Decreased Memory and Learning Ability Mediated by Bmal1/M1 Macrophages/Angptl2/Inflammatory Cytokine Pathway in Mice Exposed to Long-Term Blue Light Irradiation.

Humans are persistently exposed to massive amounts of blue light via sunlight, computers, smartphones, and similar devices. Although the positive and negative effects of blue light on living organisms have been reported, its impact on learning and memory remains unknown. Herein, we examined the effects of widespread blue light exposure on the learning and memory abilities of blue light-exposed mice. Ten-week-old male ICR mice were divided into five groups (five mice/group) and irradiated with blue light from a light-emitting diode daily for 6 months. After 6 months of blue light irradiation, mice exhibited a decline in memory and learning abilities, assessed using the Morris water maze and step-through passive avoidance paradigms. Blue light-irradiated mice exhibited a decreased expression of the clock gene brain and muscle arnt-like 1 (Bmal1). The number of microglia and levels of M1 macrophage CC-chemokine receptor 7 and inducible nitric oxide synthase were increased, accompanied by a decrease in M2 macrophage arginase-1 levels. Levels of angiopoietin-like protein 2 and inflammatory cytokines interleukin-6, tumor necrosis factor-α, and interleukin-1ß were elevated. Our findings suggest that long-term blue light exposure could reduce Bmal1 expression, activate the M1 macrophage/Angptl2/inflammatory cytokine pathway, induce neurodegeneration, and lead to a decline in memory.

The Effect of Bacillus coagulans Induced Interactions among Intestinal Bacteria, Metabolites, and Inflammatory Molecules in Improving Natural Skin Aging.

BACKGROUND: Lactic acid bacteria consumption serves several health benefits to humans. However, their effect on natural skin aging is still unclear. METHODS: This study examined the effects of skin naturalization (particularly skin drying) by administering a spore-bearing lactic acid bacteria (Bacillus coagulans) in mice for 2 years. RESULTS: B. coagulans administration improved the natural skin of mice and significantly increased proportions of the genera Bacteroides and Muribaculum, among other intestinal bacteria. As metabolites, increases in nicotinic acid, putrescin, and pantothenic acid levels and a decrease in choline levels were observed. Increased hyaluronic acid, interleukin-10, and M2 macrophage levels indicate aging-related molecules in the skin. Intestinal permeability was also suppressed. Thus, these changes together improved natural skin aging. CONCLUSIONS: This study revealed that B. coagulans administration improved the natural skin aging in mice. This enhancement might be induced by the interaction of alterations in intestinal flora, metabolites, or inflammatory substances.

Induction of Skin Cancer by Long-Term Blue Light Irradiation.

Presently, people are not only exposed to sunlight but also to a large amount of blue light from personal computers and smartphones. This blue light has various effects on the living body. However, its effect on the induction of skin cancer is unknown. In this study, we investigated the induction of skin cancer by long-term blue light irradiation. Hairless mice were irradiated with blue light (LED; peak emission 479 nm) every day for one year, and a control was irradiated with white light (LED), green light (LED; peak emission 538 nm), and red light (LED; peak emission 629 nm) for one year, respectively. Skin cancer was induced only in the mice exposed to blue light. Long-term blue light irradiation also increased the migration of neutrophils and macrophages involved in carcinogenesis in the skin. In neutrophils, an increased expression of citH3 and PAD4 was observed, suggesting the possibility of NETosis. Conversely, in macrophages, inflammatory macrophages (type 1 macrophages) increased and anti-inflammatory macrophages (type 2 macrophages) decreased due to continuous blue light irradiation. These findings suggest that long-term continuous irradiation with blue light induces neutrophil NETosis and an increase in type 1 macrophages, resulting in skin cancer.

Analgesic Mechanisms of Steroid Ointment against Oral Ulcerative Mucositis in a Rat Model.

Despite the long history of use of steroid ointments for oral mucositis, the analgesic mechanism has not been fully elucidated. In this study, we examined the effects of triamcinolone acetonide (Tmc) on oral ulcerative mucositis-induced pain in conscious rats by our proprietary assay system. Based on evaluations of the physical properties and retention periods in the oral mucosa of human volunteers and rats, we selected TRAFUL® ointment as a long-lasting base. In oral ulcerative mucositis model rats, TRAFUL® with Tmc suppressed cyclooxygenase-dependent inflammatory responses with upregulations of glucocorticoid receptor-induced anti-inflammatory genes and inhibited spontaneous nociceptive behavior. When an ointment with a shorter residual period was used, the effects of Tmc were not elicited or were induced to a lesser extent. Importantly, TRAFUL® with Tmc also improved oral ulcerative mucositis-induced mechanical allodynia, which has been reported to be independent of cyclooxygenase. Ca2+ imaging in dissociated trigeminal ganglion neurons showed that long-term preincubation with Tmc inhibited the hypertonic stimulation-induced Ca2+ response. These results suggest that the representative steroid Tmc suppresses oral ulcerative mucositis-induced pain by general anti-inflammatory actions and inhibits mechanical sensitivity in peripheral nerves. For drug delivery, long-lasting ointments such as TRAFUL® are needed to sufficiently induce the therapeutic effects.

