Search

Background The accumulation of amyloid β protein (Aβ) is an important factor in determining the onset of Alzheimer’s disease (AD) since it is related to oxidative stress, which is thought to promote neuronal cell death. In addition, the concentration of acetylcholine, a neurotransmitter, decreases. We clarified the protective effects ofHuperzia serrata, a traditional Chinese herbal medicine, as well as its components against neuronal cell death and cognitive dysfunction. Methods Extract was prepared from whole plant. Murine hippocampal cells (HT22 cells) were damaged by Aβ, hydrogen peroxide, and glutamate. Acetylcholinesterase (AChE) inhibitory activity was measured using mice brain.Huperzia serrata was administrated orally once a day and cognitive behavioural tests were performed. Results Huperzia serrata was identified caffeic acid, ferulic acid, and huperzine A as a major component. Huperzia serrata and its components prevented cell death and reduced the production of reactive oxygen species. Huperzia serrata and its components also inhibited AChE activity. Huperzia serrata (100 mg/kg, p.o.) treatment significantly improved arm alternation and latency in scopolamine-induced cognitive dysfunction. Huperzia serrata also decreased lipid peroxidation in the brain. Conclusions Huperzia serrata prevents cognitive dysfunction through its inhibitory effects on AChE activity and providing protective effects against oxidative damage.

The E/Z switching mechanism of the rotary switch ethyl-2-(2-(quinolin-8-yl)hydrazono)-2-(pyridin-2-yl)acetate was studied by NMR, UV–Vis, and ultrafast spectroscopy and modeled by advanced quantum-chemical calculations. Three possible mechanisms were considered theoretically: out-of-plane rotation, in-plane inversion and proton transfer. Neither of them correctly describes the experimental data by using implicit solvation. Taking into account the existence of water in the used solvents, which influence the measured rate constants, an explicit solvation was attempted in the quantum-chemical calculations. According to a simplified model, the water molecules form a wire, being able to transfer the proton from Z to E form of the switch. This leads to substantially lower transition states and corresponds to the experimentally observed rate constants. This information shines new light on the mechanism of isomerization in the rotary switches and on the understanding of the mechanism of hydrazone tautomerism as a whole.

Our aim was to understand better the causal relationships between material attributes (MAs), process parameters (PPs), and critical quality attributes (CQAs) using an originally created large dataset and regularized linear regression models. In this study, we focused on the following three points: (1) creation of a dataset comprising several tablet production methods, (2) the influence of interaction terms of MAs and/or PPs, and (3) comparison of regularized linear regression models with partial least squares (PLS) regression. First, we prepared 44 kinds of tablets using direct compression and five kinds of granulation methods. We then measured 12 MAs and two model CQAs (tensile strength and disintegration time of tablet). Principal component analysis showed that the constructed dataset comprised a wide variety of particles. We applied regularized linear regression models, such as ridge regression, LASSO and Elastic Net (ENET), and PLS to our dataset to predict CQAs from the MAs and PPs. As a result of external validation, the prediction performance of the models was sufficiently high, although ENET was slightly better than the other methods. Moreover, in almost all cases, the models with interaction terms showed higher predictive ability than those without interaction terms, indicating that the interaction terms of MAs and/or PPs have a strong influence on CQAs. ENET also allowed the selection of critical factors that strongly affect CQAs. The results of this study will help to understand systematically knowledge obtained in pharmaceutical development.

The purpose of this study was to explore the potential of a quantitative structure–property relationship (QSPR) model to predict tablet density. First, we calculated 3381 molecular descriptors for 81 active pharmaceutical ingredients (API). Second, we obtained data that were merged with a dataset including powder properties that we had constructed previously. Next, we prepared 81 types of tablet, each containing API, microcrystalline cellulose, and magnesium stearate using direct compression at 120, 160, and 200 MPa, and measured the tablet density. Finally, we applied the boosted-tree machine learning approach to construct a QSPR model and to identify crucial factors from the large complex dataset. The QSPR model achieved statistically good performance. A sensitivity analysis of the QSPR model revealed that molecular descriptors related to the average molecular weight and electronegativity of the API were crucial factors in tablet density, whereas the effects of powder properties were relatively insignificant. Moreover, we found that these descriptors had a positive linear relationship with tablet density. This study indicates that a QSPR approach is possibly useful for in silico prediction of tablet density for tablets prepared using more than a threshold compression pressure, and to allow a deeper understanding of tablet density.

To investigate the inhibitory effect of a commercial proton pump inhibitor (lansoprazole) on the gastric proton pump H

Chronic myelomonocytic leukemia (CMML) constitutes a hematopoietic stem cell (HSC) disorder characterized by prominent monocytosis and myelodysplasia. Although genome sequencing has revealed the CMML mutation profile, the mechanism of disease development remains unclear. Here we show that aberrant histone acetylation by nucleoporin-98 (NUP98)-HBO1, a newly identified fusion in a patient with CMML, is sufficient to generate clinically relevant CMML pathogenesis. Overexpression of NUP98-HBO1 in murine HSC/progenitors (HSC/Ps) induced diverse CMML phenotypes, such as severe leukocytosis, increased CD115+ Ly6Chigh monocytes (an equivalent subpopulation to human classical CD14+ CD16− monocytes), macrocytic anemia, thrombocytopenia, megakaryocyte-lineage dysplasia, splenomegaly, and cachexia. A NUP98-HBO1–mediated transcriptional signature in human CD34+ cells was specifically activated in HSC/Ps from a CMML patient cohort. Besides critical determinants of monocytic cell fate choice in HSC/Ps, an oncogenic HOXA9 signature was significantly activated by NUP98-HBO1 fusion through aberrant histone acetylation. Increased HOXA9 gene expression level with disease progression was confirmed in our CMML cohort. Genetic disruption of NUP98-HBO1 histone acetyltransferase activity abrogated its leukemogenic potential and disease development in human cells and a mouse model. Furthermore, treatment of azacytidine was effective in our CMML mice. The recapitulation of CMML clinical phenotypes and gene expression profile by the HBO1 fusion suggests our new model as a useful platform for elucidating the central downstream mediators underlying diverse CMML-related mutations and testing multiple compounds, providing novel therapeutic potential.

