Cytotoxicity and Apoptosis Induction of 6,7-Dehydroroyleanone from Taiwania cryptomerioides Bark Essential Oil in Hepatocellular Carcinoma Cells.

The objective of the present study is to evaluate the cytotoxicity of Taiwania cryptomerioides essential oil and its phytochemical on the Hep G2 cell line (human hepatocellular carcinoma). Bark essential oil has significant cytotoxicity to Hep G2 cells, and S3 fraction is the most active fraction in cytotoxicity to Hep G2 cells among the six fractions. The diterpenoid quinone, 6,7-dehydroroyleanone, was isolated from the active S3 fraction by bioassay-guided isolation. 6,7-Dehydroroyleanone exhibited significant cytotoxicity in Hep G2 cells, and the efficacy of 6,7-dehydroroyleanone was better than the positive control, etoposide. Apoptosis analysis of Hep G2 cells with different treatments was characterized via flow cytometry to confirm the cell death situation. Etoposide and 6,7-dehydroroyleanone could induce the apoptosis in Hep G2 cells using flow cytometric assay. Results revealed 6,7-dehydroroyleanone from T. cryptomerioides bark essential oil can be a potential phytochemical to develop the anticancer chemotherapeutic agent for the treatment of the human hepatocellular carcinoma.

Antimelanogenesis Effects of Leaf Extract and Phytochemicals from Ceylon Olive (Elaeocarpus serratus) in Zebrafish Model.

The melanogenesis inhibition effect in zebrafish (Danio rerio) and antityrosinase activity of the ethanolic extract and its phytochemicals from Ceylon olive (Elaeocarpus serratus Linn.) leaves were investigated in this study. Among the leaf extract and four soluble fractions, the ethyl acetate soluble fraction exhibits the best antityrosinase and antimelanogenesis activities. One phenolic acid, gallic acid, and two flavonoids, myricetin and mearnsetin, are isolated from the active subfractions through the bioassay-guided isolation; their structures are elucidated based on the 1D and 2D NMR, FTIR, UV, and MS spectroscopic analyses. These compounds have significant antityrosinase activity whether using l-tyrosine or l-DOPA as the substrate; mearnsetin shows the optimal activity. In the enzyme kinetic investigation, both gallic acid and mearnsetin are the competitive-type inhibitors against mushroom tyrosinase, and myricetin acts as a mixed-type tyrosinase inhibitor. Leaf extract and an ethyl acetate soluble fraction show effective performance in the inhibition of melanin formation in zebrafish embryos. Mearnsetin also possesses a promising antimelanogenesis effect, which is superior to the positive control, arbutin. Results reveal that the Ceylon olive leaf extract and its phytochemicals, especially mearnsetin, have the potential to be used as antimelanogenesis and skin-whitening ingredients.

A triterpenoid-enriched extract of bitter melon leaves alleviates hepatic fibrosis by inhibiting inflammatory responses in carbon tetrachloride-treated mice.

Liver fibrosis is a progression of chronic liver disease characterized by excess deposition of fibrillary collagen. The aim of this study was to investigate the protective effect of a triterpenoid-enriched extract (TEE) from bitter melon leaves against carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice. Male ICR mice received TEE (100 or 150 mg kg-1) by daily oral gavage for one week before starting CCl4 administration and throughout the entire experimental period. After intraperitoneal injection of CCl4 for nine weeks, serum and liver tissues of the mice were collected for biochemical, histopathological and molecular analyses. Our results showed that TEE supplementation reduced CCl4-induced serum aspartate aminotransferase and alanine aminotransferase activities. Histopathological examinations revealed that CCl4 administration results in hepatic fibrosis, while TEE supplementation significantly suppressed hepatic necroinflammation and collagen deposition. In addition, TEE supplementation decreased α-smooth muscle actin (α-SMA)-positive staining and protein levels of α-SMA and transforming growth factor-ß1. TEE-supplemented mice had lower mRNA expression levels of interleukin-6, tumor necrosis factor-α, and toll-like receptor 4. Moreover, TEE (150 mg kg-1) supplementation significantly reduced intrahepatic inflammatory Ly6C+ monocyte infiltration. We demonstrated that TEE could ameliorate hepatic fibrosis by regulating inflammatory cytokine secretion and α-SMA expression in the liver to reduce collagen accumulation.

