Relationship between Saturated Fatty Acid Intake and Blood Pressure in Healthy Japanese Women with the Fat Mass and Obesity-Associated Gene Variant.

Hypertension is one of the most prevalent diseases and a risk factor for stroke and cardiovascular disease. Our previous study indicated a negative correlation between fat intake and blood pressure in subjects with a fat mass and obesity-associated gene variant. We investigated the effects of four fatty acid groups on blood pressure in healthy Japanese women with the gene variants, including the involvement of body mass index. A total of 227 Japanese women aged 18-64 years completed a 3-day nutritional intake diary and their blood pressure was measured. The single nucleotide polymorphism rs9939609 of the gene was genotyped, and the participants were divided into two genetic groups (those with or without at least one minor allele). Spearman's rank correlation coefficient was applied to investigate the relationships between the fatty acids and blood pressure. A path analysis was performed to determine the effect of fatty acids on blood pressure including the involvement of body mass index. In the group with the gene variant, a significant negative correlation was detected between saturated fatty acid intake and systolic and diastolic blood pressures, and between monounsaturated fatty acid intake and only diastolic blood pressure. In a path analysis of both systolic and diastolic blood pressures, the path from only saturated fatty acid intake to blood pressure was significant, but the path from saturated fatty acids to body mass index was not significant. These results suggest that saturated fatty acid intake, without the involvement of body mass index, may be associated with the lower systolic and diastolic blood pressures in healthy Japanese women with a fat mass and obesity-associated gene variant.

Differential Effect of Fat Intake on Blood Pressure in Healthy Japanese Women with and without the Variant of Fat Mass and Obesity-Associated Gene.

A high-fat diet is believed to be a risk factor for hypertension through inducing obesity. It has been reported that variants of the fat mass and obesity-associated (FTO) and beta-3 adrenergic receptor (B3AR) genes are associated with obesity and blood pressure. The purpose of this study was to investigate the effect of dietary fat on blood pressure with or without the variant of the FTO and B3AR genes. A total of 227 healthy Japanese women aged 18 to 64 years were recruited for measurement of nutrient intake and blood pressure. The single nucleotide polymorphism rs9939609 of the FTO gene and rs4994 of the B3AR gene were genotyped. Spearman's rank correlation coefficient was applied to investigate the relationship between fat intake and blood pressure. A hierarchical multiple regression analysis was performed to determine whether the genotype interacts with fat intake to affect blood pressure. No significant correlations were found between fat intake and either systolic or diastolic blood pressure. A significant negative correlation was found between fat intake and both blood pressures in the FTO-gene-variant group, but not in the normal-FTO-gene group. In hierarchical multiple regression analysis, the interaction of fat intake and the gene variant showed significance, and the change in coefficient of determination (R 2) was significantly increased with increases of the interaction variable. These results indicate that the effect of fat intake on blood pressure may be modified by the variant of the FTO gene such that a high-fat diet intake may be associated with a decrease of systolic and diastolic blood pressure in healthy Japanese women with the FTO variant. Our results did not support the hypothesis that a high-fat diet increases blood pressure.

Cysteinyl Leukotriene Synthesis via Phospholipase A2 Group IV Mediates Exercise-induced Bronchoconstriction and Airway Remodeling.

It is well known that the prevalence of asthma is higher in athletes, including Olympic athletes, than in the general population. In this study, we analyzed the mechanism of exercise-induced bronchoconstriction by using animal models of athlete asthma. Mice were made to exercise on a treadmill for a total duration of 1 week, 3 weeks, or 5 weeks. We analyzed airway responsiveness, BAL fluid, lung homogenates, and tissue histology for each period. In mice that were treated (i.e., the treatment model), treatments were administered from the fourth to the fifth week. We also collected induced sputum from human athletes with asthma and analyzed the supernatants. Airway responsiveness to methacholine was enhanced with repeated exercise stimulation, although the cell composition in BAL fluid did not change. Exercise induced hypertrophy of airway smooth muscle and subepithelial collagen deposition. Cysteinyl-leukotriene (Cys-LT) levels were significantly increased with exercise duration. Montelukast treatment significantly reduced airway hyperresponsiveness (AHR) and airway remodeling. Expression of PLA2G4 (phospholipase A2 group IV) and leukotriene C4 synthase in the airway epithelium was upregulated in the exercise model, and inhibition of PLA2 ameliorated AHR and airway remodeling, with associated lower levels of Cys-LTs. The levels of Cys-LTs in sputum from athletes did not differ between those with and without sputum eosinophilia. These data suggest that AHR and airway remodeling were caused by repeated and strenuous exercise. Cys-LTs from the airway epithelium, but not inflammatory cells, may play an important role in this mouse model.

