Changes in nonalcoholic fatty liver disease and M2BPGi due to lifestyle intervention in primary healthcare.

BACKGROUND: A healthy lifestyle is the most important method for managing nonalcoholic fatty liver disease (NAFLD). Mac-2-binding protein glycosylated isomer (M2BPGi) has been suggested as a biomarker for NAFLD. This study aimed to determine the efficacy of personalized lifestyle interventions on NAFLD remission. METHODS: This single-arm intervention study recruited participants with NAFLD who underwent health checkups at seven health-promotion centers in five South Korean cities. Fatty liver diagnosis was based on ultrasonography (US). The 109 individuals were recruited for personalized lifestyle interventions of hypocaloric diets and exercise. The participants attended the lifestyle intervention programs once per month for the first 3 months, and once every 3 months for the subsequent 6 months. In addition to sessions through center visits, phone-based intervention and self-monitoring at 4-, 5-, 7-, and 8-month were provided during the 9-month intervention period. And phone-based self-monitoring were also provided monthly during the 3-month follow-up period. The primary outcome was NAFLD remission at month 12 as measured on US and magnetic resonance elastography. The secondary outcomes were the changes in metabolic factors and M2BPGi. RESULTS: The 108 individuals (62 males and 46 females; age 51.1±12.4 years, mean±standard deviation) were finally analyzed after the 12month intervention. Body mass index, waist circumference (WC), blood pressure, blood lipids (total cholesterol, triglycerides, and HDL-C), and fasting blood sugar levels were improved relative to baseline (all P<0.05). Fatty liver at or above the moderate grade according to US was decreased at month 12 relative to baseline (67.6% vs 50.9%) (P = 0.002). M2BPGi levels decreased during the 12-month study period (P<0.001). M2BPGi levels were moderately correlated with hepatic fat fraction by magnetic resonance imaging (r = 0.33, P = 0.05). WC (OR = 0.82, 95% CI = 0.67-1.00, P = 0.05) and HDL-C (OR = 1.17, 95% CI = 1.03-1.32, P = 0.014) were associated with remission of fatty liver in the multivariate analysis. CONCLUSION: The personalized lifestyle intervention was effective in improving fatty liver and metabolic factors, but not hepatic stiffness, in NAFLD. TRIAL REGISTRATION: ICTRP, cris.nih.go.kr (KCT0006380).

Reference intervals of cell population data parameters in Sysmex XN-Series and its patterns of changes from early adulthood to geriatric ages in South Korea.

INTRODUCTION: Cell population data (CPD) parameters may be putative biomarkers for the screening of various diseases including some infections and myelodysplastic syndrome. This study aimed to establish the age- and sex-specific reference intervals (RIs) for the CPD parameters in the Korean population. METHODS: The reference population for the RIs of CPD parameters comprised 124 856 subjects aged 20-99 years. CPD parameters were obtained from Sysmex XN-2000 (Kobe, Japan) datasets from 17 health promotion centers in 13 South Korean cities. We determined significant partitions for age and sex, and calculated RIs according to Clinical and Laboratory Standards Institute C28-A3 guidelines. RESULTS: The side scattered light intensity in the neutrophil area and the lymphocyte area did not require sex-related partitioning except in those over the age of 50, among whom the lower limit (LL) and upper limit (UL) were lower in females. However, the side scattered light distribution width in the lymphocyte area required age- and sex-related partitioning, in which LL and UL were higher in females. The LL and UL of the fluorescent light distribution width were higher in males in the neutrophil area and higher in females in the lymphocyte area, but age-related partitioning was not required. The forward scattered light intensity in the neutrophil area, lymphocyte area, and monocyte area did not require age-related partitioning in males. CONCLUSION: This study has determined comprehensive age- and sex-specific RIs for CPD parameters, which could help to prove the clinical significance of these parameters in the Sysmex XN-2000.

SARS-CoV-2 Antibody Response and Sustainability after a Third Dose of BNT162b2 in Healthcare Workers at Health Promotion Centers.

