Transcriptome analysis to identify the downstream genes of androgen receptor in dermal papilla cells.

BACKGROUND: Testosterone signaling mediates various diseases, such as androgenetic alopecia and prostate cancer. Testosterone signaling is mediated by the androgen receptor (AR). In this study, we fortuitously found that primary and immortalized dermal papilla cells suppressed AR expression, although dermal papilla cells express AR in vivo. To analyze the AR signaling pathway, we exogenously introduced the AR gene via a retrovirus into immortalized dermal papilla cells and comprehensively compared their expression profiles with and without AR expression. RESULTS: Whole-transcriptome profiling revealed that the focal adhesion pathway was mainly affected by the activation of AR signaling. In particular, we found that caveolin-1 gene expression was downregulated in AR-expressing cells, suggesting that caveolin-1 is controlled by AR. CONCLUSION: Our whole transcriptome data is critical resources for discovery of new therapeutic targets for testosterone-related diseases.

Detailed chromosome analysis of wild-type, immortalized fibroblasts with SV40T, E6E7, combinational introduction of cyclin dependent kinase 4, cyclin D1, telomerase reverse transcriptase.

Cell immortalization enables us to expand the cultured cell infinitely. However, the process of immortalization sometimes changes the nature of the original cell. In this study, we established immortalized embryonic fibroblasts with oncogenic SV40T and human papilla virus-derived E6E7, combinational expression of mutant cyclin-dependent kinase 4 (CDK4), cyclin D1, and telomerase reverse transcriptase (TERT) from identical primary wild-type human embryonic fibroblasts (HE16). After the establishment of immortalized cells, we compared the details of chromosome condition with the G-banding and Q-banding methods. There is no example of detailed analysis so far about chromosome abnormalities, such as trisomy, ring chromosome, reciprocal translocation, and dicentric chromosomes. The detailed chromosome analysis revealed that immortalized cells with SV40T and E6E7 showed intensive chromosome abnormalities, such as gain or loss of the chromosomes all through the genome. Furthermore, we detected that the incidence of chromosome abnormities in the immortalized cell with the combinational introduction of R24C mutant of CDK4, cyclin D1, and TERT is almost identical to that of wild-type cell. Furthermore, short tandem repeat analysis demonstrated that the origin of K4DT cell is primary HE16. These results showed that cellular immortalization with CDK4, cyclin D1, and TERT is more advantageous in keeping the chromosome's original condition than oncogenic immortalization methods.

Factors Associated with Insomnia Among Truck Drivers in Japan.

PURPOSE: Truck drivers with insomnia are at a high risk of traffic accidents. We aimed to investigate the prevalence of insomnia and identify its associated factors among truck drivers in Japan. METHODS: Participants were 2927 male truck drivers younger than 65 years old. Self-administered questionnaires were used to assess insomnia symptoms, State-Trait Anxiety Inventory, drinking, smoking habits, body mass index, caffeine intake, as well as daily driving hours, consecutive days away from home, and driving distance. Insomnia symptoms included difficulty initiating sleep, maintaining sleep and early morning awakening. Insomnia was defined when any of these symptoms were observed with daily tiredness. RESULTS: The prevalence of insomnia among the subjects was 13.3% (n=356), of which 13.5% had difficulty initiating sleep, 78% had difficulty maintaining sleep, and 26.4% had early morning awakening. After adjusting for covariates, drinking habits, daily driving hours, and STAI score were significantly and linearly associated with insomnia; the adjusted odds ratio (OR) of drinking habits for insomnia was 1.74 [95% confidence interval (CI), 1.23-2.47] for heavy drinkers compared to non-drinkers (trend p<0.001); the adjusted OR of daily driving hours was 1.87 (95% CI, 1.00-3.49) for 12 hours or longer in a day compared to <8 hours in a day (trend p<0.001); the adjusted OR of STAI quartiles was 5.30 (95% CI, 3.66-7.67) for the highest quartile compared to the lowest quartile (trend p<0.001). CONCLUSION: The present study demonstrated that insomnia is prevalent among truck drivers in Japan, and its risk factors include drinking habits, daily driving hours, and anxiety.

