A novel capsid-XL32-derived adeno-associated virus serotype prompts retinal tropism and ameliorates choroidal neovascularization.

Gene therapy has been adapted, from the laboratory to the clinic, to treat retinopathies. In contrast to subretinal route, intravitreal delivery of AAV vectors displays the advantage of bypassing surgical injuries, but the viral particles are more prone to be nullified by the host neutralizing factors. To minimize such suppression of therapeutic effect, especially in terms of AAV2 and its derivatives, we introduced three serine-to-glycine mutations, based on the phosphorylation sites identified by mass spectrum analysis, to the XL32 capsid to generate a novel serotype named AAVYC5. Via intravitreal administration, AAVYC5 was transduced more effectively into multiple retinal layers compared with AAV2 and XL32. AAVYC5 also enabled successful delivery of anti-angiogenic molecules to rescue laser-induced choroidal neovascularization and astrogliosis in mice and non-human primates. Furthermore, we detected fewer neutralizing antibodies and binding IgG in human sera against AAVYC5 than those specific for AAV2 and XL32. Our results thus implicate this capsid-optimized AAVYC5 as a promising vector suitable for a wide population, particularly those with undesirable AAV2 seroreactivity.

Revelation of AbfR in regulation of mismatch repair and energy metabolism in S. epidermidis by integrated proteomic and metabolomic analysis.

Staphylococcus epidermidis is a common causative of nosocomial infections associated with indwelling medical devices. To date, the mechanisms of the pathogenicity and drug resistance of S. epidermidis have not been clearly elucidated. AbfR has been previously identified as an oxidation-sensing regulator that regulates bacterial aggregation and biofilm formation by responding to oxidative stress in S. epidermidis; however, the regulatory pathways of AbfR are underexplored. In this study, we investigated the oxidation-sensing regulatory mechanism of AbfR using TMT10-plex labelling quantitative proteomic and untargeted metabolomic approaches. Integrated analysis of two omics datasets indicated that abfR depletion influenced nucleic acid metabolism and activated the DNA mismatch repair pathway. In addition, several energy-related metabolic pathways, including tricarboxylic acid (TCA) cycle, glycolysis, and arginine metabolism, were remarkably impacted by the deletion of abfR. This study revealed the regulatory networks of the transcription factor AbfR from a multi-omics view and demonstrated that AbfR played a broad role in not only mismatch repair but also energy metabolism, enabling S. epidermidis to constantly sense and adapt to environmental stress. SIGNIFICANCE: Staphylococcus epidermidis has emerged as a major nosocomial infection causing pathogen. AbfR, a transcription factor of S. epidermidis, plays an important role in oxidative stress, cell aggregation, and biofilm formation; however, the regulatory mechanism of AbfR is unknown. Using proteomic and metabolomic approaches, this study unveils the global regulatory networks of AbfR, and demonstrates that AbfR not only regulates the DNA mismatch repair pathway by an oxidation sensing mechanism but also affects energy metabolism. This study expands the body of knowledge related to regulatory transcription factors in staphylococci and lays a foundation for future research on clinical infections caused by S. epidermidis.

The false smut pathogen Ustilaginoidea virens requires rice stamens for false smut ball formation.

Rice false smut has emerged as a serious grain disease in rice production worldwide. The disease is characterized by the transformation of individual rice florets into false smut balls, which is caused by the fungal pathogen Ustilaginoidea virens. To date, little is known about the host factors required for false smut ball formation by U. virens. In this study, we identified histological determinants for the formation of false smut balls by inoculating U. virens into rice floral mutants defective with respect to individual floral parts. The results showed that U. virens could form mature false smut balls in rice floral mutants with defective pistils, but failed to develop false smut balls in the superwoman mutant lacking stamens, identifying that U. virens requires rice stamens to complete its infection cycle. Comparative transcriptome analysis indicated a list of candidate host genes that may facilitate nutrient acquisition by U. virens from the rice stamens, such as SWEET11, SWEET14 and SUT5, and genes involved in the biosynthesis of trehalose and raffinose family sugars. These data pinpoint rice stamens as the key target organ of U. virens infection and provide a valuable starting point for dissecting the molecular mechanism of false smut ball formation.

[Effects of micro-sprinkling hose length and width on wheat field water condition and flag leaf chlorophyll fluorescence characteristics in different sampling districts]. / å¾®å–·å¸¦é•¿å®½å¯¹ä¸åŒåŒºæ®µéº¦ç”°æ°´åˆ†å’Œå°éº¦æ——å¶å¶ç»¿ç´ è§å ‰ç‰¹æ€§çš„å½±å“.