Evaluation of Motor Coordination and Antidepressant Activities of Cinnamomum osmophloeum ct. Linalool Leaf Oil in Rodent Model.

Cinnamomum plants (Lauraceae) are a woody species native to South and Southeast Asia forests, and are widely used as food flavors and traditional medicines. This study aims to evaluate the chemical constituents of Cinnamomum osmophloeum ct. linalool leaf oil, and its antidepressant and motor coordination activities and the other behavioral evaluations in a rodent animal model. The major component of leaf oil is linalool, confirmed by GC-MS analysis. Leaf oil would not induce the extra body weight gain compared to the control mice at the examined doses after 6 weeks of oral administration. The present results provide the first evidence for motor coordination and antidepressant effects present in leaf oil. According to hypnotic, locomotor behavioral, and motor coordination evaluations, leaf oil would not cause side effects, including weight gain, drowsiness and a diminishment in the motor functions, at the examined doses. In summary, these results revealed C. osmophloeum ct. linalool leaf essential oil is of high potential as a therapeutic supplement for minor/medium depressive syndromes.

Small nucleolar RNA of silkworm can translocate from the nucleolus to the cytoplasm under abiotic stress.

Small nucleolar RNAs (snoRNAs) are thought to be exclusively nuclear and guide nucleotide modifications of ribosomal RNAs. Recently, more and more evidence has suggested that the nucleolus is a stress sensor for changes in growth status and that snoRNAs may orchestrate the response to environmental stress through molecular interactions outside of the nucleus. We previously showed that a box C/D snoRNA Bm-15 had both nuclear and cytoplasmic location in BmN4 cell line of the silkworm, Bombyx mori. To further study the functional roles of Bm-15, changes in expression level and cellular location of Bm-15 were examined in BmN4 cells subjected to serum starvation and ultraviolet (UV) ray radiation. Results indicated that total RNA level of Bm-15 was unchanged after 24 h serum starvation, but exhibited 3-fold increases in the cytoplasm, and the nuclear-to-cytosolic distribution ratio was reduced from 5:1 to 2:1. Moreover, UV radiation also causes rapid decline in nuclear Bm-15 and progressive cytoplasmic accumulation with a percentage of 22% and 57% after 6 and 24 h UV radiation. UV treatment results in a dramatic decrease in Bm-15 nuclear-to-cytosolic ratio from 7:1 to 2:1 and 2:1 to 1:20 after 6 and 24 h UV radiation, respectively. We show here for the first time that box C/D snoRNAs can translocate from the nucleus to the cytoplasm under the abiotic stress of nutritional deficiency and UV radiation. The rapid translocation of snoRNAs from nucleus to cytoplasm may slow down the maturation of rRNAs and synthesis of ribosomes to enhance the stress resistance of cells.

Infants' Vitamin D Nutritional Status in the First Year of Life in Northern Taiwan.

