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APOBEC3H is a deoxycytidine deaminase that can restrict the replication of HIV-1 in the absence of the viral protein Vif that induces APOBEC3H degradation in cells. APOBEC3H exists in humans as seven haplotypes (I-VII) with different cellular stabilities. Of the three stable APOBEC3H haplotypes (II, V, and VII), haplotypes II and V occur most frequently in the population. Despite APOBEC3H being a bona fide restriction factor, there has been no comparative biochemical characterization of APOBEC3H haplotypes. We characterized the ssDNA scanning mechanisms that haplotypes II and V use to search their ssDNA substrate for cytosine-containing deamination motifs. APOBEC3H haplotype II was able to processively deaminate multiple cytosines in a single enzyme-substrate encounter by using sliding, jumping, and intersegmental transfer movements. In contrast, APOBEC3H haplotype V exhibited diminished sliding and intersegmental transfer abilities but was able to jump along ssDNA. Due to an Asp or Glu at amino acid 178 differentiating these APOBEC3H haplotypes, the data indicated that this amino acid on helix 6 contributes to processivity. The diminished processivity ofAPOBEC3HhaplotypeVdid not result in a reduced efficiency to restrict HIV-1 replication in single-cycle infectivity assays, suggesting a redundancy in the contributions of jumping and intersegmental transfer to mutagenic efficiency. Optimal processivity on ssDNA also required dimerization of APOBEC3H through the β2 strands. The findings support a model in which jumping can compensate for deficiencies in intersegmental transfer and suggest that APOBEC3H haplotypes II and V induce HIV-1 mutagenesis efficiently but by different mechanisms. [ABSTRACT FROM AUTHOR]

The aim of the study was to investigate the prevalence, distribution, and diversity of Escherichia coli in goat-milk-powder plants in Shaanxi, China. Three plants manufacturing goat milk powder in Shaanxi province were visited once for sampling during 2012 and 2013. Samples were taken for isolation of E. coli. Isolates were characterized by antimicrobial susceptibility testing and detection of virulence genes. All isolates were further examined by pulsed-field gel electrophoresis analysis. In total, 53 E. coli strains were isolated from 32 positive samples out of 534 samples. Among E. coli isolates, resistance was most frequently observed to trimethoprim-sulfamethoxazole (75.5%), whereas all isolates were sensitive to gatifloxacin, kanamycin, amikacin, and amoxicillin-clavulanate. The 6 virulence genes of pathogenic E. coli were not detected. Pulsed-field gel electrophoresis results showed that E. coli strains in plants were genetically diverse and milk storage tank could be an important contamination source. This study could provide useful information for plants manufacturing goat milk powder to establish proper management practices that help minimize the chance of microbial contamination. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the prevalence and antimicrobial susceptibility of Escherichia coil in retail whole chicken in the People's Republic of China. Five hundred seventy-six raw whole chicken samples, randomly purchased from 146 farmers' markets or supermarkets in four provinces from March through December 2010, were analyzed for E. coli contamination, and the E. coli isolates were further tested for the presence of virulence genes and antimicrobial susceptibility. The overall positive rate for E. coli in retail chicken was 69.1%. E. coli prevalence was the highest in Beijing (86.8%), followed by Henan province (78.5%), Shaanxi province (65.3%), and the lowest prevalence was found in Sichuan province (45.8%). Among 398 isolates recovered, only the eae gene was detected in one isolate; no other virulence genes were detected. Resistance was most common to tetracycline (84.4%), followed by nalidixic acid (74.1%), ampicillin (71.1%), trimethoprim-sulfamethoxazole (70.l%), amoxicillin-clavulanic acid (68.8%), and streptomycin (58.5%). Lower resistance was detected to chloramphenicol (43.7%), kanamycin (42.7%), ciprofloxacin (30.2%), gentamicin (29.4%), cefoperazone (13.6%), amikacin (12.6%), gatifloxacin (8%), and cefoxitin (7.8%). Only 3.8% of the isolates were susceptible to all tested antimicrobials. Six percent of the isolates displayed resistance to one antimicrobial, 6.3% to two, and 83.9% to three or more of the antimicrobials. Our findings indicate that retail chicken in China was commonly contaminated with E. coli, and many E. coli strains exhibited multiple drug resistance. The implementation of good manufacturing practices throughout the poultry production chain is necessary to reduce E. coli contamination in retail chicken, and the prudent use of antibiotics is imperative in poultry production in China. [ABSTRACT FROM AUTHOR]