Higher aerodynamic performance of turbochargers has been demanded because of vehicle engine down-sizing. Centrifugal compressors for automotive turbochargers require higher efficiency and wider operating range. Meridional geometry of the centrifugal compressors is one of their design specifications and it drastically affects the aerodynamic performance of the compressors. In this study, we designed the meridional geometry by using the aerodynamic design method based on a meridional viscous flow analysis and investigated the relation between the meridional geometry and the aerodynamic performance by analyzing meridional viscous flow calculation results and three-dimensional RANS calculation results. As a result, the relation between the boundary layer development near the shroud and the mass flux at the trailing edge was found out according to the meridional viscous flow calculation results. In addition, the relation and the performance improvement were confirmed according to experimental results.

The mechanical strain distribution of scored tablets was simulated using the finite element method (FEM). The score was fabricated as a triangular runnel with the pole on the top surface of flat tablets. The effect of diametral compression on the tablet surface strain was evaluated by changing the angle between the scored line and the diametral compression axis. Ten types of granules were prepared according to an extreme vertices design. Young's modulus and the Poisson ratio for the model powder bed were measured as elastic parameters. The FEM simulation was then applied to the scored tablets represented as a continuous elastic model. Strain distributions in the inner structure of the tablets were simulated after the application of external force. The maximum principal strain (e1) value was obtained with tablets containing a large amount of corn starch, in all scored line positions. In contrast, the e1 value of the tablets containing a large amount of microcrystalline cellulose was minimal. The adequacy of the simulation was evaluated by experiments with scored tablets. The results indicated a fairly good agreement between the FEM simulation and experiments. Moreover, it was found that the e1 value correlated negatively with the value of tablet hardness. These results suggest that the FEM simulation was advantageous for designing scored tablets.

The influence of granule size on simulation parameters and residual shear stress in tablets was determined by combining the finite element method (FEM) into the design of experiments (DoE). Lactose granules were prepared using a wet granulation method with a high-shear mixer and sorted into small and large granules using sieves. To simulate the tableting process using the FEM, parameters simulating each granule were optimized using a DoE and a response surface method (RSM). The compaction behavior of each granule simulated by FEM was in reasonable agreement with the experimental findings. Higher coefficients of friction between powder and die/punch (μ) and lower by internal friction angle (αy) were generated in the case of small granules, respectively. RSM revealed that die wall force was affected by αy. On the other hand, the pressure transmissibility rate of punches value was affected not only by the αy value, but also by μ. The FEM revealed that the residual shear stress was greater for small granules than for large granules. These results suggest that the inner structure of a tablet comprising small granules was less homogeneous than that comprising large granules. To evaluate the contribution of the simulation parameters to residual stress, these parameters were assigned to the fractional factorial design and an ANOVA was applied. The result indicated that μ was the critical factor influencing residual shear stress. This study demonstrates the importance of combining simulation and statistical analysis to gain a deeper understanding of the tableting process.

The rhodium(III)-catalyzed direct alkenylation of N-phenylindole-3-carboxylic acids with alkenes including acrylate ester, acrylamide, and acrylonitrile proceeds smoothly at the C4-position through regioselective C-H bond cleavage directed by the carboxyl group. In marked contrast, the indole substrates react with diarylacetylenes accompanied by cleavage of the C2-H and C2'-H bonds and decarboxylation to produce 5,6-diarylindolo[1,2- a]quinolone derivatives. DFT calculations have suggested that the smooth insertion of an alkene to a C4-rhodated six-membered metallacycle intermediate leads to the C4 alkenylated products, while the latter annulation at the C2- and C2'-positions is attributable to facile reductive elimination in the corresponding seven-membered metallacycles formed by the double C-H bond cleavage and alkyne insertion.

Background and aims: There is no information currently available regarding the effects of bisphenol A (BPA) on testes in ruminants. Therefore, to establish and clarify the effects of BPA in ruminants, testicular tissue cultures were obtained from immature Shiba goats.

Objectives: The aim of this study was to explore the potential of boosted tree (BT) to develop a correlation model between active pharmaceutical ingredient (API) characteristics and a tensile strength (TS) of tablets as critical quality attributes. Methods: First, we evaluated 81 kinds of API characteristics, such as particle size distribution, bulk density, tapped density, Hausner ratio, moisture content, elastic recovery, molecular weight, and partition coefficient. Next, we prepared tablets containing 50% API, 49% microcrystalline cellulose, and 1% magnesium stearate using direct compression at 6, 8, and 10 kN, and measured TS. Then, we applied BT to our dataset to develop a correlation model. Finally, the constructed BT model was validated using k-fold cross-validation. Results: Results showed that the BT model achieved high-performance statistics, whereas multiple regression analysis resulted in poor estimations. Sensitivity analysis of the BT model revealed that diameter of powder particles at the 10th percentile of the cumulative percentage size distribution was the most crucial factor for TS. In addition, the influences of moisture content, partition coefficients, and modal diameter were appreciably meaningful factors. Conclusions: This study demonstrates that BT model could provide comprehensive understanding of the latent structure underlying APIs and TS of tablets.

