Construction of Cox7a2 fluorescent vector and its effect on cytochrome C oxidase activity in mouse Sertoli cell line TM4

Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. PCR product was

Effects of Different Preservative Treatments on Physiological Metabolism and Preservation of Sweet Cherry

[Objective] This study aimed to investigate the effects of different preservative treatments on physiological metabolism and preservation of sweet cherry. [Method] Sweet cherry （Prunus avium var. Summit） was soaked into benziothiazolinone （1 000 ppm）, lysozyme （500 ppm）, lysozyme （500 ppm） ＋ NPS polysaccharide （5 000 ppm） and water for 5 min, respectively. Non-treated sweet cherry was set as control. All the sweet cherries were then put into 3 mm thick PE bags and preserved at （-0.5±0.5） ℃. [Result] The results showed that the malate dehydrogenase （MDH） activity of benziothiazolinone treatment researched a significant peak on the 14 th d, while the MDH activity of Lysozyme （500 ppm）, Lysozyme （500 ppm） ＋ NPS polysaccharide （5 000 ppm） and water treatments began to increase on the 20 th d; the polyphenol oxidase （PPO） activity in various treatments showed a decreasing trend during the experiment, which researched a significant peak on the 14 th d, while that in water treatment was decreased consistently; on the 21 st d, the PPO activity in each treatment increased slowly; the titratable acid （TA） content in preservative treatments was higher than that in both water treatment and control; the soluble solid （SSC） content showed no significant difference between various treatments and between experimental treatment and control; on the 40 th d, the healthy fruit rate in preservative treatments was significantly higher than that in water treatment and control. [Conclusion] Benziothiazolinone, lysozyme and other preservatives show good effects on preservation of sweet cherry; lysozyme treatment can decrease the activity of malate dehydrogenase, maintain the relatively high content of organic acid and significantly improve the healthy fruit rate within a certain period of time.

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

Identification of Antarctic culturable bacteria able to produce diverse enzymes of potential biotechnological interest

It is estimated that more than three quarters of the Earth＇s biosphere is in perennially cold environments. Despite the extreme environmental conditions of desiccation and freezing, microbes can colonize these habitats through the adaptation of metabolic functions and the synthesis of structurally adapted enzymes. Enzymes within psychrophilic microbes exhibit high specific activity at low and moderate temperature, with low thermostability. In this study we used a classic microbiological approach to isolate Antarctic bacteria with cellulolytic, lipolytic, and ligninolytic activities. From 15 different environmental samples, we generated a collection of approximately 800 bacterial isolates that could grow on R2A or Marine medium at 4℃. This collection was then screened for the presence of the three types of activity at 4℃. We found that 47.7% of the isolates displayed lipolytic activity, 10.2% had cellulase/xylanase activity, and 7.7% showed guaiacol oxidase activity. Of these, 10% displayed two different types of activity, while 0.25% displayed all three types of activity. Our results indicate that cold environments represent outstanding resources for bioprospecting and the study of enzymatic adaptation.

Widely Untargeted Metabolome Profiling Provides Insight into Browning and Nutritional Quality Changes in Short-Term Stored Fresh-Cut Potato(Solanum tuberosum L.)Shreds

Potato(Solanum tuberosum L.)is susceptible to enzymatic browning after fresh processing,resulting in color change and potential alteration in the nutritional quality.In this study,a popular potato cultivar,Feiwuruita,was used to profile the metabolites involved in color and nutritional quality changes in fresh shreds stored at 0 and 4 h at 25°C(designated CK and CK4H,respectively).The shreds turned brown within 4 h of storage.In all,723 metabolites consisting 12 classes of compounds were detected in the samples,largely lipids,phenolic acids,alkaloids,amino acids and derivatives,flavonoids,organic acids,nucleotides and derivatives.Of these,163 metabolites accumulated differentially between CK and CK4H shreds.Polyphenolic compounds(phenolic acids and flavonoids)mostly increased in the shreds after 4 h storage.Conversely,the short-term storage drastically reduced lipid compounds(25 LysoPC and 19 LysoPE),while essential alkaloids and terpenoid compounds that are beneficial to human health increased in accumulation.The findings present global metabolome and nutritional composition changes in short-term stored shreds of Feiwuruita.This study provides important foundation for future studies on browning prevention/reduction and for better utilization of Feiwuruita.