The aim of this study was to determine the antibody response and the sustainability of immunogenicity after a third dose of BNT162b2 (BNT) in homologous [ChAdOx1 (ChAd)/ChAd, BNT/BNT, and mRNA-1273 (Moderna)/Moderna] and heterologous (ChAd/BNT) vaccinations of two primary doses with different schemes. This prospective observational study recruited consenting healthcare workers from 16 health checkup centers in 13 Korean cities. Three-point blood tests were analyzed as the antibody response after the third vaccination: T3-1 (1 month after the third dose), T3-3 (3 months after the third dose), and T3-4-10 (4-10 months after the third dose). SARS-CoV-2 antibodies were measured using a chemiluminescence microparticle immunoassay with SARS-CoV-2 IgG II Quant in the ARCHITECT system (Abbott Diagnostics). The antibody levels were significantly higher in the Moderna /Moderna and BNT/BNT groups than in the ChAd/ ChAd and ChAd/BNT groups (p < 0.05) at T3-1. At T3-3, antibody levels had decreased by 29.1% in the BNT/BNT group and by 45.3% in the ChAd/ChAd group compared with the antibody levels at T3-1. The anti-SARS-CoV-2 S-RBD IgG levels at T3-1 were significantly associated with having received mRNA vaccines as the two primary doses (p < 0.001). The third dose of BNT induced an increased humoral immune response in various vaccination schemes, which was more prominent for the two primary doses of homologous mRNA vaccines. However, this immunogenicity decreased within 3-10 months after the third dose. These results suggest that another booster dose (a fourth dose), which would be able to counteract SARS-CoV-2 variants, is needed.

Establishment and validation of reference intervals for tumor markers (AFP, CEA, CA19-9, CA15-3, CA125, PSA, HE4, Cyfra 21-1, and ProGRP) in primary care centers in Korea: A cross-sectional retrospective study.

Background and Aims: The reference interval (RI) for a tumor marker may vary between populations, detection systems, and the methods used to obtain their values. The aims of this study were to establish age- and sex-specific RIs for the following nine common tumor markers and to validate the established RIs in Korean adults: alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen (CA) 19-9, CA15-3, CA125, Human epididymis protein 4 (HE4), total prostate specific antigen, cytokeratin fragment (Cyfra) 21-1, and progastrin-releasing peptide (ProGRP). Methods: This cross-sectional study consecutively selected 214,159 individuals (aged 18-98 years) who underwent health checkups at 16 health-promotion centers in 13 Korean cities. Finally, 62,752 examinees were used to establish the RIs after removing outliers. RIs were established using an indirect method according to the CLSI EP28-A3C guideline. The established RIs were validated by calculating the proportion of individuals outside each RI. Results: Sex-related differences were observed for AFP, CEA, CA19-9, Cyfra 21-1, and ProGRP (p < 0.05): AFP, CEA and Cyfra 21-1 were higher in males, and CA19-9 and proGRP were higher in females. Most of the tumor markers except CA15-3 and CA125 increased with age: CA125 decreased at ≥50 years of age (p < 0.05), while CA15-3 did not vary with age. Less than 5% of subjects were outside all RIs (the 2.5th and 97.5th percentiles) established in the present study. Meanwhile, less than 3% of the healthy reference subjects fell outside the current and manufacturers' RIs of all tumor markers except Cyfra 21-1. Conclusion: This study has determined age- and sex-specific RIs for nine common tumor markers in the healthy Korean population, which could be useful for clinicians making clinical decisions and assessments.

Chronic kidney disease in nonalcoholic fatty liver disease at primary healthcare centers in Korea.

BACKGROUND: The prevalence rates of nonalcoholic fatty liver disease (NAFLD) and chronic kidney disease (CKD) are expected to increase with the rising trends in diabetes and obesity associated with aging populations. Considering the impacts of coexistent NAFLD and CKD on morbidity and mortality rates, screening strategies for groups at high-risk of CKD are needed in community-dwelling individuals with NAFLD. The aims of this study were to determine the prevalence and distribution of CKD in NAFLD, as well as the risk factors for CKD and the correlation with liver fibrosis in asymptomatic individuals with NAFLD at primary healthcare centers in Korea. METHODS: This retrospective cross-sectional study used data from 13 health-promotion centers in 10 Korean cities. Liver steatosis and stiffness were assessed using ultrasonography and magnetic resonance elastography (MRE), respectively. CKD was defined as an estimated glomerular filtration rate of <60 mL/min/1.73m2, and urine albumin-to-creatinine ratio or proteinuria. CKD was categorized into four stages: no CKD, mild, moderate, and severe. Comparisons according to the CKD stages in NAFLD were performed using Student's t-test or the chi-square test. Multivariable logistic regression analyses were performed to identify the risk factors for CKD and the correlation with liver fibrosis in NAFLD. RESULTS: The prevalence of CKD was 12.4% in NAFLD. Albuminuria (16.2%) and proteinuria (8.0%) were more prevalent in NAFLD. NAFLD (odd ratio = 1.27, 95% CI = 1.09-1.48, P = 0.003) was independently associated with CKD of at least mild stage. However, there was no significant association between CKD of at least moderate stage and NAFLD after adjusting for age and a metabolically unhealthy status. CKD was associated with significant liver fibrosis as measured by MRE in NAFLD. CONCLUSION: The presence of NAFLD and liver fibrosis were independent risk factors for CKD, but NAFLD was not an independent risk factor for the later stages of CKD.