Hepatic stellate cells (HSCs) play a principal role in Vitamin A metabolism and are considered the major matrix-producing cell type in the diseased liver. Rat HSCs are identified by immunohistochemistry with myogenic or mesenchymal (desmin, vimentin, and alpha-smooth muscle actin) or neural (e.g., GFAP or neuronal cell adhesion molecule) markers. Embryonic origin of rat HSCs was determined using these markers. Nestin, an intermediate filament protein originally identified in neuronal stem or progenitor cells, is widely used as a stem cell marker, including hepatic stem cells in adult rat livers. Additionally, nestin is reportedly expressed in activated HSCs during liver injury and hepatic regeneration. However, little is known about nestin expression in rat fetal liver HSCs. The present study aimed to clarify nestin-positive HSC expression during rat liver development. At embryonic day (ED) 10.5, nestin expression in mesenchymal cells adjacent to the liver bud was detected by immunohistochemistry. At ED 11.5, nestin-positive cells were also detected in desmin-positive cells appearing and increasing in intensity by ED 16.5. However, nestin-positive cells in the parenchyma decreased by ED 20.5 or later. These findings reveal that the nestin-positive HSCs during rat liver development originate from nestin-positive mesenchymal cells in the septum transversum.

The NOTCH family of membranous receptors plays key roles during development and carcinogenesis. Since NOTCH2, yet not NOTCH1 has been shown essential for murine hepatogenesis, NOTCH2 rather than NOTCH1 may be more relevant to human hepatocarcinogenesis; however, no previous studies have supported this hypothesis. We therefore assessed the role of NOTCH2 in human hepatocellular carcinoma (HCC) by immunohistochemistry and cell culture. Immunohistochemically, 19% of primary HCCs showed nuclear staining for NOTCH2, indicating activated NOTCH2 signaling. NOTCH2-positive HCCs were on average in more advanced clinical stages, and exhibited more immature cellular morphology, i.e. higher nuclear-cytoplasmic ratios and nuclear densities. Such features were not evident in NOTCH1‑positive HCCs. In human HCC cell lines, abundant NOTCH2 expression was associated with anaplasia, represented by loss of E-cadherin. When NOTCH2 signaling was stably downregulated in HLF cells, an anaplastic HCC cell line, the cells were attenuated in potential for in vitro invasiveness and migration, as well as in vivo tumorigenicity accompanied by histological maturation. Generally, inverse results were obtained for a differentiated HCC cell line, Huh7, manipulated to overexpress activated NOTCH2. These findings suggested that the NOTCH2 signaling may confer aggressive behavior and immature morphology in human HCC cells.

Made available in DSpace on 2018-11-12T17:26:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2018-11-12T17:34:42Z : No. of bitstreams: 1 S1516-18462015000300792.pdf: 393312 bytes, checksum: 0599eff354c317974a3b95eb97cd4daa (MD5) OBJETIVO: investigar o impacto da Reabilitação Vestibular personalizada e comparar os aspectos físicos, emocionais e funcionais pré e pós a aplicação do Dizziness Handicap Inventory. MÉTODOS: participaram 10 pacientes, com sintomas decorrentes de distúrbios do sistema vestibular e hipótese diagnóstica de disfunção vestibular crônica. Estes foram avaliados quanto aos aspectos físicos, emocionais e funcionais por meio do Dizziness Handicap Inventory pré e pós reabilitação vestibular personalizada. RESULTADOS: no Dizziness Handicap Inventory pré foi verificado que o aspecto físico foi o mais pontuado, seguido pelo emocional e funcional. A reabilitação vestibular foi eficaz, uma vez que houve diminuição nas queixas de qualidade de vida, e melhores resultados em todos os aspectos avaliados no Dizziness Handicap Inventory pós, apenas um paciente obteve melhora somente dos aspectos emocionais e funcionais, além de piora dos aspectos físicos. CONCLUSÃO: o Dizziness Handicap Inventory brasileiro aplicado pré e pós reabilitação vestibular personalizada mostrou-se como um teste eficaz para acompanhar pacientes submetidos a reabilitação vestibular, capaz de mostrar a melhora significante nos sintomas da vertigem crônica, além do impacto negativo na qualidade de vida dos pacientes deste estudo. PURPOSE: to describe the results obtained from the application of the DHI, pre and post customized vestibular rehabilitation in order to verify the efficiency of rehabilitation in this population. METHODS: the sample consisted of 10 patients with symptoms caused by disorders of the vestibular system and diagnosis of chronic vestibular dysfunction. These patients were evaluated for physical, emotional and functional aspects using the Dizziness Handicap Inventory pre and post customized vestibular rehabilitation. RESULTS: on the pre Dizziness Handicap Inventory it was verified that the physical aspects was the most highly scored, followed by the emotional and functional. Vestibular rehabilitation was effective, since there was a decrease in complaints concerning quality of life and better results in all the aspects evaluated in Dizziness Handicap Inventory post rehabilitation, only one patient reported improvement in emotional and functional, and worsening of the physical aspects. CONCLUSION: The Dizziness Handicap Inventory, Brazilian version, applied pre and post customized vestibular rehabilitation proved to be an efficient test to monitor patients undergoing vestibular rehabilitation, this device was able to show significant improvement in symptoms of chronic vertigo, and improved the quality of life of the patients in this study. Universidade Estadual Paulista Faculdade de Filosofia e Ciências Universidade Estadual Paulista Faculdade de Filosofia e Ciências

The physical stability of pharmaceutical emulsions is an important quality attribute to be considered. To obtain a better understanding of this issue, this study investigated the contribution of the state of water to the physical stability of pharmaceutical emulsions. The key technology to evaluate the state of water was magnetic resonance imaging (MRI). For sample preparation, model emulsions with different formulation variables (surfactant content, water content, and hydrophilic–lipophilic balance) were prepared. The T1 relaxation time, diffusion coefficient, and viscosity were measured as physical properties. The physical stability of the samples was evaluated using apparent diffusion coefficient maps acquired by MRI. Data analysis of the observed data was performed using the nonlinear response surface method and Kohonen’s self-organizing map (SOM). It was determined that, depending on the formulation variables, the state of water was substantially changed and it played a significant role in the physical stability. SOM analysis successfully classified the conditions of formulation variables into four distinct clusters in terms of the similarity of the physical properties of the resultant emulsions, and then clarified the characteristics of the stable emulsions. This study provided us with a comprehensive understanding of the formulation variables, physical properties, and stability concerning the preparation of the model emulsion.

