Microfluidic Bioreactor with Fibrous Micromixers for In Vitro mRNA Transcription.

A new type of microfluidic bioreactor with fibrous micromixers for the ingredient mixing and a long macrochannel for the in vitro transcription reaction was fabricated for the continuous production of mRNA. The diameter of the fibrous microchannels in the micromixers was tuned by using an electrospun microfibrous disc with different microfiber diameters. The micromixer with a larger diameter of fibrous microchannels exhibited a better mixing performance than the others. The mixing efficiency was increased to 0.95 while the mixture was passed through the micromixers, suggesting complete mixing. To demonstrate the continuous production of mRNA, the ingredients for in vitro transcription were introduced into the perfluoropolyether microfluidic bioreactor. The mRNA synthesized by the microfluidic bioreactor had the same sequence and in vitro/in vivo performances as those prepared by the bulk reaction. The continuous reaction in the microfluidic bioreactor with efficient mixing performance can be used as a powerful platform for various microfluidic reactions.

Seroprevalence of scrub typhus, murine typhus and spotted fever groups in North Korean refugees.

OBJECTIVES: The aim of this study was to investigate the seroprevalence of antibodies against scrub typhus, murine typhus and spotted fever groups among North Korean refugees within 1 year of their arrival in South Korea. METHODS: We recruited North Korean refugees who had settled in South Korea after a short stay in a third country and did not have any health problems. The antibody titer was measured using a commercial indirect fluorescence assay immunoglobulin G antibody kit. RESULTS: The seroprevalence of antibodies against scrub typhus, murine typhus, and spotted fever groups among the 99 participants was 22.2%, 17.2%, and 10.1%, respectively, with 8.1% of participants testing positive for both spotted fever and murine typhus. CONCLUSIONS: Refugees may be exposed to rickettsial infections in North Korea and their journey from North Korea. This study is the first to report the seroprevalence of antibodies against the 3 common rickettsial diseases among North Korean refugees. The findings suggest that rickettsial infections should be added to the list of differential diagnoses for North Koreans with fever after entering South Korea.

Characterization of physicochemical properties of casein mixture preparation extracted from organic milk for use as an emulsifier in organic processed foods.

BACKGROUND: Sodium caseinate (SC) is not considered suitable for use as an emulsifier in organic processed food in the food industry because of the use of prohibited synthetic chemical substances during its production. Casein mixture preparation (CMP), one of the permissible substances specified in the regulations, was isolated from organic milk using citric acid and dibasic potassium phosphate for organic processed foods. RESULTS: To compare CMP and SC, model emulsions stabilized with each substance were prepared at various concentrations and their physicochemical properties were analyzed. The emulsions' stability was determined using Turbiscan under various environmental stresses. The zeta potential of SC and CPM showed a high surface charge (≤ 30 mV) at all protein concentrations. Because the concentration of the protein preparation increased to 0.75% (w/v), the particle size of the CMP emulsion decreased with the surface load increased as much as that of SC. The CMP and SC emulsions were stable at neutral pH and room temperature. However, at acidic pH and high temperature, both emulsions were destabilized by creaming and flocculation and increased the creaming migration rates. CONCLUSION: Overall, our data suggest the use of CMP as an emulsifier substitute for SC in organic processed foods. © 2018 Society of Chemical Industry.

Unobtrusive Estimation of Cardiorespiratory Fitness with Daily Activity in Healthy Young Men.

Despite the importance of cardiorespiratory fitness, no practical method exists to estimate maximal oxygen consumption (VO2max) without a specific exercise protocol. We developed an estimation model of VO2max, using maximal activity energy expenditure (aEEmax) as a new feature to represent the level of physical activity. Electrocardiogram (ECG) and acceleration data were recorded for 4 days in 24 healthy young men, and reference VO2max levels were measured using the maximal exercise test. aEE was calculated using the measured acceleration data and body weight, while heart rate (HR) was extracted from the ECG signal. aEEmax was obtained using linear regression, with aEE and HR as input parameters. The VO2max was estimated from the aEEmax using multiple linear regression modeling in the training group (n = 16) and was verified in the test group (n = 8). High correlations between the estimated VO2max and the measured VO2max were identified in both groups, with a 15-hour recording being sufficient to produce a highly accurate VO2max estimate. Additional recording time did not significantly improve the accuracy of the estimation. Our VO2max estimation method provides a robust alternative to traditional approaches while only requiring minimal data acquisition time in daily life.

Dietary D-psicose reduced visceral fat mass in high-fat diet-induced obese rats.

UNLABELLED: D-Psicose, a C-3 epimer of D-fructose, has shown promise in reducing body fat accumulation in normal rats and plasma glucose level in genetic diabetic mice. Effects of D-psicose on diet-induced obesity are not clearly elucidated, and we investigated food intake, body weight, and fat accumulation in rats fed high-fat (HF) diet. Sprague-Dawley rats became obese by feeding HF diet for 4 wk, and were assigned either to normal or HF diet supplemented with or without D-psicose, sucrose, or erythritol for 8 wk. Changing HF to normal diet gained less body weight and adipose tissue due to different energy intake. D-psicose-fed rats exhibited lower weight gain, food efficiency ratio, and fat accumulation than erythritol- and sucrose-fed rats. This effect was more prominent in D-psicose-fed rats with normal diet than with HF diet, suggesting combination of psicose and calorie restriction further reduced obesity. There was no difference in serum cholesterol/high-density lipoprotein (HDL)-C and low-density lipoprotein (LDL)-C/HDL-C ratios between D-psicose group and other groups. Liver weight in 5% psicose group with normal diet was higher than in other groups, but histopathological examination did not reveal any psicose-related change. D-Psicose inhibited the differentiation of mesenchymal stem cell (MSC) to adipose tissue in a concentration-dependent manner. These results demonstrate that D-psicose produces a marked decrease, greater than erythritol, in weight gain and visceral fat in an established obesity model by inhibiting MSC differentiation to adipocyte. Thus, D-psicose can be useful in preventing and reducing obesity as a sugar substitute and food ingredient. PRACTICAL APPLICATION: We can develop D-psicose as a sugar substitute and food ingredient since it can prevent obesity in normal people, but also suppress adiposity as a sugar substitute or food ingredients with antiobesity effect in obese people. D-psicose can be unique functional sweetener because of its function of reducing visceral fat mass and weight gain.