Vitamin D deficiency (VDD) and insufficiency (VDI) are common among exclusively breastfeeding infants. However, epidemiological evidence for the prevalence of VDD in infants during their first year of life in Taiwan has never been found. This trial determined the prevalence of VDD and VDI and the association between dietary vitamin D and vitamin D nutritional status in Northern Taiwan. A cross-sectional study was conducted on infants who returned to well-baby examinations from October 2012 to January 2014 in three hospitals: Shin Kong Wu Ho-Su Memorial Hospital, Taipei Medical University Hospital, and Shuang Ho Hospital. The specific vitamin D cut-off concentrations for VDD, VDI, and VDS are 25(OH)D3 levels ≤ 20, 21-29, and ≥ 30 (ng/mL). Overall, 481 infants' parents completed a questionnaire comprising questions related to vitamin D nutritional status, including weekly time outdoors, breastfeeding status, anthropometric measurement, and assessment of dietary intake, including milk and complementary food. The results revealed that 197 (41%) and 212 (44%) of infants in their first year of life had VDI and VDD, respectively, by the Endocrine Society guidelines. Breastfed infants had a higher prevalence of VDI (86.1%) than did mixed-fed (51.9%) and formula-fed (38.5%) infants (p < 0.001). The prevalence of VDD was 55.4% in infants aged under six months but increased to 61.6% in infants aged over six months. Infants in the VDI and VDD groups had the same anthropometrics as those in the vitamin D sufficiency (VDS) group. Our results revealed that 25(OH)D3 had a negative correlation with the intact parathyroid hormone (iPTH) when the serum 25(OH)D3 level ≤ 20 ng/mL (r = -0.21, p = 0.001). The VDS group had a higher total vitamin D intake than did the VDI and VDD groups, which was mainly obtained from infant formula. Our data revealed that dietary vitamin D intake and birth season were major indicators in predicting VDD. Lower dietary vitamin D intake and born in winter and spring significantly increased the odds ratio (OR) for VDI by 1.15 (95% CI 1.09-1.20) and 2.02 (95% CI 1.10-3.70), respectively, and that for VDD by 1.23 (95% CI 1.16-1.31) and 2.37 (95% CI 1.35-4.17) without covariates adjustment, respectively. Furthermore, ORs for VDI and VDD significantly differed after adjustment for covariates. In conclusion, the prevalence of VDI and VDD were high in infants during the first year of life. Breastfeeding infants had difficulty in obtaining sufficient vitamin D from diet. In cases where the amount of sun exposure that is safe and sufficient to improve vitamin D status is unclear, breastfed infants aged below one year old are recommended to be supplemented with vitamin D.

Iron Status of Infants in the First Year of Life in Northern Taiwan.

Iron deficiency (ID) and iron deficiency anemia (IDA) typically occur in developing countries. Notably, ID and IDA can affect an infant's emotion, cognition, and development. Breast milk is considered the best food for infants. However, recent studies have indicated that breastfeeding for more than six months increases the risk of ID. This study investigated the prevalence of ID and IDA, as well as the association between feeding type and iron nutritional status in northern Taiwan. A cross-sectional study was conducted on infants who returned to the well-baby clinic for routine examination from October 2012 to January 2014. Overall, 509 infants aged 1-12 months completed the iron nutritional status analysis, anthropometric measurement, and dietary intake assessment, including milk and complementary foods. The results revealed that 49 (10%) and 21 (4%) infants in their first year of life had ID and IDA, respectively, based on the World Health Organization criteria. Breastfed infants had a higher prevalence rate of ID and IDA than mixed-fed and formula-fed infants (p < 0.001). Regarding biomarkers of iron status, plasma hemoglobin (Hb), ferritin, and transferrin saturation (%) levels were significantly lower in ID and IDA groups. The prevalence of ID and IDA were 3.7% and 2.7%, respectively, in infants under six months of age, but increased to 20.4% and 6.6%, respectively, in infants above six months of age. The healthy group had a higher total iron intake than ID and IDA groups, mainly derived from infant formula. The total dietary iron intake was positively correlated with infants' Hb levels. Compared with formula-fed infants, the logistic regression revealed that the odds ratio for ID was 2.157 (95% confidence interval [CI]: 1.369-3.399) and that for IDA was 4.196 (95% CI: 1.780-9.887) among breastfed infants (p < 0.001) after adjusted for all confounding factors (including gestational week, birthweight, sex, body weight percentile, body length percentile, age of infants, mothers' BMI, gestational weight gain, education level, and hemoglobin level before delivery). In conclusion, our results determined that breastfeeding was associated with an increased the prevalence of ID and/or IDA, especially in infants above six months. This suggests that mothers who prolonged breastfeed after six months could provide high-quality iron-rich foods to reduce the prevalence of ID and IDA.

