Cholesterol accumulation in ovarian follicles causes ovulation defects in Abca1a⁻/⁻ Japanese medaka (Oryzias latipes)

ATP-binding cassette A1 (ABCA1) is a membrane protein, which exports excess cellular cholesterol to generate HDL to reduce the risk of the onset of cardiovascular diseases (CVD). In addition, ABCA1 exerts pleiotropic effects on such as inflammation, tissue repair, and cell proliferation and migration. In this study, we explored the novel physiological roles of ABCA1 using Japanese medaka (Oryzias latipes), a small teleost fish. Three Abca1 genes were found in the medaka genome. ABCA1A and ABCA1C exported cholesterol to generate nascent HDL as human ABCA1 when expressed in HEK293 cells. To investigate their physiological roles, each Abca1-deficient fish was generated using the CRISPR-Cas9 system. Abca1a−/− female medaka was found to be infertile, while Abca1b−/− and Abca1c−/− female medaka were fertile. In vitro ovarian follicle culture suggested that Abca1a deficiency causes ovulation defects. In the ovary, ABCA1A was expressed in theca cells, an outermost layer of the ovarian follicle. Total cholesterol content of Abca1a−/− ovary was significantly higher than that of the wild-type, while estrogen and progestin contents were compatible with those of the wild-type. Furthermore, cholesterol loading to the wild-type follicles caused ovulation defects. These results suggest that ABCA1A in theca cells regulates cholesterol content in the ovarian follicles and its deficiency inhibits successful ovulation through cholesterol accumulation in the ovarian follicle

A novel A792D mutation in the CSF1R gene causes hereditary diffuse leukoencephalopathy with axonal spheroids characterized by slow progression

Hereditary diffuse leukoencephalopathy with spheroids (HDLS) is an autosomal dominant white matter disease that causes adult-onset cognitive impairment. The clinical manifestations are a variable combination of personality and behavioral changes, cognitive decline, parkinsonism, spasticity, and epilepsy. In 2012, mutations in the gene encoding colony stimulating factor 1 receptor (CSF1R) were identified as the cause of HDLS. As the numbers of reported mutations are limited, the understanding of whole pathogenesis needs accumulation of disease-causing mutations with detailed clinical descriptions. We describe a Japanese family with autosomal dominant adult-onset cognitive impairment and characteristic white matter lesions. Genetic testing revealed a novel p.A792D mutation in the tyrosine kinase domain of CSF1R in two affected family members. The symptom profile of the present cases mostly matched the previously reported cases, with the notable exceptions of late-onset and long disease duration

In vivo FRET analyses reveal a role of ATP hydrolysis–associated conformational changes in human P-glycoprotein

P-glycoprotein (P-gp; also known as MDR1 or ABCB1) is an ATP-driven multidrug transporter that extrudes various hydrophobic toxic compounds to the extracellular space. P-gp consists of two transmembrane domains (TMDs) that form the substrate translocation pathway and two nucleotide-binding domains (NBDs) that bind and hydrolyze ATP. At least two P-gp states are required for transport. In the inward-facing (pre-drug transport) conformation, the two NBDs are separated, and the two TMDs are open to the intracellular side; in the outward-facing (post-drug transport) conformation, the NBDs are dimerized, and the TMDs are slightly open to the extracellular side. ATP binding and hydrolysis cause conformational changes between the inward-facing and the outward-facing conformations, and these changes help translocate substrates across the membrane. However, how ATP hydrolysis is coupled to these conformational changes remains unclear. In this study, we used a new FRET sensor that detects conformational changes in P-gp to investigate the role of ATP binding and hydrolysis during the conformational changes of human P-gp in living HEK293 cells. We show that ATP binding causes the conformational change to the outward-facing state and that ATP hydrolysis and subsequent release of γ-phosphate from both NBDs allow the outward-facing state to return to the original inward-facing state. The findings of our study underscore the utility of using FRET analysis in living cells to elucidate the function of membrane proteins such as multidrug transporters

Sexual Experience Induces the Expression of Gastrin-Releasing Peptide and Oxytocin Receptors in the Spinal Ejaculation Generator in Rats

