Strategies for the efficient biosynthesis of ß-carotene through microbial fermentation.

ß-Carotene is an orange fat-soluble compound, which has been widely used in fields such as food, medicine and cosmetics owing to its anticancer, antioxidant and cardiovascular disease prevention properties. Currently, natural ß-carotene is mainly extracted from plants and algae, which cannot meet the growing market demand, while chemical synthesis of ß-carotene cannot satisfy the pursuit for natural products of consumers. The ß-carotene production through microbial fermentation has become a promising alternative owing to its high efficiency and environmental friendliness. With the rapid development of synthetic biology and in-depth study on the synthesis pathway of ß-carotene, microbial fermentation has shown promising applications in the ß-carotene synthesis. Accordingly, this review aims to summarize the research progress and strategies of natural carotenoid producing strain and metabolic engineering strategies in the heterologous synthesis of ß-carotene by engineered microorganisms. Moreover, it also summarizes the adoption of inexpensive carbon sources to synthesize ß-carotene as well as proposes new strategies that can further improve the ß-carotene production.

Effect of pre-drying and post-frying holding treatments on the oil absorption and quality of fried batter-coated peanuts.

In this study, the effect of pre-drying and post-frying holding treatments on the oil absorption and the quality of the fried batter-coated peanuts were explored. The results showed that hot air drying and microwave drying induced the gelatinization of starch in the batter before frying. The thermodynamic properties of starch in the batter after frying indicated that pre-drying could protect the orderliness of the starch. CLSM images showed that the pre-drying treatment reduced the number of large oil spots on the surface of batter of fried batter-coated peanuts. SEM observation revealed that the structure of the batter treated with pre-drying was denser and the number of large pores was reduced after frying. The post-frying holding treatment improved the color and texture of the batter-coated peanuts. In conclusion, the pre-drying and post-frying holding treatment can reduce the oil content and improve the fracturability of the fried batter-coated peanuts.

The addition of resistant starch and protein to the batter reduces oil uptake and improves the quality of the fried batter-coated nuts.

The batter compositions can affect the oil uptake and texture of fried batter-coated nuts. In this study, the oil uptake and quality of fried batter-coated peanuts and sunflower seeds added with resistant starch and protein were investigated. The results demonstrated that the addition of resistant starch increased the batter hardness and fracturability of the fried batter-coated peanuts by 34.36 % and 33.73 %, respectively. The oil content of fried batter-coated peanuts and sunflower seeds were decreased by 17.98 % and 15.69 %, respectively, with the addition of protein. The microstructure and roughness of the batter revealed that the batter added with protein became denser and uniform. Furthermore, the protein in the batter added with 6 % soy protein isolate had a high surface hydrophobicity. In summary, the addition of resistant starch and protein in batter will be a promising strategy for reducing the oil content and improving the quality of fried batter-coated nuts.

Chitosan-Delivered Chlorantraniliprole for Pest Control: Preparation Optimization, Deposition Behavior, and Application Potential.

Chitosan has emerged as a promising biopolymer carrier for the sustained release of pesticides owing to its good biocompatibility, biodegradability, and bioactivity. In this work, a controlled-release formulation of insecticide chlorantraniliprole was fabricated through coprecipitation-based synchronous encapsulation with chitosan, where the optimum preparation conditions, storage stability, deposition behavior, and application potential were investigated. Preparation of optimization data from response surface methodology showed high correlation coefficient (R2) of 0.9875 and adjusted coefficient (Radj2) of 0.9715. The resulting formulation displayed good loading content of 28.39%, high encapsulation efficiency of 75.71%, and good storage stability. Compared with the commercial suspension concentrate, the formulation exhibited better wettability and retention behaviors on plant leaves. Excitingly, effective control against one species of mealybug genus Paraputo Laing (outside the killing spectrum) on the Hippeastrum reticulatum plant was successfully achieved by spraying the controlled-release formulation at different time intervals. This work indicates the good potential of the developed formulation in expanding the application scope of chlorantraniliprole, which shows a new strategy for sustainable pest management.

