Silicon chambers for enhanced incubation and imaging of microfluidic droplets.

Droplet microfluidics has become a powerful tool in life sciences, underlying digital assays, single-cell sequencing or directed evolution, and it is making foray in physical sciences as well. Imaging and incubation of droplets are crucial, yet they are encumbered by the poor optical, thermal and mechanical properties of PDMS, a material commonly used in microfluidics labs. Here we show that Si is an ideal material for droplet chambers. Si chambers pack droplets in a crystalline and immobile monolayer, are immune to evaporation or sagging, boost the number of collected photons, and tightly control the temperature field sensed by droplets. We use the mechanical and optical benefits of Si chambers to image ≈1 million of droplets from a multiplexed digital assay - with an acquisition rate similar to the best in-line methods. Lastly, we demonstrate their applicability with a demanding assay that maps the thermal dependence of Michaelis-Menten constants with an array of ≈150 000 droplets. The design of the Si chambers is streamlined to avoid complicated fabrication and improve reproducibility, which makes Si a complementary material to PDMS in the toolbox of droplet microfluidics.

The Association between Habitual Green Tea Consumption and Comprehensive Frailty as Assessed by Kihon Checklist Indexes among an Older Japanese Population.

BACKGROUND: It is unclear whether habitual green tea consumption is related to comprehensive frailty. OBJECTIVES: We conducted this study to investigate this relationship among an elderly Japanese population. METHODS: This was a cross-sectional study of baseline data from 2012. The study included 5668 Japanese participants (2766 men and 2902 women aged 65 years or older). The subjects completed a validated self-administered food frequency questionnaire that included questions on their green tea consumption. We evaluated comprehensive frailty using a 25-item Kihon Checklist (KCL), which comprised seven domains (instrumental activities of daily living (IADL), physical function, malnutrition, oral or eating function, socialization and housebound, cognitive function, and depression). Frailty was defined as a KCL score greater than or equal to seven. RESULTS: We found that a higher consumption of green tea was associated with a lower prevalence of comprehensive frailty in both sexes. Further age-stratified analysis showed that a higher consumption of green tea among women was associated with a lower prevalence of comprehensive frailty, regardless of age. In men, however, this association was found only in the older age groups. An analysis of the association between green tea consumption and the frailty subdomains showed that green tea consumption was associated with a lower prevalence of oral dysfunction and cognitive problems in both sexes. In addition, only in women was higher green tea consumption found to be associated with a lower prevalence of IADL and mobility-related disability problems. CONCLUSIONS: Green tea consumption is inversely associated with the prevalence of comprehensive frailty in Japanese men and women. Longitudinal studies are required to confirm this association.

Myofascial Release of the Hamstrings Improves Physical Performance-A Study of Young Adults.

Physical performance is mainly assessed in terms of gait speed, chair rise capacity, and balance skills, and assessments are often carried out on the lower limbs. Such physical performance is largely influenced by the strength of the quadriceps and hamstrings muscles. Flexibility of the hamstrings is important because quadriceps muscle activity influences the hip flexion angle. Therefore, hamstring flexibility is essential to improve physical performance. In this study, Myofascial Release (MFR) was applied to the hamstrings to evaluate its effects. MFR on the hamstrings was performed on 17 young adults. Physical function and physical performance were measured before, immediately after, and 5 days after the MFR intervention: finger floor distance (FFD), range of motion (ROM) of the straight leg raising test (SLR), standing long jump (SLJ), squat jump (SJ), functional reach test (FRT), comfortable walking speeds (C-walking speed), and maximum walking speeds (M-walking speed). The results of the analysis show a significant increase in FFD (-2.6 ± 8.9 vs. 0.4 ± 9.4 vs. 2.4 ± 8.9, p < 0.01), SLJ (185.6 ± 44.5 vs. 185.0 ± 41.8 vs. 196.6 ± 40.1, p < 0.01), and M-walking speed (2.9 ± 0.6 vs. 3.0 ± 0.6 vs. 3.3 ± 0.6, p < 0.01). This study has shown that MFR for hamstrings not only improves flexibility but also increases M-walking speed and physical performance of the SLJ. As MFR is safe and does not involve joint movement, it may be useful for maintaining and improving performance and flexibility during inactivity and for stretching before exercise.

Health-care resource use and current treatment of adult atopic dermatitis patients in Japan: A retrospective claims database analysis.

