Switching from Tenofovir-Based Combination Therapy to Tenofovir Monotherapy in Multidrug-Experienced Chronic Hepatitis B Patients: a 5-Year Experience at Two Centers.

Patients with chronic hepatitis B (CHB) who were administered tenofovir disoproxil fumarate (TDF)-based combination therapy after receiving multiple drugs are frequently switched to TDF monotherapy in South Korea. We evaluated the efficacy and safety of switching to TDF monotherapy from TDF-based combination therapy over 5 years. This was a retrospective study of multidrug-experienced CHB patients who switched from TDF-based combination therapy to TDF monotherapy after achieving a virologic response (VR; <20 IU/mL) at Konkuk University Hospital and Sanggye Paik Hospital. The biochemical response was defined as a normalized serum ALT level during follow-up. Each patient was assessed from the date of switching to TDF monotherapy to the date of the last follow-up over 5 years. A total of 39 patients who received at least one antiviral therapy before TDF-based combination therapy were analyzed. The median duration of VR before switching to TDF monotherapy was 18 months and the median duration of TDF monotherapy was 55 months. In this study, except for one patient who had poor compliance, all patients maintained a VR. Three patients had a temporarily increased HBV DNA level and 91.2% of the patients showed a biochemical response. Switching multidrug-experienced patients to TDF monotherapy is generally safe and effective.

Enhancement of sophorolipids production in Candida batistae, an unexplored sophorolipids producer, by fed-batch fermentation.

Sophorolipids (SLs) from Candida batistae has a unique structure that contains ω-hydroxy fatty acids, which can be used as a building block in the polymer and fragrance industries. To improve the production of this industrially important SLs, we optimized the culture medium of C. batistae for the first time. Using an optimized culture medium composed of 50 g/L glucose, 50 g/L rapeseed oil, 5 g/L ammonium nitrate and 5 g/L yeast extract, SLs were produced at a concentration of 24.1 g/L in a flask culture. Sophorolipids production increased by about 19% (28.6 g/L) in a fed-batch fermentation using a 5 L fermentor. Sophorolipids production more increased by about 121% (53.2 g/L), compared with that in a flask culture, in a fed-batch fermentation using a 50 L fermentor, which was about 787% higher than that of the previously reported SLs production (6 g/L). These results indicate that a significant increase in C. batistae-derived SLs production can be achieved by optimization of the culture medium composition and fed-batch fermentation. Finally, we successfully separated and purified the SLs from the culture medium. The improved production of SLs from C. batistae in this study will help facilitate the successful development of applications for the SLs.

High-level production and high-yield recovery of lactobionic acid by the control of pH and temperature in fermentation of Pseudomonas taetrolens.

Lactobionic acid (LBA) was produced by fermentation of Pseudomonas taetrolens. First, to increase the production of LBA by P. taetrolens, we controlled the pH of culture medium by CaCO3 addition (30 g/L) and then examined the initial lactose concentration ranging from 50 to 200 g/L and the growth temperature ranging from 20 to 37 °C. Both the LBA production titer (180 g/L) and the productivity (2.5 g/L h) were highest at 200 g/L lactose concentration and 25 °C of cell growth temperature in shake-flask culture. Although the production of LBA (178 g/L) was almost similar during the batch fermentation of P. taetrolens using 5 L bioreactor, the LBA productivity highly increased to 4.9 g/L h. The method using ethanol precipitation and ion-exchange chromatography was developed to recover the pure LBA from the fermentation broth. The optimum volume of ethanol and pH of culture medium for the precipitation of Ca2+ salt form of LBA were six volume of ethanol and pH 6.5, respectively. The cation-exchange resin T42 finally showed the best recovery yield (97.6%) of LBA from the culture supernatant. The production titer (178 g/L) and the productivity (4.9 g/L h) of lactobionic acid in this study were highest among the previous studies ever reported using P. taetrolens as a production strain of LBA.

Successive complementary model-based experimental designs for parameter estimation of fed-batch bioreactors.

