Ca2+ Depletion in the ER Causes Store-Operated Ca2+ Entry via the TRPC6 Channel in Mouse Brown Adipocytes.

beta3-adrenergic activation causes Ca2+ release from the mitochondria and subsequent Ca2+ release from the endoplasmic reticulum (ER), evoking store-operated Ca2+ entry (SOCE) due to Ca2+ depletion from the ER in mouse brown adipocytes. In this study, we investigated how Ca2+ depletion from the ER elicits SOCE in mouse brown adipocytes using fluorometry of intracellular Ca2+ concentration ([Ca2+]i). The administration of cyclopiazonic acid (CPA), a reversible sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) pump blocker in the ER, caused an increase in [Ca2+]i. Moreover, CPA induced SOCE was suppressed by the administration of a Ca2+ free Krebs solution and the transient receptor potential canonical 6 (TRPC6) selective blockers 2-APB, ML-9 and GsMTx-4 but not Pico145, which blocks TRPC1/4/5. Administration of TRPC6 channel agonist 1-oleoyl-2-acetyl-sn-glycerol (OAG) and flufenamic acid elicited Ca2+ entry. Moreover, our RT-PCR analyses detected mRNAs for TRPC6 in brown adipose tissues. In addition, western blot analyses showed the expression of the TRPC6 protein. Thus, TRPC6 is one of the Ca2+ pathways involved in SOCE. These modes of Ca2+ entry provide the basis for heat production via activation of Ca2+-dependent dehydrogenase and the expression of uncoupling protein 1 (UCP1). Enhancing thermogenic metabolism in brown adipocytes may serve as broad therapeutic utility to reduce obesity and metabolic syndrome.

Rectangular tapered short stem excellently preserves proximal bone mineral density preservation than tapered wedge short stem.

Fitmore stem is a rectangular, tapered, short, cementless stem. A characteristic feature of this stem is that it provides rotational stability due to the high medullary occupancy achieved by its rectangular cross-section and thick antero- posterior width. We aimed to investigate the differences in periprosthetic bone remodelling between a rectangular- tapered short stem and a short tapered-wedge stem. Eighty patients who underwent primary total hip arthroplasty using a rectangular-tapered short stem (Fitmore) or a short tapered-wedge stem (Tri-Lock BPS) were enrolled in this study. Bone mineral densities (BMDs) in the seven Gruen zones were evaluated using dual-energy X-ray absorptiometry at baseline, and at 6 and 24 months postoperatively. Peri-prosthetic BMD and clinical factors were assessed and compared. In addition, correlations between periprosthetic BMD changes and stem anteversion error were analyzed using Pearson's correlation coefficient in the two groups. A significantly better postoperative periprosthetic BMD change was found in zones 1 and 7 in the rectangular-tapered group. Additionally, no significant correlation was observed between stem anteversion error and periprosthetic BMD changes in the rectangular-tapered groups. However, in the tapered-wedge group, there were significant negative correlations between the stem anteversion error and BMD changes at 6 months and 24 months in zones 1 and 7. In the rectangular-tapered group, a significantly better postoperative periprosthetic BMD change was found particularly in the region proximal to the stem. Rectangular-tapered short stem can be more resistant to rotation due to higher medullary occupancy and may lead to better periprosthetic BMD than the tapered-wedge short stem, especially in the proximal region of the stem.

Lipopolysaccharide structure modulates cationic biocide susceptibility and crystalline biofilm formation in Proteus mirabilis.

Chlorhexidine (CHD) is a cationic biocide used ubiquitously in healthcare settings. Proteus mirabilis, an important pathogen of the catheterized urinary tract, and isolates of this species are often described as "resistant" to CHD-containing products used for catheter infection control. To identify the mechanisms underlying reduced CHD susceptibility in P. mirabilis, we subjected the CHD tolerant clinical isolate RS47 to random transposon mutagenesis and screened for mutants with reduced CHD minimum inhibitory concentrations (MICs). One mutant recovered from these screens (designated RS47-2) exhibited ~ 8-fold reduction in CHD MIC. Complete genome sequencing of RS47-2 showed a single mini-Tn5 insert in the waaC gene involved in lipopolysaccharide (LPS) inner core biosynthesis. Phenotypic screening of RS47-2 revealed a significant increase in cell surface hydrophobicity and serum susceptibility compared to the wildtype, and confirmed defects in LPS production congruent with waaC inactivation. Disruption of waaC was also associated with increased susceptibility to a range of other cationic biocides but did not affect susceptibility to antibiotics tested. Complementation studies showed that repression of smvA efflux activity in RS47-2 further increased susceptibility to CHD and other cationic biocides, reducing CHD MICs to values comparable with the most CHD susceptible isolates characterized. The formation of crystalline biofilms and blockage of urethral catheters was also significantly attenuated in RS47-2. Taken together, these data show that aspects of LPS structure and upregulation of the smvA efflux system function in synergy to modulate susceptibility to CHD and other cationic biocides, and that LPS structure is also an important factor in P. mirabilis crystalline biofilm formation.