The real-world evidence on the profiles of patients suffering from atopic dermatitis (AD) in Japan is sparse. A retrospective claim database analysis was conducted to estimate the health-care resource use (HCRU) and current AD treatment. Data from October 2013 to September 2016 were extracted from the JMDC (Tokyo, Japan) claims database. HCRU was assessed by a comparison of AD patients and matched non-AD controls. A multivariate analysis was performed to estimate HCRU attributable to AD. AD patients (n = 39 893) have more claims of certain diagnoses such as rhinitis, viral and fungal infections, sleep disorders and conjunctivitis as well as higher HCRU (outpatient visits, prescriptions of AD-related and non-AD-related medications, phototherapy, laboratory tests) than matched non-AD controls (n = 39 893). Treatment pattern analysis included treatment-naive patients (n = 8478) and previously treated AD patients (n = 30 109). Approximately 20% of previously treated patients were on the continuous systemic treatment during 18-month follow up. Systemic corticosteroids were the most frequently used systemic treatments. Oral cyclosporin was less frequently used in both groups, but for the longest duration. Almost half of previously treated patients with oral cyclosporin continued treatment for more than 3 months. In conclusion, HCRU was higher in AD patients than non-AD controls, indicating a high burden of the disease imposed on AD patients. Continuous administration of systemic treatment, such as oral cyclosporin, systemic corticosteroids and phototherapy, observed in AD patients sheds light on the difficulties of managing AD in Japanese clinical practise.

Secukinumab improves psoriasis symptoms in patients with inadequate response to cyclosporine A: A prospective study to evaluate direct switch.

There are limited data on the safety and efficacy of switching to secukinumab from cyclosporine A (CyA) in patients with psoriasis. The purpose of the present study was to assess the efficacy and safety of secukinumab for 16 weeks after direct switching from CyA in patients with moderate-to-severe psoriasis. In this multicenter, open-label, phase IV study, 34 patients with moderate-to-severe psoriasis and inadequate response to CyA received secukinumab 300 mg s.c. at baseline and weeks 1, 2, 3, 4, 8 and 12. The primary end-point was ≥75% improvement from baseline in Psoriasis Area and Severity Index score (PASI 75) at week 16. The efficacy of secukinumab treatment was evaluated up to week 16, and adverse events (AE) were monitored during the study. The primary end-point of the PASI 75 response at week 16 was achieved by 82.4% (n = 28) of patients receiving secukinumab. Early improvements were observed with secukinumab, with PASI 50 response of 41.2% at week 2 and PASI 75 response of 44.1% at week 4. AE were observed in 70.6% (n = 24) of patients, and there were no serious AE or deaths reported in the entire study period. Secukinumab showed a favorable safety profile consistent with previous data with no new or unexpected safety signals. The results of the present study show that secukinumab is effective in patients with psoriasis enabling a smooth and safe direct switch from CyA to biological therapy.

A hair growth-promoting effect of Chinese black tea extract in mice.

Chinese black tea extract (CBTE) fermented with Aspergillus sp. significantly promoted hair growth after 2 weeks of topical application in shaved 6 week-old male C3H/He mice. The hair growth-promoting effect of CBTE was potentiated synergistically by capsaicin, which has no effect on hair growth by itself. CBTE displayed an affinity for estrogen receptor (ER)α, with an IC50 value of 74.8 µg/mL. This effect of CBTE might be mediated by the ERs, since a similar effect induced by orally administered soy isoflavone, a mixture of ERs ligands, has been reported to be synergistically potentiated by capsaicin.

Phase 2 study of arsenic trioxide followed by autologous hematopoietic cell transplantation for relapsed acute promyelocytic leukemia.

The optimal treatments for relapsed acute promyelocytic leukemia (APL) remain equivocal. We conducted a phase 2 study to evaluate the efficacy and feasibility of a sequential treatment consisting of induction and consolidation with arsenic trioxide (ATO), peripheral blood stem cell (PBSC) harvest after high-dose cytarabine chemotherapy, and autologous hematopoietic cell transplantation (HCT). Between 2005 and 2009, 35 patients (26 with hematologic and 9 with molecular relapse) were enrolled. Induction therapy resulted in complete remission in 81% of those with hematologic relapse, and most patients became negative for PML-RARα after the first ATO consolidation course, but 4 remained positive. Administration of the second ATO consolidation course further decreased the transcript levels in 3 patients. In total, 25 patients proceeded to PBSC harvest, all of whom successfully achieved the target CD34+ cell doses, and 23 underwent autologous HCT with PML-RARα-negative PBSC graft. Posttransplant relapse occurred in 3 patients, and there was no transplant-related mortality. With a median follow-up of 4.9 years, the 5-year event-free and overall survival rates were 65% and 77%, respectively. These findings demonstrate the outstanding efficacy and feasibility of the sequential treatment featuring ATO and autologous HCT for relapsed APL. This study was registered at http://www.umin.ac.jp/ctr/ as #C000000302.

