS: The cloned wild type pGAP-S was used as the DNA template to generate mutant type pGAP-MS with a single or double nucleotide changes incorporated in complementary oligonucleotide primers. The product was linearized with BspH I and transformed into Pichia pastoris strain GS115, and stable multicopy integrants were screened in medium containing different concentrations of Zeocin. RESULTS: The pGAP-MS expression vector was successfully constructed and stable numbers integrated strains with high copy number were obtained. The expression of HBsAg mutant protein was identified by SDS-PAGE and Western blot with specific polyclonal antibody. The molecular weight of recombinant HBsAg mutant was 38 kDa. AxSYM HBsAg V2(Abbott)assays demonstrated all 10 HBsAg mutants were reactive. CONCLUSION: The recombinant HBsAg mutant with immunoreactivity was successfully expressed in Pichia pastoris, and it was of practical value on the quality control and clinical applications of commercial assays.

Expression of androgen receptor coactivator ARA70/ELE1 in androgenic alopecia.

BACKGROUND: Androgens have been implicated in androgenic alopecia as evidenced by the increased cutaneous expression of androgen receptor (AR), 5alpha-reductase, and decreased aromatase. Abnormalities of the AR-signal transduction pathway probably participate in the development of androgenic alopecia. ARA70/ELE1 is an AR coactivator with two isoforms, one full-length form (ARA70alpha/ELE1alpha), and an internally deleted form (ARA70beta/ELE1beta). We decided to examine the cutaneous expression of both isoforms in male androgenic alopecia. METHODS: Formalin-fixed, paraffin-embedded tissue sections from seven subjects with androgenic alopecia with matched punch biopsies from non-balding and balding areas were examined by in situ hybridization. RESULTS: Expression of at least one of the two probes for ARA70/ELE1 was present in all phases of the hair-growth cycle in all epithelial hair structures except for the inner root sheath. The dermal papilla and hair bulb expressed only the short (beta) but not the long (alpha) form of ARA70/ELE1. In situ labeling for ARA70beta/ELE1beta was weaker in the dermal papilla of balding recipient areas than those from donor ones. CONCLUSIONS: Our data further support that the hair growth is regulated by androgens. The differential expression pattern of ARA70/ELE1 suggests that this key androgen receptor coactivator is involved in androgenic alopecia. Lee P, Zhu C-C, Sadick NS, Diwan AH, Zhang PS, Liu JS, Prieto VG. Expression of androgen receptor coactivator ARA70/ELE1 in androgenic alopecia.

Thyrotropin-releasing hormone receptor processing: role of ubiquitination and proteasomal degradation.

These studies were designed to characterize ubiquitination of the G protein-coupled TRH receptor (TRHR). TRHRs and ubiquitin coprecipitated with antibodies to either receptor or ubiquitin in Chinese hamster ovary or pituitary GHFT cells. Inhibition of the proteasome with MG-132 resulted in an accumulation of total TRHRs and the appearance of a small amount of cytosolic receptor. MG-132 caused an increase in newly synthesized receptors, detected by microscopy using a TRHR coupled to Timer, a DsRed that undergoes a spontaneous time-dependent color change. Misfolded TRHRs were particularly heavily ubiquitinated. These results show that the proteasome participates in TRHR quality control early after receptor synthesis. Under normal circumstances, most ubiquitinated TRHRs were absorbed to wheat germ agglutinin, indicating that they had undergone complex glycosylation in the Golgi apparatus. When cells were treated with tunicamycin to block glycosylation, a ladder of ubiquitinated species was detectable. Cell surface receptors, which were labeled selectively with either radioligand or antibody, showed no detectable ubiquitin modification. To determine if ubiqutination plays a role in TRH-induced receptor endocytosis, the receptor was expressed in Ts20 cells, which have a temperature-sensitive ubiquitin pathway. TRH induced a significant calcium response and rapid and extensive receptor internalization at both the permissive and nonpermissive temperatures, indicating that ligand-dependent ubiquitination of the receptor, or any other protein, is not necessary for TRHR signaling or internalization. These results show that ubiquitin modification targets misfolded receptors for degradation and suggest a possible role for ubiquitination in receptor trafficking.

The concept of bronchioloalveolar cell adenocarcinoma: redefinition, a critique of the 1999 WHO classification, and an ultrastructural analysis of 155 cases.

Bronchioloalveolar cell adenocarcinoma (BACA) is bronchioloalveolar because (1) it arises in bronchioles and alveoli and (2) differentiates into bronchiolar and alveolar cells. Every entity possesses unique characteristics that separate it from other entities. The unique characteristic of BACA is its cell type. Lepidic growth is a clue to the cell type and, even though present in the vast majority, is not unique or absolutely essential. Because of the algebraic nature of concepts, the degree of differentiation, the extent of lepidic growth, and the degree of stromal desmoplasia cannot be used as definitional requirements. Likewise, in malignant tumors, absence of stromal invasion cannot be required. An epistemologically valid definition of BACA is proposed and a study of 155 cases defined this way and examined ultrastructurally is presented.

Dimerization and phosphorylation of thyrotropin-releasing hormone receptors are modulated by agonist stimulation.

Dimerization and phosphorylation of thyrotropin-releasing hormone (TRH) receptors was characterized using HEK293 and pituitary GHFT cells expressing epitope-tagged receptors. TRH receptors tagged with FLAG and hemagglutinin epitopes were co-precipitated only if they were co-expressed, and 10-30% of receptors were isolated as hemagglutinin/FLAG-receptor dimers under basal conditions. The abundance of receptor dimers was increased when cells had been stimulated by TRH, indicating that TRH either stabilizes pre-existing dimers or increases dimer formation. TRH increased receptor dimerization and phosphorylation within 1 min in a dose-dependent manner. TRH increased phosphorylation of both receptor monomers and dimers, documented by incorporation of (32)P and an upshift in receptor mobility reversed by phosphatase treatment. The ability of TRH to increase receptor phosphorylation and dimerization did not depend on signal transduction, because it was not inhibited by the phospholipase C inhibitor. Receptor phosphorylation required an agonist but was not blocked by the casein kinase II inhibitor apigenin, the protein kinase C inhibitor GF109203X, or expression of a dominant negative form of G protein-coupled receptor kinase 2. TRH receptors lacking most of the cytoplasmic carboxyl terminus formed dimers constitutively but failed to undergo agonist-induced dimerization and phosphorylation. TRH also increased phosphorylation and dimerization of TRH receptors expressed in GHFT pre-lactotroph cells.

Preparation and Characterization of Novel Amphiphilic C(60) Derivatives.

Amphiphilic C(60) derivatives of [(ethoxycarbonyl)decylene]fullerene and bis[(ethoxycarbonyl)decylene]fullerene, which are used to obtain organized molecular films by the LB technique, were prepared by the reaction of C(60) with diazo compounds. They were characterized by IR, UV-vis, (1)H-NMR, (13)C-NMR, and FD-MS. Direct evidence confirmed that the [6,5]-fullerenoid is the thermodynamically more stable isomer. It was converted from the arrangement of a [6,6]methanofullerene rather than a reaction product in 6,5 double bonds. A carbene mechanism is suggested for the reaction.