The dinuclear palladium(I) complexes [L(Ar2 HGe)Pd(μ-GeAr2 )2 Pd(GeHAr2 )L] (Ar=Ph, p-Tol; L=PMe3 , tBuNC) contain terminal germyl and bridging germylene ligands with the experimentally observed Ge⋅⋅⋅Ge bond lengths of 2.8263(4) Å (L=PMe3 ) and 2.928(1) Å (L=tBuNC), which are close to the longest Ge-Ge bond reported to date [2.714(1) Å]. Significant Ge⋅⋅⋅Ge interactions between the germylene and germyl ligands (PMe3 complexestBuNC complexes) are supported by DFT calculations, Wiberg bond indices (WBI), and natural bond orbital (NBO) analyses. Exchanging tBuNC for PMe3 ligands increases the Ge⋅⋅⋅Ge interaction, and simultaneously activates two Pd-Ge bonds. Adding the chelating diphosphine 1,2-bis(diethylphosphino)ethane (depe) to the PMe3 complexes results in the intramolecular coupling of germyl and germylene ligands followed by extrusion of a digermane.

The stress distribution of tablets after compression was simulated using a finite element method, where the powder was defined by the Drucker-Prager cap model. The effect of tablet shape, identified by the surface curvature, on the residual stress distribution was investigated. In flat-faced tablets, weak positive shear stress remained from the top and bottom die walls toward the center of the tablet. In the case of the convexly curved tablet, strong positive shear stress remained on the upper side and in the intermediate part between the die wall and the center of the tablet. In the case of x-axial stress, negative values were observed for all tablets, suggesting that the x-axial force always acts from the die wall toward the center of the tablet. In the flat tablet, negative x-axial stress remained from the upper edge to the center bottom. The x-axial stress distribution differed between the flat and convexly curved tablets. Weak stress remained in the y-axial direction of the flat tablet, whereas an upward force remained at the center of the convexly curved tablet. By employing multiple linear regression analysis, the mechanical properties of the tablets were predicted accurately as functions of their residual stress distribution. However, the multiple linear regression prediction of the dissolution parameters of acetaminophen, used here as a model drug, was limited, suggesting that the dissolution of active ingredients is not a simple process; further investigation is needed to enable accurate predictions of dissolution parameters.

Myelodysplastic syndromes (MDS) are heterogeneous hematopoietic disorders that are incurable with conventional therapy. Their incidence is increasing with global population aging. Although many genetic, epigenetic, splicing, and metabolic aberrations have been identified in patients with MDS, their clinical features are quite similar. Here, we show that hypoxia-independent activation of hypoxia-inducible factor 1α (HIF1A) signaling is both necessary and sufficient to induce dysplastic and cytopenic MDS phenotypes. The HIF1A transcriptional signature is generally activated in MDS patient bone marrow stem/progenitors. Major MDS-associated mutations (Dnmt3a, Tet2, Asxl1, Runx1, and Mll1) activate the HIF1A signature. Although inducible activation of HIF1A signaling in hematopoietic cells is sufficient to induce MDS phenotypes, both genetic and chemical inhibition of HIF1A signaling rescues MDS phenotypes in a mouse model of MDS. These findings reveal HIF1A as a central pathobiologic mediator of MDS and as an effective therapeutic target for a broad spectrum of patients with MDS. Significance: We showed that dysregulation of HIF1A signaling could generate the clinically relevant diversity of MDS phenotypes by functioning as a signaling funnel for MDS driver mutations. This could resolve the disconnection between genotypes and phenotypes and provide a new clue as to how a variety of driver mutations cause common MDS phenotypes. Cancer Discov; 8(11); 1438–57. ©2018 AACR. See related commentary by Chen and Steidl, p. 1355. This article is highlighted in the In This Issue feature, p. 1333

Hepatocellular carcinoma (HCC) is one of the most difficult cancers to treat owing to the lack of effective chemotherapeutic methods. Sorafenib, the first-line and only available treatment for HCC, extends patient overall survival by several months, with a response rate below 10%. Thus, the identification of an agent that enhances the anticancer effect of sorafenib is critical for the development of therapeutic options for HCC. Endoplasmic reticulum (ER) stress response is one of the methods of sorafenib-induced cell death. Here we report that questiomycin A suppresses expression of GRP78, a cell-protective ER chaperone protein. Analysis of the molecular mechanisms of questiomycin A revealed that this compound stimulated GRP78 protein degradation in an ER stress response-independent manner. Cotreatment with sorafenib and questiomycin A suppressed GRP78 protein expression, which is essential for the stimulation of sorafenib-induced cell death. Moreover, our in vivo study demonstrated that the coadministration of sorafenib and questiomycin A suppressed tumor formation in HCC-induced xenograft models. These results suggest that cotreatment with sorafenib and questiomycin A is a novel therapeutic strategy for HCC by enhancing sorafenib-dependent ER stress-induced cell death, and downregulation of GRP78 is a new target for the stimulation of the therapeutic effects of sorafenib in HCC.