When a dynamic model is used for the description of (fed-)batch bioreactors, it is typical that the model parameters are highly correlated to each other. In this case, it is important to keep the parameter correlation as small as possible to obtain a reliable set of parameter estimates. In this study, we propose an anticorrelation parameter estimation scheme that can be best utilized when a number of different batch experiments are sequentially processed. The scheme iteratively performs parameter estimation and model-based design of experiment (MBDOE) at the beginning and between the batches. The important difference from the existing approaches is that the MBDOE objective is defined according to the system analysis performed a priori, so that each new batch supplements what is lacking from the previous batches combined, in terms of information. The use of the scheme is illustrated on a fed-batch bioreactor model.

Isoprene production by Escherichia coli through the exogenous mevalonate pathway with reduced formation of fermentation byproducts.

BACKGROUND: Isoprene, a volatile C5 hydrocarbon, is an important platform chemical used in the manufacturing of synthetic rubber for tires and various other applications, such as elastomers and adhesives. RESULTS: In this study, Escherichia coli MG1655 harboring Populus trichocarpa isoprene synthase (PtispS) and the exogenous mevalonate (MVA) pathway produced 80 mg/L isoprene. Codon optimization and optimal expression of the ispS gene via adjustment of the RBS strength and inducer concentration increased isoprene production to 199 and 337 mg/L, respectively. To augment expression of MVA pathway genes, the MVA pathway was cloned on a high-copy plasmid (pBR322 origin) with a strong promoter (Ptrc), which resulted in an additional increase in isoprene production up to 956 mg/L. To reduce the formation of byproducts derived from acetyl-CoA (an initial substrate of the MVA pathway), nine relevant genes were deleted to generate the E. coli AceCo strain (E. coli MG1655 ΔackA-pta, poxB, ldhA, dld, adhE, pps, and atoDA). The AceCo strain harboring the ispS gene and MVA pathway showed enhanced isoprene production of 1832 mg/L in flask culture with reduced accumulation of byproducts. CONCLUSIONS: We achieved a 23-fold increase in isoprene production by codon optimization of PtispS, augmentation of the MVA pathway, and deletion of genes involved in byproduct formation.

Synthesis of novel arylaminoquinazolinylurea derivatives and their antiproliferative activities against bladder cancer cell line.

A novel series of arylurea and arylamide derivatives 1a-z, 2a-d having aminoquinazoline scaffold was designed and synthesized. Their in vitro antiproliferative activities against RT112 bladder cancer cell line and inhibitory activities against FGFR3 kinase were tested. Most compounds showed good antiproliferative activities against RT112 bladder cancer cell line, and arylurea compounds 1a-z were more potent than arylamide compounds 2a-d. Among them, eight compounds 1a, 1d-g, 1l, 1y, and 1z showed potent activities with GI50 values below submicromolar range. Especially, arylurea compounds 1d and 1g possessing 2,3-dimethyl and 3,4-dimethyl moieties exhibited superior or similar antiproliferative activity (GI50=8.8nM and 30.2nM, respectively) to AZD4547 (GI50=29.2nM) as a reference standard.

Optimization of microalgal photobioreactor system using model predictive control with experimental validation.

To maximize biomass and lipid concentrations, various optimization methods were investigated in microalgal photobioreactor systems under mixotrophic conditions. Lipid concentration was estimated using unscented Kalman filter (UKF) with other measurable sources and subsequently used as lipid data for performing model predictive control (MPC). In addition, the maximized biomass and lipid trajectory obtained by open-loop optimization were used as target trajectory for tracking by MPC. Simulation studies and experimental validation were performed and significant improvements in biomass and lipid productivity were achieved in the case where MPC was applied. However, occurence of a lag phase was observed while manipulating the feed flow rates, which is induced by large amount of inputs. This is an important phenomenon that can lead to model-plant mismatch and requires further study for the optimization of microalgal photobioreactors.

Analysis of Chameleonic Change of Red Cabbage Depending on Broad pH Range for Dye-Sensitized Solar Cells.

Dye-sensitized solar cells (DSSCs) were assembled using natural dyes extracted from red cabbage as a sensitizer. In this work, we investigated the adsorption characteristics and the electrochemical behavior for harvesting sunlight and electron transfer in red cabbage DSSCs under different solvents and pH. For the red cabbage dye-sensitized electrode adsorbed at pH 3.5, the solar cell yields a short-circuit current density (Jsc) of 1.60 mA/cm2, a photovoltage (Vcc) of 0.46 V, and a fill factor of 0.55, corresponding to an energy conversion efficiency (η) of 0.41%.