The usefulness of the estimated average glucose/fasting blood glucose ratio for pancreatic ß-cell function assessment in hyperglycemia during health checkups.

BACKGROUND: Type 2 diabetes (T2DM) is a disease marked by inadequate insulin secretion by pancreatic beta-cell function (BCF) failure and insulin resistance (IR). Assessing and managing the BCF and IR should be started early to prevent or delay the progression of the disease. The aim of this study was to determine the usefulness of the estimated average glucose (eAG)/fasting blood glucose (FBG) ratio for pancreatic BCF in hyperglycemia. METHODS: This cross-sectional study consecutively selected 10,594 subjects who underwent a health checkup at 16 health checkup centers in 13 Korean cities between 2019 and 2021. The subjects consisted of 3003 patients with normoglycemia, 3413 with impaired fasting glucose and 4178 with T2DM. The eAG was calculated using Nathan's regression equation. BCF and IR were estimated by the homeostasis model assessment (HOMA)-ß and HOMA-IR, respectively. Multivariate (adjusted) regression analysis was performed to evaluate the association between the eAG/FBG ratio and HOMA. RESULTS: The median values among FBG groups for the eAG/FBG ratio, HOMA-ß, -IR and insulin differed significantly (p < 0.001). The second-, third- and fourth-quartile groups of the eAG/FBG ratio had positive higher correlation coefficients [9.533, 10.080 and 12.021, respectively (all p < 0.001)] for HOMA-ß than the first quartile group, and higher negative coefficients for HOMA-IR [-0.696, -0.727 and -0.598, respectively (all p = 0.001)]. CONCLUSION: The eAG/FBG ratio was significantly correlated with both HOMA-ß and -IR, which suggests that eAG/FBG ratio reveals BCF and IR in hyperglycemia. Measurement of this ratio could be useful for monitoring BCF and IR in prediabetes and T2DM.

Antibody response after two doses of homologous or heterologous SARS-CoV-2 vaccines in healthcare workers at health promotion centers: A prospective observational study.

Assaying of anti-spike-protein receptor-binding domain (S-RBD) antibodies are used to aid evaluations of the immune statuses of individuals. The aim of this study was to determine the antibody response after two doses of homologous or heterologous severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines and to identify the factors affecting this response among healthcare workers (HCWs) at health promotion centers. In this prospective observational study, 1095 consenting HCWs were recruited from 16 health checkup centers and were tested at T0 (day of first dose), T1-1 (1 month after first dose), T2-0 (day of second dose), T2-1 (1 month after second dose), and T2-3 (3 months after second dose). SARS-CoV-2 antibodies were measured using a chemiluminescence microparticle immunoassay with SARS-CoV-2 IgG II Quant in the ARCHITECT system (Abbott Diagnostics). At T1-1, anti-SARS-CoV-2 S-RBD IgG levels were significantly higher in participants who received messenger RNA (mRNA) vaccines than in those who received viral vector vaccines (p < 0.001). At T2-1, anti-SARS-CoV-2 S-RBD IgG levels were about 10 times higher than at T1-1 in participants who received homologous mRNA vaccines, which decreased to a third of those at T2-3. Anti-SARS-CoV-2 S-RBD IgG levels were highest among those who received homologous mRNA vaccines, followed by heterologous mRNA viral vector vaccines and homologous viral vector vaccines at T2-3 (p < 0.001). In a multivariable linear regression analysis, being female, taking at least one mRNA vaccine, and having a history of recovery from coronavirus disease 2019 (COVID-19) were significantly associated with anti-S-RBD levels. Anti-SARS-CoV-2 S-RBD IgG levels were decreased at 3 months after two-dose vaccinations and were associated with sex, vaccine type, and COVID-19 history.