Association of upper and lower airway eosinophilic inflammation with response to omalizumab in patients with severe asthma.

Background: The anti-immunoglobulin E monoclonal antibody, omalizumab, is used to treat severe asthma and has the potential to ameliorate airway inflammation. However, the effect of omalizumab in ameliorating upper airway inflammation has not been fully elucidated. Objective: We investigated the association of upper and lower airway inflammation with the response to omalizumab treatment. Methods: We used the Global Evaluation of Treatment Effectiveness to assess the efficacy of omalizumab in treating 16 patients with severe asthma. We also investigated the symptom score, short-acting ß-agonist inhaler use, pulmonary function, biomarkers, computed tomography scans, and nasal mucosa pathology at omalizumab initiation and after four months of treatment. Results: When the fraction of exhaled nitric oxide (FeNO) and the percentage of sputum eosinophil were used as indicators of lower airway inflammation, positive correlations were found between CD20 B-cell, mast cell, and eosinophil counts in the nasal mucosa. Improved asthma symptoms were observed in 12 of the 16 severe asthma cases. The FeNO and eosinophil levels in the nasal tissue, prior to the administration of omalizumab were predictors of the response to asthma treatment. Conclusions: These findings suggest heterogeneity among people with severe asthma. In addition, the phenotype associated with response to omalizumab, leading to improvement in asthma symptoms, comprises upper airway eosinophilia and high FeNO levels.

Supplementation with trans fatty acid at 1% energy did not increase serum cholesterol irrespective of the obesity-related genotypes in healthy adult Japanese.

BACKGROUND AND OBJECTIVES: The excessive intake of trans fatty acids increases serum low-density lipoproteincholesterol and reduces high-density lipoprotein-cholesterol. We studied the effects of 1% energy trans fatty acid supplementation on serum lipid concentrations in healthy adult Japanese with different obesity-related gene polymorphisms. METHODS AND STUDY DESIGN: A randomized, double-blind, parallel trial was conducted in 53 healthy adults. The volunteers consumed one cookie containing either 1% energy or <0.01% energy (control) of trans fatty acids every day for 4 weeks, and a blood sample was then obtained after overnight fasting. The single nucleotide polymorphisms of the fat mass- and obesity-associated gene rs9939609 and beta-3 adrenergic receptor rs4994 were genotyped. RESULTS: The mean trans fatty acid intake of the control and trans fatty acid groups corresponded to 0.28% and 1.31 % energy, respectively. There were no significant differences in serum cholesterol (total, low-density lipoprotein and high-density lipoprotein) or triacylglycerol between the control and trans fatty acid groups. The responses of serum cholesterol, triacylglycerol, glucose, insulin and hemoglobinA1c were also independent of the fat mass- and obesity-associated gene and beta-3 adrenergic receptor gene variants. CONCLUSIONS: Our findings indicate that supplementation with 1% energy trans fatty acids has little effect on serum cholesterol in healthy adult Japanese, regardless of genotype of fat mass- and obesity-associated gene or beta-3 adrenergic receptor. More systematic studies, with respect to dietary trans fatty acid intakes above those used here, may be warranted to determine the tolerable upper level of dietary trans fatty acid.

Industrial Trans Fatty Acid and Serum Cholesterol: The Allowable Dietary Level.

Trans fatty acid (TFA) from partially hydrogenated oil is regarded as the worst dietary fatty acid per gram due to its role in coronary heart disease. TFA consumption is decreasing worldwide, but some but not all observational studies indicate that TFA intake has little relevance to serum cholesterol levels in populations with low TFA intake (<1% E [percentage of total energy intake],

Acute exacerbation of interstitial pneumonia associated with rheumatoid arthritis during the course of treatment for Pneumocystis jirovecii pneumonia: a case report.