Usefulness of serial lung ultrasound for a severe COVID-19 patient on extracorporeal membrane oxygenation.

Computed tomography (CT) is the most reliable method to evaluate the progression of COVID-19 pneumonitis. However, in a pandemic, transportation of critically ill invasively ventilated patients to radiology facilities is challenging, especially for those on extracorporeal membrane oxygenation (ECMO). Notably, lung ultrasound (LUS) is a favored alternative imaging modality due to its ease of use at the point of care, which reduces the infectious risk of exposure and transmission; repeatability; absence of radiation exposure; and low cost. We demonstrated that serial LUS compares favorably with other imaging modalities in terms of usefulness for evaluating lung aeration and recovery in an ECMO-managed COVID-19 patient.

Combinatorial expression of cell cycle regulators is more suitable for immortalization than oncogenic methods in dermal papilla cells.

The immortalized cell is an essential research tool that uses robust growth properties for the functional investigation of gene products. Immortalized mammalian cells have mainly been established using three methods: expression of simian vacuolating virus 40 T antigen (the SV40 method); human papilloma virus-derived oncoprotein E6/E7 (the E6/E7 method); or combinatorial expression of R24C mutant cyclin-dependent kinase 4, cyclin D1, and telomerase reverse transcriptase (the K4DT method). However, it is unclear as to which method is optimal for an in vitro model. Here, we compared the biological characteristics and genome-wide expression profiles of immortalized human dermal papilla cells generated by the SV40, E6/E7, or K4DT method. To our knowledge, this is the first study to comprehensively compare expression profiles to determine the optimal immortalization method for maintaining the original nature of the wild-type cells. These data would be valuable to scientists aiming to establish new immortalized cell lines.

Ultimate suppression of thermal transport in amorphous silicon nitride by phononic nanostructure.

Engineering the thermal conductivity of amorphous materials is highly essential for the thermal management of future electronic devices. Here, we demonstrate the impact of ultrafine nanostructuring on the thermal conductivity reduction of amorphous silicon nitride (a-Si3N4) thin films, in which the thermal transport is inherently impeded by the atomic disorders. Ultrafine nanostructuring with feature sizes below 20 nm allows us to fully suppress contribution of the propagating vibrational modes (propagons), leaving only the diffusive vibrational modes (diffusons) to contribute to thermal transport in a-Si3N4 A combination of the phonon-gas kinetics model and the Allen-Feldmann theory reproduced the measured results without any fitting parameters. The thermal conductivity reduction was explained as extremely strong diffusive boundary scattering of both propagons and diffusons. These findings give rise to substantial tunability of thermal conductivity of amorphous materials, which enables us to provide better thermal solutions in microelectronic devices.

Enhanced Reduction of Thermal Conductivity in Amorphous Silicon Nitride-Containing Phononic Crystals Fabricated Using Directed Self-Assembly of Block Copolymers.

Studies have demonstrated that the thermal conductivity (κ) of crystalline semiconductor materials can be reduced by phonon scattering in periodic nanostructures formed using templates fabricated from self-assembled block copolymers (BCPs). Compared to crystalline materials, the heat transport mechanisms in amorphous inorganic materials differ significantly and have been explored far less extensively. However, thermal management of amorphous inorganic solids is crucial for a broad range of semiconductor devices. Here we present the fabrication of freestanding amorphous silicon nitride (SiNx) membranes for studying κ in an amorphous solid. To form a periodic nanostructure, directed self-assembly of cylinder-forming BCPs is used to pattern in the SiNx highly ordered, hexagonally close packed nanopores with pitch and neck width down to 37.5 and 12 nm, respectively. The κ of the nanoporous SiNx membranes is 60% smaller than the classically predicted value based on just the membrane porosity. In comparison, holes with much larger neck widths and pitches patterned by e-beam lithography lead to only a slight reduction in κ, which is closer to the classical porosity-based prediction. These results demonstrate that κ of amorphous SiNx can be reduced by introducing periodic nanostructures that behave as a phononic crystal, where the relationship between the smallest dimension of the nanostructure and the length scale of the mean-free paths of the dominant, heat-carrying phonons is critical. Additionally, changing the orientation of the hexagonal array of nanopores relative to the primary direction of heat flow has a smaller impact on amorphous SiNx than was previously observed in silicon.