The Reproducibility and Usefulness of Estimated Average Glucose for Hyperglycemia Management during Health Checkups: A Retrospective Cross-Sectional Study.

HbA1c reflects average glucose levels over 3 months, but it does not measure glycemic variability. This study aimed to determine the reproducibility and usefulness of HbA1c-derived estimated average glucose (eAG) and to analyze the factors associated with eAG during health checkups. This cross-sectional retrospective study consecutively selected subjects who had undergone health checkups at 16 health-promotion centers in 13 Korean cities in 2020. The subjects comprised 182,848 healthy subjects with normoglycemia, 109,555 with impaired fasting glucose (IFG), and 35,632 with diabetes. eAG was calculated using Nathan's regression equation. In all subjects, fasting plasma glucose (FPG) was found to be fairly strongly correlated with eAG (r = 0.811). When the subjects were divided into FPG subgroups, the strength of the correlation decreased among those with normoglycemia and IFG (p < 0.001). Higher eAG levels were associated with older age, females, higher FPG, and lower HDL-C and triglycerides (p < 0.05). The proportion of subjects with a higher value of FPG than eAG was 46.3% in poorly controlled diabetic patients, compared with only 1.5% in normoglycemic subjects. This suggests eAG could help patients to understand their glycemic variability intuitively and healthcare providers to identify patients who might worsen in hyperglycemia control through measuring the difference between eAG and FPG.

Relative Lean Body Mass and Waist Circumference for the Identification of Metabolic Syndrome in the Korean General Population.

Lean body mass (LBM) comprises organs and muscle, which are the primary determinants of energy expenditure and regulation of glucose and lipid metabolism. Excessive abdominal fat is associated with metabolic abnormality. Little is known about the relationship between metabolic abnormality and LBM and waist circumference (WC), especially in the Asian general population. The aim of this study was to clarify this relationship. We performed a cross-sectional study with 499,648 subjects who received health check-ups at 16 health promotion centers in 13 Korean cities between January 2018 and October 2019. The subjects were categorized into four groups: (a) High (H)-RLBM (relative lean body mass)/Normal (N)-WC, (b) High-RLBM/Abnormal (A)-WC, (c) Low (L)-RLBM/Normal-WC, and (d) Low-RLBM/Abnormal-WC. RLBM was calculated using fat mass data that were estimated via bioelectrical impedance analysis. L-RLBM/A-WC was significantly associated with metabolically unhealthy status (OR: 4.40, 95% CI: 4.326-4.475) compared to H-RLBM/N-WC. L-RLBM/N-WC (OR: 2.170, 95% CI: 2.122-2.218) and H-RLBM/A-WC (OR: 2.713, 95% CI: 2.659-2.769) were also significantly related to metabolic unhealthy status. The cut-offs of RLBM for predicting metabolic syndrome (MetS) were 74.9 in males and 66.4 in females (p < 0.001). L-RLBM and A-WC are associated with metabolic abnormality in the Korean general population. RLBM is an anthropometric index that can be used to predict MetS in primary health care.

Subclinical steatohepatitis and advanced liver fibrosis in health examinees with nonalcoholic fatty liver disease (NAFLD) in 10 South Korean cities: A retrospective cross-sectional study.

BACKGROUND: Nonalcoholic steatohepatitis (NASH) has a risk of progressing to cirrhosis. The prevalence of NASH and its associated risk factors in community populations are relatively unknown. This study aimed to determine the prevalence of NASH and advanced liver fibrosis using magnetic resonance elastography (MRE), and determine those risk factors in health examinees with asymptomatic fatty liver. METHODS: This study consecutively selected subjects who underwent health checkups at 13 health-promotion centers in 10 Korean cities between 2018 and 2020. Hepatic steatosis and stiffness were assessed using ultrasonography and MRE, respectively. Stages of liver stiffness were estimated using MRE with cutoff values for NASH and advanced liver fibrosis of 2.91 and 3.60 kPa, respectively. RESULTS: The overall prevalence of NASH and advanced liver fibrosis in the subjects with fatty liver were 8.35% and 2.04%, respectively. Multivariate logistic regression analysis indicated that central obesity (OR = 5.12, 95% CI = 2.70-9.71), increased triglyceride (OR = 3.29, 95% CI = 1.72-6.29), abnormal liver function test (OR = 3.09, 95% CI = 1.66-5.76) (all P<0.001), and decreased high-density lipoprotein cholesterol (OR = 5.18, 95% CI = 1.78-15.05) (P = 0.003) were associated with NASH. The main risk factor for advanced liver fibrosis was diabetes (OR = 4.46, 95% CI = 1.14-17.48) (P = 0.032). CONCLUSION: NASH or advanced liver fibrosis is found in one-tenth of health examinees with asymptomatic fatty liver. This suggests that early detection of NASH should be considered to allow early interventions such as lifestyle changes to prevent the adverse effects of NASH and its progression in health examinees with asymptomatic fatty liver.