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) is potentially fatal infectious complication in patients with rheumatoid arthritis (RA) during immunosuppressive therapy. Hospital survival due to human immunodeficiency virus-unrelated PCP reaches to 60%. The high mortality rate results from difficulties in establishing an early diagnosis, concurrent use of prophylactic drugs, possible bacterial coinfection. We herein report a case of PCP in RA patients who developed the architectural distortions of lung in spite of combined modality therapy. CASE PRESENTATION: A 73-year-old Japanese woman with RA was admitted with shortness of breath. Five weeks previously, she had been started on etanercept in addition to methotrexate (MTX). Chest computed tomography (CT) demonstrated diffuse ground glass opacities distributed throughout the bilateral middle to lower lung fields, and serum ß-D-glucan was elevated. Bronchoalveolar lavage fluid revealed no P. jirovecii, but the organism was detected by polymerase chain reaction method. Trimethoprim/sulfamethoxazole was administered with methylprednisolone pulse therapy. However, the follow-up chest X-ray and chest CT demonstrated aggravation of the pneumonia with architectural distortions. Additional direct hemoperfusion with polymyxin B-immobilized fibers and intravenous cyclophosphamide therapy were insufficiently effective, and the patient died on day 25. CONCLUSION: The architectural distortions of lung should be considered as a cause of death of PCP. For this reason, a high suspicion of this infectious complication must be kept in mind in order to establish an early diagnosis and treatment in patients with RA managed with MTX and biologics.

Little Effect of Supplementation with 0.6% Energy Trans Fatty Acids on Serum Cholesterol Levels in Adult Japanese Women.

The excessive intake of trans fatty acids (TFAs) increases serum LDL-cholesterol and reduces HDL-cholesterol. Limited data exist regarding the low-level intake of TFAs, and the tolerable upper-limit level remains to be fully elucidated. A randomized, double-blind, parallel trial was conducted to assess the effects of a low level of TFA supplementation on serum cholesterol levels in healthy adult Japanese women. The volunteers who participated in this examination took in approximately 0.4% of energy (%E) TFAs from daily meals. Fifty-one volunteers consumed one cookie containing 0.6%E (TFA) or 0.04%E (control) of TFAs every day for 4 wk, and blood was harvested after overnight fasting. The mean TFA intakes of the control and TFA groups during the experimental period were 0.4%E and 1.1%E, respectively. There were no significant differences in serum total, LDL- or HDL-cholesterol levels between the control and TFA groups. The serum glucose and insulin levels were not influenced by TFA supplementation. These results confirm that dietary supplementation with 0.6%E TFAs (a total TFA intake of approximately 1%E) would have little effect on serum cholesterol levels in healthy adult Japanese women.

Impact of Humidity on In Vitro Human Skin Permeation Experiments for Predicting In Vivo Permeability.

In vitro skin permeation studies have been commonly conducted to predict in vivo permeability for the development of transdermal therapeutic systems (TTSs). We clarified the impact of humidity on in vitro human skin permeation of two TTSs having different breathability and then elucidated the predictability of in vivo permeability based on in vitro experimental data. Nicotinell(®) TTS(®) 20 and Frandol(®) tape 40mg were used as model TTSs in this study. The in vitro human skin permeation experiments were conducted under humidity levels similar to those used in clinical trials (approximately 50%) as well as under higher humidity levels (approximately 95%). The skin permeability values of drugs at 95% humidity were higher than those at 50% humidity. The time profiles of the human plasma concentrations after TTS application fitted well with the clinical data when predicted based on the in vitro permeation parameters at 50% humidity. On the other hand, those profiles predicted based on the parameters at 95% humidity were overestimated. The impact of humidity was higher for the more breathable TTS; Frandol(®) tape 40mg. These results show that in vitro human skin permeation experiments should be investigated under realistic clinical humidity levels especially for breathable TTSs.

Characterization of a riboflavin non-aqueous nanosuspension prepared by bead milling for cutaneous application.