Antiobesity effects of Chinese black tea (Pu-erh tea) extract and gallic acid.

The antiobesity effects of Chinese black tea (Pu-erh tea) and of gallic acid (GA) were investigated using in vitro and in vivo assays. Chinese black tea extract (BTE) and GA inhibited pancreatic lipase activity in a dose-dependent manner in vitro; the IC(inhibitory concentration)(50) values were 101.6 and 9.2 µg/mL, respectively. Black tea extract (50, 100 mg/kg body weight (b.w.)) and GA (15, 45 mg/kg b.w.) significantly suppressed the elevation of blood triglyceride after oral administration of a corn oil emulsion (8 mL oil/kg b.w.) to male ddY mice. Moreover, the antiobesity effects of BTE and GA were also evaluated in a mouse model of diet-induced obesity. Female ddY mice were divided into seven groups; normal diet (ND) group, high fat diet (HFD) group, BTE (0.2% and 0.6% of diets) groups, and GA (0.007%, 0.02% and 0.1% of diets) groups; the experimental groups were fed the test diets for 12 weeks. The BTE 0.6% and GA 0.1% groups showed significant suppression of weight gain. The weight of parametrial adipose tissue was strongly correlated with the body weight. These results suggest that GA contributes to the antiobesity effect of BTE as an active constituent by inhibiting pancreatic lipase activity.

Improvements of mean body mass index and body weight in preobese and overweight Japanese adults with black Chinese tea (Pu-Erh) water extract.

Water-soluble black Chinese (Pu-Erh) tea extract (BTE), which contains high gallic acid content, has been demonstrated to elicit antiobese effects in animals. Because gallic acid is related with the reduction of visceral fat and cholesterol contents and improvement of obesity in animals, we investigated the effects of BTE intake on 36 preobese Japanese adults (body mass index [BMI], >25- <30 kg/m(2)) in a 12-week double-blind, randomized, placebo-controlled group comparison study using powdered barley tea with or without (placebo) BTE. A follow-up 4-week period after BTE intake termination was monitored to observe the withdrawal effect. All subjects ingested barley tea with or without BTE (333 mg) before each of the 3 daily meals. In the BTE-treated group, the mean pretreament values of body weight and BMI significantly decreased after intake and after BTE withdrawal. However, the corresponding values scored significant differences only from 8 weeks after intake (vs the placebo-treated group). The mean values of the waist circumference indicated a similar tendency. Furthermore, coronal navel section (same anatomical position) images of computed tomography of all BTE- and non-BTE-treated subjects revealed that the visceral fat areas (cm(2)) were significantly (P < .05) less in the former 12 weeks after BTE ingestion. Measured biochemical parameters did not indicate significant differences, and BTE-treated subjects did not complain of any adverse effects (abdominal distension, etc). Ingestion of BTE exhibited significant effects in reducing the mean waist circumference, BMI, and visceral fat values and might be useful for weight control and prevention of obesity development (or metabolic syndrome) in humans.

Antihypercholesterolemic effect of Chinese black tea extract in human subjects with borderline hypercholesterolemia.

A water-soluble extract of a traditional Chinese black tea (Pu-Ehr) has been shown to precipitate mixed bile salt micelles in foods. In addition, long-term ingestion of this black tea extract (BTE) significantly reduces blood cholesterol levels in rats. We investigated the effects of BTE tablets (a formula designed to enhance compliance) as a dietary supplement in a 3-month double-blind randomized group comparison study in borderline hypercholesterolemic human subjects (n = 47). All subjects ingested BTE tablets (333 mg) or placebo 3 times daily before meals for 3 months. In the BTE-treated group, the initial mean blood total (6.14 +/- 0.14 mol/L) and low-density lipoprotein (LDL) cholesterol (4.32 +/- 0.14 mol/L) levels decreased with time and were significantly (P < .01) lower (total cholesterol, 5.62 +/- 0.11; LDL cholesterol, 3.81 +/- 0.13 mol/L) after 3 months of ingestion. Furthermore, the mean body weights (P < .05) and triacylglycerol levels (P < .01) were also significantly reduced after 3 months of BTE intake compared with the baseline levels. Significant improvements in the mean LDL cholesterol, body weight, and triacylglycerol values were not accompanied with undesirable changes in other biochemical parameters measured in the subjects. None of the subjects complained of any adverse effects (eg, abdominal distension). The results indicate that BTE intake elicited a significant antihypercholesterolemic effect and might be useful for improving blood cholesterol levels in subjects at risk for heart disease or obesity.