APOBEC3G is a retroviral restriction factor that can inhibit the replication of human immunodeficiency virus, type 1 (HIV-1) in the absence of the viral infectivity factor (Vif) protein. Virion-encapsidated APOBEC3G can deaminate cytosine to uracil in viral (-)DNA, which leads to hypermutation and inactivation of the provirus. APOBEC3G catalyzes these deaminations processively on single-stranded DNA using sliding and jumping movements. Vif is thought to primarily overcome APOBEC3G through an interaction that mediates APOBEC3G ubiquitination and results in its proteasomal degradation. However, Vif may also inhibit APOBEC3G mRNA translation, virion encapsidation, and deamination activity. Here we investigated the molecular mechanism of VifIIIB- and VifHXB2-mediated inhibition of APOBEC3G deamination activity. Biochemical assays using a model HIV-1 replication assay and synthetic single-stranded or partially double-stranded DNA substrates demonstrated that APOBEC3G has an altered processive mechanism in the presence of Vif. Specifically, VifHXB2 inhibited the jumping and VifIIIB inhibited the sliding movements of APOBEC3G. The absence of such an effect by Vif on degradation-resistant APOBEC3G D128K indicates that a Vif- APOBEC3G interaction mediates this effect. That the partially processive APOBEC3G was less effective at inducing mutagenesis in a model HIV-1 replication assay suggests that Vif co-encapsidation with APOBEC3G can promote sublethal mutagenesis of HIV-1 proviral DNA. [ABSTRACT FROM AUTHOR]

APOBEC3G is a single-stranded (ss) DNA deaminase that restricts replication of HIV-1 by inducing viral genome mutagenesis through deamination of cytosine to uracil on HIV-1 cDNA. APOBEC3G has polydisperse oligomeric states and deaminates ssDNA processively through jumping and sliding. APOBEC3G has a catalytically inactive N-terminal CD1 domain that mediates processivity and an active C-terminal CD2 domain that catalyzes deaminations. Here, we assess the determinants of APOBEC3G deamination efficiency mediated by the CD1 domain by comparing native APOBEC3G and two CD1 mutants, a monomeric mutant (F126A/W127A) and a clinical mutant associated with high viral loads (H186R). Biochemical assays on ssDNA or partially dsDNA and with a reconstituted HIV replication system demonstrate that both mutants of Apo3G have altered DNA scanning properties in either jumping (F126A/W127A) or sliding (H186R), which results in decreased abilities to induce mutagenesis during reverse transcription. The data reveal a functionality for Apo3G oligomers in deamination and provide the first biochemical characterization of the clinical mutant H186R. The data demonstrate that the balance between the jumping and sliding of Apo3G is needed for efficient mutational inactivation of HIV-1. [ABSTRACT FROM AUTHOR]

Numerical simulations using two-fluid model incorporating the kinetic theory of granular flow (KTGF) are conducted to study the behavior of bed particles in a two dimensional baffle type internally circulating fluidized bed (ICFB). Complete knowledge of the solid circulation flux in an ICFB is very important as the bed particles are continuously circulating between different chambers above and below the central baffle. The effects of the gas distributor types, superficial fluidizing velocity and chambers height elevation difference are evaluated through computer simulations and the results are quantified in terms of solid circulation flux. By increasing the gas velocity (U f ) in the RC chamber, solid circulation flux from the heat exchange chamber (HEC) to the reaction chamber (RC) through the slot under the baffle is increased regardless of the distributor design. From further investigations, the solid circulation flux and particles radial velocity through the slot in the tubular gas distributor ICFB are found to be considerably increased as compared to the other configurations in our case. The elevated RC distributor is found to have a little impact on the solid circulation flux as a function of U f . By varying RC distributor elevation while keeping HEC constant, the circulation flux is also changing and the highest flux is observed at an elevation level of 90 mm. [ABSTRACT FROM AUTHOR]

Processing of minerals and metals involves phenomena functioning over a wide range of length scales. Equipment is generally large in order to process the required large flow rates, yet achieving the functional requirements of the operation requires successful execution of micro-scale processes. Multi-scale modelling approaches to process simulation are firstly reviewed, and similarities and differences compared with the more common multi-scale approaches to materials modelling are given. The sequential multi-scale method is then illustrated with reference to the mineral flotation process. In this case, multi-phase CFD (computational fluid dynamics) models of large-scale cells has been complemented by micro-scale CFD simulations of bubble-particle collision, and experimental and modelling studies of the bubble-particle attachment process itself. Finally, other examples of sequential multi-scale modelling are summarised, highlighting progress on unit operations including aluminium reduction cells, leaching heaps, copper solvent-extraction settlers, and fluidised beds. [ABSTRACT FROM AUTHOR]