The purpose of this study was to characterize the non-aqueous nanosuspension of a hydrophilic drug prepared by bead milling for cutaneous application. Riboflavin was used as the model hydrophilic drug. The non-aqueous nanosuspensions were prepared by grinding riboflavin with zirconia beads using eight non-aqueous bases. The mean particle size of riboflavin in the suspensions ranged from 206 to 469 nm, as determined by the dynamic light scattering method. Among the well-dispersed samples, riboflavin nanosuspension prepared in oleic acid was selected for evaluation of the drug permeability through rat skin. The cumulative amount and permeation rate of riboflavin from the nanosuspension were approximately three times higher than those for unprocessed riboflavin in oleic acid. Fluorescence imaging of the riboflavin nanosuspension suggested improved penetration of riboflavin into the stratum corneum. Furthermore, the addition of polysorbate 65 or polyglyceryl-6 polyricinoleate to the nanosuspension prepared in oleic acid markedly improved the riboflavin dispersibility. These results show that the preparation of a nanosuspension in a non-aqueous base by bead milling is one of the simple methods to improve the skin permeability of hydrophilic drugs.

Lowering effect of firefly squid powder on triacylglycerol content and glucose-6-phosphate dehydrogenase activity in rat liver.

Effects of dietary firefly squid on serum and liver lipid levels were investigated. Male Wistar rats were fed a diet containing 5% freeze-dried firefly squid or Japanese flying squid for 2 weeks. There was no significant difference in the liver triacylglycerol level between the control and Japanese flying squid groups, but the rats fed the firefly squid diet had a significantly lower liver triacylglycerol content than those fed the control diet. No significant difference was observed in serum triacylglycerol levels between the control and firefly squid groups. The rats fed the firefly squid had a significantly lower activity of liver glucose-6-phosphate dehydrogenase compared to the rats fed the control diet. There was no significant difference in liver fatty acid synthetase activity among the three groups. Hepatic gene expression and lipogenic enzyme activity were investigated; a DNA microarray showed that the significantly enriched gene ontology category of down-regulated genes in the firefly squid group was "lipid metabolic process". The firefly squid group had lower mRNA level of glucose-6-phosphate dehydrogenase compared to the controls. These results suggest that an intake of firefly squid decreases hepatic triacylglycerol in rats, and the reduction of mRNA level and enzyme activity of glucose-6-phosphate dehydrogenase might be related to the mechanisms.

Supplementation with 1% energy trans fatty acids had little effect on serum cholesterol levels in healthy young Japanese women.

The World Health Organization (WHO) has recommended that trans fatty acid (TFA) intake should be less than 1% of total energy intake, but few data are available as to the influence of energy TFA intake of as low as 1% on blood cholesterol levels. A randomized, double-blind, parallel trial was conducted to assess the effects of 1% TFA dietary supplementation on serum cholesterol levels in healthy young women. Sixty-five volunteers consumed cookies containing 1% (TFA) or 0.04% (control) energy of TFA for 4 weeks and blood was harvested after overnight fasting. There were no significant differences in serum LDL- or HDL-cholesterol levels between the two groups. The hemoglobin A1c level was not influenced by dietary TFA. These results suggest that energy of TFAs at less than 1% has little effect on serum cholesterol or hemoglobin A1c levels in healthy young women. This confirms the correctness of the WHO recommendation.

Effects of dietary trans fatty acids on fat accumulation and metabolic rate in rat.

Dietary intake of high trans-fatty acids (TFAs) is well known to increase the risk of cardiovascular diseases. However, few reports demonstrated definitive relationships between dietary TFAs and obesity. In addition, the difference in the gastrointestinal absorption rate of TFAs containing oil from that of cis-FAs containing oil was not taken into consideration in many rat studies. In experiment A, we investigated the difference in the apparent absorption rate of TFAs containing oil from control oil. Hydrogenated rapeseed oil and a mixture of camellia oil and tristearin (90:10 [w/w]) were used as TFA-containing test oil and control oil, respectively. Ten Wistar rats were divided into the control group or TFA group, and fed the respective diet containing the control oil or the test oil for 1 week. The apparent absorption rates of these oils were measured by fecal fat excretion rate and dietary fat intake. The results showed a significantly lower gastrointestinal apparent absorption rate of the test oil (93.1%) than that of the control oil (96.2%). In consideration of the apparent absorption rate of these dietary oils, the effects of dietary TFAs on body fat accumulation and energy metabolism were investigated in rats. Twenty-eight Wistar rats were divided into the control group or the TFA group. Each group received an isoenergetic diet containing the control oil or the test oil for 8 weeks. Pre- and postprandial metabolic rates were measured between weeks 7 and 8. The test oil-based diet did not significantly influence body weight gain, fat accumulation, and metabolic rate. In contrast, liver weight, hepatic triglyceride content, and serum non-high density lipoprotein (HDL)-cholesterol (CHO)/HDL-CHO ratio were significantly higher in the TFA group than in the control group. In conclusion, these findings suggest that dietary TFAs did not influence body fat accumulation but increased the levels of risk markers of cardiovascular diseases.