Efficacy and safety of Chinese black tea (Pu-Ehr) extract in healthy and hypercholesterolemic subjects.

BACKGROUND/AIMS: Water-extracted Chinese black tea (Pu-Ehr) exerts a precipitating effect on mixed bile salt micelles in foods. The amount of black tea extract (BTE) effective for hypercholesterolemia (HC) was examined in humans. MATERIALS AND METHODS: Animals (test 1) and humans (tests 2-5) were given BTE in the following studies: (test 1) an acute 2-week toxicity test conducted in mice with 2,000 mg/kg BTE or saline; (test 2) a dose-finding 8-week toxicity study with excessive BTE ingestion (0.75 or 1 g/day) in 10 healthy and 10 borderline HC subjects; (test 3) a 1-day acute toxicity test using a (10 g) single-bolus study (n = 10); (test 4) a 5-week long-term safety test (5 g/day, n = 11), and (test 5) a 4-month noncomparative study in 21 HC patients ingesting 1 g/day. RESULTS: The safety study showed no changes in hematological or relevant biochemical parameters in both mice and humans in the acute and long-term toxicity tests. In test 5, significant reductions in total and low-density lipoprotein cholesterol levels accompanied by significant decreases in body weight were observed without affecting other biochemical parameters. CONCLUSIONS: BTE significantly reduced blood cholesterol levels in humans and may prove safe and useful in preventing and improving metabolic syndrome-induced arteriosclerosis and/or obesity.

Extract of black tea (pu-ehr) inhibits postprandial rise in serum cholesterol in mice, and with long term use reduces serum cholesterol and low density lipoprotein levels and renal fat weight in rats.

A water-soluble extract of a traditional Chinese fermented black tea, pu-ehr, decomposes bile acid cholesterol micelles. This black tea extract (BTE) was studied to see if it could decrease the postprandial elevation of blood cholesterol levels after a single administration in ddY mice. It was found that BTE (0.3 g/kg) significantly decreased the postprandial rise in blood cholesterol levels after oral administration of cholesterol (130 mg/kg). A non-fermented tea (i.e. green tea) extract did not prevent the postprandial increase in blood cholesterol. In a subsequent study, 5-week-old Sprague-Dawley (SD) rats were fed BTE for 3 weeks, following which a dose-dependent and significant decrease in serum total cholesterol levels (1.36 mmol/L, 0.1% BTE, p < 0.05) was found and also in renal fat weight (0.3% BTE, p < 0.05). LDL cholesterol levels (0.51 mmol/L, 0.1% BTE, p < 0.05) were also significantly decreased. There were no significant changes in the weights of other organs or in the serum levels of other clinical markers. Thus, BTE has a specific antihypercholesterol effect in rodents, which might potentially aid in the management of hyperlipidaemia in man.

Absence of mutagenicity, genotoxicity, and subchronic oral toxicity of Touchi extract.

Touchi, a traditional Chinese food used mainly for seasoning is obtained by first steaming soybeans followed by fermentation with Aspergillus oryzae (koji). A series of toxicological studies was conducted to evaluate the mutagenic and genotoxic potential and subchronic toxicity of a water extract of Touchi, a known inhibitor of alpha -glucosidase activity. Touchi extract (TE) did not induce reverse mutations in Salmonella typhimurium strains TA98, TA1537, TA100, TA1535, and Escherichia coli WP2uvrA at concentrations up to 5000 microg/plate, in either the absence or presence of exogenous metabolic activation. No deaths occurred and no abnormal clinical signs were observed in any animal in any group in an in vivo micronucleus test, and TE was devoid of clastogenic activity when administered orally to mice at doses up to 2000 mg/kg/day. Thus, TE was evaluated as negative in the bacterial reverse mutation and mouse bone marrow micronucleus tests under the conditions of these assays. To evaluate its subchronic toxicity, SPF rats were administered TE at doses of 0, 250, 1000, and 2500 mg/kg/day via oral gastric intubation. No treatment-related toxic changes were seen in clinical signs, body weight, food consumption, urinalysis, hematology, blood chemistry, necropsy, organ weight, or histopathology. The no observed adverse effect level for TE was thus considered to be more than 2500 mg/kg/day in both males and females. These results are consistent with Touchi's status as a traditional Chinese food derived from fermented soybeans and its purported long history of use. Specifically, these data are consistent with the expected safety of human consumption of TE up to at least 5 g/day.