Association between Postpartum Nutritional Status and Postpartum Depression Symptoms.

Taiwanese women may practice traditional confinement after childbirth, and no study has investigated the nutritional status and the effects of postpartum depression on such women. The aim of this study was to investigate the association between nutritional status and postpartum depression at 6-8 weeks postpartum. A cross-sectional study was conducted on postpartum women who returned to the obstetrics and gynecology clinic for routine examination from January 2016 to September 2017. A total of 344 women received assessments based on the Edinburgh Postnatal Depression Scale (EPDS). An EPDS score of ≥10 indicated the presence of postpartum depressive symptoms (PPDS). A total of 97 women without such symptoms and 23 with PPDS completed nutritional parameter analyses and questionnaires. The results showed that the prevalence of postpartum depression (PPD) was 8.4%. The proportion was 70% for those who practiced confinement at home, significantly higher than for those in the non-PPDS group (45%). The overall psychological stress score was significantly higher and the postpartum care satisfaction score was significantly lower in those with PPDS compared to those without. In terms of nutritional biomarkers, the plasma riboflavin levels in the PPDS group were significantly lower than those in their symptomless counterparts (13.9%). The vitamin D insufficiency and deficiency rates in the non-PPD and PPDS groups were 35%, 41%, 48%, 26%, respectively. However, compared with those in the non-PPDS group, those with PPDS had significantly higher ratios of Σn-6/Σn-3, C20:3n-6/C18:3n-6, and C20:4n-6/(C20:5n-3 + C22:6n-3) (by 8.2%, 79.7%, and 8.8%, respectively), whereas they had lower ratios of C22:6n-3/C22:5n-6 (by 15.5%). Higher plasma riboflavin and erythrocyte C16:1n-9, C24:1n-9, C18:3n-6, and C20:5n-3 levels and lower Σn-6 fatty acid and C22:5n-6 levels decreased the risk of PPD after type of confinement, overall mental stress scores, and postpartum care satisfaction scores were adjusted for the logistic regression analysis. In conclusion, the plasma riboflavin level and erythrocyte fatty acid composition are potentially major contributors to PPD development.

Ethanolic Extract of Origanum vulgare Suppresses Propionibacterium acnes-Induced Inflammatory Responses in Human Monocyte and Mouse Ear Edema Models.

Acne vulgaris (acne) is a common inflammatory skin disorder, and Propionibacterium acnes plays a major role in the development and progression of acne inflammation. Herbs possessing antimicrobial and anti-inflammatory activity have been applied as a medical option for centuries. In this study, we examined the suppressive effect of ethanolic oregano (Origanum vulgare) extract on live P. acnes-induced in vivo and in vitro inflammation. Following ethanol extraction of oregano leaves, four compounds with strong antioxidant activity, including rosmarinic acid, quercetin, apigenin, and carvacrol, were identified by high-performance liquid chromatography. Using the mouse ear edema model, we demonstrated that ethanol oregano extracts (EOE) significantly suppressed P. acnes-induced skin inflammation, as measured by ear thickness (32%) and biopsy weight (37%). In a separate study, using the co-culture of P. acnes and human THP-1 monocytes, EOE reduced the production of interleukin (IL)-8, IL-1ß and tumor necrosis factor (TNF)-α up to 40%, 37%, and 18%, respectively, as well as the expression of these three pro-inflammatory mediators at the transcriptional level. Furthermore, EOE inhibited the translocation of nuclear factor-kappa B (NF-κB) into the nucleus possibly by inactivating toll-like receptor-2 (TLR2). The suppressive effect of EOE on live P. acnes-induced inflammatory responses could be due, in part, to the anti-inflammatory and antioxidant properties, but not the anti-microbial effect of EOE.

Taiwanese Green Propolis Ethanol Extract Delays the Progression of Type 2 Diabetes Mellitus in Rats Treated with Streptozotocin/High-Fat Diet.

