Rapid identification and antimicrobial susceptibility testing directly from blood cultures of gram-negative and gram-positive isolates.

BACKGROUND: Bloodstream infection is still an important cause for morbidity and mortality. In order to reduce the turnaround time for laboratory diagnosis of bacteremia, the efficacy of identification and antimicrobial susceptibility testing using samples taken directly from positive culture bottles by Vitek 2 compact and Kirby-Bauer disk diffusion method was evaluated. METHODS: A total of 218 monomicrobial blood cultures, including 81 Gram-negative and 137 Gram-positive isolates, were detected by a direct (that is, the organism was taken from the positive blood culture bottle) and standardized (taken from the overnight agar medium subculture) method on Vitek 2 compact ID cards for identification and Kirby-Bauer disk diffusion method for antimicrobial susceptibility testing. RESULTS: 79 of 81 Gram-negative rods (97.5%) and 84 of 137 Gram-positive cocci (61.3%) were correctly identified to the species level. Among 41 strains of wrongly identified Gram-positive cocci, 30 strains (73.2%) were misidentified as Kocuria. For antimicrobial susceptibility testing, the direct method had an overall error rate of 2.1% for Gram-negative rods, with 0.1% very major, 0.4% major, and 1.6% minor discrepancies compared with the standard method. The overall error rate for Gram-positive cocci was 4.4%, with 0.2% very major, 1.3% major, and 2.9% minor discrepancies. CONCLUSIONS: Direct identification and antibiotic susceptibility testing with samples taken directly from blood cultures provided excellent results for Gram-negative rods and decreased turnaround time, while it would be less reliable for identification of Gram-positive cocci, although their antibiotic susceptibility testing yielded very good results.

Combined gas well hydrate prevention and control technology and its application.

The high pressure in some gas wells, such as those in the Xushen gas field in Daqing, China, makes them susceptible to freezing and hydrate blockages. Downhole throttling technology is widely used to reduce costs during well construction, however, due to the limitations of temperature, pressure and depth structure, this technology is sometime applied independently in some gas wells in which freezing and blockages are a frequent problem that can seriously affect production capacity. Moreover, artificial alcohol injection of 'passive plugging' to prevent hydrate formation not only consumes significant amounts of methanol but its efficiency is also dependent on factors such as weather, personnel and equipment, so it is not a continuous solution. In order to solve the above problems, the mechanism of hydrate formation was analyzed in this study, from which a combined mechanical and chemical hydrate control process was developed. OLGA software was used to design the process parameters of the novel mechanical and chemical inhibition technology for hydrate prevention and control, and also to simulate and analyze the wellhead temperature, pressure and hydrate generation once the process was implemented. Based on the results of the parameters calculation, the downhole throttle and hydrate inhibitor automatic filling device are used to realize the functions of downhole throttle depressurization and hydrate inhibitor continuous filling, reduce the wellhead pressure and hydrate generation temperature, and ensure the continuous production of gas well. This novel combination process was subsequently tested in three wells in the Daqing gas oilfield. Measurements showed that the average daily gas increase from a single well was 0.5×104m3, methanol consumption was reduced from the original maximum daily amount of 1750 kg to just 60 kg, the manual maintenance workload was reduced by 80%, and the rate of the well openings was increased from 45% to 100%. These results proved that this technology is feasible and efficient for applications in gas wells with high downhole pressure and low wellhead temperature, and, thus, provides important technical support for the prevention of gas hydrate and improvement of gas well production.

Accessibility of Park Green Space in Wuhan, China: Implications for Spatial Equity in the Post-COVID-19 Era.

During the COVID-19 pandemic, people have seen the precious value of park green space for health. In the post-COVID-19 Era, it is essential to understand the different needs and expectations of different communities for the use of park green space. A myriad of previous studies focused on the whole city's demand for park green space, while few studies examined spatial equity from a supply-demand perspective. This paper aims to investigate the differences in park green space accessibility among people of different ages at a community scale. Specifically, to better evaluate the accessibility of park green space and account for the travel choice, we compared the effects of the two-step floating catchment area (2SFCA) method containing different distance decay functions (i.e., the improved 2SFCA methods) by considering the traffic network and the scale of park green space. In addition, we compared the improved 2SFCA methods with the traditional 2SFCA. This study investigated the spatial equity of park green space accessibility in 1184 communities with a total population of 6,468,612 in the central urban districts of Wuhan. The results showed that the high accessible communities were concentrated in the urban center along the Yangtze River. The improved 2SFCA methods outperformed the traditional 2SFCA, and presented smoother gradient information. It was revealed that over half of communities' park green space accessibility levels did not match their population density. Inequality of accessibility to park green space was found in people of different ages, especially for the youth (Gini coefficient was as high as 0.83). The difference in the accessibility of urban park green space among different age structures implies the need to integrate community green space planning into urban planning in the post-COVID-19 Era.

