Enhanced production of a novel cytotoxic chromone oxalicumone A by marine-derived mutant Penicillium oxalicum SCSIO 24–2

Many marine natural products hold great potential for the development of new and much needed drugs. However, the production of active metabolites by marine-derived microorganisms is usually very low, and large-scale culture has to be involved to meet the need of chemical structural modification and deep pharmacy study. In order to enhance the production of a novel cytotoxic sulfur-containing chromone oxalicumone A (OA), germinating spores of a marine-derived wild strain Penicillium oxalicum SCSGAF 0023 were mutated by microwave and ultraviolet light irradiation, which led to the obtainment of a mutant P. oxalicum SCSIO 24–2 that could produce fivefold increase in OA production (3.42?±?0.21 mg/l) as compared to the wild strain. This is the first report that germinating spores are applied in marine-derived Penicillium sp. mutating to enhance the production of OA. Further, Plackett–Burman design and central composite design were adopted to optimize the basic medium components for increasing OA production by the mutant SCSIO 24–2 in shake flasks. The results indicated that three medium components including mannitol, maltose, and l-cysteine had significant effects on OA production, and their concentrations were optimized as 36, 27.9, and 0.99 g/l, respectively. In the optimized medium, the OA production (18.31?±?0.27 mg/l) by mutant SCSIO 24–2 was 4.4-fold higher than that in the basic medium. These results of this work promise to improve the present production of OA and may be adopted to enhance other objective products' production by marine-derived fungi.     

Genotype distribution and evolutionary analysis of rotavirus associated with acute diarrhea outpatients in Hubei,China, 2013–2016

Group A human rotaviruses (RVAs) annually cause the deaths of 215,000 infants and young children. To understand the epidemiological characteristics and genetic evolution of RVAs, we performed sentinel surveillance on RVA prevalence in a rotavirus-surveillance network in Hubei, China. From 2013 to 2016, a total of 2007 fecal samples from hospital outpatients with acute gastroenteritis were collected from four cities of Hubei Province. Of the 2007 samples, 153 (7.62%) were identified positive for RVA by real-time RT-PCR. RVA infection in Hubei mainly occurred in autumn and winter. The highest detection rate of RVA infection was in 1–2 years old of outpatients (16.97%). No significant difference of RVA positive rate was observed between females and males. We performed a phylogenetic analysis of the G/P genotypes based on the partial VP7/VP4 gene sequences of RVAs. G9P[8] was the most predominant strain in all four years but the prevalence of G2P[4] genotype increased rapidly since 2014. We reconstructed the evolutionary time scale of RVAs in Hubei, and found that the evolutionary rates of the G9, G2, P[8], and P[4] genotypes of RVA were 1.069×10^-3, 1.029×10^-3, 1.283×10^-3 and 1.172×10^-3 nucleotide substitutions/site/year, respectively. Importantly, using a molecular clock model, we showed that most G9, G2, P[8], and P[4] genotype strains dated from the recent ancestor in 2005, 2005, 1993, and 2013, respectively. The finding of the distribution of RVAs in infants and young children in Hubei Province will contribute to the understanding of the epidemiological characteristics and genetic evolution of RVAs in China.     

Intraductal Radiofrequency Ablation Followed by Locoregional Tumor Treatments for Treating Occluded Biliary Stents in Non-Resectable Malignant Biliary Obstruction: A Single-Institution Experience

Objectives

To determine the safety and feasibility of intraductal radiofrequency ablation (RFA) followed by locoregional tumor treatments in patients with non-resectable malignant biliary obstruction and stent re-occlusion.

Methods

Fourteen patients with malignant biliary obstruction and blocked metal stents were studied retrospectively. All had intraductal RFA followed by locoregional tumor treatments and were monitored clinically and radiologically. The practicality, safety, postoperative complications, jaundice remission, stent patency and survival time were analyzed.

Results

Combination treatment was successful for all patients. There were no severe complications during RFA or local treatments. All patients had stent patency restored, with a decline in serum bilirubin. Three patients had recurrent jaundice by 195, 237 and 357 days; two patients underwent repeat intraductal RFA; and one required an internal-external biliary drain. The average stent patency time was 234 days (range 187-544 days). With a median follow-up of 384 days (range 187-544 days), six patients were alive, while eight had died. There was no mortality at 30 days. The 3, 6, 12 and 18 month survival rates were 100%, 100%, 64.3% and 42.9%, respectively.

