Protein Tyrosine Phosphatase Inhibitor, Orthovanadate, Induces Contraction via Rho Kinase Activation in Mouse Thoracic Aortas.

Orthovanadate (OVA), a protein tyrosine phosphatase inhibitor, induces contraction in endothelium-denuded mouse thoracic aortas. OVA-induced contraction was significantly (vs. control rings) suppressed by Rho kinase (Y-27632, 10 µM), extracellular signal-regulated kinase 1 and 2 (Erk1/2, FR180204, 10 µM), Erk1/2 kinase (MEK, PD98059, 10 µM), epidermal growth factor receptor (EGFR, AG1478, 10 µM), and Src inhibitors, and was partially suppressed by c-Jun N-terminal kinase (JNK, AS601245, 10 µM) and p38 (SB203580, 10 µM) inhibitors. However, a myosin light chain kinase inhibitor (ML-7, 10 µM) and a metalloproteinase inhibitor (TAPI-0, 10 µM) had no effect on OVA-induced contraction in mouse thoracic aortas. Phosphorylation of myosin phosphatase target subunit 1 (MYPT1) was abolished by inhibitors of Src, EGFR, MEK, Erk1/2, and Rho kinase, but not by inhibitors of JNK and p38. Erk1/2 phosphorylation by OVA was blocked by inhibitors of EGFR, Src, MEK, and Erk1/2, but not by Rho kinase inhibition. Src phosphorylation at Tyr-416 was abrogated by only Src inhibitor. EGFR phosphorylation at Tyr-1173 was suppressed by a Src inhibitor. These findings suggest that OVA induces contraction via activation of Src, EGFR, MEK, Erk1/2, and Rho kinase, leading to inactivation of myosin light chain phosphatase via MYPT1 phosphorylation.

Structure and dynamics of room temperature ionic liquids with bromide anion: results from 81Br NMR spectroscopy.

We report the results of a comprehensive (81)Br NMR spectroscopic study of the structure and dynamics of two room temperature ionic liquids (RTILs), 1-butyl-3-methylimidazolium bromide ([C(4)mim]Br) and 1-butyl-2,3-dimethylimidazolium bromide ([C(4)C(1)mim]Br), in both liquid and crystalline states. NMR parameters in the gas phase are also simulated for stable ion pairs using quantum chemical calculations. The combination of (81)Br spin-lattice and spin-spin relaxation measurements in the motionally narrowed region of the stable liquid state provides information on the correlation time of the translational motion of the cation. (81) Br quadrupolar coupling constants (C(Q)) of the two RTILs were estimated to be 6.22 and 6.52 MHz in the crystalline state which were reduced by nearly 50% in the liquid state, although in the gas phase, the values are higher and span the range of 7-53 MHz depending on ion pair structure. The C(Q) can be correlated with the distance between the cation-anion pairs in all the three states. The (81)Br C(Q) values of the bromide anion in the liquid state indicate the presence of some structural order in these RTILs, the degree of which decreases with increasing temperature. On the other hand, the ionicity of these RTILs is estimated from the combined knowledge of the isotropic chemical shift and the appropriate mean energy of the excited state. [C(4)C(1)mim]Br has higher ionicity than [C(4)mim]Br in the gas phase, while the situation is reverse for the liquid and the crystalline states.

Measurement of Femoral Trochlear Morphology in Dogs Using Ultrasonography.

OBJECTIVE: The aim of this study was to examine a method for measuring femoral trochlear morphology in dogs using ultrasonography, and survey femoral trochlear morphology in skeletally normal dogs. STUDY DESIGN: To examine the validity of ultrasonography, three raters measured the sulcus angle (SA; the angle between the medial and lateral facets) five times in five skeletally normal dogs. Intra- and interrater reliabilities were then calculated. We also calculated the correlation coefficient between SAs measured using ultrasonography and computed tomography (CT) or magnetic resonance imaging (MRI). Then, we surveyed the femoral trochlear morphology in skeletally normal dogs using ultrasonography. RESULTS: The intra- and interrater reliability for SAs measured using ultrasonography was good, and the correlation coefficient between SAs measured using ultrasonography and CT, or MRI, was strong. In skeletally normal dogs, the SAs for osseous contours decreased until 6 months of age. However, the SAs for cartilaginous contours did not change during the growth period. CONCLUSION: The ultrasonographic measurement method is reliable for the evaluation of femoral trochlear morphology in dogs. The contour of the articular cartilage surface of the femoral trochlea is already determined early in life, and ossification of the articular cartilage of the femoral trochlea proceeds until 6 months of age. The results of this study will provide crucial information for the evaluation of femoral trochlear morphology in dogs.

