Efficiency of biochar,nitrogen addition,and microbial agent amendments in remediation of soil properties and microbial community in Qilian Mountains mine soils

Lacking systematic evaluations in soil quality and microbial community recovery after different amendments addition limits optimization of amendments combination in coal mine soils. We performed a short‐term incubation experiment with a varying temperature over 12 weeks to assess the effects of three amendments (biochar: C; nitrogen fertilizer at three levels: N‐N1~N3; microbial agent at two levels: M‐M1~M2) based on C/N ratio (regulated by biochar and N level: 35:1, 25:1, 12.5:1) on mine soil properties and microbial community in the Qilian Mountains, China. Over the incubation period, soil pH and MBC/MBN were significantly lower than unamended treatment in N addition and C + M + N treatments, respectively. Soil organic carbon (SOC), total nitrogen (TN), available nitrogen (AN), available phosphorus (AP), available potassium (AK), microbial biomass carbon (MBC), and nitrogen (MBN) contents increased significantly in all amended treatments (p < .001). Higher AP, AK, MBC, MBN, and lower MBC/MBN were observed in N2‐treated soil (corresponding to C/N ratio of 25:1). Meanwhile, N2‐treated soil significantly increased species richness and diversity of soil bacterial community (p < .05). Principal coordinate analysis further showed that soil bacterial community compositions were significantly separated by N level. C‐M‐N treatments significantly increased the relative abundance (>1%) of the bacterial phyla Bacteroidetes and Firmicutes, and decreased the relative abundance of fungal phyla Chytridiomycota (p < .05). Redundancy analysis illustrated the importance of soil nutrients in explaining variability in bacterial community composition (74.73%) than fungal composition (35.0%). Our results indicated that N addition based on biochar and M can improve soil quality by neutralizing soil pH and increasing soil nutrient contents in short‐term, and the appropriate C/N ratio (25:1) can better promote microbial mass, richness, and diversity of soil bacterial community. Our study provided a new insight for achieving restoration of damaged habitats by changing microbial structure, diversity, and mass by regulating C/N ratio of amendments.     

β(1) -adrenergic receptor autoantibodies from heart failure patients enhanced TNF-α secretion in RAW264.7 macrophages in a largely PKA-dependent fashion

Autoantibodies against the second extracellular loop of β₁‐adrenergic receptor (β₁‐AA) not only contribute to increased susceptibility to heart failure, but also play a causative role in myocardial remodeling through their catecholamine‐like effects via binding with the β₁‐adrenergic receptor. The current study was designed to determine whether β₁‐AA isolated from the sera of heart failure patients could cause TNF‐α secretion from the murine macrophage‐like cell line RAW264.7. Blood samples were collected from 40 patients who had suffered heart failure, as well as from 40 healthy subjects. The titer of β₁‐AA and the level of TNF‐α were detected using ELISA. The effect of β₁‐AA on murine macrophage‐like cell line RAW264.7 proliferation was detected by CCK‐8 kits and CFSE assay. Western blot assay was used to analyze the expression of phospho‐VASP. β₁‐AA appeared more frequently in patients with heart failure than in healthy subjects. The β₁‐AA isolated from heart failure patients promoted an increase of TNF‐α levels, which could be completely blocked by the selective β₁‐adrenergic receptor antagonist metoprolol and the second extracellular loop of β₁‐adrenergic receptor (β₁‐AR‐EC_II), but only partially inhibited by PKA inhibitor H89. Furthermore, the β₁‐AA could enhance the proliferation of RAW264.7 cells in vitro. Meanwhile, the expression of phospho‐VASP was markedly increased in the presence of β₁‐AA. These results demonstrate for the first time that the β₁‐AA isolated from heart failure patients could bind with β₁‐AR on the surface of RAW264.7 cells, causing the release of TNF‐α largely in a PKA‐dependent fashion. J. Cell. Biochem. 113: 3218–3228, 2012. © 2012 Wiley Periodicals, Inc.     

Mutation identification of the DSPP in a Chinese family with DGI-II and an up-to-date bioinformatic analysis

In this study, through linkage analysis of a four-generation Chinese family with multiple members afflicted with DGI (type II), we identified a novel missense mutation in DSPP. The mutation was located in exon 2 at the second nucleotide position of the last codon and resulted in a substitution of a proline with a leucine residue (c.50C>T, p.P17L, g.50C>T). To assess the potential effects of this novel mutation, we utilized various bioinformatics analysis programs. The results indicate that the mutation likely affects protein cleavage/trafficking. We also analyzed previously reported mutations of DSPP. In summary, our finding supports that the genomic sequence that corresponds to the P17 residue of DSPP is a mutational hotspot and P17 may be critical for the function of DSPP.     

Identification and Molecular Analysis of Pathogenic Yeasts in Droppings of Domestic Pigeons in Beijing, China

Feral pigeons are known as reservoirs of pathogenic yeasts that cause opportunistic infections in human. In the outskirts of Beijing, China, pigeons are more frequently raised at homes than are encountered in public areas. Many studies have focused on the presence of pathogenic yeasts in the excreta (fresh or withered) of a variety kinds of birds, pigeon crop and cloacae. One hundred and forty-three samples of fresh droppings were collected from three suburban pigeon-raising homes in an area of northern Beijing, China. The internal transcribed sequences (ITS) of all strains (except for 8 strains of Rhodotorula sp. ) were sequenced and compared with those of the databases of the National Center for Biotechnology Information website ( http://www.ncbi.nlm.nih.gov ) using the Basic Local Alignment Search Tool (BLAST). Yeasts representing 8 genera, Cryptococcus, Filobasidium, Rhodotorula, Candida, Debaryomyces, Saccaromyces, Trichosporon and Sporidiobolus, were identified from 120 isolates. Cryptococcus was the most prolific genera represented by eight species. The populations of yeast species isolated from fresh pigeon droppings were different among homes. Although it is well established that Cryptococcus neoformans exists mainly in old pigeon guano, several C. neoformans strains were still isolated from fresh pigeon excreta, providing a clue that live cryptococcal cells could move through the gastrointestinal tract of the pigeons. Eight genera identified from fresh droppings of domestic pigeons further confirm that pigeons serve as reservoirs, carriers and even spreaders of Cryptococcus species and other medically significant yeasts. The proportion of pathogenic yeasts in all isolates is more than 90 %.     

