Wavefront restoration from lateral shearing data using spectral interpolation.

Although a lateral-shear interferometer is robust against optical component vibrations, its interferogram provides information about differential wavefronts rather than the wavefronts themselves, resulting in the loss of specific frequency components. Previous studies have addressed this limitation by measuring four interferograms with different shear amounts to accurately restore the two-dimensional wavefront. This study proposes a technique that employs spectral interpolation to reduce the number of required interferograms. The proposed approach introduces an origin-shift technique for accurate spectral interpolation, which in turn is implemented by combining two methods: natural extension and least-squares determination of ambiguities in uniform biases. Numerical simulations confirmed that the proposed method accurately restored a two-dimensional wavefront from just two interferograms, thereby indicating its potential to address the limitations of the lateral-shear interferometer.

Reduction Mechanism of Transition Metal Oxide Particles in Thermally Induced Nanobubbles during Pulsed Laser Melting in Ethanol.

Pulsed laser melting in liquid (PLML) is a technique to fabricate spherical submicrometer particles (SMPs) wherein nanosecond pulsed laser (several tens to several hundreds of mJ pulse-1 cm-2 ) irradiates raw particles dispersed in liquid. Raw particles are transiently heated above the melting point to form spherical particles, which enables pulsed heating of surrounding liquid to form thermally induced bubbles by liquid vaporization. These transient bubbles play an important role as a thermal barrier to rapidly heat the particle. Reduced SMPs are generated from raw metal-oxide nanoparticles by PLML process in ethanol. This reduction cannot be explained by high-temperature thermal decomposition, but by mediation of molecules decomposed from ethanol. Computational simulations of ethanol decomposition by pulsed heating for 100 ns at the temperature 1000-4000 K revealed that ethylene is generated as the main product. Gibbs free energies of oxide reduction reactions mediated by ethylene greatly decreased compared to those without ethylene mediation. This explanation can be applied to reductive SMP formation from various transition metal oxides by PLML.

Behavior of Thermally Induced Nanobubbles during Instantaneous Particle Heating by Pulsed Laser Melting in Liquid.

Pulsed laser melting in liquid (PLML) is a technique to produce submicrometer spherical particles (SMPs). In this process, raw particles dispersed in liquid are selectively heated, and thermally induced nanobubbles (TINBs) at the particle surface are generated and act as a thermal barrier to enhance the temperature increase during heating. However, monitoring TINBs is difficult since PLML is a low-temperature, nonplasma process. Simple transmittance measurements of monodisperse Au SMP (250 nm) colloidal solutions on a transient time scale were used to monitor the temporal dependence of the TINB thickness and the pressure within the bubble. By applying this technique for ZnO and Sn SMP formation, TINBs in the PLML process are important in promoting the formation of large particles via particle merging during laser heating.

Phase III long-term study to evaluate the efficacy and safety of ulipristal acetate in Japanese patients with uterine fibroids.

AIM: To assess the efficacy and safety of long-term intermittent administration of 10-mg ulipristal acetate (UPA) for symptomatic uterine fibroids in Japanese women. METHODS: Open-label, noncomparative study (Japan Primary Registries Network identifier: JapicCTI-173737) conducted at 32 gynecological centers (November 2017-December 2019). Premenopausal women diagnosed with uterine fibroids associated with heavy menstrual bleeding received three 12-week courses of 10-mg UPA once daily. Amenorrhea, fibroid volume, endometrial histology, and safety were assessed. RESULTS: Of 155 patients enrolled, 140 received ≥1 dose of UPA and were analyzed. Across all courses, the rates of patients with amenorrhea for 35 days were >90%, and >99% of patients achieved uterine bleeding normalization. Median time to amenorrhea after each course started was 4-5 days; menstruation returned after treatment within a median of 25-27 days. Mean changes in fibroid volume from baseline were -21.5%, -31.4%, and -35.0% for Courses 1, 2, and 3, respectively. Patients experienced sustained improvements in anemia, pain, and quality of life during treatment. Most adverse events were mild/moderate in severity and decreased in frequency with each course. Seven serious adverse events (six patients) were reported; anemia, embolic cerebral infarction, and pituitary apoplexy (one patient each) were considered UPA-related. Nonphysiological changes in endometrial histology were transient and benign. No safety concerns were detected in hormone concentrations or liver function tests. CONCLUSIONS: Long-term administration of 10-mg UPA is effective for reducing symptoms associated with uterine fibroids in Japanese women. UPA was well tolerated and few safety concerns were reported.