Computed Tomographic Measurements of the Sulcus Angle of the Femoral Trochlea in Small-Breed Dogs with and without Medial Patellar Luxation.

Trochleoplasty is often performed in dogs with medial patellar luxation (MPL); however, the current guidelines on when to perform a trochleoplasty in dogs are vague. The sulcus angle (SA) is used to assess the femoral trochlear morphology in humans. The aim of this study is to describe a method to measure the SA and other parameters of trochlea morphology in dogs using computed tomography. First, we searched for a suitable measuring location for the SA. Transverse images of the femurs were obtained as perpendicular planes to the tangent of the femoral trochlea which was 0 to 60 degrees (every 5 degrees) to the anatomical axis of the femur. The deepest point of the femoral trochlea was found in the transverse images perpendicular to the tangent of the femoral trochlea which was at 15 degrees to the anatomical axis of the femur. The SA and the other parameters of femoral trochlea morphology were measured at the deepest point of the femoral trochlea. The SA of the stifle joints with grade 3 and 4 MPL was significantly higher than the SA of stifle joints not affected by MPL. There was no significant difference in the SA between dogs affected by grade 1 and 2 MPL and dogs not affected by MPL. Further studies are needed to establish whether the SA can be used as selection criteria for trochleoplasty.

Effect of lactate administration on mouse skeletal muscle under calorie restriction.

Calorie restriction (CR) involves a reductions of calorie intake without altering the nutritional balance, and has many beneficial effects, such as improving oxidative metabolism and extending lifespan. However, CR decreases in skeletal muscle mass and fat mass in correlation with the reduction in food intake. Lactate is known to have potential as a signaling molecule rather than a metabolite during exercise. In this study, we examined the effects of the combination of caloric restriction and lactate administration on skeletal muscle adaptation in order to elucidate a novel role of lactate. We first demonstrated that daily lactate administration (equivalent to 1 g/kg of body weight) for 2 weeks suppressed CR-induced muscle atrophy by activating mammalian/mechanistic target of rapamycin (mTOR) signaling, a muscle protein synthesis pathway, and inhibited autophagy-induced muscle degradation. Next, we found that lactate administration under calorie restriction enhanced mitochondrial enzyme activity (citrate synthase and succinate dehydrogenase) and the expression of oxidative phosphorylation (OXPHOS) protein expression. Our results suggest that lactate administration under caloric restriction not only suppresses muscle atrophy but also improves mitochondrial function.

Glucose levels between the anterior chamber of the eye and blood are correlated based on blood glucose dynamics.

Glycemic control is essential to manage metabolic diseases such as diabetes. Frequent measurements of systemic glucose levels with prompt managements can prevent organ damages. The eye is a glucose highly demanding organ in our body, and the anterior chamber (AC) in the eye has been suggested for a noninvasive blood glucose monitoring site. However, calculating blood glucose levels from measuring glucose levels in AC has been difficult and unclear. In this study, we aimed to examine glucose levels from AC and find a correlation with blood glucose levels. A total of 30 patients with cataracts (men and women, study 1; 7 and 3, study 2; 9 and 11) who visited Keio University Hospital from 2015 to 2018 and agreed to participate in this study were recruited. Glucose levels from AC and the blood were examined by a UV-hexokinase or H2O2-electrode method before/during the cataract surgery. These values were analyzed with regression analyses depending on the groups (blood glucose-ascending and descending groups). In the blood glucose-descending group, glucose levels from AC were strongly correlated with blood glucose levels (a high R2 value, 0.8636). However, the relatively moderate correlation was seen in the blood glucose-ascending group (a low R2 value, 0.5228). Taken together, we showed different correlation ratios on glucose levels between AC and the blood, based on blood glucose dynamics. Stacking data regarding this issue would enable establishing noninvasive blood glucose monitoring from measuring glucose levels in AC more correctly, which will be helpful for proper and prompt managements for glucose-mediated complications.

Effects of a heat- and steam-generating sheet on relieving symptoms of primary dysmenorrhea in young women.