Relationship of familial cytochrome P450 4V2 gene mutation with liver cirrhosis:A case report and review of the literature

BACKGROUND Many genetic and metabolic diseases affect the liver,but diagnosis can be difficult because these diseases may have complex clinical manifestations and diverse clinical patterns.There is also incomplete clinical knowledge of these many different diseases and limitations of current testing methods.CASE SUMMARY We report a 53-year-old female from a rural area in China who was hospitalized for lower limb edema,abdominal distension,cirrhosis,and hypothyroidism.We excluded the common causes of liver disease(drinking alcohol,using traditional Chinese medicines,hepatitis virus infection,autoimmunity,and hepatolenticular degeneration).When she was 23-years-old,she developed night-blindness that worsened to complete blindness,with no obvious cause.Her parents were first cousins,and both were alive.Analysis of the patient’s family history indicated that all 5 siblings had night blindness and impaired vision;one sister was completely blind;and another sister had night-blindness complicated with cirrhosis and subclinical hypothyroidism.Entire exome sequencing showed that the patient,parents,and siblings all had mutations in the cytochrome P450 4V2gene(CYP4V2).The CYP4V2 mutations of the parents and two sisters were heterozygous,and the others were homozygous.Two siblings also had heterozygous dual oxidase activator 2(DUOXA2) mutations.CONCLUSION Mutations in the CYP4V2 gene may affect lipid metabolism and lead to chronic liver injury,fibrosis,and cirrhosis.

Improving the xanthine oxidase and adenosine deaminase inhibitory activities of Flos Sophorae Immaturus by ultrasound-assisted heating treatments

Flos Sophorae Immaturus(FSI)is consumed as a natural food rich in flavonoids and exhibits potential xanthine oxidase(XOD)and adenosine deaminase(ADA)inhibitory activities.Therefore,ultrasound and heating treatments were applied to improve the XOD and ADA inhibitory effects of FSI,and the mechanisms were revealed by tracking the changes of flavones.Results showed that ultrasound(480 W,60 min)assisted heating treatments(180℃,60 min)significantly improved the XOD(from 40.97%to 83.47%)and ADA(6.64%-23.28%)inhibition rates.Meanwhile,ultrasound and heat treatment caused significant variations in total polyphenols and flavonoid contents and altered the flavonoid composition of FSI.Specifically,rutin,kaempferol-3-o-rutinoside,and narcissoside of FSI converted to quercetin,kaempferol,and isorhamnetin during heating treatments.However,the model experiment revealed that ultrasound treatment did not cause the conversion of flavones and just promoted the dissolution of FSI flavones.Furthermore,quercetin,kaempferol,and isorhamnetin possessed better XOD and ADA inhibitory activities than rutin,kaempferol-3-o-rutinoside,and narcissoside at a concentration from 0.025 mg/mL to 1.0 mg/mL,implying de-glycosylation is a feasible approach for enhancing XOD and ADA activity.The present study provides a promising strategy for the processing of FSI.

Effects of Earthworms and Ryegrass on the Removal of Fluoranthene from Soil

Earthworms can promote the bioremediation of contaminated soils through enhancing plant growth and microorganism development. The individual and combined effects of earthworms and ryegrass （Lolium multifloram Lain.） on the removM of fiuoranthene from a sandy-loam alluvial soil were investigated in a 70-d microcosm experiment. The experiment was set up in a complete factorial design with treatments in four replicates： without earthworms or ryegrass （control, CK）, with earthworms only （E）, with ryegrass only （P）, and with both earthworms and ryegrass （EP）. The residual fluoranthene, microbial biomass C, and polyphenol oxidase activity in the soil changed significantly （P 〈 0.01） with time. In general, the residual concentration of fluoranthene in the soil decreased sharply from 71.8-88.7 to 31.7-37.4 mg kg-1 in 14 d, and then decreased gradually to 19.7-30.5 mg kg-1 on the 70th d. The flu- oranthene concentration left in the soil was the least with both earthworms and ryegrass, compared to the other treatments at the end of the experiment. Half-life times of fluoranthene in the E, P, and EP treatments were 17.8%-36.3% smaller than that of CK. More fluoranthene was absorbed by earthworms than ryegrass. However, the total amounts of fluoranthene accumulated in both the ryegrass and earthworms were small, only accounting for 0.01%-1.20% of the lost fluoranthene. Therefore, we assumed that microbial degradation would play a dominant functional role in fluoranthene removal from soil. We found that earthworms significantly increased microbial biomass C and polyphenol oxidase activity （P 〈 0.01） in the presence of ryegrass at the end of the experiment. Furthermore, microbial biomass C and polyphenol oxidase activity were significantly （P 〈 0.05） and negatively related to the residual fluoranthene concentration. This implied that earthworms might promote the removal of fluoranthene from soil via stimulating microbial biomass C and polyphenol oxidase activity.