Conclusion

Intraductal RFA followed by locoregional tumor treatments for occluded metal stents is safe and practically feasible and potential increase stent patency and survival times.     

A Chinese Creutzfeldt-Jakob disease patient with E196K mutation in PRNP

Genetic Creutzfeldt-Jakob disease (gCJD) is caused by a range of mutations in the prion protein gene (PRNP) and account for approximately 10–15% of overall human prion diseases worldwide. They are different with disease onset, disease duration, clinical signs and diagnostic findings. Here we reported a 71 year-old female with an E196K mutation in one PRNP allele, while the codon 129 was a methionine homozygous genotype. The patient started with non-specific symptoms, but displayed rapidly progressive disturbances of speech, memory, cognitive and physical movement. No periodic activity was recorded at electroencephalography (EEG) during the entire disease course. Retrospective investigation of her family members did not reveal similar neurological disorders. Total clinical course was about seven months.Key words: Creutzfeldt-Jakob disease, PRNP, E196K     

棉花根际亲和性高效促生细菌的分离筛选

为了从棉花根际土壤筛选能与棉花凝集素具有亲和作用的高效促生细菌,以选择性培养基从棉花根部初步筛选具有固氮能力、解磷能力及解钾能力的促生细菌,再以异硫氰酸磺(FITC)标记的棉花凝集素为复筛工具,从棉花根际促生细菌中筛选能与棉花凝集素结合的亲和性菌株,分别挑选2株固氮菌、2株解磷细菌和2株解钾细菌作为微生物肥料接种到棉花根部进行盆栽试验.观察其在根部定殖情况.结果是在选择性平板上有20%～30%的菌株具有凝集素染色阳性.盆栽试验显示,接种的6株亲和性菌株能在棉花根部成功定殖,根际细菌数量约是灭活对照的`0倍.通过初步鉴定,固氮菌株N1111为固氮菌属(Azotobacter),N2121属于德克斯氏菌属(Derxia);解磷菌株P2126属于黄单胞菌属(Xanthomonas),P1108菌株为假单胞菌属(Pseudomonas);解钾菌株K2204和K2116属于芽孢杆菌属(Bacillus).     

Metabolic engineering of <Emphasis Type="Italic">Methylobacterium extorquens</Emphasis> AM1 for the production of butadiene precursor

Background

Butadiene is a platform chemical used as an industrial feedstock for the manufacture of automobile tires, synthetic resins, latex and engineering plastics. Currently, butadiene is predominantly synthesized as a byproduct of ethylene production from non-renewable petroleum resources. Although the idea of biological synthesis of butadiene from sugars has been discussed in the literature, success for that goal has so far not been reported. As a model system for methanol assimilation, Methylobacterium extorquens AM1 can produce several unique metabolic intermediates for the production of value-added chemicals, including crotonyl-CoA as a potential precursor for butadiene synthesis.

Results

In this work, we focused on constructing a metabolic pathway to convert crotonyl-CoA into crotyl diphosphate, a direct precursor of butadiene. The engineered pathway consists of three identified enzymes, a hydroxyethylthiazole kinase (THK) from Escherichia coli, an isopentenyl phosphate kinase (IPK) from Methanothermobacter thermautotrophicus and an aldehyde/alcohol dehydrogenase (ADHE2) from Clostridium acetobutylicum. The K_m and k_cat of THK, IPK and ADHE2 were determined as 8.35 mM and 1.24 s^?1, 1.28 mM and 153.14 s^?1, and 2.34 mM and 1.15 s^?1 towards crotonol, crotyl monophosphate and crotonyl-CoA, respectively. Then, the activity of one of rate-limiting enzymes, THK, was optimized by random mutagenesis coupled with a developed high-throughput screening colorimetric assay. The resulting variant (THK^M82V) isolated from over 3000 colonies showed 8.6-fold higher activity than wild-type, which helped increase the titer of crotyl diphosphate to 0.76 mM, corresponding to a 7.6% conversion from crotonol in the one-pot in vitro reaction. Overexpression of native ADHE2, IPK with THK^M82V under a strong promoter mxaF in M. extorquens AM1 did not produce crotyl diphosphate from crotonyl-CoA, but the engineered strain did generate 0.60 μg/mL of intracellular crotyl diphosphate from exogenously supplied crotonol at mid-exponential phase.