Food intake survey of kindergarten children in Korea: Part 2 increased dietary intake of tin possibly associated with canned foods.

OBJECTIVES: Dietary intake of tin (Sn) may be increased in some children in kindergartens in Korea. The present study was intended to examine this possibility and clarify the extent of the elevation. METHODS: 24-hour food duplicate and spot urine samples were collected in 2003-2004 from 108 4-6-year-old children (boys and girls combined) in 4 kindergartens (1 in Seoul and 3 in Jeju Island), as reported in a previous publication. These samples were employed in the present analyses to examine tin levels in the diet (including beverages) (Sn-D). A portion of the samples were wet-ashed, and the liquid samples were analyzed for Sn by the ICP-MS method. For statistical evaluation, χ (2) method and Smirnov's test for extreme value were used. RESULTS: Sn-D in the 108 cases distributed as extremely biased, and could be divided into two groups, i.e., those with <10 µg Sn/day (accounting for 90% of the cases), and those with >10 µg/day (for 10%). Sn-D in the former group was distributed quasi-normally with an AM (median) of 2.9 (2.5) µg/day. The maximum in the latter group was 3012 µg/day. No correlation was detected between Sn-D and Sn in urine (Sn-U). Comparison of the findings with published articles strongly suggested that the high Sn-D was due to consumption of foods (including beverages) preserved in tin-plated cans. No positive confirmation was however possible due to insufficient information on food records. CONCLUSIONS: About 10% of children surveyed had elevated Sn-D (up to 3 mg/day). It was quite possible that high Sn-D was associated with tin-canned food intake.

Synthetic biology and metabolic engineering for marine carotenoids: new opportunities and future prospects.

Carotenoids are a class of diverse pigments with important biological roles such as light capture and antioxidative activities. Many novel carotenoids have been isolated from marine organisms to date and have shown various utilizations as nutraceuticals and pharmaceuticals. In this review, we summarize the pathways and enzymes of carotenoid synthesis and discuss various modifications of marine carotenoids. The advances in metabolic engineering and synthetic biology for carotenoid production are also reviewed, in hopes that this review will promote the exploration of marine carotenoid for their utilizations.

Conversion of toxic pyrogenic products into syngas through catalytic pyrolysis of insulation material waste under the presence of CO2.

Plastic-based insulation materials have been widely employed owing to their exceptional durability, cost-effectiveness, low weight, and low thermal conductivity. Nevertheless, the disposal of the insulation material waste (IMW) within construction waste and its recycling and recovery are challenging. Meanwhile, landfilling or incineration methods can release toxic chemicals into the environment. Consequently, the accumulation of IMW in construction waste has become a pressing environmental concern. To address this issue, this paper proposes a pyrolysis platform as a disposal management method for IMW that employs CO2 as a reactive medium. IMW composed of polystyrene in the form of extruded polystyrene underwent pyrolysis to yield pyrogenic products containing toxic chemicals. These toxic chemicals were subsequently transformed into syngas via homogeneous reactions with CO2 under certain thermal conditions and Ni/Al2O3 catalyst. This resulted in a significant reduction in the total peak areas of toxic substances in the pyrogenic oil compared with that obtained using N2 as a medium. Furthermore, the efficacy of CO2 was demonstrated to increase with an increase in the atmospheric concentration. This study implied that catalytic pyrolysis under CO2 conditions is a potential platform for converting toxic chemicals from IMW into syngas through homogeneous reactions with CO2.

Functional use of carbon dioxide for the sustainable valorization of orange peel in the pyrolysis process.