Ulipristal acetate for Japanese women with symptomatic uterine fibroids: A double-blind, randomized, phase II dose-finding study.

PURPOSE: A multicenter, randomized, double-blind, placebo-controlled trial was conducted to evaluate the efficacy, safety, and appropriate dose of ulipristal acetate (UPA) in Japanese women with symptomatic uterine fibroids (UFs). METHODS: A total of 121 premenopausal women with UFs were enrolled to receive either placebo, UPA-2.5 mg, UPA-5 mg, UPA-10 mg, or leuprorelin acetate (LEU), a reference drug, for 12 weeks. The primary end point was the rate of patients having achieved amenorrhea for 35 days at Week 12. RESULTS: The rates for amenorrhea were 4.5%, 60.0%, 72.7%, 88.0%, and 76.2% in the placebo, UPA-2.5 mg, UPA-5 mg, UPA-10 mg, and LEU groups, respectively. The median times to amenorrhea were 20.0, 5.0, 5.0, and 23.0 days for treatment with UPA-2.5 mg, UPA-5 mg, UPA-10 mg, and LEU, respectively. A significant dose-response of UPA for the rate of amenorrhea was observed. The overall incidence rates of adverse events were 45.8% in the placebo group, 56.5%-80.0% in the UPA groups, and 100.0% in the LEU group. There were no notable safety issues with UPA. CONCLUSIONS: Ulipristal acetate was effective and well tolerated in Japanese women with UFs. The recommended dose of UPA is considered to be 10 mg.

Interplay between microbial trait dynamics and population dynamics revealed by the combination of laboratory experiment and computational approaches.

Filament formation is a common bacterial defense mechanism and possibly has a broad impact on microbial community dynamics. In order to examine the impact of filament formation on population dynamics, we developed an experimental system with a filamentous bacterium Flectobacillus sp. MWH38 and a ciliate predator Tetrahymena pyriformis. In this system, the effective defense of Flectobacillus resulted in the extinction of Tetrahymena by allowing almost no population growth. The result of a kairomone experiment suggested the existence of chemical signals for filament formation. To examine the mechanism further, we developed a quantitative mechanistic model and optimized the model for the experimental result using the simulated annealing method. We also performed a global parameter sensitivity analysis using an approximated Bayesian computation based on the sequential Monte Carlo method to reveal parameters to which the model behavior is sensitive to. Our model reproduced the population dynamics, as well as the cell size dynamics of Flectobacillus. The model behavior is sensitive to the nutrient uptake of Flectobacillus and the propensity of filament formation. It robustly predicts the extinction of Tetrahymena at the condition used in the experiment and predicts the transition from equilibrium to population cycle at higher nutrient conditions. Contrary to the previous study that disproved the presence of chemical signals for filament formation, our result suggested the importance of chemical signals at low predator density, suggesting the variety in bacterial resistance mechanisms that act at different stages of predator-prey interactions.

Design, synthesis, and structure-activity relationships of a series of 4-benzyl-5-isopropyl-1H-pyrazol-3-yl ß-D-glycopyranosides substituted with novel hydrophilic groups as highly potent inhibitors of sodium glucose co-transporter 1 (SGLT1).

Sodium glucose co-transporter 1 (SGLT1) plays a dominant role in the absorption of glucose in the gut and is considered a promising target in the development of therapeutic options for postprandial hyperglycemia. Previously, we reported potent and selective SGLT1 inhibitors 1 and 2 showing efficacy in oral carbohydrate tolerance tests in diabetic rat models. In a pharmacokinetic (PK) study of 2, excessive systemic exposure to metabolites of 2 was observed, presumably due to the high permeability of its aglycone (2a). To further improve SGLT1 inhibitory activity and reduce aglycone permeability, a series of 4-benzyl-5-isopropyl-1H-pyrazol-3-yl ß-D-glycopyranoside derivatives bearing novel hydrophilic substitution groups on the phenyl ring were synthesized and their inhibitory activity toward SGLTs was evaluated. Optimized compound 14c showed an improved profile satisfying both higher activity and lower permeability of its aglycone (22f) compared with initial leads 1 and 2. Moreover, the superior efficacy of 14c in various carbohydrate tolerance tests in diabetic rat models was confirmed compared with acarbose, an α-glucosidase inhibitor (α-GI) widely used in the clinic.

Evaluation of a library of loxP variants with a wide range of recombination efficiencies by Cre.