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor.

Androgenetic alopecia (AGA) is the most common type of hair loss, and is mainly caused by the biological effects of testosterone on dermal papilla cells (DPCs). In vitro culturing of DPCs might be a useful tool for the screening of target molecule of AGA. However, primary DPCs cannot continuously proliferate owing to cellular senescence and cell culture stress. In this study, we introduced mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomerase reverse transcriptase (TERT) into DPCs. We confirmed protein expression of CDK4 and Cyclin D1, and enzymatic activity of TERT. Furthermore, we found the established cell line was free from cellular senescence. We also introduced the androgen receptor gene using a recombinant retrovirus, to compensate the transcriptional suppressed endogenous androgen receptor in the process of cell proliferation. Furthermore, we detected the efficient nuclear translocation of androgen receptor into the nucleus after the treatment of dihydrotestosterone, indicating the functionality of our introduced receptor. Our established cell line is a useful tool to identify the downstream signaling pathway, which activated by the testosterone.

Mechanical Thrombectomy for Postoperative Stroke in a Patient with Acute Aortic Dissection Type A.

Mechanical thrombectomy has been proposed to expand the treatment time window and enhance revascularization. However, it is unclear whether its use can be extended to patients with occlusions in acute aortic dissection, especially the thoracic aorta. A 55-year-old man underwent graft replacement for acute aortic dissection type A. On postoperative day 2, he developed stroke and computed tomography showed occlusion of the right middle cerebral artery. Mechanical thrombectomy was performed by transbrachial approach. Although successful recanalization was achieved, he suffered hemorrhagic stroke. Since there is no other effective treatment and the neurologic outcome with conservative management is poor, we consider mechanical thrombectomy to be a viable therapeutic option for the treatment of postoperative stroke in patients with acute aortic dissection type A. However, further study is warranted regarding the safety of this technique.

Global transcriptome analysis of pig induced pluripotent stem cells derived from six and four reprogramming factors.

Pigs are important, both for agriculture and as animal models for human diseases. However, due to the lack of embryonic stem cells, the possibility of genetic modification is quite limited. To overcome this limitation, induced pluripotent stem (iPS) cells have been derived from pigs. Despite the public availability of a large number of expression datasets from mice, rats, and primates-derived iPS cells, the expression profile of pig-derived iPS cells is quite limited. Furthermore, there is no dataset focused on the profiling of pig-derived iPS cell with six reprogramming factors (Oct3/4, Sox2, Klf4, c-Myc, Lin28, and Nanog). Here, we used Illumina RNA sequencing platform to characterize the mRNA expression of four-factor derived and six-factor derived pig iPS cells. We observed that the expression levels of whole genes in our established six factors derived iPS cells and parent fibroblast, and compared with that of iPS cells with four factors in public database. These data are valuable in understanding species difference in the reprogramming process of stem cells, and could help identify the key regulating genes involved in the process.

Efficient immortalization of cells derived from critically endangered Tsushima leopard cat (Prionailurus bengalensis euptilurus) with expression of mutant CDK4, Cyclin D1, and telomerase reverse transcriptase.

Tsushima leopard cat is the subspecies of Amur cats, and it is classified as the most highest class of critically endangered animals. Although the protection activity is highly recognized, the number of animals is decreasing due to the human activity and invasion of domestic cats and infectious disease. In this study, we succeeded primary culture of normal fibroblasts derived from the Tsushima leopard cat (Prionailurus bengalensis euptilurus). Furthermore, we introduced the human derived mutant Cyclin Dependent Kinase 4, Cyclin D1, and telomere reverse transcriptase. We showed that the expression of these three genes efficiently immortalized cells derived from Tsushima leopard cat. Furthermore, we showed that the chromosome pattern of the established cells is identical with the original one. These data indicate that our method of immortalization is useful to establish cell lines from critically endangered cats, which potentially contributes to the re-generation of critically endangered animals from cultured cell with reproductive technique, such as somatic cloning.