A two-year field experiment was conducted in 2014-2015 and 2015-2016 wheat growing seasons to study the effects of micro-sprinkling hose length and width on field water condition, and flag leaf chlorophyll fluorescence characteristics in different sampling districts (D1 to D6 along with the hose laying direction). Six micro-sprinkling hose treatments were set: 60 m (T1), 80 m (T2) and 100 m (T3) lengths under 65 mm width; 60 m (T4), 80 m (T5) and 100 m (T6) lengths under 80 mm width. The results showed that after irrigation at jointing, the Christiansen uniformity coefficient (Cu) of T1 was significantly higher than T2 and T3 under 65 mm hose width. Under 80 mm hose width, T4 and T5 had the highest Cu compared to T6. After irrigation at anthesis, the Cu showed T1>T2>T3 under 65 mm hose width, and T4>T5>T6 under 80 mm hose width. Under 65 mm hose width, the average relative soil water content of 0-40 cm soil layers after irrigation at anthesis, flag leaf ΦPSII, NPQ and ETR at 20 and 30 d after anthesis and the grain yield of different sampling district did not differ in T1; T2 showed the order of D1, D2>D3>D4>D5; T3 showed D1, D2>D3>D4>D5, D6. The average ΦPSII, NPQ and ETR at 20 and 30 d after anthesis, and the average dry matter at maturity of different sampling districts were presented as T1>T2, T3. Under 85 mm hose width, no significant differences were observed in the average relative soil water content of 0-40 cm soil layers after irrigation at ahthesis, flag leaf ΦPSII, NPQ and ETR at 20 and 30 d after anthesis and the grain yield of different sampling districts in T4; in T5, the indexes mentioned above in D1, D2 and D3 sampling districts were significantly higher than those in D4 and D5; in T6, the decreasing order was D1, D2, D3>D4>D5>T6. The average ΦPSII, NPQ and ETR at 20 and 30 d after anthesis, and the average dry matter at maturity of different districts showed the order of T4, T5>T6. The ave-rage grain yield and water use efficiency of T1, T4 and T5 were significantly higher than those in T2, T3 and T6, T1 and T4 had a better irrigation benefit than T5. Under this experimental condition, T1 treatment under 65 mm hose width, T4 treatment under 80 mm hose width were the most recommendable treatments considering high yield and water saving, and T5 treatment was also recommendable under 80 mm hose width.

Current understanding on Villosiclava virens, a unique flower-infecting fungus causing rice false smut disease.

Villosiclava virens (Vv) is an ascomycete fungal pathogen that causes false smut disease in rice. Recent reports have revealed some interesting aspects of the enigmatic pathogen to address the question of why it specifically infects rice flowers and converts a grain into a false smut ball. Comparative and functional genomics have suggested specific adaptation of Vv in the colonization of rice flowers. Anatomical studies have disclosed that Vv specifically infects rice stamen filaments before heading and intercepts seed formation. In addition, Vv can occupy the whole inner space of a spikelet embracing all floral organs and activate the rice grain-filling network, presumably for nutrient acquisition to support the development of the false smut ball. This profile provides a general overview of the rice false smut pathogen, and summarizes advances in the Vv life cycle, genomics and genetics, and the molecular Vv-rice interaction. Current understandings of the Vv-rice pathosystem indicate that it is a unique and interesting system which can enrich the study of plant-pathogen interactions. Taxonomy: Ustilaginoidea virens is the anamorph form of the pathogen (Kingdom Fungi; Phylum Ascomycota; Class Ascomycetes; Subclass Incertae sedis; Order Incertae sedis; Family Incertae sedis; Genus Ustilaginoidea). The teleomorph form is Villosiclava virens (Kingdom Fungi; Phylum Ascomycota; Class Ascomycetes; Subclass Sordariomycetes; Order Hypocreales; Family Clavicipitaceae; Genus Villosiclava). Disease symptoms: The only visible symptom is the replacement of rice grains by ball-shaped fungal mycelia, namely false smut balls. When maturing, the false smut ball is covered with powdery chlamydospores, and the colour changes to yellowish, yellowish orange, green, olive green and, finally, to greenish black. Sclerotia are often formed on the false smut balls in autumn. Identification and detection: Vv conidia are round to elliptical, measuring 3-5 µm in diameter. Chlamydospores are ornamented with prominent irregularly curved spines, which are 200-500 nm in length. The sclerotia are black, horseshoe-shaped and irregular oblong or flat, ranging from 2 to 20 mm. Nested polymerase chain reaction (PCR) and quantitative PCR have been developed to specifically detect Vv presence in rice tissues and other biotic and abiotic samples in fields. Host range: Rice is the primary host for Vv. Natural infection by Vv has been found on several paddy field weeds, including Digitaria marginata, Panicum trypheron, Echinochloa crusgalli and Imperata cylindrica. However, the occurrence of infection in these potential alternative hosts is very rare. Life cycle: Vv infects rice spikelets at the late rice booting stage, and produces false smut balls covered with dark-green chlamydospores. Occasionally, sclerotia form on the surface of false smut balls in late autumn when the temperature fluctuates greatly between day and night. Both chlamydospores and sclerotia may serve as primary infection sources. Rainfall at the rice booting stage is a major environmental factor resulting in epidemics of rice false smut disease. Disease control: The use of fungicides is the major approach for the control of Vv. Several fungicides, such as cuproxat SC, copper oxychloride, tebuconazole, propiconazole, difenoconazole and validamycin, are often applied. However, the employment of resistant rice cultivars and genes has been limited, because of the poor understanding of rice resistance to Vv. Useful websites: Villosiclava virens genome sequence: http://www.ncbi.nlm.nih.gov/Traces/wgs/?val=JHTR01#contigs.