An optimum aerodynamic design method for centrifugal compressor impeller has been developed. The present optimum design method is using a genetic algorithm (GA) and a two-dimensional inverse blade design method based on a meridional viscous flow analysis. In the meridional viscous flow analysis, an axisymmetric viscous flow is numerically analyzed on a two-dimensional meridional grid to determine the flow distribution around the impeller. Full and splitter blade effects to the flow field are successfully evaluated in the meridional viscous flow analysis by a blade force modeling. In the inverse blade design procedure, blade loading distribution is given as the design variable. In the optimization procedure, the total pressure rise and adiabatic efficiency obtained from the meridional viscous flow analysis are employed as objective functions. Aerodynamic performance and three-dimensional flow fields in the Pareto-optimum design and conventional design cases have been investigated by three-dimensional Reynolds averaged Navier-Stokes (3D-RANS) and experimental analyses. The analyses results show performance improvements and suppressions of flow separations on the suction surfaces in the optimum design cases. Therefore, the present aerodynamic optimization using the inverse method based on the meridional viscous flow analysis is successfully achieved.

Simple 2D scaling of semiconductor devices by “Moore's law” will not work well soon to improve the performances and power efficiencies due to tight physical directional limits. System integrations, however, might continue to advance further by 3D structural evolutions either in monolithic on-chip integrations or heterogeneous off-chip stacks. Accelerated implementations of new architectures and new functional materials would be key factors, especially to execute machine learnings more efficiently for ΑΙ-based smart applications. Just now, we have established SDRJ (The System Device Roadmap Committee of Japan, https://www.sdrj.jp/) in JSAP collaborated with IEEE IRDS (International Roadmap for Devices and Systems, http://irds.ieee.org/) to discuss the roadmaps internationally toward the year of 2030.

In this study, we evaluated the correlation between the response surfaces for the tablet characteristics of placebo and active pharmaceutical ingredient (API)-containing tablets. The quantities of lactose, cornstarch, and microcrystalline cellulose were chosen as the formulation factors. Ten tablet formulations were prepared. The tensile strength (TS) and disintegration time (DT) of tablets were measured as tablet characteristics. The response surfaces for TS and DT were estimated using a nonlinear response surface method incorporating multivariate spline interpolation, and were then compared with those of placebo tablets. A correlation was clearly observed for TS and DT of all APIs, although the value of the response surfaces for TS and DT was highly dependent on the type of API used. Based on this knowledge, the response surfaces for TS and DT of API-containing tablets were predicted from only two and four formulations using regression expression and placebo tablet data, respectively. The results from the evaluation of prediction accuracy showed that this method accurately predicted TS and DT, suggesting that it could construct a reliable response surface for TS and DT with a small number of samples. This technique assists in the effective estimation of the relationships between design variables and pharmaceutical responses during pharmaceutical development.

Familial platelet disorder with propensity to myeloid malignancies (FPD/AML) is an autosomal dominant disorder characterized by quantitative and/or qualitative platelet defects with a tendency to develop a variety of hematological malignancies. Heterozygous germ line mutations in the RUNX1 gene are responsible genetic events for FPD/AML. Notably, about half of individuals in the family with germ line mutations in RUNX1 develop overt hematological malignancies. The latency is also relatively long as an average age at diagnosis is more than 30 years. Similar to what is observed in sporadic hematological malignancies, acquired additional genetic events cooperate with inherited RUNX1 mutations to progress the overt malignant phase. Reflecting recent increased awareness of hematological malignancies with germ line mutations, FPD/AML was added in the revised WHO 2016 classification. In this review, we provide an update on FPD/AML with recent clinical and experimental findings.

A diagnostic ultrasound (US) system transmits acoustic waves at several to tens of MHz into the human body for clinical purposes and detects the reflected waves to observe the internal organs without having a medical operation or radiation exposure. The system is composed of a main unit and probe connected via coaxial cables. The probe is very small because medical technicians laboriously grab and manipulate it for a long time. To avoid image obscurity depending on medical technicians, high-speed and high-resolution 3D/4D imaging is necessary. For this reason, several thousands of lead bulk piezoelectric material transducers (TD) need to be squeezed into the small probe. Since the number of cables is limited to several hundreds, the probe needs to include beamforming functionality and a 2D array IC [1–6], which includes thousands of US transceivers.

Rasilez(®) tablets (RTs) contain the active ingredient aliskiren, which is a direct renin inhibitor of the renin-angiotensin system and used for the treatment of hypertension. We examined the influence of temperature and humidity on the physico-pharmaceutical characteristics (mass, volume, hardness, elution) of RTs. The RTs were preserved under conditions in which the temperature and humidity were altered using the second-order spherical composite experimental design for multi-objective problems. The characteristics of RTs were influenced more by the humidity than temperature, and differed markedly with over 55% relative humidity (RH). The mass and volume were increased with increasing humidity, and the tablets swelled. The hardness after vacuum-drying of the tablets, which preserved moisture conditions, was increased. Semitransparent particles were observed in the cross-section of the drying tablets in which aliskiren crystal forms were changed to amorphous forms. The mean dissolution time (MDT) of tablets was reduced with increasing humidity. The critical relative humidity (CRH) of the tablets was 36.1%RH at 30°C. These results suggest that RTs, on moisture absorption, showed changes in not their appearance and hardness, but also in crystal forms and the elution characteristics of aliskiren.