Taiwanese green propolis ethanol extract (TGPE) is produced only in Taiwan and has a different composition from other types of propolis. TGPE is known for its anti-inflammation, anti-oxidation, and anti-microbial properties, but the effects and mechanisms of TGPE in the modulation of diabetes are unclear. In this study, we investigated the effects of TGPE on type 2 diabetes mellitus (T2DM) in a streptozotocin/high-fat-diet (STZ/HFD)-induced T2DM rat model. The results revealed that TGPE delayed the development and progression of T2DM and reduced the severity of β-cell failure. TGPE also attenuated inflammation and reactive oxygen species ROS in the rats. Moreover, there were higher levels of oxidant cytokines, leptin, and adiponectin in the serum of the TGPE-treated group. Unlike Brazilian propolis, TGPE promoted hepatic genes PPAR-α and CYP7A1, which were related to lipid catabolism and removal. TGPE may thus delay the progression of T2DM through anti-inflammation effects, anti-oxidation effects, and balancing lipid metabolism. It is suggested that TGPE can be a potential alternative medicine for T2DM.

Almonds ameliorate glycemic control in Chinese patients with better controlled type 2 diabetes: a randomized, crossover, controlled feeding trial.

BACKGROUND: Almonds can decrease glycemic index of co-consumed foods and are a rich source for oleic acid and α-tocopherol. The aim of the randomized, crossover, controlled feeding trial was to examine whether as compared to NCEP step II diet as control (CON), ~60 g/d almonds (ALM) added to CON would improve glucoregulation and cardiovascular disease (CVD) risk factors in 33 Chinese T2DM patients. METHODS: Forty T2DM patients were enrolled and randomly assigned to receive CON or ALM for 12 wks after a 2-wk. run-in period. Blood and urine samples were collected in the beginning and at the end of each dietary intervention phase for the assessment of biomarkers of glucoregulation, lipid profile, inflammation, and oxidative stress. RESULTS: While ALM had a better overall nutritional quality than CON, neither ALM nor CON improved the glycemic status as the primary study outcome and other CVD risk factors, except the circulating nitric oxide being decreased by ALM compared to CON. Among 27 of 33 patients with the baseline HbA1c ≤8, ALM decreased post-interventional fasting serum glucose and HbA1c by 5.9% and 3.0% as compared to that of CON, respectively (P = 0.01 and 0.04). Mean total and LDL-cholesterol concentrations were not changed by both diets. CONCLUSIONS: These results suggest almonds incorporated into healthful diets can improve glycemic status in diabetic patients with a better glycemic control. TRIAL REGISTRATION: NCT01656850, registered 13 January 2012.

Low-Level MHC Class II Expression Leads to Suboptimal Th Cell Response, Increased Autoaggression, and Heightened Cytokine Inducibility.

The development and activation of MHC class II (MHC-II)-restricted CD4+ T cells are distinct immunological processes that are strictly MHC-II-dependent. To address their relative dependence on MHC-II, we established a novel ENU-induced mutant mouse on the C57BL/6 background, named I-A12%, with ∼8-fold reduced I-A expression on the surface of B cells, dendritic cells, cortical thymic epithelial cells, and medullary thymic epithelial cells. I-A100% and I-A12% mice are highly similar with respect to the numbers of double-positive thymocytes, CD4+CD8- T cells, regulatory T cells, CD4+ T cell marker expression, lifespan, and Th/regulatory T cell function. Despite the demonstration of functional intrathymic negative selection in I-A12% mice, transfer of I-A12% CD25-CD4+ T cells into RAG-knockout hosts revealed increased autoaggression activity against the liver. Compared to I-A100% mice, infection of I-A12% mice with graded doses of Listeria monotcytogenes or influenza virus revealed comparable and significantly reduced generation of Ag-specific CD4+ T cells at high and low infection doses, respectively. A significantly weakened Ag-specific recall cytokine production response was also found for I-A12% mice previously infected with a relative low dose of L. monocytogenes CD44hiCD4+ T cells from I-A100% and I-A12% mice previously infected with a relatively high L. monocytogenes dose displayed highly similar Ag-specific multicytokine production profiles. In contrast, polyclonal activation of endogenous memory-like I-A12% CD44hiCD4+ T cells revealed highly elevated production of multiple cytokines. Our results demonstrate that there exist distinct thresholds for different MHC-II-dependent immunological processes. The I-A12% mutant mouse model we describe in the present study is a valuable tool for investigations on the quantitative cause-effect relationship in MHC-II-dependent normal and autoimmune responses.