AIM: To test the efficacy of heat- and steam-generating (HSG) sheets for the relief of symptoms of primary dysmenorrhea in young women. MATERIALS & METHODS: Thirty-four female university students were enrolled in this study. HSG sheets generate moist heat to keep the attached body area at 38.5 degrees C for 8 h. Subjects attached the HSG sheet to the lower abdominal or lumbar region for 5 to 8 h once a day on the first, second and third days of menstruation. Subjects documented symptoms of dysmenorrhea (abdominal pain, lumbago and lumbar dullness) on a self-recording form using a 4-score scale of 0 (mild) to 3 (severe) just before applying and after removing the HSG sheet. Either a small (54 cm(2)) or large (164 cm(2)) HSG sheet was used for warming. RESULTS: By applying HSG sheets on the abdomen or lumbar region, 57 and 63% of subjects felt relief of abdominal pains, and 54 and 61% of subjects felt relief from lumbago on the first and second days of menstruation, respectively. Applying the HSG sheets was significantly effective to relieve symptoms compared to the control. Small and large HSG sheets were equally effective. Applying HSG sheets to the abdomen was as effective as that to the lumbar region except for cases of lumbago on the second day of menstruation. Applying HSG sheets two days prior to the onset of menstruation was more effective in relieving lumbar dullness on the second day of menstruation than those just before its onset. CONCLUSION: HSG sheets are useful as non-pharmacological methods to relieve symptoms of primary dysmenorrhea.

Salivary melatonin suppression under 100-Hz flickering blue light and non-flickering blue light conditions.

Bright light at night has been known to suppress melatonin secretion. Photoreceptors, known as intrinsically photosensitive retinal ganglion cells (ipRGCs), project dark/bright information into the superchiasmatic nucleus, which regulates the circadian system. Electroretinograms of ipRGCs show fluctuation that is synchronized with light ON-OFF stimulation. This finding suggests that the flickering condition of light may have an impact on our circadian system. In this study, we evaluate light-induced melatonin suppression under flickering and non-flickering light conditions. Fifteen male subjects between the ages of 20 and 23 years (mean ± SD, 21.9 ± 1.9) were exposed to three light conditions (dim, 100-Hz flickering and non-flickering light) from 1:00 a.m. to 2:30 a.m. Saliva samples were taken just before 1:00 and at 1:15, 1:30, 2:00, and 2:30 a.m. Repeated-measure t-test with Bonferroni correction showed a significant decrease in melatonin levels under both 100-Hz and non-flickering light conditions compared to dim light conditions after 2:00 a.m. Moreover, at 2:30 a.m., the rate of change in melatonin level under 100 Hz of flickering light was significantly lower than that under non-flickering light. Our present findings suggest that 100-Hz flickering light may suppress melatonin secretion more than non-flickering light.

Effects of a single-bolus bupivacaine injection into the coccygeal spinal canal of rabbits.

It has been reported that drugs intended for epidural administration through the lumbosacral junction are accidentally administered into the subarachnoid space frequently in rabbits. Therefore, we evaluated the epidural single-bolus injection technique for the administration of bupivacaine into the coccygeal spinal canal of rabbits. After epidural distribution was confirmed by the injection of iohexol into the coccygeal spinal canal, 0.3 ml/kg 0.5% bupivacaine or 0.3 ml/kg normal saline was injected via the same needle. After the first attempt of iohexol injection, although the contrast was found in the epidural space in all rabbits, the additional contrast was also found in blood vessel in 3 rabbits and in muscular layer in 1 rabbit. Subarachnoid distribution was not observed in any of the rabbits. The time taken to regain normal anal reflex, movement of the hind limbs during walking, conscious proprioception of the hind limbs, and pain sensation of the tail and left hind limb, following coccygeal spinal canal injection, were significantly longer in the bupivacaine group than in the normal saline group. These findings indicated that coccygeal epidural injection of bupivacaine in rabbits may provide anesthesia for the hind limbs, perineum, and tail, but inadvertent vascular entry of the epidural drug may occur.

Beneficial Effects of Fermented Green Tea Extract in a Rat Model of Non-alcoholic Steatohepatitis.

Oxidative stress is frequently considered as a central mechanism of hepatocellular injury in non-alcoholic steatohepatitis (NASH). The aim of this study was to investigate the effects of fermented green tea extracts (FGTE) on NASH. Rats were fed a choline-deficient high-fat diet for 4 weeks to nutritionally generate fatty livers. NASH was induced chemically by oxidative stress using repeated intraperitoneal injections of nitrite. Rats with NASH developed steatohepatitis and liver fibrosis after 6-week of such treatment. At 10 weeks, blood and liver samples were collected from anesthetized animals and assessed for extent of OS injury and effects of FGTE, by biochemical, histological and histochemical analyses. FGTE reduced serum levels of liver enzymes, lipid peroxidation and production of mitochondrial reactive oxygen species. In addition, FGTE showed inhibition of progressions of cirrhosis. Our findings suggest that our FGTE have strong radical scavenging activity and may be beneficial in the prevention of NASH progression.

Shortened Measurement Time of Functional Visual Acuity for Screening Visual Function.