Coprecipitation-based synchronous chlorantraniliprole encapsulation with chitosan: carrier-pesticide interactions and release behavior.

BACKGROUND: Controlled-release pesticide formulations have emerged as a promising approach towards sustainable pest control. Herein, an environment-friendly formulation of insecticide chlorantraniliprole (CAP) was fabricated through a simple approach of coprecipitation-based synchronous encapsulation by chitosan (CTS), with carrier-pesticide interaction mechanism and release behavior investigated. RESULTS: The resulting CAP/CTS controlled-release formulation (CCF) showed a good loading content of 28.1% and a high encapsulation efficiency of 75.6%. Instrument determination in combination with molecular dynamics (MD) simulations displayed that the primary interactions between CAP and CTS were physical adsorption and complicated hydrogen (H)-bonds, which formed dominantly between NH in amides [or nitrogen (N) in ring structures] of CAP and hydroxyl (or amino) groups of CTS, as well as oxygen (O) in CAP with hydrogen in CTS or H2 O molecules. The in vitro release tests exhibited obvious pH/temperature sensitivity, with release dynamics following the first-order or Ritger-Peppas model. As the temperature increased, the CAP release process of the Ritger-Peppas model changed from Case-II to anomalous transport, and ultimately to a Fickian diffusion mechanism. The control effect against Plutella xylostella larvae also was evaluated by toxicity tests, where comparable efficacy of CCF to the commercial suspension concentrate was obtained. CONCLUSION: The innovative, easy-to-prepare CCF can be used as a formulation with obvious pH/temperature sensitivity and good efficacy on target pests. This work contributes to the development of efficient and safe pesticide delivery systems, especially using the natural polymer materials as carriers. © 2023 Society of Chemical Industry.

Evaluation of a Spinosad Controlled-Release Formulation Based on Chitosan Carrier: Insecticidal Activity against Plutella xylostella (L.) Larvae and Dissipation Behavior in Soil.

Controlled-release pesticide formulations using natural polymers as carriers are highly desirable owing to their good biocompatibility, biodegradability, and improved pesticide utilization. In this study, the application potential of our previously prepared spinosad/chitosan controlled-release suspension (SCCS) was evaluated through both toxicity and dissipation tests. A comparison with the spinosad suspension concentrate and the commercial spinosad emulsion in water showed that the insecticidal activity of SCCS against Plutella xylostella larvae displayed the best quick-acting performance as well as long-term efficacy of more than 20 days. The 48 h LC50 for a 20-day efficacy was calculated to be 29.36 mg/L. The dissipation behavior of spinosad in the spinosad/chitosan microparticles in soil was found to follow the first-order kinetics, with a relatively shorter half-life (2.1 days) than that observed for the unformulated spinosad (3.1 days). This work showed the positive effect of chitosan on spinosad in improving insecticidal activity and reducing environmental risks in soil, which provided useful information on the application potential of pesticide-carrier systems based on natural polymer materials in crop protection and food safety.

[Biosynthesis of spherical selenium nanoparticles with halophilic Bacillus subtilis subspecies stercoris strain XP for inhibition of strawberry pathogens].