Sparse labeling of individual cells is an important approach in neuroscience and many other fields of research. Various methods have been developed to sparsely label only a small population of cells; however, there is no simple and reproducible strategy for managing the probability of sparse labeling at desired levels. Here, we aimed to develop a novel methodology based on the Cre-lox system to regulate sparseness at desired levels, and we purely analyzed cleavage efficiencies of loxP mutants by Cre. We hypothesized that mutations in the loxP sequence reduce the recognition efficiency by Cre, which enables the regulation of the sparseness level of gene expression. In this research, we mutagenized the loxP sequence and analyzed a library of loxP variants. We evaluated more than 1000 mutant loxP sequences, including mutants with reduced excision efficiencies by Cre ranging from 0.51% to 59%. This result suggests that these mutant loxP sequences can be useful in regulating the sparseness of genetic labeling at desired levels.

Ulipristal acetate compared with leuprorelin acetate for Japanese women with symptomatic uterine fibroids: a phase III randomized controlled trial.

OBJECTIVE: To evaluate the efficacy and safety of ulipristal acetate (UPA) for uterine fibroids (UFs), a phase III study was conducted with leuprorelin (LEU) as a comparator. This is the first confirmatory trial of UPA for UFs among Asians. DESIGN: Multicenter, randomized, double-blind, double-dummy, parallel-group study. SETTING: Thirty-two sites in Japan. PATIENT(S): Patients were assigned to 2 arms, with 82 patients in the UPA group and 79 patients in the LEU group. INTERVENTION(S): In the UPA group, 10 mg of UPA was orally administered once a day for 12 weeks. In the LEU group, 1.88 mg or 3.75 mg of LEU was subcutaneously administered at weeks 0, 4, and 8. MAIN OUTCOME MEASURE(S): The primary endpoint was the percentage of patients with amenorrhea for 35 days. For safety evaluation, adverse events (AEs) were recorded. RESULT(S): The percentage of patients with amenorrhea for 35 days was 87.0% in the UPA group and 81.8% in the LEU group, and the efficacy of UPA for causing amenorrhea for 35 days was confirmed to be noninferior to that of LEU. AEs occurred in 78.0% of the patients in the UPA group and 88.8% of the patients in the LEU group. CONCLUSION(S): The effect of UPA on heavy menstrual bleeding was shown to be comparable with that of LEU in Japanese patients with symptomatic UFs. No notable AEs occurred because of the UPA treatment, and the incidence of AEs in the UPA group was comparable with that of AEs in the LEU group. This result demonstrates the clinical utility of UPA for Asians.

Advanced esophagogastric junction cancer with brain, bone and gastric intramural metastases responding to combined modality therapy.

A 63-year-old man was admitted to our hospital in March 2017 with dysphagia and right homonymous hemianopsia. We diagnosed him with esophagogastric junction cancer (adenocarcinoma) with metastases to the cerebral occipital lobe, bone, and lymph nodes. After one cycle of 5FU + cisplatin (FP), the brain metastasis was resected because of the hemiplegic symptoms he developed. Histology of the resected tissue showed no viable tumor cells. After three cycles of FP, the primary lesion and metastases were resolved. Upper gastrointestinal endoscopy revealed a scar at the primary site. This was considered a complete response (CR). In April 2018, CT revealed a mass at the cardia, which was considered as lymph node metastases with gastric wall invasion. Although two additional cycles of FP were administered for recurrent tumors, the efficacy was progressive. In August 2018, proximal gastrectomy and D1 + lymph node dissection were performed. The pathological diagnosis was gastric intramural metastases and lymph node metastases (ypN1 [2/22]). Weekly paclitaxel therapy was administered for three months after surgery. Two years have passed since the last surgery without recurrence. We report a rare case of esophagogastric junction cancer with brain, bone, and gastric intramural metastases that responded to combined modality therapy.

Neuronal subclass-selective proteomic analysis in Caenorhabditis elegans.