The functional visual acuity test which is the average of the visual acuities measured during a specific time frame (standard, 60 seconds) has been used recently to assess the visual function in various conditions. The availability of a shorter version of the functional visual acuity test promises to be patient friendly in that it is a simple screening test performed in a shorter period of time than the standard test. The results of measurements of the FVA test between the 30-second measurement time (short-version FVA test) and the standard 60-second measurement are compared, and the feasibility of the short-version FVA test instead of the standard FVA test is investigated. Twenty-eight healthy volunteers (25 men and 3 women) were enrolled in this prospective observational study. All subjects underwent measurement of the binocular distance-corrected visual acuity and the binocular distance-corrected FVA with the 60-second and 30-second measurement times. The interchangeability of the corrected-distance FVA, maximal VA, visual maintenance ratio, and average response time in the short-version and the standard FVA tests was evaluated using the Bland-Altman method, and the results showed agreements of the two tests except for the minimal VA. The short-version FVA test is equivalent to the standard method except for evaluating the visual acuity fluctuations and promises to be a simple visual screening test that can be performed in a shorter time.

Non-cotton swab sample collection may not affect salivary melatonin assay results.

BACKGROUND: Salivary melatonin levels have been analyzed in many research fields, including physiological anthropology. Although various devices have been utilized for saliva collection, cotton swabs are among the most common. However, previous studies have reported that cotton swabs may interfere with melatonin assay results, whereas synthetic swabs may not. These studies compared only mean melatonin levels between passive and synthetic-polymer swab collection methods but did not evaluate relative and proportional biases. Our study examines the effects of using swabs made of materials other than cotton, such as polypropylene-polyethylene polymer, on salivary melatonin assay results using a Bland-Altman (BA) plot. The effects of the saliva collection method were analyzed using two concentrations of melatonin, lower (< 6 pg/ml) and higher (> 6 pg/ml), because the threshold of dim light melatonin onset was lower than 6 pg/ml in many studies. RESULTS: Differences detected between passive and polypropylene-polyethylene polymer swab methods of saliva collection were not significant in both lower (< 6 pg/ml) and higher (> 6 pg/ml) melatonin levels detected. All correlations between the collection methods were significant, and 95% confidence intervals for differences in melatonin levels in all samples detected using passive and non-cotton swab saliva collection methods included zero in the BA plots. Averages and differences between non-cotton and passive saliva collection obtained from the BA plots were not significantly correlated at lower and higher melatonin levels. CONCLUSIONS: Our findings demonstrate that swabbing methods, including the use of polypropylene-polyethylene polymer, do not affect salivary melatonin assay results. Therefore, the authors suggest that polypropylene-polyethylene polymer swab methods are appropriate for the assessment of dim light melatonin onset and dose response of the circadian system to light.

Suppression of salivary melatonin secretion under 100-Hz flickering and non-flickering blue light.

BACKGROUND: Bright light at night is known to suppress melatonin secretion. Novel photoreceptors named intrinsically photosensitive retinal ganglion cells (ipRGCs) are mainly responsible for projecting dark/bright information to the suprachiasmatic nucleus and thus regulating the circadian system. However, it has been shown that the amplitude of the electroretinogram of ipRGCs is considerably lower under flickering light at 100 Hz than at 1-5 Hz, suggesting that flickering light may also affect the circadian system. Therefore, in this study, we evaluated light-induced melatonin suppression under flickering and non-flickering light. METHODS: Twelve male participants between the ages of 20 and 23 years (mean ± S.D. = 21.6 ± 1.5 years) were exposed to three light conditions (dim, 100-Hz flickering, and non-flickering blue light) from 1:00 A.M. to 2:30 A.M., and saliva samples were obtained just before 1:00 A.M. and at 1:15, 1:30, 2:00, and 2:30 A.M. RESULTS: A repeated measures t test with Bonferroni correction showed that at 1:15 A.M., melatonin concentrations were significantly lower following exposure to non-flickering light compared with dim light, whereas there was no significant difference between the dim and 100-Hz flickering light conditions. By contrast, after 1:30 A.M., the mean melatonin concentrations were significantly lower under both 100-Hz flickering and non-flickering light than under dim light. CONCLUSION: Although melatonin suppression rate tended to be lower under 100-Hz flickering light than under non-flickering light at the initial 15 min of the light exposure, the present study suggests that 100-Hz flickering light may have the same impact on melatonin secretion as non-flickering light.

Changes in corneal aberrations after cataract surgery.