Porcine reproductive and respiratory syndrome virus NADC30-like strain accelerates Streptococcus suis serotype 2 infection in vivo and in vitro.

Porcine reproductive and respiratory syndrome (PRRS), an economically significant pandemic disease, commonly results in increased impact of bacterial infections, including those by Streptococcus suis (S. suis). In recent years, PRRS virus (PRRSV) NADC30-like strain has emerged in different regions of China, and coinfected with S. suis and PRRSV has also gradually increased in clinical performance. However, the mechanisms involved in host innate responses towards S. suis and their implications of coinfection with NADC30-like strain remain unknown. Therefore, the pathogenicity of NADC30-like strain and S. suis serotype 2 (SS2) coinfection in vivo and in vitro was investigated in this study. The results showed that NADC30-like increased the invasion and proliferation of SS2 in blood and tissues, resulting in more severe pneumonia, myocarditis, and peritonitisas well as higher mortality rate in pigs. In vitro, NADC30-like strain increased the invasion and survival of SS2 in porcine alveolar macrophages (PAM) cells, causing more drastic expression of inflammatory cytokines and activation of NF-ĸB signalling. These results pave the way for understanding the interaction of S. suis with the swine immune system and their modulation in a viral coinfection.

Porcine Reproductive and Respiratory Syndrome Virus strains with Higher Virulence Cause Marked Protein Profile Changes in MARC-145 Cells.

Porcine reproductive and respiratory syndrome is an infectious disease that causes serious economic losses to the swine industry worldwide. To better understand the pathogenesis of the porcine reproductive and respiratory syndrome virus (PRRSV), three PRRSV strains with different molecular markers and virulence were used to infect MARC-145 cells. A total of 1804 proteins were identified, and 233 altered proteins and 72 signaling pathways involved in the proteomic profiling of virus-infected MARC-145 cells increased with the virulence of the PRRSV strain. The three types of viral strains shared a common pathway-the electron transport reaction in mitochondria-in the infected-MARC-145 cells. Moreover, the antisense pathway was the most variable of all significant signaling pathways for the highly virulent SX-1 strain, indicating that this unique pathway may be connected to the high virulence of the SX-1 strain. Our study is the first attempt to provide a proteome profile of MARC-145 cells infected with PRRSV strains with different virulence, and these findings will facilitate a deep understanding of the interactions between this virus and its host.

Cd accumulation and subcellular distribution in two ecotypes of Kyllinga brevifolia Rottb as affected by Cd treatments.

The Cadmium (Cd) accumulation capacity and subcellular distribution in the mining ecotype (ME) and non-mining ecotype (NME) of Kyllinga brevifolia Rottb were investigated in pot experiments. The results showed that average Cd contents in shoots of the two ecotypes of K. brevifolia were higher than those in roots, whereas Cd concentrations in roots were greater than those in shoots. Also, shoot Cd contents in NME of K. brevifolia were 1.65-45.45 times greater than those in ME when the plants were grown at 5, 25, 50, and 100 mg Cd kg(-1) soil. Moreover, Cd contents in the roots in NME were 1.75-45.45 times higher than those in ME. Subcellular distribution of Cd demonstrated that the majority of Cd in the two ecotypes of K. brevifolia was distributed in the cell walls and soluble fraction, and a small percentage of Cd existed in organelle fraction. In addition, proportions of Cd distributed in shoots and roots cell walls of NME were greater than those in ME. It could be assumed that compared with ME, NME of K. brevifolia has better Cd accumulation capacity, and the subcellular distribution of Cd might be one of the mechanisms to explain such phenomena.

Changes in pH, dissolved organic matter and Cd species in the rhizosphere soils of Cd phytostabilizer Athyrium wardii (Hook.) Makino involved in Cd tolerance and accumulation.

Phytostabilization has great practical significance and flexibility in the ecological restoration of mining tailings and remediation of heavy metals polluted soils. However, potential use of metallophytes in phytostabilization is limited by a lack of knowledge of many basic plant processes. A mining ecotype (ME) Athyrium wardii, Pb/Cd phytostabilizer, and a non-mining ecotype (NME) A. wardii were grown in a pot experiment to investigate the chemical characteristics of the rhizosphere when exposed to the Cd polluted soils. Rhizobags were used to collect rhizosphere and bulk soils, separately. The results indicated that the ME A. wardii was more efficient in Cd accumulation in the root than NME after growing in Cd polluted soils for 50 days in a green house. Soil solution pH and dissolved organic carbon (DOC) concentration in the rhizosphere of ME A. wardii were higher than in the bulk soil and initial values (before planting), whereas the increment in the ME A. wardii were greater than NME. Owing to the increasing of rhizosphere soil pH, exchangeable Cd significantly decreased, whereas the other Cd species were increased with increasing soil DOC values. It is assumed that the ME A. wardii was effective in stabilizing Cd from the mobile fraction to non-mobile fractions. Results from this study suggest that rhizosphere alkalinization and the exudation of high amounts of dissolved organic matter (DOM) to reduce heavy metal mobility might be the two important mechanisms involved in the metal tolerance/accumulation of ME A. wardii.