Laboratory studies assessing the microbial biocontrol potential of diverse strains of Beauveria bassiana isolated from coffee berry borer, with emphasis on strains from Hawai'i Island and comparisons to commercial strain GHA.

A series of experiments investigated genetically diverse strains of Beauveria bassiana (Bb) isolated from coffee berry borer (CBB). Objectives included assessment of their biocontrol potential, particularly in comparison to Bb commercial strain GHA currently applied for CBB control, and identification of various attributes potentially contributing to their comparatively greater epizootic potential in CBB populations. Bioassays identified one strain from Hawai'i Island and one from Puerto Rico with virulence greater than GHA based on equal weights of unformulated conidial powder (CP); however, the greater potency of the CPs was ultimately explained by their 2.4-fold greater conidial densities (ca. 3.1 vs 1.3 × 1011 viable conidia/g CP). Density was explained, in large part, by conidial size, but not by size alone. Bb-inoculated CBB held on moist filter paper were more susceptible to infection than those held on cooked green coffee bean (CGCB). A Bb strain representative of the most common Hawaiian haplotype produced 2.6x more conidia after death of CGCB-held beetles than GHA (19.1 vs 7.3 x106 conidia/cadaver). Following host death, no difference was observed in time to emergence and initial conidial production by GHA and a selected group of Hawaiian strains; however, mass sporulation was initiated within 2 days by strain GHA compared to 4-5 days by the Hawaiian strains. In a preliminary evaluation of conidial mass-production potential, CP yields of several strains were comparable to GHA on a weight basis and significantly greater than GHA on a conidial basis (1.3-1.6 vs 0.7 × 1013 viable conidia/kg barley substrate).

Pre-operative endovascular coil embolisation for chronic pulmonary aspergillosis.

OBJECTIVE: To retrospectively evaluate the clinical outcomes of pre-operative endovascular coil embolisation (ECE) for chronic pulmonary aspergillosis (CPA).METHODS: We evaluated surgical patients with CPA between November 2016 and April 2020. Pre-operative ECE for CPA with severe adhesions was selectively performed to reduce intra-operative blood loss. ECE procedures, operative procedures, intra-operative blood loss and complications were evaluated.RESULTS: Twenty-eight patients (21 males and 7 females; median age: 55 years) were included in the study. Of the 28 patients, 8 (28.6%) underwent pre-operative ECE. Technical success rate in pre-operative ECE was 100%. The median time required for ECE procedures was 123 min. The median number of vessels embolised per procedure was 2.5. The median period between embolisation and surgery was 5 days. Major complications were observed in three patients (10.7%). There were no significant differences between patients with and without pre-operative ECE in operative time (284 vs. 365 min, respectively, P = 0.7602) and intra-operative blood loss (294 vs. 228 mL, respectively, P = 0.8987).CONCLUSIONS: Pre-operative ECE for CPA appears to be feasible and safe; however, its role in reducing intra-operative blood loss needs further investigation.

MHD mode identification by higher order singular value decomposition of C-2W Mirnov probe data.

The C-2W device (also known as "Norman") at TAE Technologies has proven successful at generating stable, long-lived field-reversed configuration (FRC) plasmas with record temperatures. The largest Mirnov probe array in C-2W measures three components of the magnetic field just inside the vessel wall at 64 locations distributed approximately evenly in the cylindrical vessel's azimuthal and axial dimensions. This nearly rectangular array of probes creates a unique opportunity to apply higher order singular value decomposition (HOSVD) to efficiently analyze the external magnetic field data for the purposes of reconstructing the magnetohydrodynamic mode structures in the FRC. In the first application of this method for this purpose, HOSVD is shown to quickly and effectively detect and separate toroidal modes while indicating longitudinal dependence of mode phases and amplitudes, enhancing the coherence and utility of the vast quantity of data produced by this array. Analysis of the data from the entire array at once via HOSVD proves not only computationally more efficient than methods that separately analyze groups of probes at different axial locations but also leads to improved mode resolution at axial locations where these modes are weaker.