Biosynthesis of nanomaterials has attracted much attention for its excellent characteristics such as low energy consumption, high safety, and environmental friendliness. As we all know, the toxic selenite can be transformed into higher-value nanomaterials by using bacteria. In this study, nano-selenium was synthesized by halophilic Bacillus subtilis subspecies stercoris strain XP in LB medium supplemented with selenite (electron acceptor). The physicochemical characteristics of nano-selenium were analyzed by scanning electron microscope (SEM), X-ray energy dispersive spectral analysis (EDAX), X-ray diffraction (XRD), and fourier transform infrared spectroscopy (FTIR). Meanwhile, the antifungal activity of nano-selenium to strawberry pathogens (fusarium wilt, erythema, and purple spot fungi) was determined. The products from reduction of selenite by strain XP was amorphous spherical selenium nanoparticles (SeNPs) with a diameter range of 135-165 nm. The production of SeNPs was positively correlated with time (0-48 h) and no changes were observed on cell morphology. Selenium was dominant in the surface of SeNPs where the organic elements (C, O, N, and S) existed at the same time. SeNPs were coated with biomolecules containing functional groups (such as -OH, C=O, N-H, and C-H) which were associated with the stability and bioactivity of particles. Although the highest concentration of SeNPs had significant (P<0.05) inhibitory effects on three strains of strawberry pathogens, antifungal activity to erythema and fusarium wilt pathogenic fungi was higher than that to purple spot pathogenic fungi from strawberry. In conclusion, strain XP not only has strong tolerance to high salt stress, but can be also used to synthesize biological SeNPs with good stability and biological activity. Thus, the strain XP has bright perspectives and great potential advantage in pathogens control and green selenium-rich strawberry planting as well as other fields.

Multiple Roles of Mesoporous Silica in Safe Pesticide Application by Nanotechnology: A Review.

Pollution related to pesticides has become a global problem due to their low utilization and non-targeting application, and nanotechnology has shown great potential in promoting sustainable agriculture. Nowadays, mesoporous silica-based nanomaterials have garnered immense attention for improving the efficacy and safety of pesticides due to their distinctive advantages of low toxicity, high thermal and chemical stability, and particularly size tunability and versatile functionality. Based on the introduction of the structure and synthesis of different types of mesoporous silica nanoparticles (MSNs), the multiple roles of mesoporous silica in safe pesticide application using nanotechnology are discussed in this Review: (i) as nanocarrier for sustained/controlled delivery of pesticides, (ii) as adsorbent for enrichment or removal of pesticides in aqueous media, (iii) as support of catalysts for degradation of pesticide contaminants, and (iv) as support of sensors for detection of pesticides. Several scientific issues, strategies, and mechanisms regarding the application of MSNs in the pesticide field are presented, with their future directions discussed in terms of their environmental risk assessment, in-depth mechanism exploration, and cost-benefit consideration for their continuous development. This Review will provide critical information to related researchers and may open up their minds to develop new advances in pesticide application.

Enhanced Anaerobic Biogas Production From Wheat Straw by Herbal-Extraction Process Residues Supplementation.

Trace metals are essential constituents of cofactors and enzymes and that their addition to anaerobic digesters increases methane production. Many trace elements are contained in herbal-extraction process residues (HPR). The present study concerns the effect of six kinds of HPR [Danshen root (Dr), Astragalus membranaceus (Am), Isatis root (Ir), Angelica sinensis (As), and Pseudo-ginseng (Pg)] that were used as additives, respectively, in the anaerobic digestion of wheat straw on biogas and methane production. The ratios of HPR residues/wheat straw [based on total solids (TS), of wheat straw] were 3, 5, and 10%, respectively. The digesters were at 37 ± 1°C of water bath during 30 days of anaerobic digestion. The results showed that HPR had significant effects on the anaerobic co-digestion. The highest biogas productivity was achieved when treated with 10% Pseudo-ginseng residues (PGR), which yielded 337 ml/g TS of biogas and 178 ml/g TS of methane. Cumulative production of biogas and methane increased by 28 and 37% compared to the production achieved in the control. These results suggest that PGR is an effective HPR to enhance the production of methane.

Relieving ammonia nitrogen inhibition in high concentration anaerobic digestion of rural organic household waste by Prussian blue analogue nanoparticles addition.

The application of Prussian blue analogue nanoparticles in anaerobic digestion was firstly used to evaluate the removal effect of ammonia nitrogen inhibition in anaerobic digestion. We have successfully prepared Prussian blue analogue nanoparticles, which has a high adsorption capacity of ammonia nitrogen in anaerobic digestion is 71.09 mg/g. The high concentration anaerobic digestion of rural organic household waste was not successful because of the serious inhibition of ammonia nitrogen. After adding Prussian blue analogue nanoparticles, the methane production of each group increased greatly, up to 302.22 ml/gVS. The concentration of ammonia nitrogen in anaerobic digestion decreased to 1700.77 mg/l. Prussian blue analogue nanoparticles have a good application prospect in high concentration anaerobic digestion of rural organic household waste enriched with a high concentration of ammonia nitrogen.