Cd accumulation and phytostabilization potential of dominant plants surrounding mining tailings.

The objectives of the present study were to compare nine dominant plant species growing in mine tailings and nonmining areas in terms of biomass and Cd concentrations and to search for Cd accumulation and tolerance. Also, more detailed experiments were conducted on Athyrium wardii using a pot experiment to assure its Cd-accumulation ability and tolerance as a potential phytostabilizer of Cd-polluted soils. Nine dominant plant species growing on Pb/Zn mine tailings and their corresponding nonmining ecotypes were investigated for their potential to phytostabilize Cd. The performance of A. wardii exposed to high levels of Cd was investigated under controlled conditions. A field study revealed that the Cd concentrations in the roots of these plants ranged from 0.21 to 251.07 mg kg(-1), and the highest concentrations were found in A. wardii, which reached a concentration of 69.78, 251.07, and 126.35 mg kg(-1) during the early growth stage (May), vigorous growth stage (August), and late growth stage (October), respectively. The Cd concentrations of roots among the nine mining ecotypes were positively correlated with available content of Cd in the rhizosphere soils, whereas a negative correlation was observed in the nonmining ecotypes. A pot experiment showed that the mining ecotype of A. wardii had a higher biomass production and Cd retention capacity in roots than that of the nonmining ecotype. Due to the relatively high tolerance to Cd and the capacity of roots to retain this metal, A. wardii may be useful for the phytostabilization of soils contaminated by Cd.

[Quantitative analysis of telomerase reverse transcriptase gene expression in goat reprogramming cells].

Currently, animal somatic cell reprogramming into the induced pluripotent stem cell (iPS) is one of the hottest research target in the field of cell biology. We focused on the analysis of telomerase reverse transcriptase (TERT) gene expression during goat somatic fibroblasts reprogramming, and investigated the relationship between the expression of TERT and the pluripotency of reprogrammed cells. RNA samples of fetal tissues isolated from Guanzhong milk goat fetus, and the induced goat reprogramming cell clones were used to determine the relative expression levels of TERT by the real-time RT-PCR method. Goat embryonic fibroblasts (GEF) collected from the Guanzhong milk goat with normal karyotype were induced by 4 transcription factors to become reprogramming cells. The expression of TERT in reprogramming cells was detected by Real-time RT-PCR. The results showed that the expression of TERT in testis tissue was higher than that in epithelial tissues (P < 0.01). The expression level of TERT was higher in AP staining positive cells than that in AP staining negative cells (P < 0.01). This result indicated that TERT activity played an important role in cell reprogramming.

[Over-expression of phospholipase D3 inhibits Akt phosphorylation in C2C12 myoblasts].

Phospholipase D (PLD) hydrolyzes phosphocholine into choline and phosphatide acid, and these metabolites play an important role in regulating cell physiology and biochemistry. To study the biological function of phospholipase D3 (PLD3) during the insulin stimulation in C2C12 myoblasts, we constructed PLD3 over-expressed cell lines (C2C12/pPLD3) and investigated the phosphorylation of Akt. The results showed that the level of phosphorylated Akt (P-Akt) was significantly increased in control C2C12 cells when insulin concentration was elevated during cell treatment, whereas the level of P-Akt in C2C12/pPLD3 cells was not changed. When extending the time of insulin treatment, P-Akt level in C2C12/pPLD3 cells was increased around 2 folds, but the total level of P-Akt in C2C12/pPLD3 was still lower than that in control group. 1-Butanol, a PLD inhibitor, could completely block Akt phosphorylation in C2C12 cells that even stimulated by insulin. However, 1-Butanol did not inhibit the Akt phosphorylation in C2C12/pPLD3 cells, but increased the phosphorylation up to 6 folds higher than control cells. The level of Akt phosphorylation in control C2C12 cells was increased significantly when stimulated by phosphatidic acid (PA), while there was no change in C2C12/pPLD3 cells with the similar treatment. When cells simulated by both PA and insulin, P-Akt level in both C2C12/pPLD3 cells and C2C12 cells were down regulated. Our observations indicated that PLD3 over expression may inhibit Akt phosphorylation and further block the transduction of insulin signaling in C2C12 cells.