Antilipotoxicity Activity of Osmanthus fragrans and Chrysanthemum morifolium Flower Extracts in Hepatocytes and Renal Glomerular Mesangial Cells.

The excess influx of free fatty acids (FFAs) into nonadipose tissues, such as those of liver and kidney, induces lipotoxicity leading to hepatic steatosis and renal dysfunction. The aim of this study was to investigate the protective effects of methanolic flower extracts of Osmanthus fragrans (OF) and Chrysanthemum morifolium (CM) against FFA-induced lipotoxicity in hepatocytes (human HepG2 cells) and renal glomerular mesangial cells (mouse SV40-Mes13 cells). The results showed that OF and CM significantly suppressed FFA-induced intracellular triacylglycerol accumulation via partially inhibiting the gene expression of sterol regulatory element-binding protein-1c (SREBP-1c) and glycerol-3-phosphate acyltransferase (GPAT) in HepG2 cells. Both extracts inhibited reactive oxygen species (ROS) generation by FFA-stimulated HepG2 cells. OF and CM also suppressed the mRNA expression of interleukin- (IL-) 1ß, IL-6, IL-8, tumor necrosis factor- (TNF-) α, and transforming growth factor- (TGF-) ß by HepG2 cells treated with conditioned medium derived from lipopolysaccharide-treated THP-1 monocytes. Furthermore, OF and CM effectively inhibited oleate-induced cellular lipid accumulation, TGF-ß secretion, and overexpression of fibronectin in mesangial cells. In conclusion, OF and CM possess hepatoprotective activity by inhibiting hepatic fat load and inflammation and renal protection by preventing FFA-induced mesangial extracellular matrix formation.

An alternatively spliced IL-15 isoform modulates abrasion-induced keratinocyte activation.

In a routine phenotype-driven screen, we identified a point mutation in exon 7 of the IL-15 gene in Pedigree 191 (deficient memory (DM)) of N-ethyl-N-nitrosourea mutagenized mice. The DM epidermis expressed an alternatively spliced IL-15 mRNA isoform, IL-15ΔE7, and a wild-type (WT) IL-15 isoform at comparable levels. Mechanical stimulation of DM skin or DM skin graft transplanted onto the WT host resulted in reduced keratinocyte activation and inhibition of neutrophil infiltration into the dermis, demonstrating that DM keratinocytes produced less inflammatory response to external stimulation. Ectopic expression of IL-15ΔE7 in WT skin prevented abrasion-induced epidermal thickening, blocked the accumulation of nuclear antigen Ki67(+) cells in the basal and the suprabasal cell layers, increased loricrin expression, and also increased keratinocyte CXCL1 and G-CSF production. IL-15ΔE7 also profoundly blocked neutrophil infiltration in SDS- or immiquimod (IMQ)-treated WT skin. Recombinant IL-15ΔE7 failed to activate STAT-5 and its downstream target bcl-2 expression. Our study points to IL-15ΔE7 as a potential therapeutic agent for treating neutrophilia-associated inflammatory skin disorders.

Carotid intima-media thickness and plaque occurrence in predicting stable angiographic coronary artery disease.