Prevalence of liver fibrosis and associated risk factors in the Korean general population: a retrospective cross-sectional study.

OBJECTIVES: The health burden of chronic liver disease is increasing worldwide. Its main histological consequence is liver fibrosis, and eventually cirrhosis. This process is rarely diagnosed at the pre-cirrhotic stage due to it being asymptomatic. Little is known about the prevalence of liver fibrosis and associated risk factors in the general population. The aims of this study were to determine the prevalence and distribution of liver fibrosis using magnetic resonance elastography (MRE), as well as the risk factors associated with liver fibrosis in the asymptomatic general population. DESIGN, SETTING AND PARTICIPANTS: This cross-sectional retrospective study consecutively selected subjects who underwent health check-ups including MRE at 13 health promotion centres in Korea between 2018 and 2020. Liver fibrosis was estimated using MRE with cut-off values for significant and advanced liver fibrosis of 2.90 and 3.60 kPa, respectively. PRIMARY AND SECONDARY OUTCOME MEASURES: The Χ2 test was used to compare the prevalence of liver fibrosis according to sex and age groups. Multivariable logistic regression analyses were performed to identify the factors for significant and advanced liver fibrosis. RESULTS: Among the 8183 subjects, 778 (9.5%) had ≥significant fibrosis (≥2.9 kPa), which included 214 (2.6%) subjects with ≥advanced fibrosis (≥3.6 kPa). Multivariable analysis revealed that liver fibrosis was associated with age (OR=1.34, 95% CI=1.18 to 1.51), male sex (OR=3.18, 95% CI=1.97 to 5.13), diabetes (OR=2.43, 95% CI=1.8 to 3.28), HBsAg positivity (OR=3.49, 95% CI=2.55 to 4.79), abnormal liver function test (OR=1.9, 95% CI=1.49 to 2.42) and obesity (OR=1.77, 95% CI=1.35 to 2.32) (all p<0.001), as well as metabolic syndrome (OR=1.4, 95% CI=1.05 to 1.87) (p=0.024). CONCLUSIONS: The prevalence of significant or more liver fibrosis was high in the Korean general population and much higher among individuals with risk factors. This suggests that screening of liver fibrosis should be considered in general population, especially among high-risk groups.

Distribution of hemoglobin levels and prevalence of anemia according to sex, age group, and region in 13 Korean cities.

INTRODUCTION: The distribution of hemoglobin (Hb) levels and the prevalence of anemia are significant public health indicators. The aims of this study were to determine the distribution of Hb levels and the prevalence of anemia according to sex, age group, and region throughout Korea. METHODS: The study analyzed data on 1 159 298 subjects who received health checkups at 16 health-promotion centers in 13 Korean cities during 2018. Anemia and its severity were defined according to the World Health Organization classification for Hb levels as follows: mild anemia (11-12.9 g/dL in males and 11-11.9 g/dL in females), moderate anemia (10-10.9 g/dL in both sexes), and severe anemia (<10.0 g/dL in both sexes). RESULTS: The Hb level in the general sample was lower in females (13.25 ± 1.13 g/dL, mean ± SD) than in males (15.29 ± 1.22 g/dL). The overall prevalence of anemia was 6.0% (2.98% in males and 8.56% in females), and the prevalence of severe anemia was 0.92% (0.23% in males and 1.51% in females). While the prevalence of anemia increased monotonically with age in males, it was bimodal in females with two peaks at 40-49 years and ≥80 years. The highest prevalence of anemia in females aged 40-49 years was attributed to microcytic anemia, while increases in anemia prevalence in males aged ≥50 years and females aged ≥70 years were attributed to macrocytic anemia. CONCLUSION: The distribution of Hb levels and the prevalence of anemia overall and by severity differ according to sex, age group, and region throughout Korea.