Trans fatty acid intake and serum cholesterol levels in young Japanese women.

There are very limited data concerning the influence of low-level trans fatty acid (TFA) intake on blood lipid levels. In this study, correlation of total and diene TFA intake with serum cholesterol levels was studied in young Japanese women. The mean intakes of total and diene TFAs were 0.36% and 0.05% of energy, respectively. There was a significant correlation between total fat intake and TFA intake. TFA intake was significantly correlated with erythrocyte TFA content. Total TFA intake was not correlated with total, LDL- or HDL-cholesterol levels. No correlatuon was found between diene TFA intake and cholesterol level. Total and diene TFA intake were not correlated with hemoglobin A1c or C-reactive protein levels. These results suggest that the average TFA intake of young Japanese women does not adversely affect serum cholesterol levels.

Influence of skin thickness on the in vitro permeabilities of drugs through Sprague-Dawley rat or Yucatan micropig skin.

The purpose of this study was to clarify the influence of skin thickness on the in vitro permeabilities of 3 model drugs with different physicochemical properties (nicorandil (NR), isosorbide dinitrate (ISDN) and flurbiprofen (FP)) through Sprague-Dawley rat (rat) or Yucatan micropig (YMP) skin. Intact, dermis-split, stratum corneum-stripped or stratum corneum-stripped and dermis-split rat or YMP skin (rat skin thickness: approximately 0.4, 0.9 or 1.2 mm; YMP skin thickness: approximately 0.4, 0.9, 1.8 or 2.8 mm) were set in Franz-type diffusion cells to determine the permeation rate, lag time and resistance ratio of the viable epidermis and dermis against whole skin (R(ved)/R(tot)) of the drugs. The YMP skin permeabilities of the drugs decreased with an increase in the skin thickness, and significant differences were observed in the permeation rates and lag times between intact and dermis-split (0.4 mm) YMP skins. The decreases in the permeabilities of the drugs through the YMP skin were larger than those through the rat skin. The influence of resistances of ISDN and FP through the dermis-split rat or YMP skin was greater at 0.9 mm skin thickness than 0.4 mm skin thickness. The R(ved)/R(tot) values for the YMP skins were relatively large for lipophilic drugs (ISDN and FP), and these ratios increased with an increase in the dermis thickness. These results suggest that in vitro skin permeation studies must be done using dermis-split (0.4 mm) skin with the thinnest dermis for predicting in vivo human percutaneous absorption rate.

Effect of a 0.6% energy trans fatty acid intake on serum cholesterol concentrations in healthy young Japanese subjects.

A randomized crossover study in healthy young Japanese showed no significant effects of a 0.6% energy trans fatty acid (TFA) intake on the serum cholesterol concentrations and parameters of glucose metabolism. The results indicate that TFAs at this dietary level may have no adverse metabolic effects on healthy young Japanese.

Usefulness of rat skin as a substitute for human skin in the in vitro skin permeation study.

