Effect of Air Pollutants and Meteorological Factors on Daily Outpatient Visits of Allergic Rhinitis in Hohhot, China.

Objective: There is limited evidence that atmospheric pollutants are associated with the number of allergic rhinitis (AR) visits. This study aimed to investigate the relationship between atmospheric pollutants and meteorological factors and the number of daily visits for AR in Hohhot City, providing a theoretical basis for further research on the effects of atmospheric pollutants on AR. Methods: Microsoft Excel 2016 was used to collect and organize the AR outpatient consultation data, air pollution and meteorological data in Hohhot City during 2018-2020, and the data were descriptively analyzed and Spearman correlation test was performed using SPSS22.0; A generalized additive model was built in R4.2.3 to analyze the effect of air pollution on the number of AR visits and its lagged and cumulative effects, while the robustness of the model was tested using a two-pollutant analysis. Finally, we analyzed the effects of the interaction of temperature, relative humidity and pollutants on the number of AR visits. Results: The daily average concentrations of PM2.5, PM10, SO2, NO2, CO, and O3 in Hohhot during 2018-2020 were 38.1 µg/m3, 83.3 µg/m3, 36.1 µg/m3, 15.1 µg/m3, 14.2 mg/m3, and 99.95 µg/m3, respectively, with the concentrations of PM2.5 and PM10 exceeding the secondary limit of the national standard. The results of the generalized additive model analysis showed that the RR and 95% CI of the effect of each 10 µg/m3 increase in pollutant concentration (1 mg/m3 increase in CO) on the number of AR outpatient clinics were 1.008 (1.001-1.016), 1.002 (1-1.005), 1.072 (1.033-1.113), 1.020 (1007-1.034), 1.033 (1.014-1.052), 0.987 (0.9804-0.9936). Conclusion: Short-term exposure to PM2.5, PM10, SO2, NO2, and CO was significantly associated with an increase in AR clinic visits, and short-term exposure to O3 was significantly associated with a decrease in allergic rhinitis clinic visits.

Relationship Between Outdoor Air Pollutant Exposure and Premature Delivery in China- Systematic Review and Meta-Analysis.

Objective: Preterm birth (PTB) is considered as a public health problem and one of the main risk factors related to the global disease burden. The purpose of this study aims to explore the influence of exposure to major air pollutants at different pregnancies on PTB. Methods: The relationship between air pollutants and PTB in China was collected from cohort studies and case-control studies published before 30 April 2022. Meta-analysis was carried out with STATA 15.0 software. Results: A total of 2,115 papers were retrieved, of which 18 papers met the inclusion criteria. The comprehensive effect of pollutant exposure and PTB were calculated. PM2.5 during entire pregnancy and O3 exposure during third trimester were positively associated with preterm birth. Every 10 µg/m3 increase in the average concentration of PM2.5 during the whole pregnancy will increase the risk of premature delivery by 4%, and every 10 µg/m3 increase in the average concentration of O3 in the third trimester will increase the risk of premature delivery by 1%. Conclusion: Exposure to PM2.5 entire prenatal pregnancy and O3 in third trimester is associated with an increased risk of preterm birth occurrence.

A Fungal P450 Enzyme from Fusarium graminearum with Unique 12ß-Steroid Hydroxylation Activity.

In this study, a new cytochrome P450 enzyme, namely, CYP68J5_Fusarium graminearum (CYP68J5_fg), was identified from Fusarium graminearum via a combination of transcriptome sequencing and heterologous expression in Saccharomyces cerevisiae. The biotransformation of progesterone by whole-cells of S. cerevisiae expressing CYP68J5_fg revealed that the CYP68J5_fg possessed steroidal 12ß- and 15α-hydroxylase activities. To the best of our knowledge, this is the first time that a fungal P450 enzyme with 12ß-hydroxylase activity has been identified. This advance offers opportunities to boost the efficiency and selectivity of the CYP68J5_fg hydroxylating system and thus shows great potential for further applications of this enzyme for the synthesis of steroid drugs. IMPORTANCE Regioselective and stereoselective hydroxylation is of vital importance in the functionalization of steroids, which remains challenging in organic synthesis. In particular, the C12-hydroxy steroids play a significant role in the synthesis of many important steroidal drugs. In this study, a novel fungal P450 enzyme with 12ß-hydroxylation activity was identified, and it shows different substrate specificity and regioselectivity, compared to the bacterial and fungal steroidal hydroxylases that are known to date. This lays the foundation for the creation of effective biocatalysts for the process of 12ß-hydroxylation, although further understanding of the molecular structural basis of this fungal P450 is needed to facilitate the engineering of this enzyme for industrial applications.