Molecular Studies of the Protein Complexes Involving Cis-Prenyltransferase in Guayule (Parthenium argentatum), an Alternative Rubber-Producing Plant.

Guayule (Parthenium argentatum) is a perennial shrub in the Asteraceae family and synthesizes a high quality, hypoallergenic cis-1,4-polyisoprene (or natural rubber; NR). Despite its potential to be an alternative NR supplier, the enzymes for cis-polyisoprene biosynthesis have not been comprehensively studied in guayule. Recently, implications of the protein complex involving cis-prenyltransferases (CPTs) and CPT-Binding Proteins (CBPs) in NR biosynthesis were shown in lettuce and dandelion, but such protein complexes have yet to be examined in guayule. Here, we identified four guayule genes - three PaCPTs (PaCPT1-3) and one PaCBP, whose protein products organize PaCPT/PaCBP complexes. Co-expression of both PaCBP and each of the PaCPTs could complemented the dolichol (a short cis-polyisoprene)-deficient yeast, whereas the individual expressions could not. Microsomes from the PaCPT/PaCBP-expressing yeast efficiently incorporated 14C-isopentenyl diphosphate into dehydrodolichyl diphosphates; however, NR with high molecular weight could not be synthesized in in vitro assays. Furthermore, co-immunoprecipitation and split-ubiquitin yeast 2-hybrid assays using PaCPTs and PaCBP confirmed the formation of protein complexes. Of the three PaCPTs, guayule transcriptomics analysis indicated that the PaCPT3 is predominantly expressed in stem and induced by cold-stress, suggesting its involvement in NR biosynthesis. The comprehensive analyses of these PaCPTs and PaCBP here provide the foundational knowledge to generate a high NR-yielding guayule.

Efficacy of lifestyle interventions in the reversion to normoglycemia in Korean prediabetics: One-year results from a randomised controlled trial.

AIM: This study aimed to determine the efficacy of personalized lifestyle interventions on the reversion of a prediabetic state to normoglycemia compared with regular blood glucose testing alone in prediabetes. METHODS: A randomized, multicenter trial was conducted in prediabetes aged 30-70 with fasting blood glucose level of 5.6-6.9mmol/L (100-125mg/dL) and/or HbA1c level of 39-46mmol/mol (5.7-6.4%) recruited from health checkups at 16 health-promotion centers in Korea. The 799 recruited individuals were randomized to either the personalized lifestyle intervention group (LIG) or the control group (CG) by a computer generated random number list prepared by an independent statistician. The CG was provided with fasting blood glucose and HbA1c tests alone every 3months during the first year. The LIG was provided not only blood glucose test but five sessions of personalized lifestyle counseling by nutritionists every 3months during the first year aimed at improving the diet, alcohol and exercise behaviors. Data from lifestyle assesments and laboratory measurements were analyzed at 1-year after baseline. The primary outcome was the reversion rate from prediabetes to normoglycemia. Additional outcome include the effect of the lifestyle intervention program on lifestyle changes in the LIG to support primary outcome. RESULTS: The 799 participants randomly allocated to the LIG (n=398) or the CG (n=401). For the analyses of outcomes, 629 participants (313 men and 316 women; mean age, 53.7±9.4years; mean body mass index (BMI), 24.7kg/m2) were included: 325 in the LIG; 304 in the CG. Diet (7.03, 95% CI=4.56-10.86, P<0.001), alcohol (2.24, 95% CI=1.48-3.41, P<0.001), and exercise behaviors (1.85, 95% CI=1.31-2.63, P<0.001) were improved relative to baseline by the personalized lifestyle intervention in the LIG after adjusting age, sex, and family history of diabetes. In terms of main outcome, the cumulative incidence of reversion from prediabetes to normoglycemia at the first year was 37.9% (123/325) [95% CI=32.6-43.1%] in the LIG and 29.6% (90/304) (95% CI=24.5-34.7%) in the CG. After adjustment for age, sex, family history of diabetes, BMI, blood pressure, and lipids, the hazard ratio for reverting to normoglycemia remained significantly higher in the LIG (1.40, 95% CI=1.06-1.83, P=0.017) than in the CG. CONCLUSION: Personalized lifestyle intervention could be more effective compared with regular blood glucose testing alone in the reversion of a prediabetic state to normoglycemia in Korean prediabetics. This finding suggests that diabetes prevention care would be benefited by incorporating personalized lifestyle counseling. This study was registered at cris.nih.go.kr (KCT0001580).