Although biomass is carbon-neutral, its use as a primary feedstock faces challenges arising from inconsistent supply chains. Therefore, it becomes crucial to explore alternatives with reliable availability. This study proposes a strategic approach for the thermochemical valorization of food processing waste, which is abundantly generated at single sites within large-scale processing plants. As a model biomass waste from the food industry, orange peel waste was particularly chosen considering its substantial consumption. To impart sustainability to the pyrolysis system, CO2, a key greenhouse gas, was introduced. As such, this study highlights elucidating the functionality of CO2 as a reactive feedstock. Specifically, CO2 has the potential to react with volatile pyrolysates evolved from orange peel waste, leading to CO formation at ≥490 °C. The formation of chemical constituents, encompassing acids, ketones, furans, phenols, and aromatics, simultaneously decreased by 15.1 area% in the presence of CO2. To activate the efficacy of CO2 at the broader temperature spectrum, supplementary measures, such as an additional heating element (700 °C) and a nickel-based catalyst (Ni/Al2O3), were implemented. These configurations promote thermal cracking of the volatiles and their reaction kinetics with CO2, representing an opportunity for enhanced carbon utilization in the form of CO. Finally, the integrated process of CO2-assisted catalytic pyrolysis and water-gas shift reaction was proposed. A potential revenue when maximizing the productivity of H2 was estimated as 2.62 billion USD, equivalent to 1.11 times higher than the results from the inert (N2) environment. Therefore, utilizing CO2 in the pyrolysis system creates a promising approach for enhancing the sustainability of the thermochemical valorization platform while maximizing carbon utilization in the form of CO.

Effect of age on the prognosis of intrahepatic cholangiocarcinoma.

BACKGROUND/AIMS: Intrahepatic cholangiocarcinoma (iCCA) is a subgroup of cholangiocarcinoma and is the second- most-common primary hepatic tumor. Several predictive and prognostic factors have been analyzed; however, in this study we focused on the influence of age. Our aim was to use real-world results to determine the influence of age in iCCA patients. METHODS: A retrospective analysis of patients treated between 2005 and 2016 at Konkuk University Medical Center. In total, 133 patients with iCCA were identified. The mass-forming, periductal-infiltrating, and intraductal-growth types were included; patients with extrahepatic or hilar-type cholangiocarcinoma were excluded. We defined two groups: a younger group, age < 65 years, and an older group, age ≥ 65 years. Statistical analyses using univariate and multivariate Cox regression analyses, including the Kaplan-Meier method, were conducted. RESULTS: In total, 114 patients were enrolled. The two groups differed with regard to treatment options such as surgery with adjuvant chemotherapy or palliative chemotherapy (p = 0.012, p < 0.001). The younger group had significantly longer survival than the older group (p = 0.017). In the younger group, patients who received therapy had longer survival than those who did not (hazard ratio, 3.942; 95% confidence interval, 2.053 to 7.569; p < 0.001). Multivariate analysis indicated that younger age, lower bilirubin, low CA 19-9, and no lymph-node involvement were independent factors for improved survival. CONCLUSION: Younger patients and those who underwent surgery with adjuvant chemotherapy had longer survival. The younger the patient, the more treatments received, including palliative chemotherapy.

Intravesical bacillus Calmette-Guérin-induced myopathy presenting as rhabdomyolysis: a case report.

Intravesical bacillus Calmette-Guérin (BCG) instillation is an adjuvant treatment for non-muscle-invasive urinary bladder cancer. Although most complications associated with BCG immunotherapy are mild and self-limiting, rare albeit serious complications have been reported. Only a few cases of BCG-related rhabdomyolysis have been reported. In this study, we present the case of a 72-year-old woman who developed severe weakness and hyperCKemia following intravesical BCG instillation. A muscle biopsy was performed, and a diagnosis of drug-induced myopathy was made.

Subacute toxicity and stability of Soshiho-tang, a traditional herbal formula, in Sprague-Dawley rats.

BACKGROUND: Soshiho-tang (SST, Xiao-chai-hu-tang in Chinese and Sho-saiko-to in Japanese), an oriental herbal formula, is used for treatment of chronic liver diseases. Although many researchers have studied the pharmacological properties of SST, information about its safety and toxicity is limited. Therefore, we evaluated the potential safety of SST in Sprague-Dawley rats over a period of 4-weeks. METHODS: The SST was administered once daily by gavage to male and female rats at doses of 0, 500, 1000 and 2000 mg/kg/day for 4 weeks. We measured the body weight, mortality, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross pathological findings, absolute/relative organ weights and histopathology. In addition, we analyzed the component of SST and measured the stability of its component in SST according to study periods using high performance liquid chromatography. RESULTS: The SST treatment did not result in any toxicologically significant changes in mortality, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross pathological findings, absolute/relative organ weights and histopathology, except for salivation and reduction in body weight in the 2000 mg/kg/day male group. These findings in the 2000 mg/kg/day male group are considered toxicologically insignificant because they are not accompanied by other pathological findings, including in hematology, serum biochemistry and histopatholgy, and they do not exhibit a dose-response relationship. SST is detected three components including liquiritin, baicalin, and glycyrrhizin. In addition, there were not observed the significant differences among the contents of three components in SST according to storage periods. CONCLUSION: These results indicate that SST may be a safe material. Based on these results, the no-observed-adverse-effect level was more than 2000 mg/kg for both genders.