Effects of air pollutants and temperature on the number of asthma outpatient visits in Hohhot, China.

Although numerous studies have linked asthma to air temperature and pollution, few studies have examined their interactive effects on asthma outpatient visits. This study investigated how air pollutants and their interactions with temperature affect asthma outpatient visits in a city in northern Chinaduring the time period 2018 - 2020 . . As the results, 24,163 asthma outpatients were recorded, a 10-µg/m3 increase in PM10, PM2.5, SO2, and NO2 concentrations was associated with significant increases in visits of 3.47% (95% CI: 2.35%-4.60%), 0.83% (95% CI: 0.36%-1.30%), 3.17% (95% CI: 1.47%-4.90%), and 8.90% (95% CI: 6.09%-11.79%), respectively. The effect was stronger in females than males, and stronger in the elderly (≥65 years) than among the young. The interaction between low temperatures and high air pollution levels significantly increased the number of asthma outpatient visits. This study emphasizesthe importance of reducing air pollution in order to lessen the effects of cold.

Exploring the Potential of Cytochrome P450 CYP109B1 Catalyzed Regio-and Stereoselective Steroid Hydroxylation.

Cytochrome P450 enzyme CYP109B1 is a versatile biocatalyst exhibiting hydroxylation activities toward various substrates. However, the regio- and stereoselective steroid hydroxylation by CYP109B1 is far less explored. In this study, the oxidizing activity of CYP109B1 is reconstituted by coupling redox pairs from different sources, or by fusing it to the reductase domain of two self-sufficient P450 enzymes P450RhF and P450BM3 to generate the fused enzyme. The recombinant Escherichia coli expressing necessary proteins are individually constructed and compared in steroid hydroxylation. The ferredoxin reductase (Fdr_0978) and ferredoxin (Fdx_1499) from Synechococcus elongates is found to be the best redox pair for CYP109B1, which gives above 99% conversion with 73% 15ß selectivity for testosterone. By contrast, the rest ones and the fused enzymes show much less or negligible activity. With the aid of redox pair of Fdr_0978/Fdx_1499, CYP109B1 is used for hydroxylating different steroids. The results show that CYP109B1 displayed good to excellent activity and selectivity toward four testosterone derivatives, giving all 15ß-hydroxylated steroids as main products except for 9 (10)-dehydronandrolone, for which the selectivity is shifted to 16ß. While for substrates bearing bulky substitutions at C17 position, the activity is essentially lost. Finally, the origin of activity and selectivity for CYP109B1 catalyzed steroid hydroxylation is revealed by computational analysis, thus providing theoretical basis for directed evolution to further improve its catalytic properties.

Regio- and Stereoselective Steroid Hydroxylation at C7 by Cytochrome†P450 Monooxygenase Mutants.

Steroidal C7ß alcohols and their respective esters have shown significant promise as neuroprotective and anti-inflammatory agents to treat chronic neuronal damage like stroke, brain trauma, and cerebral ischemia. Since C7 is spatially far away from any functional groups that could direct C-H activation, these transformations are not readily accessible using modern synthetic organic techniques. Reported here are P450-BM3 mutants that catalyze the oxidative hydroxylation of six different steroids with pronounced C7 regioselectivities and ß stereoselectivities, as well as high activities. These challenging transformations were achieved by a focused mutagenesis strategy and application of a novel technology for protein library construction based on DNA assembly and USER (Uracil-Specific Excision Reagent) cloning. Upscaling reactions enabled the purification of the respective steroidal alcohols in moderate to excellent yields. The high-resolution X-ray structure and molecular dynamics simulations of the best mutant unveil the origin of regio- and stereoselectivity.

[Construction of expression vectors for alkaliphilic Bacillus sp. N16-5].