THOC4 regulates energy homeostasis by stabilizing TFEB mRNA during prolonged starvation.

TFEB, a basic helix-loop-helix transcription factor, is a master regulator of autophagy, lysosome biogenesis and lipid catabolism. Compared to posttranslational regulation of TFEB, the regulation of TFEB mRNA stability remains relatively uncharacterized. In this study, we identified the mRNA-binding protein THOC4 as a novel regulator of TFEB. In mammalian cells, siRNA-mediated knockdown of THOC4 decreased the level of TFEB protein to a greater extent than other bHLH transcription factors. THOC4 bound to TFEB mRNA and stabilized it after transcription by maintaining poly(A) tail length. We further found that this mode of regulation was conserved in Caenorhabditiselegans and was essential for TFEB-mediated lipid breakdown, which becomes over-represented during prolonged starvation. Taken together, our findings reveal the presence of an additional layer of TFEB regulation by THOC4 and provide novel insights into the function of TFEB in mediating autophagy and lipid metabolism.

CD44v8-10 and CD44s Are Age-dependently Expressed in Primary Cultured Papillary Thyroid Carcinoma Cells and Are Associated with Cell Proliferation.

Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy, arising from follicular cells, and accounts for more than 80% of all thyroid malignant tumors. Although age is the strongest prognostic factor of PTC, and various cut-off ages (40-55 years) were suggested in previous studies, the molecular mechanisms causing age-related changes of PTC cell proliferation remain unclear. CD44 is a major cell surface receptor for hyaluronate and is known as a cancer stem cell marker. However, the association between CD44 and PTC is still unknown. Therefore, we determined the proliferation of primary cultured cells obtained from patients with PTC, and the CD44 mRNA expression profile to elucidate age-related association of CD44 with PTC. The results showed that cell proliferation was significantly decreased according to age. We also found that CD44v8-10 and CD44 splice variants were expressed dominantly in patients with PTC. Moreover, the CD44v8-10/CD44s mRNA expression ratio was significantly increased according to age, and there was a significant negative correlation between this expression ratio and cell proliferation. Our findings suggest that the CD44v8-10/CD44s expression ratio in PTC cells is useful for screening for aggressive PTC and may provide clinically valuable information.

Suppression of autophagic activity by Rubicon is a signature of aging.

Autophagy, an evolutionarily conserved cytoplasmic degradation system, has been implicated as a convergent mechanism in various longevity pathways. Autophagic activity decreases with age in several organisms, but the underlying mechanism is unclear. Here, we show that the expression of Rubicon, a negative regulator of autophagy, increases in aged worm, fly and mouse tissues at transcript and/or protein levels, suggesting that an age-dependent increase in Rubicon impairs autophagy over time, and thereby curtails animal healthspan. Consistent with this idea, knockdown of Rubicon extends worm and fly lifespan and ameliorates several age-associated phenotypes. Tissue-specific experiments reveal that Rubicon knockdown in neurons has the greatest effect on lifespan. Rubicon knockout mice exhibits reductions in interstitial fibrosis in kidney and reduced α-synuclein accumulation in the brain. Rubicon is suppressed in several long-lived worms and calorie restricted mice. Taken together, our results suggest that suppression of autophagic activity by Rubicon is one of signatures of aging.

Increase in the apparent intercalation ability of a platinum complex via multivalency by installation into the sidechain of a graft copolymer and observation of structural changes in the intercalated DNA.

Metal complexes with planar structures have been utilized as DNA intercalators that can be inserted into the base pairs of DNA strands, and have potential applications in DNA-targeting drug therapies. When designing the intercalator metal complexes, controlling their interactions with DNA is important, and has been performed by modifying the chemical structure of the metal ligand. Herein, we designed a graft copolymer segment having Pt complexes with bipyridine and poly(ethylene glycol) (p(PEGMA-co-BPyMA-Pt)) as another strategy to control the interaction with DNA via a multivalent effect. The p(PEGMA-co-BPyMA-Pt) increased not only the binding constant as one macromolecule but also the apparent binding constant per intercalator unit compared to the Pt complex with bipyridine (BPy-Pt). Moreover, p(PEGMA-co-BPyMA-Pt) induced a larger change in DNA structure using lower amounts of Pt than BPy-Pt. These observed properties of p(PEGMA-co-BPyMA-Pt) suggest that grafting intercalators on polymer segments is a promising approach for designing novel types of intercalators.