[Research on land use structure optimization based on nonpoint source dissolved nitrogen load estimation in Shuaishui watershed].

Regional Nutrient Management (ReNuMa) was applied to estimate dissolved nitrogen (DN) load and perform source apportionment in Shuaishui watershed during 2000-2010. Satisfactory performance of ReNuMa was revealed by the E(ns) and R2 of greater than 0.9 in calibrating and validating streamflow and DN. The average nonpoint DN load in this watershed was 1.11 x 10(3) t x a(-1), with the load intensity of (0.75 +/- 0.22) t x km(-2). Among all the land uses, paddy field had the largest DN load intensity [28.60 kg x (hm2 x a)(-1)], while forest had the least [2.71 kg x (hm2 x a)(-1)]. Agricultural land (including paddy, grain, cash crop, tea plant and orchard) contributed most to DN load in Shuaishui watershed, indicating that the human dominated agricultural activities was the major contributor of nonpoint source pollution. Land use structure optimization for Shuaishui watershed in 2015 was conducted under the rule of reducing pollutants loads and maximizing the agricultural output value. The results demonstrated that agricultural monetary growth was accompanied with the increasing DN load at the optimal level, although output increment was higher than that of DN load.

[Effects of land use structure on water quality in Xin'anjiang River].

Take Xin'anjiang upstream watershed as a case study. Based on data of interpreting TM orthophoto images and water quality monitoring in May 2010, the land use map of Xin'anjiang River, which was categorized to cultivated land, forestland, grassland, water body, building site, was obtained. Using ArcGIS hydrological and spatial analysis function, Xin'anjiang River was divided into eight sub-watersheds, and its watershed land use structure was analyzed. The water quality parameters such as TN, TP, permanganate index, fecal coliform bacteria were monitored from Jan 2010 to Dec 2010. The relations between water quality and land use were analyzed. The results showed that TN and NH4(+) -N had a significant temporal variation: dry season > wet season > normal river flow period, but other parameters did not vary significantly. In the space, Yuliang and Pukou were the most serious pollution sites. Cultivated land, water body, building site had a positive impact on water quality parameters, while there were negative correlation between the forestland and grassland. Annually, cultivated land had the most significantly important effect on TN, NH4(+) -N and permanganate index, and grassland had the most significantly important effect on TP. Cultivated land had the most prominently important impact on water quality parameters in dry season and wet season. What's more, in the normal river flow, cultivated land, grassland and forestland had the most remarkably important influence on TN, TP and fecal coliform bacteria respectively.

Relationships between PAHs and PCBs, and quantitative source apportionment of PAHs toxicity in sediments from Fenhe reservoir and watershed.