[Successful Repair of Chronic Expanding Hematoma after Coronary Artery Bypass Grafting by Lower Partial Sternotomy Approach;Report of a Case].

Chronic expanding hematoma (CEH) after coronary artery bypass grafting (CABG) is extremely rare. We reported a case of successful surgical repair of CEH through lower partial sternotomy approach. The patient was a 73-year-old man who underwent CABG. Fifteen years after the operation, he was admitted to referral hospital because of dyspnea on exertion. Echocardiography and computed tomography revealed a giant intrapericardial mass, which was pressing on the inferior wall of the ventricle. To avoid injury of the aorta and the patent bypass graft in the reentry of the chest, we selected lower partial sternotomy approach. The mass was encapsulated with thick fibrous membrane containing old degenerated coagula and was histopathologically diagnosed as CEH. Postoperative course was uneventful and diastolic heart failure was relieved.

[Successful Emergency Pulmonary Embolectomy in a Patient with Shock；Report of a Case].

The patient was a 48-year-old man, who had hemodynamic shock after orthopedic surgery. He was then diagnosed with right pulmonary artery embolism. Circulatory collapse remained, despite cardiac resuscitation. Therefore, he was treated with percutaneous cardiopulmonary support (PCPS) and intra-aortic balloon pumping (IABP). He was then transferred to our hospital and emergency pulmonary embolectomy was performed under cardiopulmonary bypass. The postoperative course was uneventful, and he was discharged without complications. Early diagnosis and early placement of PCPS and IABP allows safe transfer of patients and successful performance of emergency pulmonary embolectomy.

Breaking the trade-off between thermal and electrical conductivities in the thermoelectric material of an artificially tilted multilayer.

Breaking the trade-off between thermoelectric (TE) parameters has long been demanded in order to highly enhance its performance. Here, we report the 'trade-off-free' interdependence between thermal conductivity (κ) and resistivity (ρ) in a TE/metal tilted multilayer and significant enhancement of TE power generation based on the off-diagonal thermoelectric (ODTE) effect, which generates transverse electrical current in response to vertical thermal current. ρ and κ can be simultaneously decreased by setting charge flow along more-electrically conductive layer and thermal flow across less-thermally conductive perpendicular direction by decreasing the tilting angle. Moreover, introducing porosity in the metal layer enables to decrease in κ without changing ρ, because the macroscopic ρ and κ of the tilted multilayer is respectively governed by the properties of the TE material and the metal with large dissimilarity. The obtained results reveal new strategies for developing trade-off-free TE materials, which will stimulate practical use of TE conversion for waste-heat recovery.

Hybrid thoracic endovascular aortic repair of ascending aortic pseudoaneurysm.

We report the successful thoracic endovascular aortic repair of an ascending aortic pseudoaneurysm. Because the risk of resternotomy after conventional surgery was very high, endovascular repair with extra-anatomic reconstruction of the supra-aortic vessels was performed using the femoral artery as the donor artery. Complete exclusion of the pseudoaneurysm was achieved, with no postoperative complications or neurologic sequelae.

Bifunctional thermoelectric tube made of tilted multilayer material as an alternative to standard heat exchangers.

Enormously large amount of heat produced by human activities is now mostly wasted into the environment without use. To realize a sustainable society, it is important to develop practical solutions for waste heat recovery. Here, we demonstrate that a tubular thermoelectric device made of tilted multilayer of Bi(0.5)Sb(1.5)Te3/Ni provides a promising solution. The Bi(0.5)Sb(1.5)Te3/Ni tube allows tightly sealed fluid flow inside itself, and operates in analogy with the standard shell and tube heat exchanger. We show that it achieves perfect balance between efficient heat exchange and high-power generation with a heat transfer coefficient of 4.0 kW/m(2)K and a volume power density of 10 kW/m(3) using low-grade heat sources below 100°C. The Bi(0.5)Sb(1.5)Te3/Ni tube thus serves as a power generator and a heat exchanger within a single unit, which is advantageous for developing new cogeneration systems in factories, vessels, and automobiles where cooling of excess heat is routinely carried out.

