Diversity and Dynamics of Soil Free-Living Nematode Populations in a Mediterranean Agroecosystem

To determine the effect of agricultural management on the dynamics and functional diversity of soil nematode communities in a carrot field at Kibbutz Ramat Hakovesh, Israel, soil samples from 0-10 cm and 10-20 cm depths were collected during the growing season of carrot. Indices were used to compare and assess the response of soil free-living nernatode communities to agricultural management. Eighteen nematode families and 20 genera were observed during the growing period, with Cephalobus, Rhabditidae, Aphelenchus, Tylenchus, and Dorylaimus being the dominant genera/families. During the planting, mid-season and post-harvest periods the total number of nematodes at both depths was significantly lower (P < 0.01) in the carrot treatment than in the control plots, while during the harvest period at both depths total nematodes and bacterivores were significantly higher in the treatment plots (P < 0.01). The values of the maturity index (MI) at both depths were found to be significantly lower in the treatment plots than in the control plots during the pre-planting period (P < 0.05). Overall, WI, MI and PPI were found to be more sensitive indicators than other ecological indices for assessing the response of nematode communities to agricultural management in a Mediterranean agroecosystem

The spatial-temporal changes of the land use/cover in the Dongting Lake area during the last decade

The research on the land use/cover change is one of the frontiers and the hot spots in the global change research. Based on the Chinese resource and environment spatial-temporal database, and using the Landsat TM and ETM data of 1990 and 2000 respectively, we analyzed the spatial-temporal characteristics of land use/cover changes in the Dongting Lake area during the last decade. The result shows that during the last ten years there were three land-use types that had changed remarkably. The cultivated land decreased by 0.57% of the total cultivated land. The built-up land and water area expanded, with an increase of 8.97% and 0.43% respectively. The conversion between land use types mostly happened among these three land-use types, especially frequently between cultivated land and water area. The land-use change speed of land-use type is different. Three cities experienced the greatest degree of land-use change among all the administrative districts, which means that the land use in these cities changed much quickly. The following changed area was the west and south of the Dongting Lake area. The slowest changed area is the north and east area.

Copy number changes of target genes in chromosome 3q25.3-qter of esophageal squamous cell carcinoma: TP63is amplified in early carcinogenesis but down-regulated as disease progressed

AIM: By using comparative genomic hybridization, gain of 3q was found in 45-86% cases of esophageal squamous cell carcinoma (EC-SCC). Chromosome 3q25.3-qter is the minimal common region with several oncogenes found within this region. However, amplification patterns of these genes in EC-SCC have never been reported. The possible association of copy number changes of these genes with pathologic characteristics is still not clear. METHODS: Real-time quantitative PCR (Q-PCR) was performed to analyze the copy number changes of 13 candidate genes within this region in 60 primary tumors of EC-SCC, and possible association of copy number changes with pathologic characteristics was analyzed by statistics. Immunohistochemistry (IHC) study was also performed on another set of 111 primary tumors of EC-SCC to verify the association between TP63 expression change and lymph node metastasis status. RESULTS: The average copy numbers (±SE) per haploid genome of individual genes in 60 samples were (from centromere to telomere): SSR3: 4.19 (±0.69); CCNL1: 5.24 (±0.67); SMC4L1: 2.01 (±0.16); EVI1: 2.02 (±0.12); hTERC. 5.28 (±0.54); SKIL 2.71 (±0.14); EIF5A2. 1.95 (±0.12); ECT2: 9.18 (±1.68); PIK3CA: 8.13 (±1.17); EIF4G1: 1.07 (±0.05); 557: 3.07 (±0.25); TP63: 2.51 (±0.22); TFRC. 2.42 (±0.19). Four clusters of amplification were found: SSR3 and CCLN1 at 3q25.31; hTERC and SKIL at 3q26.2; ECT2 and PIK3CA at 3q26.31-q26.32; and 55T, TP63 and TFRC at 3q27.3-q29. Patients with lymph node metastasis had significantly lower copy number of TP63 in the primary tumor than those without lymph node metastasis. IHC study on tissue arrays also showed that patients with lymph node metastasis have significantly lower TP63 staining score in the primary tumor than those without lymph node metastasis. CONCLUSION: This study showed that different amplification patterns were seen among different genes within 3q25.3-qter in EC-SCC, and several novel candidate oncogenes (SSR3, SMC4L1, ECT2, and SST) were identified. TP63 is amplified in early stage of EC-SCC carcinogenesis but down-regulated in advanced stage of disease.