Conserved and Diverged Functions of the Calcineurin-Activated Prz1 Transcription Factor in Fission Yeast.

Gene regulation in response to intracellular calcium is mediated by the calcineurin-activated transcription factor Prz1 in the fission yeast Schizosaccharomyces pombe Genome-wide studies of the Crz1 and CrzA fungal orthologs have uncovered numerous target genes involved in conserved and species-specific cellular processes. In contrast, very few target genes of Prz1 have been published. This article identifies an extensive list of genes using transcriptome and ChIP-chip analyses under inducing conditions of Prz1, including CaCl2 and tunicamycin treatment, as well as a ∆pmr1 genetic background. We identified 165 upregulated putative target genes of Prz1 in which the majority contained a calcium-dependent response element in their promoters, similar to that of the Saccharomyces cerevisiae ortholog Crz1 These genes were functionally enriched for Crz1-conserved processes such as cell-wall biosynthesis. Overexpression of prz1(+)increased resistance to the cell-wall degradation enzyme zymolyase, likely from upregulation of theO-mannosyltransferase encoding gene omh1(+) Loss of omh1(+)abrogates this phenotype. We uncovered a novel inhibitory role in flocculation for Prz1. Loss of prz1(+)resulted in constitutive flocculation and upregulation of genes encoding the flocculins Gsf2 and Pfl3, as well as the transcription factor Cbf12. The constitutive flocculation of the ∆prz1 strain was abrogated by the loss of gsf2(+) or cbf12(+) This study reveals that Prz1 functions as a positive and negative transcriptional regulator of genes involved in cell-wall biosynthesis and flocculation, respectively. Moreover, comparison of target genes between Crz1/CrzA and Prz1 indicate some conservation in DNA-binding specificity, but also substantial rewiring of the calcineurin-mediated transcriptional regulatory network.

A lettuce (Lactuca sativa) homolog of human Nogo-B receptor interacts with cis-prenyltransferase and is necessary for natural rubber biosynthesis.

Natural rubber (cis-1,4-polyisoprene) is an indispensable biopolymer used to manufacture diverse consumer products. Although a major source of natural rubber is the rubber tree (Hevea brasiliensis), lettuce (Lactuca sativa) is also known to synthesize natural rubber. Here, we report that an unusual cis-prenyltransferase-like 2 (CPTL2) that lacks the conserved motifs of conventional cis-prenyltransferase is required for natural rubber biosynthesis in lettuce. CPTL2, identified from the lettuce rubber particle proteome, displays homology to a human NogoB receptor and is predominantly expressed in latex. Multiple transgenic lettuces expressing CPTL2-RNAi constructs showed that a decrease of CPTL2 transcripts (3-15% CPTL2 expression relative to controls) coincided with the reduction of natural rubber as low as 5%. We also identified a conventional cis-prenyltransferase 3 (CPT3), exclusively expressed in latex. In subcellular localization studies using fluorescent proteins, cytosolic CPT3 was relocalized to endoplasmic reticulum by co-occurrence of CPTL2 in tobacco and yeast at the log phase. Furthermore, yeast two-hybrid data showed that CPTL2 and CPT3 interact. Yeast microsomes containing CPTL2/CPT3 showed enhanced synthesis of short cis-polyisoprenes, but natural rubber could not be synthesized in vitro. Intriguingly, a homologous pair CPTL1/CPT1, which displays ubiquitous expressions in lettuce, showed a potent dolichol biosynthetic activity in vitro. Taken together, our data suggest that CPTL2 is a scaffolding protein that tethers CPT3 on endoplasmic reticulum and is necessary for natural rubber biosynthesis in planta, but yeast-expressed CPTL2 and CPT3 alone could not synthesize high molecular weight natural rubber in vitro.

Functional characterization of fission yeast transcription factors by overexpression analysis.