The transcription factor CCAAT/enhancer-binding protein β (C/EBPβ) regulates the differentiation of a variety of cell types. Here, the role of C/EBPβ expressed by bone marrow mesenchymal stromal cells (BMMSCs) in B-cell lymphopoiesis was examined. The size of the precursor B-cell population in bone marrow was reduced in C/EBPβ-knockout (KO) mice. When bone marrow cells from C/EBPβ-KO mice were transplanted into lethally irradiated wild-type (WT) mice, which provide a normal bone marrow microenvironment, the size of the precursor B-cell population was restored to a level equivalent to that generated by WT bone marrow cells. In coculture experiments, BMMSCs from C/EBPβ-KO mice did not support the differentiation of WT c-Kit+ Sca-1+ Lineage− hematopoietic stem cells (KSL cells) into precursor B cells, whereas BMMSCs from WT mice did. The impaired differentiation of KSL cells correlated with the reduced production of CXCL12/stromal cell-derived factor-1 by the cocultured C/EBPβ-deficient BMMSCs. The ability of C/EBPβ-deficient BMMSCs to undergo osteogenic and adipogenic differentiation was also defective. The survival of leukemic precursor B cells was poorer when they were cocultured with C/EBPβ-deficient BMMSCs than when they were cocultured with WT BMMSCs. These results indicate that C/EBPβ expressed by BMMSCs plays a crucial role in early B-cell lymphopoiesis. Stem Cells 2014;32:730–740

Thermal reactions of benzodisilacyclobutene (1) and disilacyclobutene (2) with acetylene were investigated theoretically. The reactions are thought to proceed via the conventional Diels–Alder reaction of disilabutadiene, the conrotatory ring-opening product of disilacyclobutene, with acetylene. However, this mechanism is incompatible with the observed similar reactivities of 1 and 2 with acetylene and the retention of stereochemistry during the reaction. In our previous paper, we proposed an alternative [2 + 1] cycloaddition pathway that involved the direct addition of acetylene to the Si–Si σ–bond of 1 without ring opening. In this study, we extensively investigated the reaction pathways for both 1 and 2 on a theoretical basis. We found that charge transfer (CT) played a key role in the [2 + 1] cycloaddition pathway. On the basis of natural bond orbital (NBO) analysis, the interaction of the Si–Si σ-bond orbital (donor) with the π* orbital (acceptor) of the acetylene was attributed mainly to the CT proce...

To clarify the effect of N-methylation to aromatic oligoamides on their conformations, we investigated Nmethylated aromatic amides theoretically, using the density functional theory calculations. It is well-known that Nmethylbenzanilide adopts a cis-conformation rather than a trans-conformation, in contrast to benzanilide. This is ascribed to large destabilization of trans-N-methylbenzanilide and large stabilization by the orbital interaction from nitrogen lone pair to anti-bondingπ* orbital of carbonyl group in cis-N-methylbenzanilide. N-methylated para-linked aromatic diamide has two preferable conformations and two of the conformations of the N-methylated para-linked aromatic oligoamidesexhibit two distinct helical structures. The potential for helix control by solvent is presented.

Under stress or regenerative conditions, HSCs rapidly enter into cell cycle and are reprogrammed toward myeloid-biased hematopoiesis to meet the increasing demand of myeloid cells. We have previously shown that the transcription factor C/EBPβ plays critical roles at the level of HSPCs under stress conditions (Nat Immunol 2006, J Immunol 2012, Leukemia 2013 and Blood Adv 2019). However, underlying molecular mechanisms of action remain largely unknown. In this study, we have investigated the detailed function of C/EBPβ in regulation of HSPCs. We first evaluated the impact of C/EBPβ on the cell cycle status of LT-HSCs. To exclude the cell-extrinsic contribution of C/EBPβ, CD45.2+ BM cells from WT or Cebpb-/- mice were transplanted into lethally irradiated CD45.1+ WT mice, and these "BM-replaced" recipients were subjected to the following experiments. At steady state, the cell cycle statuses and the numbers of HSPCs did not significantly differ between the recipients of WT cells and those of Cebpb-/- cells. Immediately after 5-FU treatment, WT LT-HSCs entered the cell cycle, as revealed by the decreased percentage of cells in G0 phase and the increased percentage of cells in S/G2M phase. All these parameters of cell cycle acceleration were observed prior to the nadir of LT-HSCs induced by 5-FU and were significantly attenuated in Cebpb-/- LT-HSCs. Next, we assessed the numbers of LT-HSCs, KSL cells, and KL cells after 5-FU treatment. Following the nadir, the recovery of LT-HSCs preceded that of KSL and KL cells, suggesting the differentiation of LT-HSCs to KSL and KL cells. In the recipients of Cebpb-/- cells, the recovery of KSL and KL cells was delayed significantly. Collectively, cell cycle acceleration and subsequent differentiation of LT-HSCs under stress conditions were impaired in the absence of Cebpb. The Cebpb is a single exon gene, and three isoforms, namely, LAP*, LAP and LIP which lacks N-terminus, are translated from its unique mRNA. Due to their structural difference, they should have distinct functions. Here, we focused on expression and functions of these isoforms in regenerating HSPCs. To monitor expression of these isoforms in small numbers of HSCs, we devised a novel intracellular double staining method for flow cytometric analysis using two distinct anti-C/EBPβ antibodies. An antibody against the C-terminus of C/EBPβ recognized all three isoforms, while an antibody against the N-terminus of C/EBPβ only recognized LAP* and LAP. Thus, simultaneous staining with both antibodies should enable us to distinguish cells that dominantly expressed LIP (C-term+ N-term-) from those that expressed all three isoforms (C-term+ N-term+). Using this method, we monitored the expression patterns of these isoforms in LT-HSCs after 5-FU treatment. LT-HSCs initially became C-term single positive in response to 5-FU and subsequently changed to C- and N-term double positive, suggesting that LIP was upregulated prior to LAP/LAP* under stress conditions. These results suggest that phase-specific upregulation of LIP and LAP/LAP* is strongly associated with phase-specific functions of C/EBPβ in cell cycle activation and differentiation, respectively. Indeed, when EML cells, a mouse HSC line, were retrovirally transduced with LIP, the transduced cells were more proliferative and actively cycling than those transduced with the control vector, whereas proliferation and cell cycle were markedly suppressed in LAP*- and LAP-expressing EML cells. LIP-expressing cells remained undifferentiated, while LAP*- and LAP-expressing cells rapidly differentiated into CD11b+ myeloid cells and eventually stopped proliferating. In summary, our findings clearly suggest that sequential upregulation of C/EBPβ isoforms is critical for the regulation of HSCs under stress conditions. LIP amplifies the "reservoir" of HSPCs by accelerating the proliferation of HSCs during the early phase of regeneration, while LAP*/LAP induce their myeloid differentiation at a later phase. These findings should facilitate our understanding of the pathophysiology of infection, inflammation, and regenerating hematopoiesis in response to myeloablative chemotherapies or hematopoietic stem cell transplantation, all of which increase the hematopoietic demands. Disclosures Hirai: Kyowa Kirin: Research Funding.