Male sexual function in mammals is controlled by the brain neural circuits and the spinal cord centers located in the lamina X of the lumbar spinal cord (L3-L4). Recently, we reported that hypothalamic oxytocin neurons project to the lumbar spinal cord to activate the neurons located in the dorsal lamina X of the lumbar spinal cord (dXL) via oxytocin receptors, thereby facilitating male sexual activity. Sexual experiences can influence male sexual activity in rats. However, how this experience affects the brain-spinal cord neural circuits underlying male sexual activity remains unknown. Focusing on dXL neurons that are innervated by hypothalamic oxytocinergic neurons controlling male sexual function, we examined whether sexual experience affects such neural circuits. We found that >50% of dXL neurons were activated in the first ejaculation group and similar to 30% in the control and intromission groups in sexually naive males. In contrast, in sexually experienced males, similar to 50% of dXL neurons were activated in both the intromission and ejaculation groups, compared to similar to 30% in the control group. Furthermore, sexual experience induced expressions of gastrin-releasing peptide and oxytocin receptors in the lumbar spinal cord. This is the first demonstration of the effects of sexual experience on molecular expressions in the neural circuits controlling male sexual activity in the spinal cord

Deuterium Isotope Separation by Combined Electrolysis Fuel Cell

The framework about combined electrolysis fuel cell (CEFC) was reported previously [H. Matsushima et al., Energy, 2005; 30; 2413]. The purpose of the present study focused on measuring the separation factor and the energy reduction by assembling CEFC system. The separation of deuterium was studied with a 1-M KOH electrolyte containing 10 at% deuterium. Polarization plots of alkaline water electrolysis (AWE) revealed relationships between the catalytic activity of the hydrogen evolution reaction and the deuterium separation factor. The power loss was mainly attributed to gas bubble evolution. For polymer electrolyte fuel cells (PEFCs) with a Pt catalyst, approximately 21% of the electrical energy could be recovered by reusing hydrogen gas produced by the AWE. Furthermore, the PEFC could efficiently dilute protium in the gas phase, resulting in a high separation factor of 30.2 for the CEFC

Influence of unique layered microstructure on fatigue properties of Ti-48Al-2Cr-2Nb alloys fabricated by electron beam melting

The influence of a unique layered microstructure consisting of duplex-like region and equiaxed γ grain layers (γ bands) on the fatigue properties of Ti-48Al-2Cr-2Nb alloy bars fabricated by electron beam melting (EBM) was investigated at room temperature (RT) and 1023 K focusing on the angle (θ) between the building direction and cylinder (loading) axis. We found for the first time the fatigue strengths of the alloy bars with the layered microstructure depend strongly on the angle θ. Particularly, the fatigue strength of the alloy bars fabricated at θ = 45° is comparable to that of the hot isostatic pressing (HIP) treated cast alloys, even without HIP treatment. We also found the alloy bars fabricated at θ = 0° and 45° exhibit high fatigue strengths in the low-cycle fatigue life region at 1023 K similar to θ = 45° alloy bars at RT. These high fatigue strengths are caused by inhibition of the brittle main crack initiation by stress relaxation due to shear deformation at the γ bands and large plasticity of the alloys. These findings indicate that the alloys fabricated by EBM at θ = 45° with the unique layered microstructure have a great potential for aerospace and automobile applications.Cho K., Kobayashi R., Oh J.Y., et al. Influence of unique layered microstructure on fatigue properties of Ti-48Al-2Cr-2Nb alloys fabricated by electron beam melting. Intermetallics 95, 1 (2018); https://doi.org/https://doi.org/10.1016/j.intermet.2018.01.009

Diagnostic value of computed high b-value whole-body diffusion-weighted imaging for primary prostate cancer

Purpose: To investigate the utility of post-acquisition computed diffusion-weighted imaging (cDWI) for primary prostate cancer (PCa) evaluation in biparametric whole-body MRI (bpWB-MRI). Methods: Patients who underwent pelvic MRI for PCa screening and subsequent bpWB-MRI for staging were included. Two radiologists assessed the diagnostic performance of the following datasets for clinically significant PCa diagnosis (grade group >= 2 according to the Prostate Imaging-Reporting and Data System, version 2.1): bpMRI(2000) (axial DWI scans with a b-value of 2,000 s/mm(2) + axial T2WI scans from pre-biopsy pelvic MRI), computed bpWB-MRI2000 (computed WB-DWI scans with a b-value of 2,000 s/mm(2) + axial WB-T2WI scans), and native bpWB-MRI1000 (native axial WB-DWI scans with a b-value of 1,000 s/mm(2) + axial WB-T2WI scans). Systemic biopsy was used as reference standard. Results: Fifty-one patients with PCa were included. The areas under the curve (AUCs) of bpMRI(2000) (0.89 for reader 1 and 0.86 for reader 2) and computed bpWB-MRI2000 (0.86 for reader 1 and 0.83 for reader 2) were significantly higher (p < 0.001) than those of native bpWB-MRI1000 (0.67 for both readers). No significant difference was observed between the AUCs of bpMRI(2000) and computed bpWB-MRI2000 (p = 0.10 for reader 1 and p = 0.25 for reader 2). Conclusions: The diagnostic performance of computed bpWB-MRI2000 was similar to that of dedicated pelvic bpMRI(2000) for primary PCa evaluation. cDWI can be recommended for implementation in standard WB-MRI protocols to facilitate a one-step evaluation for concurrent detection of primary and metastatic PCa