Major educational factors associated with nursing adverse events by nursing students undergoing clinical practice: A descriptive study.

BACKGROUND: As the main group of healthcare providers in hospitals, nurses have more frequent contacts than any other clinician and thus are in a better position to improve patient safety. With the purpose of cultivating competent nurses, nursing educators have the responsibility to promote patient safety. A better understanding of educational factors affecting nursing adverse events by nursing students undergoing clinical practice can help nursing educators find appropriate ways to fulfil their duty. OBJECTIVE: To examine the status quo of nursing adverse events and to discuss the major educational factors concerned in different regions of China. DESIGN: A descriptive study design was undertaken in 2018. PARTICIPANTS AND SETTING: A convenience sample of 1173 nursing students undergoing clinical practice was recruited from 22 hospitals in different regions of China. METHODS: The Chinese version of the Medical Student Safety Attitudes and Professionalism Survey (MSSAPS) was administered to and demographic and professional data were collected from clinical nursing students after obtaining informed consent. RESULTS: The incidence of nursing adverse events in clinical student nurses was 17.8%. Approximately 87.01% of nursing adverse events were near miss. The positive response rate of safety attitudes and professionalism by clinical nursing students ranged from 57.5% to 96.9%. Logistic analysis indicated that gender, educational level, hospital regions, safety culture and professional behavior experience dimensions were the major factors influencing nursing adverse events. CONCLUSION: Attention should be paid to the situation in which clinical nursing students are prone to nursing adverse events. Cooperation between nursing colleges and hospitals should be strengthened to promote patient safety in clinical nursing students. We suggest that nursing educators implement patient safety education in both theoretical and practical teaching and use multiple forms, especially simulation-based training, to strengthen safe nursing behavior to reduce the incidence of nursing adverse events.

Coprecipitation-based synchronous pesticide encapsulation with chitosan for controlled spinosad release.

Controlled release pesticide formulation with long-term efficacy is highly desirable for improving pesticide utilization and reducing adverse effects. In this study, an environment-friendly controlled release formulation of biological insecticide spinosad was fabricated through the coprecipitation-based synchronous encapsulation with chitosan. The resulted spinosad/chitosan formulation showed an excellent encapsulation efficiency of 60 %, obvious pH and temperature sensitivity, and outstanding ultraviolet shielding ability. The in vitro release tests displayed long sustained-release time (>18 d) and high cumulative release (>80 %) under neutral and weakly alkaline conditions. The primary interaction between spinosad and chitosan has been found to be physical adsorption and adhesion, with the release dynamics following Fickian diffusion mechanism of Ritger-Peppas model. This work contributes to the development of pesticide carrier system of natural polymer materials, with a great potential for insect pest control.

Improving the fermentable sugar yields of wheat straw by high-temperature pre-hydrolysis with thermophilic enzymes of Malbranchea cinnamomea.