The thermal reaction of cis-3,4-benzo-1,2-diisopropyl-1,2-dimethyl-1,2-disilacyclobut-3-ene (1a) with tert-butyl alcohol at 300 °C for 24 h proceeded with high stereospecificity to give erythro-1-(...

Tablet characteristics of tensile strength and disintegration time were predicted using residual stress distribution, simulated by the finite element method (FEM). The Drucker–Prager Cap (DPC) model was selected as the method for modeling the mechanical behavior of pharmaceutical powders composed of lactose (LAC), cornstarch (CS), and microcrystalline cellulose (MCC). The DPC model was calibrated using a direct shear test and analysis of the hardening law of the powder. The constructed DPC model was fed into the analysis using the FEM, and the mechanical behavior of pharmaceutical powders during compaction was analyzed using the FEM. The results revealed that the residual stress distribution of the tablets was uniform when the compression force increased. In particular, the residual stress distribution of tablets composed of equal amounts of LAC, CS, and MCC was more uniform than the tablets composed of 67% LAC and 33% CS, with no MCC. The tensile strength and disintegration time were predicted accurately from the residual stress distribution of tablets using multiple linear regression analysis and partial least squares regression analysis. This suggests that the residual stress distribution of tablets is related closely to the tensile strength and disintegration time.

Reliability of a quad-flat-package (QFP) with a circuit-under-pad (CUP) structure is investigated for Cu interconnects with porous low dielectric constant (low-k) films in scaled-down ultralarge-scale integrations. The following experimental factors are discussed: 1) low-k material properties and their stacking structures; 2) CUP structure; and 3) mold compound material properties. The QFP characteristics are analyzed after chip dicing and Ag wire bonding, as well as after molding. Higher adhesion strength of porous low-k film to SiCN cap dielectrics and rigid Cu-anchored CUP structure can achieve highly reliable QFP packaging. A lower coefficient of thermal expansion (CTE) of the molding compound is also found to be effective in eliminating low-k delamination during thermal cycle test because it can reduce the stress at the cracking position. The adhesion-promoting porous SiOCH film with the Cu-anchored CUP in a low-CTE mold is a promising system to realize a reliable QFP with no low-k delamination, passing electrical tests after the pressure-cooker test and high-temperature storage test.

Yariv phenylglycosides [1,3,5-tri(p-glycosyloxyphenylazo)-2,4,6-trihydroxybenzene] are a group of chemical compounds that selectively bind to arabinogalactan proteins (AGPs), a type of plant proteoglycan. Yariv phenylglycosides are widely used as cytochemical reagents to perturb the molecular functions of AGPs as well as for the detection, quantification, purification, and staining of AGPs. However, the target structure in AGPs to which Yariv phenylglycosides bind has not been determined. Here, we identify the structural element of AGPs required for the interaction with Yariv phenylglycosides by stepwise trimming of the arabinogalactan moieties using combinations of specific glycoside hydrolases. Whereas the precipitation with Yariv phenylglycosides (Yariv reactivity) of radish (Raphanus sativus) root AGP was not reduced after enzyme treatment to remove α-l-arabinofuranosyl and β-glucuronosyl residues and β-1,6-galactan side chains, it was completely lost after degradation of the β-1,3-galactan main chains. In addition, Yariv reactivity of gum arabic, a commercial product of acacia (Acacia senegal) AGPs, increased rather than decreased during the repeated degradation of β-1,6-galactan side chains by Smith degradation. Among various oligosaccharides corresponding to partial structures of AGPs, β-1,3-galactooligosaccharides longer than β-1,3-galactoheptaose exhibited significant precipitation with Yariv in a radial diffusion assay on agar. A pull-down assay using oligosaccharides cross linked to hydrazine beads detected an interaction of β-1,3-galactooligosaccharides longer than β-1,3-galactopentaose with Yariv phenylglycoside. To the contrary, no interaction with Yariv was detected for β-1,6-galactooligosaccharides of any length. Therefore, we conclude that Yariv phenylglycosides should be considered specific binding reagents for β-1,3-galactan chains longer than five residues, and seven residues are sufficient for cross linking, leading to precipitation of the Yariv phenylglycosides.

The purpose of this study was to quantitatively evaluate the water content of granules drying in a fluidised bed by near infrared (NIR) spectroscopy in real time and estimate a constant drying rate. Riboflavin granules were prepared by extrusion based on a standard formulation. NIR spectra collected during drying and the water content of granules was correlated by partial least squares (PLS) regression. To consider variability among batches, the leave-one-batch-out cross-validation procedure was performed. The PLS analysis showed that the plots of predicted vs actual values were linearly correlated with a coefficient content which can be continuously predicted with small errors. The difference between batches was clarified by results of score plots. In each batch, water content was predicted accurately using the PLS model. Based on water content values predicted in real time, constant drying rate was estimated. This result suggested constant drying rate might increase as the granule size decreases. This technique provides a robust calibration model for a better understanding and control of the drying process.