Post-processing calcium subtraction method to minimize stenosis-overestimation by blooming artifact.

BACKGROUND: CT images are often affected by blooming artifacts during the diagnosis that facilitate an overestimation of the expression of calcification stenosis, thereby impeding the accurate diagnosis of this condition. OBJECTIVE: Arterial calcification can act as a blooming artifact in computed tomography (CT) images, leading to overestimations of the blood vessel and the size of calcified plaque. This study proposes an improved CT post-processing method that accurately measures calcium and lumen size in blood vessels. METHODS: Six hundred and thirty calcium datasets were obtained from 63 patients diagnosed with a vascular disease. Patients were grouped into three sets corresponding to each image acquisition method used: G1, for the invasive coronary angiography (ICA); G2, for multiplanar reconstruction (MPR) imaging and post-processing; and G3, for the novel method of mixed Gaussian filter and K-mean clustering (GK). Results of GK were generated by adding Gaussian and k-mean clustering algorithms to the MPR post-processing procedure. The analysis of variance (ANOVA), linear regression, and intraclass correlation coefficient (ICC) were used to compare the accuracy and sensitivity of the different methods. All measurements were performed multiple times to mitigate human error. RESULTS: The ANOVA test revealed no significant differences between the G1 and G3 groups. Hence, linear regression was used to analyze the correlation between the G1 and G3 groups (p< 0.05, R2 = 0.885), and a higher correlation than G1 and G2 was reported (p> 0.05, R2 = 0.432). ICC was performed for reproducibility, wherein high correlation was identified among all groups. CONCLUSIONS: Results of the study indicate that the GK method yields images that are very similar to ICA image measurements. This suggests that the GK can be used as a more effective post-processing method over the inaccurate MPR while remaining non-intrusive when determining the arterial stenosis degree, unlike the ICA.

Valorizing plastic toy wastes to flammable gases through CO2-mediated pyrolysis with a Co-based catalyst.

Toys are discarded due to their short life cycle. Unfortunately, development of sustainable disposal platform for toy has not gained particular concern. To establish a reliable disposal platform, this study employed a pyrolysis platform to valorize plastics into value-added fuels. To confer more environmentally resilient process, CO2 was used as a feedstock to enhance the process efficiency from a perspective of the yield of flammable gases. To this end, waste toy brick (WTB) was used as a model compound. The exact types of plastics (polyacrylonitrile, polybutadiene, polystyrene, and polymethyl methacrylate) in WTB were experimentally determined. In pyrolysis of WTB, the complicated mixture of benzene derivatives was inevitably generated. To detoxify them by means of syngas (H2/CO) production, catalytic pyrolysis was performed. Co catalyst effectively induced chemical bond scissions, leading to substantially enhanced H2 formation. Also, the gas phase reactions (GPRs) between CO2 and volatile compounds over Co catalyst expedited the production rate of CO, and such CO enhancement effectively offered a chance to mitigate toxic chemical generations. The synergistic contribution of CO2 and Co catalyst enhanced syngas formation more than 25 times in reference to pyrolysis of WTB without Co catalyst. The GPRs also greatly prevented catalyst deactivation.

Retinoid production using metabolically engineered Escherichia coli with a two-phase culture system.