BACKGROUND: Enzymatic hydrolysis is a key step in the conversion of lignocellulosic polysaccharides to fermentable sugars for the production of biofuels and high-value chemicals. However, current enzyme preparations from mesophilic fungi are deficient in their thermostability and biomass-hydrolyzing efficiency at high temperatures. Thermophilic fungi represent promising sources of thermostable and highly active enzymes for improving the biomass-to-sugar conversion process. Here we present a comprehensive study on the lignocellulosic biomass-degrading ability and enzyme system of thermophilic fungus Malbranchea cinnamomea N12 and the application of its enzymes in the synergistic hydrolysis of lignocellulosic biomass. RESULTS: Malbranchea cinnamomea N12 was capable of utilizing untreated wheat straw to produce high levels of xylanases and efficiently degrading lignocellulose under thermophilic conditions. Temporal analysis of the wheat straw-induced secretome revealed that M. cinnamomea N12 successively degraded the lignocellulosic polysaccharides through sequential secretion of enzymes targeting xylan and cellulose. Xylanase-enriched cocktail from M. cinnamomea N12 was more active on native and alkali­pretreated wheat straw than the commercial xylanases from Trichoderma reesei over temperatures ranging from 40 to 75 °C. Integration of M. cinnamomea N12 enzymes with the commercial cellulase preparation increased the glucose and xylose yields of alkali­pretreated wheat straw by 32 and 166%, respectively, with pronounced effects at elevated temperature. CONCLUSIONS: This study demonstrated the remarkable xylanase-producing ability and strategy of sequential lignocellulose breakdown of M. cinnamomea N12. A new process for the hydrolysis of lignocellulosic biomass was proposed, comprising thermophilic enzymolysis by enzymes of M. cinnamomea N12 followed with mesophilic enzymolysis by commercial cellulases. Developing M. cinnamomea N12 as platforms for thermophilic enzyme mixture production will provide new perspectives for improved conversion yields for current biomass saccharification schemes.

Parvibaculum sedimenti sp. nov., A Novel Soil Bacterium Isolated from Sediment.

A Gram-stain-negative, aerobic bacterium, designated HXT-9T, was isolated from a river sediment. Cells were rod-shaped and non-motile. Growth occurred at 10-42 ℃ (optimum 30 ℃), at pH 5.0-8.0 (optimum pH 7.0) and with NaCl concentrations of 0-2.5% (optimum 0.5%). Phylogenetic trees based on 16S rRNA gene sequences showed that strain HXT-9T belonged to the genus Parvibaculum, and showed highest similarities to P. lavamentivorans KCTC 22775T (96.1%), followed by P. hydrocarboniclasticum EPR 92T (94.8%) and P. indicum P31T (93.6%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain HXT-9T and P. lavamentivorans KCTC 22775T were 75.2% and 20.8%, respectively. The G + C content of strain HXT-9T genome was 62.1 mol%. The major fatty acids (> 10%) were summed feature 8 (C18:1ω7c and/or C18:1ω6c), C19:0 cyclo ω8c and C16:0 3-OH. The major respiratory quinone was ubiquinone 11 (Q-11). The major polar lipids were DPG (diphosphatidylglycerol), PG (phosphatidylglycerol), PME (phosphatidylmonomethylethanolamine), PE (phosphatidylethanolamine), PC (phosphatidylcholine) and AL (unidentified aminolipids). The phylogenetic analysis and physiological and biochemical data showed that strain HXT-9T represents a novel species in the genus Parvibaculum, for which the name P. sedimenti sp. nov. is proposed. The type strain is HXT-9T (= CCTCC AB 2019273T = KCTC 72547T).

High nursing workload is a main associated factor of poor hand hygiene adherence in Beijing, China: An observational study.

AIM: The aim of the study was to explore the impact of nurse workloads on adherence to hand hygiene. BACKGROUND: Adherence to hand hygiene and nursing workloads have been linked to quality of patient care. Therefore, it was important to understand the relationship to safe patient care. DESIGN: This cross-sectional study was performed from January 2016 to June 2016. METHODS: Workloads and adherence to hand hygiene for nurses on 3-day shifts in a tertiary hospital were investigated in 2016. Actual hours worked per shift were timed using a stopwatch to assess nursing workloads. Descriptive and inferential statistics and multiple variable regression analysis were used to analyse the data. RESULTS: Sixty-four nurses from four wards were observed. The average adherence rate of hand hygiene was 26.6% and the average nursing workload per shift was 6.7 hours. Multiple regression revealed that nursing workload was negatively related to adherence rate of hand hygiene. CONCLUSION: Nurses in this study that had a low rate of adherence with hand hygiene frequently had high workloads. Adherence to hand hygiene was independently associated with actual hours worked per shift.

Enhanced methane production from wheat straw with the assistance of lignocellulolytic microbial consortium TC-5.