Dietary intake of nitrate relative to antioxidant vitamin in relation to breast cancer risk: a case-control study.

Nitrate is a precursor in the endogenous formation of N-nitroso compounds, which are potent animal carcinogens, whereas antioxidant vitamins have been suggested to protect against carcinogenesis. Interestingly, nitrate and antioxidant vitamins stem from the same dietary sources. We investigated whether the intake of nitrate relative to antioxidant vitamins is associated with the risk of breast cancer. A total of 362 breast cancer cases were matched to the 362 controls by age and menopausal status. Dietary intake was assessed using a quantitative food frequency questionnaire with 121 food items by trained interviewers. The nitrate to antioxidant vitamin consumption ratio was then calculated. Conditional logistic regression analysis was used to obtain odds ratios (ORs) and corresponding 95% confidence intervals (CI). Mean intakes of nitrate for cases and controls were 421 mg/day and 424 mg/day, respectively. Intakes of nitrate, nitrate/beta-carotene, nitrate/vitamin C, and nitrate/vitamin E were not associated with breast cancer risk. However, higher breast cancer risk was observed with higher intake of nitrate/folate (OR = 2.03, 95% CI = 1.16-3.54, P for trend = 0.052). Our results suggest that lowering the ratio of nitrate to folate intake may be effective in reducing breast cancer risk.

Relative validities of 3-day food records and the food frequency questionnaire.

The food frequency questionnaire (FFQ) has been used as an important dietary assessment tool in epidemiologic studies, but the usefulness of the FFQ has been debated in recent years. This study was performed to evaluate the relative validities of 3-day food records and the semi-quantitative FFQ. A total of 124 subjects finished 3-day food records (FRs) during each of the four seasons, as well as the FFQ from December 2002 to May 2004. The FFQ was a food based semi-quantitative FFQ including 103 items. Three-day FRs from each season and a randomly selected season were compared with the remaining 9-day FRs. The remaining 9-day FRs, as a reference measurement, were also compared with the FFQ. Pearson's correlation coefficients between the 3-day FRs and the 9-day FRs were between 0.14 and 0.56. Pearson's correlation coefficients between the FFQ and the 9-day FRs ranged between 0.07 and 0.41. Average proportions of classification into the same quartiles, adjacent quartiles, and distant quartiles between the 3-day FRs and the 9-day FRs were 35.8%, 40.5%, and 5.2%, respectively. On average, the proportions of classification into the same quartiles, adjacent quartiles, and distant quartiles between the FFQ and the 9-day FRs were 31.1%, 39.4%, and 6.9%, respectively. Three-day FRs showed higher correlations and higher agreement proportions of quartile classification with the 9-day FRs than did the FFQ, but both relative validities of 3-day FRs and the FFQ appear to be acceptable as dietary assessment tools. Further studies for validating food intake by reliable biomarkers are necessary.

Microarray analysis of gene expression profile by treatment of Cinnamomi Ramulus in lipopolysaccharide-stimulated BV-2 cells.

It has been reported that components of Cinnamomi Ramulus (CR) demonstrate an anti-inflammatory effect by inhibiting the expression of inducible nitric oxide synthesis (iNOS) and cyclooxygenase-2 (COX-2) and by suppressing nitric oxide (NO) production in the central nervous system (CNS) as well as in the periphery. In this study, microarray analysis was performed to investigate the effect of CR on the gene expression and associated pathways of lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Microglia plays an important role in the processes of several inflammation-mediated neurodegenerative diseases. Activated microglia can produce various pro-inflammatory cytokines, chemokines, and toxic mediators, which may initiate or amplify the inflammatory responses in the CNS. In the present study, the negative control group was cultured in normal medium, the positive control group was activated with 1 microg/ml of LPS, and the CR group was previously treated with 10 microg/ml of CR before LPS stimulation. With the cutoff value of 1.5-fold change in the expression, 341 genes including pro-inflammatory cytokines, chemokines, and transcription factors were found to be up-regulated in LPS-stimulated BV-2 cells (Supplemental Table 2). CR reduced the LPS-induced up-regulation of such inflammatory genes as Ccl5, Cd80, Cxcl10, Grin2a, Ifi203, Ifit1, Il1alpha, Il6, Lilrb3, Nos2 (iNOS), Rab2b, Rsad2 and Vpreb1. This resulted in a full list of 38 and 37 annotated genes whose expression is up- and down-regulated by CR respectively (Supplemental Table 3). RT-PCR analysis showed that the expression of LPS-induced TNF-alpha, IL-1beta, IL-6, and iNOS mRNAs were attenuated in the presence of the CR extract. The results imply that CR has the anti-inflammatory effect of down-regulating the expression of various genes related to inflammatory responses in LPS-stimulated BV-2 cells, and that CR could be a candidate for the prevention or therapeutic treatment of inflammation-mediated neurodegenerative diseases.