[The number of cardiovascular surgeons is decreasing in Nagano].

Shortage of medical doctors has been conspicuous since the residency program has been changed in 2004. We investigated the changes in the number of cardiovascular surgeons, surgical cases and institutes of Nagano from 1996 to 2008. The number of cardiovascular surgeons decreased from 28 to 19. The number of newly joined cardiovascular surgeons were 2.7 +/- 1.4/year from 1996 to 2001 and 1.1 +/- 0.4/year from 2002 to 2008. The number of retirement were 0.7 +/- 0.8/year and 2.3 +/- 1.0/year from 1996 to 2001 and from 2002 to 2008, respectively. The number of surgery cases increased from 340 in 1996 to 771 in 2008. Kaplan-Mayer analysis showed the 50% of cardiovascular surgeons retired in 19 years of their career, and the only 20% of surgeons survived after 29 years. The number of newly joined general surgeons was the only 3 since 2004. We are consolidating the institutes of cardiovascular surgery in Nagano to perform more surgery by fewer surgeons. It is important that working condition of cardiovascular surgeon adapt to young doctor's favor, and new positions should be created for elder cardiovascular surgeons to prevent early retirement.

Multifrequency high-speed phase-modulation atomic force microscopy in liquids.

We have developed a new technique, called multifrequency high-speed phase-modulation atomic force microscopy (PM-AFM) in constant-amplitude (CA) mode based on the simultaneous excitation of the first two flexural modes of a cantilever. By performing a theoretical investigation, we have found that this technique enables the simultaneous imaging of the surface topography, energy dissipation and elasticity (nonlinear mapping) of materials. We experimentally demonstrated high-speed imaging at a scan speed of 5 frames/s for a polystyrene (PS) and polyisobutylene (PIB) polymer-blend thin-film surface in water.

Short-term effects of dietary fat on intramyocellular lipid in sprinters and endurance runners.

The effect of short-term fat loading on intramyocellular lipid (IMCL) in different types of muscle in endurance runners and sprinters has not been fully elucidated yet. The purpose of this study was to investigate the effect of dietary lipid on IMCL in soleus muscle (SOL) and tibialis anterior muscle (TA) during training period in endurance runners and sprinters. Seven male endurance runners and 7 male sprinters were selected to participate in the study. We measured TA- and SOL-IMCL levels after 3-day course of isocaloric normal- (25%), high- (60%), and low-fat (10%) diet during training period by (1)H-magnetic resonance spectroscopy in each subject. In sprinters, TA- and SOL-IMCL levels were comparable after each diet protocol. However, in endurance runners, TA-IMCL levels after normal-fat and high-fat diets were 1.7 times and 3.0 times higher than that after low-fat diet, respectively. The SOL-IMCL values after normal-fat diet and high-fat diet were 1.5 times and 1.6 times higher than that after low-fat diet, respectively. In addition, the TA-IMCL level after high-fat diet, but not SOL-IMCL, was significantly higher compared with that after normal-fat diet. Our data suggested that short-term dietary fat challenge during training period significantly altered IMCL level in endurance runners, but not in sprinters. In addition, response to fat loading on IMCL was influenced by variation of muscle type in endurance runners. These phenotypic and regional differences might be explained by differences in type of exercise training and muscle fiber composition.

Terahertz radiation by an ultrafast spontaneous polarization modulation of multiferroic BiFeO3 thin films.

Terahertz (THz) radiation has been observed from multiferroic BiFeO3 thin films via ultrafast modulation of spontaneous polarization upon carrier excitation with illumination of femtosecond laser pulses. The radiated THz pulses from BiFeO3 thin films were clarified to directly reflect the spontaneous polarization state, giving rise to a memory effect in a unique style and enabling THz radiation even at zero-bias electric field. On the basis of our findings, we demonstrate potential approaches to ferroelectric nonvolatile random access memory with nondestructive readability and ferroelectric domain imaging microscopy using THz radiation as a sensitive probe.