PURPOSE: To study the changes in corneal aberrations after cataract surgery. METHODS: This study comprised 36 eyes of 36 patients (mean age, 67.3 years) who underwent cataract surgery through a 2.2-mm superior sclerocorneal incision. The anterior, posterior, and total corneal wavefront aberrations for the 6.0-mm pupillary diameter were measured preoperatively and 1 month postoperatively. The postoperative changes in corneal aberrations were evaluated for individual Zernike terms (Z 2 (-2) , Z 2 (0) , Z 2 (2) , Z 3 (-1) , Z 3 (1) , and Z 4 (0) ) and the root mean squares of the third, fourth, and total higher-order aberrations (S3, S4, and THOA). RESULTS: No significant postoperative changes were observed in S3 and S4 of the total and anterior cornea, although significant changes were observed in S4 and THOA of the posterior cornea. Regarding the preoperative and postoperative Zernike terms of the total cornea, the oblique astigmatism (Z 2 (-2) ) changed significantly (P = 0.02). Significant positive correlations were observed in the Z 2 (-2) , Z 2 (0) , Z 2 (2) Z 3 (-1) , Z 3 (1) and Z 4 (0) between the changes in the total and the anterior cornea (P < 0.001 for all comparisons). However, no correlations were observed in the changes in the Zernike terms between the total and the posterior cornea, except for the Z 2 (0) . CONCLUSIONS: The oblique astigmatism (Z 2 (-2) ) of the total cornea significantly changed after 2.2-mm-incision cataract surgery. The changes in the lower- and higher-order aberrations of the anterior cornea mainly contributed to the changes in the total cornea.

A monomeric [Zr(CO3)4](4-) complex in an ammonium zirconium carbonate aqueous solution studied by extended X-ray absorption fine structure, Raman and nuclear magnetic resonance spectroscopy.

The speciation of zirconium complexes in ammonium zirconium carbonate (AZC) solutions should be further studied in order to develop high-performance AZC complexes for use in elaborate industrial materials. Thus, the speciation and the structure of various zirconium species in ammonium zirconium carbonate solutions were investigated over a wide range of total carbonate and ammonium concentrations. By employing extended X-ray absorption fine structure analysis, and Raman and (13)C NMR spectroscopy, it was found that four carbonate ions coordinate bidentately to the zirconium ion. In addition, the exact concentration ranges of carbonate and ammonium in AZC necessary to generate the monomeric zirconium carbonate complex [Zr(CO3)4](4-) were determined.

Integrated transcriptomic and metabolomic analysis of the central metabolism of Synechocystis sp. PCC 6803 under different trophic conditions.

Cyanobacteria have received considerable attention as a sustainable energy resource because of their organic material production capacity using light energy and CO2 as a carbon source. Therefore, it is important to understand the cellular metabolism of cyanobacteria for metabolic engineering. In this study, to shed light on the central metabolism of cyanobacteria, we performed transcriptomic and metabolomic analyses of a glucose-tolerant strain of the cyanobacterium Synechocystis sp. PCC 6803, which was cultured under auto- and mixotrophic conditions. Our results indicate that the oxidative pentose phosphate pathway and glycolysis are activated under mixotrophic conditions rather than autotrophic conditions. Moreover, we examined the effect of atrazine, a photosynthesis inhibitor, on the metabolism of PCC 6803 under mixotrophic conditions, which was defined as photoheterotrophic conditions, by transcriptomics and metabolomics. We demonstrated that the activity of the glycolytic pathway decreased due to the indirect effect of atrazine. In addition, the difference in transcriptional and metabolic changes between auto- and photoheterotrophic conditions could also be captured. The omics dataset reported herein provides clues for understanding the metabolism of cyanobacteria.

Purification and characterization of a novel fungal alpha-glucosidase from Mortierella alliacea with high starch-hydrolytic activity.

The fungal strain Mortierella alliacea YN-15 is an arachidonic acid producer that assimilates soluble starch despite having undetectable alpha-amylase activity. Here, a alpha-glucosidase responsible for the starch hydrolysis was purified from the culture broth through four-step column chromatography. Maltose and other oligosaccharides were less preferentially hydrolyzed and were used as a glucosyl donor for transglucosylation by the enzyme, demonstrating distinct substrate specificity as a fungal alpha-glucosidase. The purified enzyme consisted of two heterosubunits of 61 and 31 kDa that were not linked by a covalent bond but stably aggregated to each other even at a high salt concentration (0.5 M), and behaved like a single 92-kDa component in gel-filtration chromatography. The hydrolytic activity on maltose reached a maximum at 55 degrees C and in a pH range of 5.0-6.0, and in the presence of ethanol, the transglucosylation reaction to form ethyl-alpha-D-glucoside was optimal at pH 5.0 and a temperature range of 45-50 degrees C.