BACKGROUND: Retinoids are lipophilic isoprenoids composed of a cyclic group and a linear chain with a hydrophilic end group. These compounds include retinol, retinal, retinoic acid, retinyl esters, and various derivatives of these structures. Retinoids are used as cosmetic agents and effective pharmaceuticals for skin diseases. Retinal, an immediate precursor of retinoids, is derived by ß-carotene 15,15'-mono(di)oxygenase (BCM(D)O) from ß-carotene, which is synthesized from the isoprenoid building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Retinoids are chemically unstable and biologically degraded via retinoic acid. Although extensive studies have been performed on the microbial production of carotenoids, retinoid production using microbial metabolic engineering has not been reported. Here, we report retinoid production using engineered Escherichia coli that express exogenous BCM(D)O and the mevalonate (MVA) pathway for the building blocks synthesis in combination with a two-phase culture system using a dodecane overlay. RESULTS: Among the BCM(D)O tested in E. coli, the synthetic retinoid synthesis protein (SR), based on bacteriorhodopsin-related protein-like homolog (Blh) of the uncultured marine bacteria 66A03, showed the highest ß-carotene cleavage activity with no residual intracellular ß-carotene. By introducing the exogenous MVA pathway, 8.7 mg/L of retinal was produced, which is 4-fold higher production than that of augmenting the MEP pathway (dxs overexpression). There was a large gap between retinal production and ß-carotene consumption using the exogenous MVA pathway; therefore, the retinal derivatives were analyzed. The derivatives, except for retinoic acid, that formed were identified, and the levels of retinal, retinol, and retinyl acetate were measured. Amounts as high as 95 mg/L retinoids were obtained from engineered E. coli DH5α harboring the synthetic SR gene and the exogenous MVA pathway in addition to dxs overexpression, which were cultured at 29°C for 72 hours with 2YT medium containing 2.0% (w/v) glycerol as the main carbon source. However, a significant level of intracellular degradation of the retinoids was also observed in the culture. To prevent degradation of the intracellular retinoids through in situ extraction from the cells, a two-phase culture system with dodecane was used. The highest level of retinoid production (136 mg/L) was obtained after 72 hours with 5 mL of dodecane overlaid on a 5 mL culture. CONCLUSIONS: In this study, we successfully produced 136 mg/L retinoids, which were composed of 67 mg/L retinal, 54 mg/L retinol, and 15 mg/L retinyl acetate, using a two-phase culture system with dodecane, which produced 68-fold more retinoids than the initial level of production (2.2 mg/L). Our results demonstrate the potential use of E. coli as a promising microbial cell factory for retinoid production.

Comparative study on carbon dioxide-cofed catalytic pyrolysis of grass and woody biomass.

This study investigated the mechanistic functions of CO2 on the pyrolysis of two different biomasses to elucidate the effect of CO2 on syngas formations during pyrolysis. To this end, CO2-assisted pyrolysis of cellulosic biomass (barnyard grass, Echinochloa) and lignin-rich woody biomass (retinispora, Chamaecyparis obtusa) were compared. The confirmed mechanistic effectiveness of CO2 on pyrolysis of biomass was gas phase reactions between CO2 and volatile matters from biomass pyrolysis. Lignin-rich biomass had more CO2 susceptibility, resulting in more enhanced CO formation via the gas phase reactions. To expedite the slow reaction rate of the gas phase reactions during biomass pyrolysis, earth-abundant catalysts (Co/SiO2 and Ni/SiO2) were employed for pyrolysis of two biomass substrates. With Co and Ni catalysts, the syngas formations were 2 and 3 times higher comparing to the pyrolysis of without catalyst. The cumulative formations of syngas from lignin-rich biomass was nearly doubled than that from cellulosic biomass.

Valorization of carbon dioxide and waste (Derived from the site of Eutrophication) into syngas using a catalytic thermo-chemical platform.

Global warming increases a chance of eutrophication, and such fact offers that unhygienic organic waste materials (OWMs) in water must be treated. Hence, this study laid emphasis on the thermal-chemical (pyrolysis) process to establish a rapid valorization platform for OWMs. Indeed, OWMs were collected from the eutrophication site, and OWMs were mainly comprised of lignocellulosic biomass, microalgae (cyanobacteria) and the diverse types of bacteria (commonly observed from livestock waste). In an attempt to offer more sustainable valorization route for OWMs, CO2 was used as a raw material in pyrolysis process. From the CO2-assisted pyrolysis, the conversion of CO2 and OWMs into gaseous fuel (CO) was observed. A cheap Ni-based catalyst was used in pyrolysis of OWMs as a strategic practice to promote conversion of CO2 into CO. Indeed, syngas production (38 %) was enhanced from catalytic pyrolysis over Ni/SiO2 under CO2 condition as compared to inert condition (N2).