Neurons are categorised into many subclasses, and each subclass displays different morphology, expression patterns, connectivity and function. Changes in protein synthesis are critical for neuronal function. Therefore, analysing protein expression patterns in individual neuronal subclass will elucidate molecular mechanisms for memory and other functions. In this study, we used neuronal subclass-selective proteomic analysis with cell-selective bio-orthogonal non-canonical amino acid tagging. We selected Caenorhabditis elegans as a model organism because it shows diverse neuronal functions and simple neural circuitry. We performed proteomic analysis of all neurons or AFD subclass neurons that regulate thermotaxis in C. elegans. Mutant phenylalanyl tRNA synthetase (MuPheRS) was selectively expressed in all neurons or AFD subclass neurons, and azido-phenylalanine was incorporated into proteins in cells of interest. Azide-labelled proteins were enriched and proteomic analysis was performed. We identified 4,412 and 1,834 proteins from strains producing MuPheRS in all neurons and AFD subclass neurons, respectively. F23B2.10 (RING-type domain-containing protein) was identified only in neuronal cell-enriched proteomic analysis. We expressed GFP under the control of the 5' regulatory region of F23B2.10 and found GFP expression in neurons. We expect that more single-neuron specific proteomic data will clarify how protein composition and abundance affect characteristics of neuronal subclasses.

TGF-beta is specifically expressed in human dermal papilla cells and modulates hair folliculogenesis.

Dermal papilla cells (DPCs) in the mammalian hair follicle have been shown to develop hair follicles through epithelial-mesenchymal interactions. A cell therapy to regenerate human hair is theoretically possible by expanding autologous human DPCs (hDPCs) and transplanting them into bald skin, though much remains to be overcome before clinical success. In this study, we compared gene signatures of hDPCs at different passages and human dermal fibroblasts, and found transforming growth factor (TGF)-beta(2) to be highly expressed in cultured hDPCs. Keratinocyte conditioned medium, which is known to help preserve the hair-inducing capacity of hDPCs, up-regulated TGF-beta(2) expression of hDPCs and also enhanced their alkaline phosphatase (ALP) activity, a known index for hair-inductive capacity. Through screening of components secreted from keratinocytes, the vitamin D(3) analogue was found to promote TGF-beta(2) expression and ALP activity of hDPCs. In animal hair folliculogenesis models using rat epidermis and expanded hDPCs, inhibition of TGF-beta(2) signalling at the ligand or receptor level significantly impaired hair folliculogenesis and maturation. These results suggest an important role for TGF-beta(2) in hair follicle morphogenesis and provide insights into the establishment of future cell therapies for hair regrowth by transplanting expanded DPCs.

Differential expression of stem-cell-associated markers in human hair follicle epithelial cells.

Several putative biomarkers have been suggested for identifying murine follicular stem cells; however, human hair follicles have a different pattern of biomarker expression, and follicular stem cell isolation methods have not been established. To isolate a stem cell population applicable to clinical settings, we conducted a comprehensive survey of the expression of stem-cell-associated (K15, CD200, CD34, and CD271) and other biomarkers (K1, K14, CD29, and CD49f) in immunohistological sections of the human epidermis and follicular outer root sheath (ORS). We also examined freshly isolated and cultured epidermal or follicular cells with single- and multicolor flow cytometry or immunocytochemistry. After sorting cells by CD200, CD34, and forward scatter (FSC) values (cell size), colony-forming assays were performed. We found that biomarkers were differentially expressed in the epidermis and ORS. Basal bulge cells were mainly K15+CD200+CD34(-)CD271(-), and suprabasal cells were K15(-)CD200+CD34(-)CD271(-). We categorized follicular cells into nine subpopulations according to biomarker expression profiles. The CD200+CD34(-) bulge cells had much higher colony-forming abilities than the CD34+ population, and were divided into two subpopulations: a CD200+CD34(-)FSC(high) (K15-rich, basal) and a CD200+CD34(-)FSC(low) (K15-poor, suprabasal) population. The former formed fewer but larger-sized colonies than the latter. Follicular epithelial cell cultivation resulted in loss of K15, CD200, CD34, and CD271 expression, but maintenance of K14, CD29, and CD49f expression. We found that the bulge contained two populations with different localizations, cell sizes, and colony-forming abilities. We showed that K15, CD200, CD34, and CD271 were useful biomarkers for characterizing freshly isolated human follicular epithelial cells in diverse stages of differentiation.

Quasi-flow injection analysis for rapid determination of caffeine in tea using the sample pre-treatment method with a cartridge column filled with polyvinylpolypyrrolidone.

The sample pre-treatment method using a polyvinylpolypyrrolidone (PVPP) cartridge column combined with a quasi-flow injection analysis (quasi-FIA) system realized the rapid determination of caffeine in three types of tea, green, oolong and black tea. The measurement time for each tea sample pre-treated using a PVPP cartridge column was shortened to 20s. In the present system, the limits of detection and quantification were 0.3 microM (1.5 pmol injected) and 0.7 microM (3.5 pmol injected), respectively, and a linear calibration curve was afforded up to 800 microM (4 nmol injected). Within-run precision of analysis of standard solutions of 10 and 100 microM caffeine was 0.11 and 0.16% (n=6), respectively. Between-run precision of analysis of the same solutions over 6 days was 0.78 and 0.74% (n=6), respectively. Comparison with the conventional HPLC method indicated that the present quasi-FIA method using sample pre-treatment with a PVPP cartridge column was useful for the simple and precise determination of caffeine in green, oolong and black tea samples.