The major obstacle of methane production from lignocellulose lies in the inefficient deconstruction of biomass. In this study, an anaerobic microbial consortium TC-5 was enriched with high lignocellulose-degradation capacity to enhance methane production from wheat straw. High degradation ratio of 45.7% of un-pretreated wheat straw was achieved due to a multi-species lignocellulolytic enzyme presented in the crude culture supernatant. The specific activity of xylanase, xylan esterase and ß-xylosidase reached the highest level of 4.23, 0.15 and 0.48 U/mg, while cellobiohydrolase, endoglucanase and ß-glucosidase showed the highest specific activity of 0.36, 0.22 and 0.41 U/mg during 9 days' degradation. Inoculation of TC-5 in digestion sludge during anaerobic digestion of wheat straw resulted in remarkable enhancement of 22.2% and 36.6% in methane yield under mesophilic and thermophilic conditions, respectively. This work demonstrates the potential of TC-5 for enhancing the production of biogas and other chemicals through biomass based biorefinery.

Enhanced isopropanol-butanol-ethanol mixture production through manipulation of intracellular NAD(P)H level in the recombinant Clostridium acetobutylicum XY16.

BACKGROUND: The formation of by-products, mainly acetone in acetone-butanol-ethanol (ABE) fermentation, significantly affects the solvent yield and downstream separation process. In this study, we genetically engineered Clostridium acetobutylicum XY16 isolated by our lab to eliminate acetone production and altered ABE to isopropanol-butanol-ethanol (IBE). Meanwhile, process optimization under pH control strategies and supplementation of calcium carbonate were adopted to investigate the interaction between the reducing force of the metabolic networks and IBE production. RESULTS: After successful introduction of secondary alcohol dehydrogenase into C. acetobutylicum XY16, the recombinant XY16 harboring pSADH could completely eliminate acetone production and convert it into isopropanol, indicating great potential for large-scale production of IBE mixtures. Especially, pH could significantly improve final solvent titer through regulation of NADH and NADPH levels in vivo. Under the optimal pH level of 4.8, the total IBE production was significantly increased from 3.88 to 16.09 g/L with final 9.97, 4.98 and 1.14 g/L of butanol, isopropanol, and ethanol. Meanwhile, NADH and NADPH levels were maintained at optimal levels for IBE formation compared to the control one without pH adjustment. Furthermore, calcium carbonate could play dual roles as both buffering agency and activator for NAD kinase (NADK), and supplementation of 10 g/L calcium carbonate could finally improve the IBE production to 17.77 g/L with 10.51, 6.02, and 1.24 g/L of butanol, isopropanol, and ethanol. CONCLUSION: The complete conversion of acetone into isopropanol in the recombinant C. acetobutylicum XY16 harboring pSADH could alter ABE to IBE. pH control strategies and supplementation of calcium carbonate were effective in obtaining high IBE titer with high isopropanol production. The analysis of redox cofactor perturbation indicates that the availability of NAD(P)H is the main driving force for the improvement of IBE production.

A mouse model with age-dependent immune response and immune-tolerance for HBV infection.

BACKGROUND: Viral clearance of human HBV infection largely depends on the age of exposure. Thus, a mouse model with age-dependent immune response and immune-tolerance for HBV infection was established. METHODS: HBVRag1 mice were generated by crossing Rag1-/- mice with HBV-Tg mice. Following adoptive transfer of splenocytes adult (8-9 weeks old) and young (3 weeks old) HBVRag1 mice were named as HBVRag-ReA and HBVRag-ReY mice respectively. The biochemical parameters that were associated with viral load and immune function, as well as the histological evaluation of the liver tissues between the two mouse models were detected. The immune tolerance of HBVRag-ReY mice that were reconstituted at the early stages of life was evaluated by quantitative hepatitis B core antibody assay, adoptive transfer, and modulation of gut microbiota with the addition of antibiotics. RESULTS: HBVRag-ReA mice indicated apparent hepatocytes damage, clearance of HBsAg and production of HBsAb and HBcAb. HBVRag-ReY mice did not develop ALT elevation, and produced HBcAb and HBsAg. A higher number of hepatic CD8+ T and B cells promoted clearance of HBsAg in HBVRag-ReA mice following 30 days of lymphocyte transfer. In contrast to HBVRag-ReA mice, HBVRag-ReY mice exhibited higher levels of Th1/Th2 cytokines. HBVRag-ReY mice exhibited significantly higher (P < .01, approximately 10-fold) serum quantitative anti-HBc levels than HBV-Tg mice, which might be similar to the phase of immune clearance and immune tolerance in human HBV infection. Furthermore, the age-related tolerance in HBVRag-ReY mice that were sensitive to antibiotic treatment was different from that noted in HBV-Tg mice. GS-9620 could inhibit the production of HBsAg, whereas HBV vaccination could induce sustained seroconversion in HBVRag-ReY mice with low levels of HBsAg. CONCLUSIONS: The present study described a mouse model with age-dependent immunity and immune-tolerance for HBV infection in vivo, which may mimic chronic HBV infection in humans.