Scored tablets can be divided into equal halves for individual treatment of patients. However, the relationships between scored shapes and tablet characteristics such as the dividing strength, halving equality, and breaking strength are poorly understood. The purpose of this study was to simulate the mechanical stress distribution of scored tablets by using the finite element method (FEM). A runnel of triangle pole on the top surface of flat tablets was fabricated as the score shape. The depth and angle of the scores were selected as design variables. Elastic parameters such as a Young's modulus and a Poisson ratio for the model powder bed were measured. FEM simulation was then applied to the scored tablets, represented as a continuum elastic model. Stress distributions in the inner structure of the tablets were simulated after applying external force. The adequacy of the simulation was evaluated in experiments using scored tablets. As a result, we observed a relatively good agreement between the FEM simulation and the experiments, suggesting that FEM simulation is advantageous for designing scored tablets.

Native α-cyclodextrin- (α-CD) and permethylated α-CD (PMeCD)-based rotaxanes with various short alkylene chains as axles can be synthesized through a urea end-capping method. Native α-CD tends to form [3]- or [5]pseudorotaxanes and not [2]- or [4]pseudorotaxanes, which indicates that the coupled CDs act as a single fragment. End-capping reactions of the pseudorotaxanes with C18 and C24 axle lengths do not occur because the axle termini are covered by the densely stacked CDs. The number of PMeCDs on the pseudorotaxane is flexible and mainly depends on the axle length. Peracetylated α-CD (PAcCD)-based rotaxanes are synthesized through O-acetylation of the α-CD-based rotaxanes without any decomposition of the rotaxanated structures. The structures of PMeCD-based [3]- and [4]rotaxanes, and the molecular dynamics calculations on [3]pseudorotaxanes, indicate that the tail face of PMeCDs is regularly directed toward the axle termini. On the basis of the results obtained, it can be concluded that the directions and numbers of CDs in rotaxanes containing short alkylene chains depend on 1) the interactions between CDs, 2) the length of the alkylene axle, and 3) the interactions between the axle end and tail face of the CD.

Aromatase is an enzyme catalyzing the final step of 17β-estradiol (E2) biosynthesis. Aromatase-deficient (ArKO) mice displayed vital roles of E2 at various tissue sites, including ovary. Here, we report attenuated responses of ArKO ovary to equine chorionic gonadotropin (eCG), an alternative to FSH. Ovarian contents of cAMP and anti-Müllerian hormone (AMH), putative factors reducing sensitivity to gonadotropins, were significantly elevated in ArKO mice compared with those in wild type (WT) mice in the basal state. Accordingly, eCG-induced ovarian alterations in cAMP contents, phosphorylation levels of signaling molecules, and mRNA expression of eCG-targeted genes were blunted in ArKO mice compared with those in WT mice. Treatment of ArKO mice with E2 decreased ovarian cAMP and AMH contents to the WT levels but did not restore the sensitivity. Microarray analysis coupled with quantitative RT-PCR analysis identified 7 genes of which the mRNA expression levels in ArKO ovaries were significantly different from those in the WT ovaries in the basal state and were not normalized by E2 supplementation, indicating possible involvement of these gene products in the determination of ovarian sensitivity to eCG. Thus, present analyses revealed that estrogen deficiency attenuates sensitivity of the ovary to gonadotropin, which might be associated with alterations in the ovarian contents of multiple molecules including cAMP and AMH. Given the importance of the ovarian responses to gonadotropins in reproductive function, detailed knowledge about the underlying mechanisms of abnormalities in the ArKO ovary might help to develop potential targets for infertility treatments.

Granulopoiesis is tightly regulated to meet host demands during both “steady-state” and “emergency” situations, such as infections. The transcription factor CCAAT/enhancer binding protein β (C/EBPβ) plays critical roles in emergency granulopoiesis, but the precise developmental stages in which C/EBPβ is required are unknown. In this study, a novel flow cytometric method was developed that successfully dissected mouse bone marrow cells undergoing granulopoiesis into five distinct subpopulations (#1–5) according to their levels of c-Kit and Ly-6G expression. After the induction of candidemia, rapid mobilization of mature granulocytes and an increase in early granulocyte precursors accompanied by cell cycle acceleration was followed by a gradual increase in granulocytes originating from the immature populations. Upon infection, C/EBPβ was upregulated at the protein level in all the granulopoietic subpopulations. The rapid increase in immature subpopulations #1 and #2 observed in C/EBPβ knockout mice at 1 d postinfection was attenuated. Candidemia-induced cell cycle acceleration and proliferation of hematopoietic stem/progenitors were also impaired. Taken together, these data suggest that C/EBPβ is involved in the efficient amplification of early granulocyte precursors during candidemia-induced emergency granulopoiesis.

Planar chiral [2]- and [3]rotaxanes constructed from pillar[5]arenes as wheels and pyridinium derivatives as axles were obtained in high yield using click reactions. The process of rotaxane formation was diastereoselective; the obtained [2]rotaxane was a racemic mixture consisting of (pS, pS, pS, pS, pS) and (pR, pR, pR, pR, pR) forms of the per-ethylated pillar[5]arene (C2) wheel, and other possible types of the [2]rotaxane did not form. Isolation of the enantiopure [2]rotaxanes with one axle through (pS, pS, pS, pS, pS)-C2 or (pR, pR, pR, pR, pR)-C2 wheels was accomplished. Furthermore, pillar[5]arene-based [3]rotaxane was successfully synthesized by attachment of two pseudo [2]rotaxanes onto a bifunctional linker. [3]Rotaxane formed in a 1:2:1 mixture with one axle threaded through two (pS, pS, pS, pS, pS)-C2, one (pS, pS, pS, pS, pS)-C2 and one (pR, pR, pR, pR, pR)-C2 (meso form), or two (pR, pR, pR, pR, pR)-C2 wheels. The [3]rotaxane enantiomers and the meso form were successfully isolated using appropriate chiral HPLC column chromatography. The procedure developed in this study is the starting point for the creation of pillar[5]arene-based interlocked molecules.