Sprague-Dawley (SD) rats are broadly used in preclinical studies for drug development, so a lot of information for the rats can be obtained especially from pharmacokinetic, pharmacological and toxicological studies. The purpose of this study was to clarify whether SD rat skin can be used to predict human skin permeability. In vitro permeation studies of the three model drugs, nicorandil, isosorbide dinitrate, and flurbiprofen, through human skin and SD rat skin were performed using Franz-type diffusion cells. The permeation rates of the three model drugs through human skin and SD rat skin were determined, and their variations were evaluated. The inter-individual variations in SD rat skin permeability of the three model drugs were much lower than that in human skin permeability, although the permeation rates of the three model drugs through the SD rat skin were about twice those through human skin. In addition, no difference in the skin permeability coefficients of the three model drugs was obtained between fresh SD rat skin and frozen SD rat skin. The markedly smaller variation in the permeability through SD rat skin compared with that through human skin indicated that in vitro permeation studies using SD rat skin would be especially useful for evaluating differences in the skin permeability of the three model drugs as well as for predicting human skin permeability.

Variation assessment for in vitro permeabilities through Yucatan micropig skin.

The purpose of this study was to evaluate the variations in the in vitro Yucatan micropig (YMP) skin permeabilities of drugs and to clarify whether YMP skin can be used to predict human skin permeability. In vitro permeation studies of the three model drugs, nicorandil, isosorbide dinitrate and flurbiprofen, through YMP skin were performed using Franz-type diffusion cells. The permeation rates of the three model drugs were determined, and their variations were evaluated. The inter-individual variations in YMP skin permeability for the three model drugs were smaller than that in human skin permeability, and the permeation rates of the three model drugs through the YMP skin were approximately half that through human skin. In addition, the intra-individual variations in YMP skin permeability for nicorandil and flurbiprofen were much smaller than the inter-individual variations in YMP skin. The inter- and intra-regional variations in YMP skin permeability were very small. The markedly smaller variation in the permeability through YMP skin as compared with that through human skin indicated that in vitro permeation studies using YMP skin would be particularly useful for evaluating differences in the skin permeability of the three model drugs as well as for predicting human skin permeability.

Dietary soybean phospholipids increase the oxidative stability of lipids extracted from fish fillets.

We previously reported that the feeding of soybean phospholipids to fish increased the storage stability of n-3 polyunsaturated fatty acids (PUFA)-rich fish fillets. In this study, we examined the storage stability of lipids extracted from fish fed a diet containing soybean phospholipids and fish oil. Rainbow trout were divided into two groups, and were fed an either 2.5% soybean phospholipids (test) or no phospholipids (control) containing diet for 4 weeks. Lipids were extracted from fish fillets after the feeding period, and were subjected to an oxidation test. Lipids extracted from the fillets of fish in the test group exhibited lower values of oxygen absorption than those in the control group, and the degradation of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) was inhibited. Higher percentages of DHA and EPA were bound to phosphatidylcholine and phosphatidylethanolamine in the extracted lipids in the test group than in the control group. These results indicate that the oxidative stability of lipids extracted from fish fed soybean phospholipids is high, and that the higher percentages of DHA and EPA in PC and PE may have resulted in the higher stability of the lipids extracted from fish fillet.

(1S)-1,5-anhydro-1-[5-(4-ethoxybenzyl)-2-methoxy-4-methylphenyl]-1-thio-D-glucitol (TS-071) is a potent, selective sodium-dependent glucose cotransporter 2 (SGLT2) inhibitor for type 2 diabetes treatment.

Derivatives of a novel scaffold, C-phenyl 1-thio-D-glucitol, were prepared and evaluated for sodium-dependent glucose cotransporter (SGLT) 2 and SGLT1 inhibition activities. Optimization of substituents on the aromatic rings afforded five compounds with potent and selective SGLT2 inhibition activities. The compounds were evaluated for in vitro human metabolic stability, human serum protein binding (SPB), and Caco-2 permeability. Of them, (1S)-1,5-anhydro-1-[5-(4-ethoxybenzyl)-2-methoxy-4-methylphenyl]-1-thio-D-glucitol (3p) exhibited potent SGLT2 inhibition activity (IC(50) = 2.26 nM), with 1650-fold selectivity over SGLT1. Compound 3p showed good metabolic stability toward cryo-preserved human hepatic clearance, lower SPB, and moderate Caco-2 permeability. Since 3p should have acceptable human pharmacokinetics (PK) properties, it could be a clinical candidate for treating type 2 diabetes. We observed that compound 3p exhibits a blood glucose lowering effect, excellent urinary glucose excretion properties, and promising PK profiles in animals. Phase II clinical trials of 3p (TS-071) are currently ongoing.