Gender differences in muscle blood volume reduction in the tibialis anterior muscle during passive plantarflexion.

Physical flexibility, such as joint range of motion and muscle extension, may influence muscle blood volume. Women have been shown to have a greater degree of flexibility than men. We examined whether there is a gender difference in the relationship between fascicle length and muscle blood volume or oxygenation in untrained men and women. In 16 untrained men and thirteen untrained women, we measured the total-[haemoglobin (Hb) + myoglobin (Mb)] (total-[Hb + Mb]) and relative oxy-[Hb + Mb] after calibrating baseline and arterial occlusion deoxygenation levels with near-infrared spectroscopy. Also, fascicle length was measured with B-mode ultrasonography at the tibialis anterior muscle during passive plantarflexion. Increases in fascicle length from baseline (ankle joint angle 120°, composed from the caput fibulae, the malleolus (pivot), and the distal epiphysis of the fifth metatarsal bone) were greater in women than in men during plantarflexion of 140° and 160° and the maximal angle without pain. However, the decreases in total-[Hb + Mb] and relative oxy-[Hb + Mb] from baseline were not different between women and men at any degree of plantarflexion. Moreover, fascicle length and total-[Hb + Mb]/muscle thickness (men > women) showed a similar relationship, with muscle thickness increasing capillary compression. These findings indicate the possibility of a mechanical function underlying muscle blood volume during muscle stretching, which is greater in women than in men.

Superior thyroid artery mean peak systolic velocity for the diagnosis of thyrotoxicosis in Japanese patients.

Thyrotoxicosis with diffuse thyroid disease can be caused by Graves' disease (GD) or destructive thyroiditis (DT). Optimal treatment of the underlying condition requires a prompt and accurate method for the diagnosis of thyrotoxicosis. This study evaluated measurement of the mean peak systolic velocity of the superior thyroid artery (STA-PSV) by ultrasonography in detecting thyrotoxicosis in Japanese patients. We recruited 44 patients with untreated GD, 13 with DT, 55 with treated GD, and 49 subjects without thyroid disease. Blood samples were taken to analyze thyroid function and STA-PSV was measured by ultrasonography. The mean STA-PSV was the highest in the untreated GD group, followed by treated GD patients and then those with DT. Receiver operating characteristic curves of the STA-PSV values demonstrated that the area under the curve required discriminating untreated GD from DT was 0.941. The optimal sensitivity and specificity were 83.7% and 92.3%, respectively, using 45 cm/sec as the cutoff value. In conclusion measurement of STA-PSV by ultrasonography is useful for the diagnosis of thyrotoxicosis in Japanese patients.

Superior thyroid artery mean peak systolic velocity for the diagnosis of thyrotoxicosis in Japanese patients.

Thyrotoxicosis with diffuse thyroid disease can be caused by Graves' disease (GD) or destructive thyroiditis (DT). Optimal treatment of the underlying condition requires a prompt and accurate method for the diagnosis of thyrotoxicosis. This study evaluated measurement of the mean peak systolic velocity of the superior thyroid artery (STA-PSV) by ultrasonography in detecting thyrotoxicosis in Japanese patients. We recruited 44 patients with untreated GD, 13 with DT, 55 with treated GD, and 49 subjects without thyroid disease. Blood samples were taken to analyze thyroid function and STA-PSV was measured by ultrasonography. The mean STA-PSV was the highest in the untreated GD group, followed by treated GD patients and then those with DT. Receiver operating characteristic curves of the STA-PSV values demonstrated that the area under the curve required discriminating untreated GD from DT was 0.941. The optimal sensitivity and specificity were 83.7% and 92.3%, respectively, using 45 cm/sec as the cutoff value. In conclusion measurement of STA-PSV by ultrasonography is useful for the diagnosis of thyrotoxicosis in Japanese patients.

Combination therapy of miglitol and insulin in type 1 diabetes mellitus patients.