BACKGROUND: Carotid intima-media thickness (CIMT) and plaque formation have been used as surrogate end-points for evaluating the regression and/or progression of atherosclerotic cardiovascular disease, but their predictive value for stable coronary artery disease (CAD) is inconclusive. METHODS: Carotid ultrasonography was performed in patients who underwent noninvasive multislice computed tomography (MSCT) angiography for CAD suspected, due to chest pain. CIMT and plaque formation on the left and right common carotid arteries (CCAs), carotid bulb (CB), and proximal internal carotid arteries (ICAs) were evaluated, and the relationship between angiographic CAD, CIMT, and plaque formation was determined. RESULTS: 120 patients (95 male; 25 female), with a mean age ± standard deviation of 61 ± 11 years (range: 35-89 years) were recruited. Because age had a significant impact on CAD (r = 0.191; P = 0.036), CCA plaques (r = 0.368; P = 0.001), ICA plaques (r = 0.334; P = 0.004), and mean CIMT (r = 0.436; P = 0.001), patients were divided into two groups aged <60 years and ≥60 years. In the <60 years group, CIMT-CB was significantly higher in patients with CAD (P = 0.041), while in the ≥60 years group, mean CIMT, CIMT-CCA, and CIMT-CB were significantly higher in patients with CAD (P < 0.05, for each). In both groups, the occurrence of carotid plaques was significantly higher in patients with CAD than in those without CAD (P < 0.007, for each). After controlling for other risk factors, carotid plaques were an independent predictor of CAD in both groups (P < 0.05, for each), while CIMT-CB could independently predict CAD only in patients ≥60 years old (P = 0.031). CONCLUSION: Our findings suggest that carotid plaques are a strong predictor of stable CAD. However, CIMT-CB could predict stable CAD only in patients over 60 years of age.

Plasma B-type natriuretic peptide in predicting outcomes of elective coronary artery bypass surgery.

The risks of surgery and its clinical outcome are of great importance for both patients and physicians when choosing coronary artery bypass (CABG) surgery for coronary artery disease. The purpose of the current study was to clarify the relationship between serum B-type natriuretic peptide (BNP) and patient clinical outcome. Seventy-six eligible patients who underwent CABG were enrolled into the prospective study. Venous blood samples were drawn for serum BNP and N-terminal (NT)-proBNP levels measurement on preoperative Day 1, postoperative Day 1, and postoperative Day 7. Clinical end points were: (1) intensive care unit (ICU) stay longer than 4 days postoperatively and/or hospital stay longer than 13 days postoperatively; (2) major complications and poor outcomes. Patients who had prolonged ICU stay and hospitalization had significantly higher postoperative Day 1 BNP and postoperative Day 1 NT-proBNP level (p = 0.02 and 0.005, respectively). Age was significantly older in patients with prolonged ICU stay and hospitalization than those without prolonged ICU stay and hospitalization (p = 0.03). Serum creatinine level was also significantly increased in patients with prolonged ICU stay and hospitalization (p = 0.009). However, age was the only remaining factor that correlated with prolonged ICU stay and hospitalization in the multivariate logistic regression model. These results suggest that research using BNP and NT-proBNP for predicting ICU stay and hospitalization in patients who have undergone CABG must adjust risk factors to present a more appropriate estimation of its clinical outcome.

Viral load in blood is correlated with disease severity of neonatal coxsackievirus B3 infection: early diagnosis and predicting disease severity is possible in severe neonatal enterovirus infection.

We conducted a study during an outbreak of coxsackievirus B3 infection in 2005 and found that viral RNA could be detected in patients' blood specimens soon after the onset of fever, and the level of viral RNA was positively correlated with disease severity. Timely diagnosis is possible in severe neonatal enterovirus infection.

Coxsackievirus B3 in human milk.

Coxsackievirus B3 can cause severe neonatal disease with high mortality. We present the first report of detection of coxsackievirus B3 in the mothers' milk of 2 severely infected neonates by culture and reverse transcriptase-polymerase chain reaction. Reverse transcriptase-polymerase chain reaction is a rapid and sensitive tool to detect coxsackievirus B3 in breast milk within few hours. By 12 to 14 days after onset of symptoms, coxsackievirus B3 could not be detected in the breast milk of the symptomatic mothers.