Conclusions

These results represent the first step in producing a butadiene precursor in recombinant M. extorquens AM1. It not only demonstrates the feasibility of converting crotonol to key intermediates for butadiene biosynthesis, it also suggests future directions for improving catalytic efficiency of aldehyde/alcohol dehydrogenase to produce butadiene precursor from methanol.

     

低磷胁迫水稻根部基因表达谱研究

磷是植物体内重要的营养元素．土壤中含磷总量丰富,但能被植物直接吸收利用的可溶性磷含量却很低,这成为制约农作物产量的重要因素．本研究利用水稻寡核苷酸芯片分析了水稻根部在正常营养条件和低磷胁迫6,24,72h3个时间点的全基因组表达谱．和正常营养条件下相比,低磷胁迫水稻根部共发现795个差异表达基因．差异表达基因功能分析发现：（1）磷酸盐转运蛋白、酸性磷酸酶、RNA酶等基因上升表达;（2）糖酵解等与能量代谢相关基因先上升后下降表达;（3）氮吸收和脂代谢相关基因改变其表达;（4）蛋白质降解、细胞衰老相关基因上升表达;（5）部分跨膜转运蛋白基因表达上调．研究结果为进一步揭示植物低磷胁迫反应机制,改善作物对磷吸收利用效率提供了有用的信息．     

Risk Factors for Bunyavirus-Associated Severe Fever with Thrombocytopenia Syndrome,China

Background

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that is caused by a novel bunyavirus, referred to as SFTS virus. During January 2011 to December 2011 we conducted a case-control study in Henan, Hubei and Shandong Provinces of China to determine the risk factors for SFTS.

Methods

Case-patients were identified in hospitals and reported to provincial Centers for Disease Control and Prevention while being notified electronically to the National Surveillance System. Controls were randomly selected from a pool of patients admitted to the same hospital ward within one week of the inclusion of the cases. They were matched by age (+/−5 years) and gender.

Results

A total of 422 patients participated in the study including 134 cases and 288 matched controls. The median age of the cases was 58.8 years, ranging from 47.6 to 70.1 years; 54.5% were male. No differences in demographics were observed between cases and controls; however, farmers were frequent and more common among cases (88.8%) than controls (58.7%). In multivariate analysis, the odds for SFTS was 2.4∼4.5 fold higher with patients who reported tick bites or presence of tick in the living area. Other independent risk factors included cat or cattle ownership and reported presence of weeds and shrubs in the working environment.

Conclusions

Our findings support the hypothesis that ticks are important vectors of SFTS virus. Further investigations are warranted to understand the detailed modes of transmission of SFTS virus while vector management, education on tick bites prevention and personal hygiene management should be implemented for high-risk groups in high incidence areas.     

Antifouling potentials of eight deep-sea-derived fungi from the South China Sea

Marine-derived microbial secondary metabolites are promising potential sources of nontoxic antifouling agents. The search for environmentally friendly and low-toxic antifouling components guided us to investigate the antifouling potentials of eight novel fungal isolates from deep-sea sediments of the South China Sea. Sixteen crude ethyl acetate extracts of the eight fungal isolates showed distinct antibacterial activity against three marine bacteria (Loktanella hongkongensis UST950701–009, Micrococcus luteus UST950701–006 and Pseudoalteromonas piscida UST010620–005), or significant antilarval activity against larval settlement of bryozoan Bugula neritina. Furthermore, the extract of Aspergillus westerdijkiae DFFSCS013 displayed strong antifouling activity in a field trial lasting 4 months. By further bioassay-guided isolation, five antifouling alkaloids including brevianamide F, circumdatin F and L, notoamide C, and 5-chlorosclerotiamide were isolated from the extract of A. westerdijkiae DFFSCS013. This is the first report about the antifouling potentials of metabolites of the deep-sea-derived fungi from the South China Sea, and the first stage towards the development of non- or low-toxic antifouling agents from deep-sea-derived fungi.