Sedimentary samples from 28 sites throughout the Fenhe reservoir and upstream watershed in China were measured, to study the relationships between PAHs and PCBs as well as perform quantitative source apportionment of toxicity. The distributions of ∑16PAHs and ∑123PCBs showed high correlation. The ∑7C-PAHs (total concentrations of seven carcinogenic PAHs) varied from 87.7 to 2005.0ngg(-1)dw, and the ∑6DL-PCBs (total concentrations of six dioxin-like PCB congeners) were n.d.-5.96ngg(-1)dw. Toxicity and biological risk were assessed using toxic equivalent quantity (TEQ) and sediment quality guideline quotient (SQGQ). The BaP played the dominant role for TEQPAH at most sites (37.17-89.40%), although the CHR showed the highest concentration level. PCB-81 contributed the highest TEQPCB. High correlations were observed between ∑16PAHs and ∑7C-PAHs, ∑123PCBs and ∑6DL-PCBs, ∑7C-PAHs and ∑6DL-PCBs as well as TEQPAH and TEQPCB. Furthermore, quantitative source apportionments for PAHs concentrations and TEQ were carried out, combining the positive matrix factorization (PMF) with the formula of TEQ. For most sites, the vehicular sources were the largest contributors to TEQPAH (46.58%), while coal combustion sources were the highest contributor for PAHs concentrations (43.31%). The relatively higher correlations between PCBs and contributions of coal combustion to PAHs might imply the similar distribution of industrial sources for PAHs and PCBs.

[Combined effects of typical natural estrogens with MCF-7 proliferation assay].

OBJECTIVE: To study 17ß-estradiol (E2), ethinylestradiol (EE2), estriol (E3), estrone (E1) on MCF-7 proliferation effects, and compare the effects of independent action (IA) model with concentration addition (CA) model in assessing the combined effects of estrogen. METHODS: The combinations of E2 + EE2, E2 + E3 and E2 + E1 were chosen and the cellular proliferation effects were examined by MTT assay. RESULTS: The maximum proliferation effects at dose of 10⁻9 mol/L was 325.48% for E2, 330.34% for EE2, 255.22% for E3, and 199.61% for E1. In the E2 + EE2, E2 + E3, E2 + E1 groups, the results of IA model analysis were very close to the experimental results. The IA model tend to overestimated the experimental results, while the CA model often underestimated the experimental results. In the EC (E2, 30) + C (EE2, 70) group, the results exceed the maximum estrogen effects of E2, while in other groups, the results were lower. CONCLUSIONS: The estrogenic effects of the four tested substances from high to low efficiency were that: EE2 > E2 > E3 > E1. The effect of IA model in predicting the combined effects of binary mixture was better than CA model. A small proportion of binary mixture showed synergy.

Use of a nonnegative constrained principal component regression chemical mass balance model to study the contributions of nearly collinear sources.

In this study, a nonnegative constrained principal component regression chemical mass balance (NCPCRCMB) model was used to solve the near collinearity problem among source profiles for source apportionment. The NCPCRCMB model added the principle component regression route into the CMB model iteration. The model was tested with the synthetic data sets, which involved contributions from eleven actual sources, with a serious near collinearity problem among them. The actual source profiles were randomly perturbed and then applied to create the synthetic receptor. The resulting synthetic receptor concentrations were also randomly perturbed to simulate measurement errors. The synthetic receptors were separately apportioned by CMB and NCPCRCMB model. The result showed that source contributions estimated by the NCPCRCMB model were much closer to the true values than those estimated by the CMB model. Next, five real ambient data sets from five cities in China were analyzed using the NCPCRCMB model to test the model practicability. Reasonable results were obtained in all cases. It is shown that the NCPCRCMB model has an advantage over the traditional CMB model when dealing with near collinearity problems in source apportionment studies.

Influence of ethanol-diesel blended fuels on diesel exhaust emissions and mutagenic and genotoxic activities of particulate extracts.

This study was aimed at evaluating the influence of ethanol addition on diesel exhaust emissions and the toxicity of particulate extracts. The experiments were conducted on a heavy-duty diesel engine and five fuels were used, namely: E0 (base diesel fuel), E5 (5%), E10 (10%), E15 (15%) and E20 (20%), respectively. The regulated emissions (THC, CO, NOx, PM) and polycyclic aromatic hydrocarbon (PAH) emissions were measured, and Ames test and Comet assay, respectively, were used to investigate the mutagenicity and genotoxicity of particulate extracts. From the point of exhaust emissions, the introduction of ethanol to diesel fuel could result in higher brake specific THC (BSTHC) and CO (BSCO) emissions and lower smoke emissions, while the effects on the brake specific NOx (BSNOx) and particulate matters (BSPM) were not obvious. The PAH emissions showed an increasing trend with a growth of ethanol content in the ethanol-diesel blends. As to the biotoxicity, E20 always had the highest brake specific revertants (BSR) in both TA98 and TA100 with or without metabolizing enzymes (S9), while the lowest BSR were found in E5 except that of TA98-S9. DNA damage data showed a lower genotoxic potency of E10 and E15 as a whole.