In Schizosaccharomyces pombe, over 90% of transcription factor genes are nonessential. Moreover, the majority do not exhibit significant growth defects under optimal conditions when deleted, complicating their functional characterization and target gene identification. Here, we systematically overexpressed 99 transcription factor genes with the nmt1 promoter and found that 64 transcription factor genes exhibited reduced fitness when ectopically expressed. Cell cycle defects were also often observed. We further investigated three uncharacterized transcription factor genes (toe1(+)-toe3(+)) that displayed cell elongation when overexpressed. Ectopic expression of toe1(+) resulted in a G1 delay while toe2(+) and toe3(+) overexpression produced an accumulation of septated cells with abnormalities in septum formation and nuclear segregation, respectively. Transcriptome profiling and ChIP-chip analysis of the transcription factor overexpression strains indicated that Toe1 activates target genes of the pyrimidine-salvage pathway, while Toe3 regulates target genes involved in polyamine synthesis. We also found that ectopic expression of the putative target genes SPBC3H7.05c, and dad5(+) and SPAC11D3.06 could recapitulate the cell cycle phenotypes of toe2(+) and toe3(+) overexpression, respectively. Furthermore, single deletions of the putative target genes urg2(+) and SPAC1399.04c, and SPBC3H7.05c, SPACUNK4.15, and rds1(+), could suppress the phenotypes of toe1(+) and toe2(+) overexpression, respectively. This study implicates new transcription factors and metabolism genes in cell cycle regulation and demonstrates the potential of systematic overexpression analysis to elucidate the function and target genes of transcription factors in S. pombe.

Deciphering the transcriptional-regulatory network of flocculation in Schizosaccharomyces pombe.

In the fission yeast Schizosaccharomyces pombe, the transcriptional-regulatory network that governs flocculation remains poorly understood. Here, we systematically screened an array of transcription factor deletion and overexpression strains for flocculation and performed microarray expression profiling and ChIP-chip analysis to identify the flocculin target genes. We identified five transcription factors that displayed novel roles in the activation or inhibition of flocculation (Rfl1, Adn2, Adn3, Sre2, and Yox1), in addition to the previously-known Mbx2, Cbf11, and Cbf12 regulators. Overexpression of mbx2(+) and deletion of rfl1(+) resulted in strong flocculation and transcriptional upregulation of gsf2(+)/pfl1(+) and several other putative flocculin genes (pfl2(+)-pfl9(+)). Overexpression of the pfl(+) genes singly was sufficient to trigger flocculation, and enhanced flocculation was observed in several combinations of double pfl(+) overexpression. Among the pfl1(+) genes, only loss of gsf2(+) abrogated the flocculent phenotype of all the transcription factor mutants and prevented flocculation when cells were grown in inducing medium containing glycerol and ethanol as the carbon source, thereby indicating that Gsf2 is the dominant flocculin. In contrast, the mild flocculation of adn2(+) or adn3(+) overexpression was likely mediated by the transcriptional activation of cell wall-remodeling genes including gas2(+), psu1(+), and SPAC4H3.03c. We also discovered that Mbx2 and Cbf12 displayed transcriptional autoregulation, and Rfl1 repressed gsf2(+) expression in an inhibitory feed-forward loop involving mbx2(+). These results reveal that flocculation in S. pombe is regulated by a complex network of multiple transcription factors and target genes encoding flocculins and cell wall-remodeling enzymes. Moreover, comparisons between the flocculation transcriptional-regulatory networks of Saccharomyces cerevisiae and S. pombe indicate substantial rewiring of transcription factors and cis-regulatory sequences.

Preferential extravasation and accumulation of liposomal vincristine in tumor comparing to normal tissue enhances antitumor activity.

To quantitatively evaluate the extravasation, accumulation and selectivity to tumor tissues of liposomal vincristine (LV), dorsal skin-fold window chambers on athymic mice with or without LX-1, a human small cell lung cancer, xenograft implants and fluorescent intravital microscopy imaging were used. In vitro studies show that minimal loss of fluorescence marker DiI from liposomes occurs after 4 days of inoculation in murine plasma, and the release profiles of DiI-LV and LV were essentially the same with approximately 40% of the encapsulated vincristine sulfate (VCR) released after 26 h. Significantly faster extravasation of DiI-LV from tumor vessels was shown compared to non-tumor tissue after single dose i.v. administration. The relative interstitial amounts at 60 min (RIA(60)) for tumor and non-tumor tissues were 0.837+/-0.314 and 0.012+/-0.091, respectively (P=0.01). DiI-LV accumulation was significantly higher in tumor than in normal tissue, which continued beyond 48 h. Both DiI-LV and LV showed significant antitumor effects in window chambers and in flank tumors, compared with controls and VLS alone. The preferential extravasation of DiI-LV from tumor vasculature as well as its differential retention in tumor tissue provides the basis for the enhancement in antitumor activity of LV over VCR.