Comparative studies on the biochemical characteristics of natural actomyosin isolated from PSE and normal pork

Biochemical changes of natural actomyosin from fresh pale, soft, exudative (PSE) and normal pork were studied,and the effects of different storage temperatures and different incubation temperature and times on sample superprecipitation,total sulfhydryl (-SH) content, and ATP (adenosine triphosphate) sensitivity were investigated. The results demonstrated that ATPase activity and thermal stability of PSE actomyosin were lower than those of normal pork; and that PSE actomyosin had higher -SH content than that of normal pork at all incubation temperatures and times tested.

A High-affinity Activator of G551D-CFTR Chloride Channel Identified By High Throughput Screening

A stably transfected CHO cell line coexpressing G551D-CFTR and iodide-sensitive yellow fluorescent protein mutant EYFP-H148Q-I152L was successfully established and used as assay model to identify small-molecule activators of G551D-CFTR chloride channel from 100000 diverse combinatorial compounds by high throughput screening on a customized Beckman robotic system. A bicyclooctane compound was identified to activate G551D-CFTR chloride channel with high-affinity(K d=1.8 μmol/L). The activity of the bicyclooctane compound is G551D-CFTR-specific, reversible and non-toxic. The G551D-CFTR activator may be useful as a tool to study the mutant G551D-CFTR chloride channel structure and transport properties and as a candidate drug to cure cystic fibrosis caused by G551D-CFTR mutation.

Heterologous Expression of Mycobacterium tuberculosis Ag85B in Pichia pastoris

The DNA fragment encoding mature Mycobacterium tuberculosis major secretory protein Ag85B was inserted into the Pichia pastoris secretory expression vector pHBM905A, under the control of the AOX1 promoter. The recombinant plasmid pHBM905A-85B linearized by Sal I was introduced into Pichia pastoris strain GS115 by PEG1000 transformation method. After phenotype screening and PCR identification, the resulting GS115-pHBM905A-85B strain was cultivated and induced with methanol. The recombinant Ag85B protein in secreted form was attained with molecular weight of 35 × 10^3 approximately detected by SDS-PAGE and Western blot. EI,ISA experiment proved that the protein had good antigen specificity. Secretory expression of recombinant M. tuberculosis Ag85B in P. pastoris will open a door to mass production of the protein in heterologous host and allow ready evaluation of its immunological function.

A Class of High-affinity Bicyclooctane G551D-CFTR Activators Identified by High Throughput Screening

The glycine-to-aspartic acid missense mutation at the codon 551(G551D) of the cystic fibrosis transmembrane conductance regulator(CFTR) is one of the five most frequent cystic fibrosis(CF) mutations associated with a severe CF phenotype. To explore the feasibility of pharmacological correction of disrupted activation of CFTR chloride channel caused by G551D mutation, we developed a halide-sensitive fluorescence miniassay for G551D-CFTR in Fisher rat thyroid(FRT) epithelial cells for the discovery of novel activators of G551D-CFTR. A class of bicyclooctane small molecule compounds that efficiently stimulate G551D-CFTR chloride channel activity was identified by high throughput screening via the FRT cell-based assay. This class of compounds selectively activates G551D-CFTR with a high affinity, whereas little effect of the compounds on wildtype CFTR can be seen. The discovery of a class of bicyclooctane G551D-CFTR activators will permit the analysis of structure-activity relationship of the compounds to identify ideal leads for in vivo therapeutic studies.