Cellomics approach for high-throughput functional annotation of Caenorhabditis elegans neural network.

In Caenorhabditis elegans, which has only 302 neurons, relationships between behaviors and neural networks are not easily elucidated. In this study, we proposed a novel cellomics approach enabling high-throughput and comprehensive exploration of the functions of a single neuron or a subset of neurons in a complex neural network on a particular behavior. To realize this, we combined optogenetics and Brainbow technologies. Using these technologies, we established a C. elegans library where opsin is labeled in a randomized pattern. Behavioral analysis on this library under light illumination enabled high-throughput annotation of neurons affecting target behaviors. We applied this approach to the egg-laying behavior of C. elegans and succeeded in high-throughput confirmation that hermaphrodite-specific neurons play an important role in the egg-laying behavior. This cellomics approach will lead to the accumulation of neurophysiological and behavioral data of the C. elegans neural network, which is necessary for constructing neuroanatomically grounded models of behavior.

Identification of Adenine and Benzimidazole Nucleosides as Potent Human Concentrative Nucleoside Transporter 2 Inhibitors: Potential Treatment for Hyperuricemia and Gout.

To test the hypothesis that inhibitors of human concentrative nucleoside transporter 2 (hCNT2) suppress increases in serum urate levels derived from dietary purines, we previously identified adenosine derivative 1 as a potent hCNT2 inhibitor (IC50 = 0.64 µM), but further study was hampered due to its poor solubility. Here we describe the results of subsequent research to identify more soluble and more potent hCNT2 inhibitors, leading to the discovery of the benzimidazole nucleoside 22, which is the most potent hCNT2 inhibitor (IC50 = 0.062 µM) reported to date. Compound 22 significantly suppressed the increase in plasma uric acid levels after oral administration of purine nucleosides in rats. Because compound 22 was poorly absorbed orally in rats (F = 0.51%), its pharmacologic action was mostly limited to the gastrointestinal tract. These findings suggest that inhibition of hCNT2 in the gastrointestinal tract can be a promising approach for the treatment of hyperuricemia.

High performance liquid chromatography equipped with a cathodic detector and column-switching device as a high-throughput method for a phosphatase assay with p-nitrophenyl phosphate.

LC coupled to an electrochemical detector (LC-EC) operating in cathodic mode with a column-switching system realizes a high-throughput detection of p-nitrophenol (NP). The measurement-time for each NP sample was shortened to 20 s, and the successive analyses of 39 samples was completed within 13 min. In the present system, the limits of detection and quantification were 0.15 and 0.20 microM, respectively, and further, up to 25 microM, a linear calibration curve was afforded. Relative standard deviations for standard solutions of 0.20, 1.0, and 25 microM NP were 4.3, 2.0, and 1.1% (n = 5), respectively. Between-run precisions of the analysis of 5.0 and 25 microM NP over 6 days were 4.8 and 1.3%, respectively. A comparison with the commonly used Bessey-Lowry-Brock method indicates that the present LC-EC is useful for the high-throughput assay of acid and alkaline phosphatases in urine and blood samples with a p-nitrophenyl phosphate substrate.

Duration of drug action of dopamine D2 agonists in mice with 6-hydroxydopamine-induced lesions.

Although 6-hydroxydopamine-induced (6-OHDA-induced) rats are a well-known Parkinson's disease model, the effects of dopamine D2 agonists in mice with 6-OHDA-induced lesions are not completely understood. We produced mice with 6-OHDA-induced lesions and measured their total locomotion counts following administration of several dopamine D2 agonists (pramipexole, ropinirole, cabergoline, rotigotine, apomorphine, talipexole, and quinelorane). Cabergoline showed the longest duration of drug action, which was in agreement with its long-lived anti-Parkinson effects in rats and humans. In contrast, pramipexole and ropinirole had notably short durations of drug action. We demonstrated that mice with 6-OHDA-induced lesions accompanied with significant lesions in the striatum may be reasonable models to predict the action duration of anti-Parkinson drug candidates in humans.