High-yield production of mannitol by Leuconostoc pseudomesenteroides CTCC G123 from chicory-derived inulin hydrolysate.

Chicory is an agricultural plant with considerable potential as a carbohydrate substrate for low-cost production of biochemicals. In this work, the production of mannitol by Leuconostoc pseudomesenteroides CTCC G123 from chicory-derived inulin hydrolysate was investigated. The bioconversion process initially suffered from the leakage of fructose to the phosphoketolase pathway, resulting in a low mannitol yield. When inulin hydrolysate was supplemented with glucose as a substrate for mannitol production in combination with aeration induction and nicotinic acid induced redox modulation strategies, the mannitol yield greatly improved. Under these conditions, significant improvement in the glucose consumption rate, intracellular NADH levels and mannitol dehydrogenase specific activity were observed, with mannitol production increasing from 64.6 to 88.1 g/L and overall yield increase from 0.69 to 0.94 g/g. This work demonstrated an efficient method for the production of mannitol from inulin hydrolysate with a high overall yield.

A Novel Hydrodynamic Injection Mouse Model of HBV Genotype C for the Study of HBV Biology and the Anti-Viral Activity of Lamivudine.

BACKGROUND: Absence of an immunocompetent mouse model of persistent hepatitis B virus (HBV) infection has hindered the research of HBV infection and the development of antiviral medications. OBJECTIVES: In the present study, we aimed to develop a novel HBV genotype C mouse model by hydrodynamic injection (HI) and then used it to evaluate the antiviral activity of lamivudine. MATERIALS AND METHODS: A quantity of 15 µg of HBV plasmid [pcDNA3.1 (+)-HBV1.3C], adeno-associated virus-HBV1.3C (pAAV-HBV1.3C) or pAAV-HBV1.2A) were injected into male C57BL/6 mice, by HI, accounting for a total of 13 mice per group. Then, lamivudine was administered to mice with sustained HBV viremia, for 4 weeks. Real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry methods were used to detect HBsAg, HBeAg, HBsAb, HBcAg and HBV DNA, in serum or liver of the mice, at indicated time points. RESULTS: In 60% of the mice injected with pcDNA3.1 (+)-HBV1.3C, HBsAg, HBeAg, HBcAg and HBV DNA persisted for > 20 weeks in liver, post-injection, with no HBsAb appearance. Meanwhile, no significant inflammation was observed in these mice. Compared with pAAV-HBV1.2A and pAAV-HBV1.3C, pcDNA3.1 (+)-HBV1.3C administration led to higher and longer HBV viremia. Furthermore, serum HBV DNA was significantly reduced by lamivudine, after 4 weeks administration, and returned to the original level, after ceasing administration for 1 week, in the mice. CONCLUSIONS: In conclusion, our observations indicated that pcDNA3.1 (+)-HBV1.3C was superior to AAV/HBV plasmid for establishment of persistent HBV infection by HI, in vivo, and this mouse model could be useful for studies of hepatitis virology and for the development of innovatory treatments for HBV infections.