UNLABELLED: Aims/Introduction: Patients with type 1 diabetes mellitus often show a precipitous postprandial rise in blood glucose that cannot be controlled, even by intensive insulin therapy. The combined use of an α-glucosidase inhibitor with insulin seems to be highly beneficial in such cases. MATERIALS AND METHODS: We investigated the efficacy and safety of miglitol, an α-glucosidase inhibitor, for 12 weeks in 43 type 1 diabetes patients on intensive insulin therapy. RESULTS: Co-administration of miglitol resulted in only a modest and temporal decrease in HbA1c level. However, it resulted in a significant reduction of plasma glucose level after breakfast (250.7 ± 102.0 mg/dL at 2 h after breakfast before treatment; 212.0 ± 95.8 mg/dL at 2 h after breakfast after treatment for 12 weeks, P = 0.01) and a significant reduction of insulin dosage (41.6 ± 17.1 U/day before treatment; 39.8 ± 17.4 U/day 12 weeks after treatment, P < 0.001). During the study period, 88.4% (38/43) of subjects experienced hypoglycemia, but all events were mild except for one case, which was considered to be moderate. No unexpected adverse events were observed during the study period. CONCLUSIONS: Co-administration of miglitol in type 1 diabetes patients on intensive insulin therapy resulted in an improvement of postprandial hyperglycemia with the reduction of insulin dosage. Considering the importance of postprandial hyperglycemia in the onset of cardiovascular disease, the combination therapy of miglitol and insulin could be advantageous in type 1 diabetes mellitus patients.

Purification by p-aminobenzoic acid (PABA)-affinity chromatography and the functional reconstitution of the nateglinide/H+ cotransport system in the rat intestinal brush-border membrane.

(-)-N-(trans-4-isopropylcyclohexanecarbonyl)-D-phenylalanine (nateglinide) is a novel oral hypoglycemic agent possessing a peptide-type bond and a carboxyl group in its structure. Recently, we have shown that nateglinide transport occurs via the ceftibuten/H+ cotransport system, which is distinct from PepT1, and that the fluorescein/H+ cotransport system is involved in the uptake of nateglinide. The aim of this study was to characterize the functional properties of the intestinal nateglinide transporter. In the first part of this study, we demonstrated that the ceftibuten/H+ cotransport system is identical to the fluorescein/H+ cotransport system. We succeeded in purification of the nateglinide transporter from brush-border membranes of the rat small intestine using p-aminobenzoic acid (PABA)-affinity chromatography. We then investigated the functional properties of the nateglinide transporter using proteoliposomes prepared from the PABA-affinity chromatography elute. We demonstrated that nateglinide, ceftibuten, and fluorescein are transported by the same transporter in the intestine.

Food-drug interaction between ferulic acid and nateglinide involving the fluorescein/H+ cotransport system.

In clinical, patients usually take many kinds of drugs at the same time. Thus, drug-drug interactions involving transporters can often directly affect the therapeutic safety and efficacy of many drugs. However, there have been few studies on food-drug interactions involving transporters. Dietary polyphenols have been widely assumed to be beneficial to human health. Polyphenols are commercially prepared and used as functional foods. We report here for the first time that ferulic acid, which is widely used as a functional food, affects the transport of clinical agents. It is important to be aware of the potential of food-drug interactions and to act in order to prevent undesirable and harmful clinical consequences.

[Depressive symptomatology and some relevant factors. Investigations at a junior college in Hokkaido, Japan].

PURPOSES: Recently in Japan, major depression, a depressive state, and suicide are increasingly becoming social problems requiring preventive intervention. However, only few investigations have been performed of actual conditions of mental health. The present study was therefore planned and carried out to examine relations between stress and relevant factors in adolescents and to search for clues for preventive intervention. METHODS: A total of 184 students in a junior college in Hokkaido were the subjects of this cross-sectional survey. In October 2003, age, sex, health-related factors, social supports, stressors, loci of control, coping styles, and depressive status were investigated. After excluding data from incomplete responses, we ultimately analyzed 153 students (mean age = 19.9, 141 females). RESULTS: Regardless of the CES-D categorization, after conducting univariate logistic regression analysis, we found that cognitive style and consciousness of stress were significantly associated with depressive symptomatology. The association remained significant even after multivariate logistic regression analysis. CONCLUSIONS: Cognitive style may influence depression in adolescents. We now need to consider preventive interventions applying the theory of cognitive-behavioral therapy.

Intestinal uptake of nateglinide by an intestinal fluorescein transporter.