OBJECTIVE: To construct an expression vector for alkaliphilic Bacillus sp. N16-5. METHODS: Bacillus subtilis expression vector pHCMC04 was used as a backbone. Its xylose-inducible promoter cassette was replaced by the constitutive promoters P43 (from B. subtilis) and P(EF) (from Bacillus sp. N165), separately, resulting in two expression vectors pABN165P43 and pABN165P(EF). Green fluorescent protein gene gfp was linked to the two vectors as a reporter gene. Fluorescence microscope and multifunctional fluorescent reader were used to test the expression efficiency of the system. RESULTS: Green fluorescence was visualized in Bacillus sp. N16-5 with pABN165P(EF)-gfp or pABN165P43-gfp. Quantitative data analysis revealed that fluorescence was first detected around the 7th hour. The fluorescence intensity increased rapidly from the 7th hour to the 12th hour and reached the maximum at about the 12th hour. CONCLUSION: Two expression vectors for Bacillus sp. N16-5 have been constructed, allowing expression of exogenous protein in alkaliphilic Bacillus sp. N165.

Efficient production of L-lactic acid with high optical purity by alkaliphilic Bacillus sp. WL-S20.

Highly efficient polymer-grade L-lactic acid production was achieved by an alkaliphilic strain Bacillus sp. WL-S20 using inexpensive peanut meal as nitrogen source and sodium hydroxide as neutralizing agent. In multi-pulse fed-batch fermentation of Bacillus sp. WL-S20, a L-lactic acid concentration of 225 g/l with a yield of 99.3% was obtained. In single-pulse fed-batch fermentation, a concentration of 180 g/l was obtained with a yield of 98.6%. No D-isomers of lactic acid were detected. The production of a high concentration of optically pure L-lactic acid by alkaliphilic Bacillus sp. WL-S20, combined with a low-cost nutrient and environment-friendly NaOH-based process, represent a potentially novel way for L-lactic acid production at an industrial scale.

Protoplast transformation of recalcitrant alkaliphilic Bacillus sp. with methylated plasmid DNA and a developed hard agar regeneration medium.

Among the diverse alkaliphilic Bacillus strains, only a little have been reported to be genetically transformed. In this study, an efficient protoplast transformation procedure was developed for recalcitrant alkaliphilic Bacillus sp. N16-5. The procedure involved polyethylene glycol-induced DNA uptake by the protoplasts and subsequent protoplast regeneration with a developed hard agar regeneration medium. An in vivo methylation strategy was introduced to methylate the exogenous plasmid DNA for improving the transformation efficiency. The transformation efficiency reached to 1.1×10(5) transformants per µg plasmid DNA with methylated plasmid pHCMC04 and the developed hard agar regeneration medium. This procedure might also be applicable to the genetic transformation of other Bacillus strains.

[Purification and characterization of mannanase from an alkaliphilic mannanase producing bacterium HMTS15].

OBJECTIVE: To isolate and identify an alkaliphilic mannanase producing bacterium, purify and characterize mannanase thereof. METHODS: Mannanase-producing alkaliphilic bacterium HMTS15 was isolated by alkaline agar with konjak from water sample of Hamatai Lake in Inner Mongolia, China. The morphological, biochemical and physiological characteristics and 16S rRNA gene were analyzed to identify the taxonomic position of strain HMTS15. Mannanase produced by strain HMTS15 was purified by four steps including (NH4)2SO4 precipitation, cellulose DEAE-sepharose, twice Superdex 200. The enzyme properties including optimal temperature, optimal pH, thermal stability, pH stability, NaCl tolerance, metal ion tolerance, EDTA and SDS tolerance were tested. RESULTS: Strain HMTS15 was Gram-positive rod. Its growth pH ranged from 7.0 to 11.0 and growth temperature ranged from 10 degrees C to 45 degrees C. The G + C content of the DNA was 40 mol%. Phylogenetic analyses based on 16S rRNA gene sequence comparisons indicated that strain HMTS15 was a member of Bacillus. The extracellular mannanase from strain HMTS15 was purified as a single band with molecular weight of about 45 kD on SDS-PAGE. The optimal catalytic activity was showed at 75 degrees C and pH 10. The mananase was stable up to 60 degrees C and retained about 60% residual activity at 65 degrees C for 30 min. The ions Fe2+, Mn2+, Co2+, Zn2+, Ag+, Hg2+ and EDTA inhibited the acitivity of the mannanase. CONCLUSION: Polyphasic taxonomy revealed that strain HTMS15 was a new member of Bacillus agaradhaerens. The alkaline mannanase produced by strain HMTS15 hold the valuable property in stability at high temperature and broad range of pH.