Pseudomonas nitritireducens sp. nov., a nitrite reduction bacterium isolated from wheat soil

A Gram-negative, non-mobile, polar single flagellum, rod-shaped bacterium WZBFD3-5A2^T was isolated from a wheat soil subjected to herbicides for several years. Cells of strain WZBFD3-5A2^T grow optimally on Luria-Bertani agar medium at 30?°C in the presence of 0–4.0?% (w/v) NaCl and pH 8.0. 16S rRNA gene sequence analysis revealed that strain WZBFD3-5A2^T belongs to the genus Pseudomonas. Physiological and biochemical tests supported the phylogenetic affiliation. Strain WZBFD3-5A2^T is closely related to Pseudomonas nitroreducens IAM1439^T, sharing 99.7?% sequence similarity. DNA–DNA hybridization experiments between the two strains showed only moderate reassociation similarity (33.92?±?1.0?%). The DNA G+C content is 62.0?mol%. The predominant respiratory quinine is Q-9. The major cellular fatty acids present are C_16:0 (28.55?%), C_16:1ω6c or C_16:1ω7c (20.94?%), C_18:1ω7c (17.21?%) and C_18:0 (13.73?%). The isolate is distinguishable from other related members of the genus Pseudomonas on the basis of phenotypic and biochemical characteristics. From the genotypic, chemotaxonomic and phenotypic data, it is evident that strain WZBFD3-5A2^T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas nitritereducens sp. nov. is proposed. The type strain is WZBFD3-5A2^T (=CGMCC 1.10702^T?=?LMG 25966^T).     

Genetic variation in IL28B is associated with the development of hepatitis B-related hepatocellular carcinoma

To evaluate the role of host IL28B (interleukin 28B; interferon lambda 3) single nucleotide polymorphisms (SNPs) in predicting hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) susceptibility, three SNPs in the IL28B gene (rs12979860C/T, rs8099917G/T and rs12980275G/A) were examined in 330 subjects (including 154 HBV-related HCC patients, 86 non-HCC patients with chronic hepatitis B (CHB), 43 HBV self-limited infections and 47 healthy controls). Notably, the frequency of CC homozygosity was 91.5% in healthy controls and 72.9% in CHB, the difference being statistically significant (χ(2) = 6.40, P = 0.01). The statistically difference was seen between healthy controls (91.5%) and HCC (74.7%) (χ(2) = 6.05, P = 0.01). However, this significant finding was not seen between HBV self-limited and healthy controls. Carriers of the minor T allele in rs12979860 had a higher risk of HCC compared with non-carriers (χ(2) = 4.44, P = 0.04). Haplotype analyses revealed significant association between haplotype C-T-A and healthy controls, but not with the HCC group (96.6 vs. 82.0%, χ(2) = 6.08, P = 0.01). Analyses of genotype combination and gene-gene interaction showed that there was a positive interaction between rs12979860 and rs12980275, with an OR rate of 11.79 (likelihood test, P = 0.04). Our results suggest that the IL28B rs12979860 C/T polymorphism might affect susceptibility to the chronic HBV infection and progression of HCC. Of note, the T allele and non-CC genotypes have strong predictive effect of increasing susceptibility of chronic HBV infection and HCC.     

Role of autophagy in angiogenesis in aortic endothelial cells

Angiogenesis plays critical roles in the recovery phase of ischemic heart disease and peripheral vascular disease. An increase in autophagy is protective under hypoxic and chronic ischemic conditions. In the present study we determined the role of autophagy in angiogenesis. 3-Methyladenine (3-MA) and small interfering RNA (siRNA) against ATG5 were used to inhibit autophagy induced by nutrient deprivation of cultured bovine aortic endothelial cells (BAECs). Assays of BAECs tube formation and cell migration revealed that inhibition of autophagy by 3-MA or siRNA against ATG5 reduced angiogenesis. In contrast, induction of autophagy by overexpression of ATG5 increased BAECs tube formation and migration. Additionally, inhibiting autophagy impaired vascular endothelial growth factor (VEGF)-induced angiogenesis. However, inhibition of autophagy did not alter the expression of pro-angiogenesis factors such as VEGF, platelet-derived growth factor, or integrin αV. Furthermore, autophagy increased reactive oxygen species (ROS) formation and activated AKT phosphorylation. Inhibition of autophagy significantly decreased the production of ROS and activation of AKT but not of extracellular regulated kinase, whereas overexpression of ATG5 increased cellular ROS production and AKT activation in BAECs. Inhibition of AKT activation or ROS production significantly decreased the tube formation induced by ATG5 overexpression. Here we report a novel observation that autophagy plays an important role in angiogenesis in BAECs. Induction of autophagy promotes angiogenesis while inhibition of autophagy suppresses angiogenesis, including VEGF-induced angiogenesis. ROS production and AKT activation might be important mechanisms for mediating angiogenesis induced by autophagy. Our findings indicate that targeting autophagy may provide an important new tool for treating cardiovascular disease.