Nateglinide, a novel oral hypoglycemic agent, rapidly reaches its maximum serum concentration after oral administration, suggesting that it is rapidly absorbed in the intestine. However, nateglinide itself is not transported by MCT1 or PEPT1. The aim of this study was to characterize the transporters on the apical side of the small intestine that are responsible for the rapid absorption of nateglinide. It has been reported that the uptake of fluorescein by Caco-2 cells occurs via an H+-driven transporter and that the intestinal fluorescein transporter is probably not MCT1. We examined the contribution of the fluorescein transporter to the uptake of nateglinide by Caco-2 cells. Fluorescein competitively inhibited H+-dependent nateglinide uptake. All of fluorescein transporter inhibitors examined reduced the uptake of nateglinide. Furthermore, nateglinide inhibited fluorescein uptake. We conclude that the intestinal nateglinide/H+ cotransport system is identical to the intestinal fluorescein/H+ cotransport system.

H+-dependent transport mechanism of nateglinide in the brush-border membrane of the rat intestine.

(-)-N-(trans-4-Isopropylcyclohexanecarbonyl)-D-phenylalanine (nateglinide) is a novel oral hypoglycemic agent possessing a carboxyl group and a peptide-type bond in its structure. Although nateglinide quickly reaches the maximal serum concentration after oral administration, nateglinide itself is not transported by PepT1 or MCT1. The aim of this study was to characterize the transporters on the apical side of the small intestine that are responsible for the rapid absorption of nateglinide. The uptake of nateglinide by rat intestinal brush-border membrane vesicles is associated with a proton-coupled transport system. Ceftibuten competitively inhibited H(+)-dependent nateglinide uptake. Glycylsarcosine (Gly-Sar), cephradine, and cephalexin did not significantly inhibit the uptake of nateglinide. The combination of Gly-Sar and nateglinide greatly reduced the uptake of ceftibuten. The effect of the combined treatment was significantly greater than that of Gly-Sar alone. Furthermore, nateglinide competitively inhibited H(+)-driven ceftibuten transporter-mediated ceftibuten uptake. Ceftibuten transport occurs via at least two H(+)-dependent transport systems: one is PepT1, and the other is the ceftibuten/H(+) cotransport system. On the other hand, we demonstrated that nateglinide transport occurs via a single system that is H(+) dependent but is distinct from PepT1 and may be identical to the ceftibuten/H(+) cotransport system.

Ankle brachial pressure index and carotid intima-media thickness as atherosclerosis markers in Japanese diabetics.

The aim of this study was to assess the clinical significance of ankle brachial pressure index (ABI) and carotid intima-media thickness (IMT) in Japanese patients with type 2 diabetes. ABI and ultrasonographic carotid IMT measurements were made in 1311 patients and the relationships between ABI, IMT, and cardiovascular diseases were examined. Patients were assigned to one of three groups depending on their ABI index: (i) ABI > or = 1.0; (ii) ABI from > or = 0.9 to < 1.0; and (iii) ABI < 0.9. The odds ratios (ORs) for groups (ii) and (iii) compared with (i) for the prevalence of coronary heart disease (CHD), cerebrovascular disorder (CVD), and carotid atherosclerosis (mean carotid IMT > or = 1.1mm) demonstrated that these conditions were inversely related to ABI. As the combined ABI/carotid IMT score increased, each OR for the prevalence of CHD and CVD increased significantly. This suggests that a lower ABI is associated with generalized atherosclerosis. Measurements of the ABI and carotid IMT might provide a good prognostic indicator, and both should be assessed during screening for atherosclerosis in Japanese patients with type 2 diabetes.

Effect of cilostazol, a phosphodiesterase inhibitor, on carotid IMT in Japanese type 2 diabetic patients.

The aim of this study was to investigate the effect of cilostazol, a cAMP phosphodiesterase inhibitor, on carotid artery intima-media thickness (IMT) and on the incidence of cardiovascular events in Japanese subjects with type 2 diabetes. A total of 62 type 2 diabetic subjects were allocated equally to the cilostazol treatment group (n = 31) and the control group (n = 31). Carotid IMT was evaluated before and after treatment using B-mode ultrasonography. After the study period (mean +/- SD: 2.6 +/- 0.17 years), carotid IMT showed a significantly greater increase in the control group than in the cilostazol group (0.12 +/- 0.14 mm vs. 0.04 +/- 0.02 mm, p < 0.05). In the control group, 1 out of 31 patients suffered from symptomatic cerebral infarction and 1 had angina pectoris during the observation period. On the other hand, no subject in the cilostazol group developed cardiovascular events during the study period. At baseline, the diabetic patients given cilostazol had a significantly lower HbA1c level than the control subjects, but the other atherosclerotic risk factors (BMI, blood pressure, and serum lipids) and the duration of diabetes did not differ between the two groups. These results indicate that cilostazol therapy can attenuate the increase of carotid artery IMT in Japanese subjects with type 2 diabetes.