Oxytocin Influences Male Sexual Activity via Non-synaptic Axonal Release in the Spinal Cord

Oxytocinergic neurons in the paraventricular nucleus of the hypothalamus that project to extrahypothalamic brain areas and the lumbar spinal cord play an important role in the control of erectile function and male sexual behavior in mammals. The gastrin-releasing peptide (GRP) system in the lumbosacral spinal cord is an important component of the neural circuits that control penile reflexes in rats, circuits that are commonly referred to as the “spinal ejaculation generator (SEG).” We have examined the functional interaction between the SEG neurons and the hypothalamo-spinal oxytocin system in rats. Here, we show that SEG/GRP neurons express oxytocin receptors and are activated by oxytocin during male sexual behavior. Intrathecal injection of oxytocin receptor antagonist not only attenuates ejaculation but also affects pre-ejaculatory behavior during normal sexual activity. Electron microscopy of potassium-stimulated acute slices of the lumbar cord showed that oxytocin-neurophysin-immunoreactivity was detected in large numbers of neurosecretory dense-cored vesicles, many of which are located close to the plasmalemma of axonal varicosities in which no electron-lucent microvesicles or synaptic membrane thickenings were visible. These results suggested that, in rats, release of oxytocin in the lumbar spinal cord is not limited to conventional synapses but occurs by exocytosis of the dense-cored vesicles from axonal varicosities and acts by diffusion—a localized volume transmission—to reach oxytocin receptors on GRP neurons and facilitate male sexual function

Clinical utility of the Bosniak classification version 2019:Diagnostic value of adding magnetic resonance imaging to computed tomography examination

Purpose: To assess the impact of the updated Bosniak classification (BC2019) for cystic renal masses (CRMs) on interobserver agreement between radiologists and urologists and the diagnostic value of adding MRI to CT examination (combined CT/MRI). Method: This study included 103 CRMs from 83 consecutive patients assessed using contrast-enhanced CT and MRI between 2010 and 2016. Nine readers in three groups (three radiologists, three radiology residents, and three urologists) reviewed CT alone and the combined CT/MRI using BC2019. Bosniak category was determined by consensus in each group for diagnosing malignancy, with a cut-off category of ?>= III. Interobserver agreement was assessed using Fleiss' kappa values. The effect of CT or combined CT/MRI on the diagnosis of malignancy was assessed using McNemar's test. Results: Interobserver agreement of BC2019 for CT alone was substantial for radiologists and residents, moderate for urologists (0.77, 0.63, and 0.58, respectively). Interobserver agreement of BC2019 for combined CT/MRI was substantial for all three groups (radiologists: 0.78; residents: 0.65; and urologists: 0.61). Among residents, the sensitivity/specificity/accuracy rates of combined CT/MRI vs. CT alone were 82.1/74.7/76.7% vs. 75.0/66.7/68.9%, and specificity and accuracy were significantly higher for combined CT/MRI than that for CT alone (p = 0.03 and 0.008, respectively). Similarly, sensitivity/specificity/accuracy values were significantly higher for combined CT/MRI among urologists (78.6/73.3/74.8% vs. 64.3/64.0/64.1%, p = 0.04/0.04/0.008). However, sensitivity/specificity/accuracy did not significantly differ between the two among radiologists (89.3/74.7/78.6% vs. 85.7/73.3/76.7%, p = 0.32/0.56/0.32). Conclusions: Combined CT/MRI is useful for diagnosing malignancy in patients with CRMs using BC2019, especially for non-expert readers