Chest complications in immunocompromised patients without acquired immunodeficiency syndrome (AIDS): differentiation between infectious and non-infectious diseases using high-resolution CT findings.

AIM: To differentiate between infectious and non-infectious diseases occurring in immunocompromised patients without acquired immunodeficiency syndrome (AIDS) using high-resolution computed tomography (HRCT). MATERIALS AND METHODS: HRCT images of 555 patients with chest complications were reviewed retrospectively. Infectious diseases (n=341) included bacterial pneumonia (n=123), fungal infection (n=80), septic emboli (n=11), tuberculosis (n=15), pneumocystis pneumonia (n=101), and cytomegalovirus pneumonia (n=11), while non-infectious diseases (n=214) included drug toxicity (n=84), infiltration of underlying diseases (n=83), idiopathic pneumonia syndrome (n=34), diffuse alveolar haemorrhage (n=8), and pulmonary oedema (n=5). Lung parenchymal abnormalities were compared between the two groups using the χ2 test and multiple logistic regression analysis. RESULTS: The χ2 test results showed significant differences in many HRCT findings between the two groups. Multiple logistic regression analysis results indicated the presence of nodules with a halo and the absence of interlobular septal (ILS) thickening were the significant indicators that could differentiate infectious from non-infectious diseases. ILS thickening was generally less frequent among most infectious diseases and more frequent among most non-infectious diseases, with a good odds ratio (7.887, p<0.001). The sensitivity and accuracy for infectious diseases in the absence of ILS thickening were better (70% and 73%, respectively) than those of nodules with a halo (19% and 48%, respectively), while the specificity in the nodules with a halo was better (93%) than that of ILS thickening (78%). CONCLUSIONS: The presence of nodules with a halo or the absence of ILS thickening tends to suggest infectious disease. Specifically, ILS thickening seems to be a more reliable indicator.

Effect of Biomechanical Environment on Degeneration of Meckel's Cartilage.

During orofacial tissue development, the anterior and posterior regions of the Meckel's cartilage undergo mineralization, while the middle region undergoes degeneration. Despite the interesting and particular phenomena, the mechanisms that regulate the different fates of Meckel's cartilage, including the effects of biomechanical cues, are still unclear. Therefore, the purpose of this study was to systematically investigate the course of Meckel's cartilage during embryonic development from a biomechanical perspective. Histomorphological and biomechanical (stiffness) changes in the Meckel's cartilage were analyzed from embryonic day 12 to postnatal day 0. The results revealed remarkable changes in the morphology and size of chondrocytes, as well as the occurrence of chondrocyte burst in the vicinity of the mineralization site, an often-seen phenomenon preceding endochondral ossification. To understand the effect of biomechanical cues on Meckel's cartilage fate, a mechanically tuned 3-dimensional hydrogel culture system was used. At the anterior region, a moderately soft environment (10-kPa hydrogel) promoted chondrocyte burst and ossification. On the contrary, at the middle region, a more rigid environment (40-kPa hydrogel) enhanced cartilage degradation by inducing a higher expression of MMP-1 and MMP-13. These results indicate that differences in the biomechanical properties of the surrounding environment are essential factors that distinctly guide the mineralization and degradation of Meckel's cartilage and would be valuable tools for modulating in vitro cartilage and bone tissue engineering.

Determination of Lycopene Concentration in Fresh Tomatoes by Spectrophotometry: A Collaborative Study.

BACKGROUND: Lycopene has been the object of considerable research attention recently, and the effects of the intake of lycopene, or of tomato products, have been studied in various ways. In Japan, interest in the health-promoting function of food components has increased. OBJECTIVE: Developing a method to determine lycopene contents in tomato that meets the Japanese Agricultural Standard (JAS). METHOD: In the proposed JAS method, the test sample consists of fresh tomatoes; a hexane-acetone mixture is utilized as the extraction solvent. A collaborative study was conducted to evaluate the interlaboratory performance of the method. RESULTS: Ten laboratories participated and analyzed six test materials characterized by a lycopene content between 39 and 170 mg/kg as blind duplicates. After removing statistical outliers, RSDr ranged from 1.2 to 3.0% and RSDR ranged from 2.4 to 4.2%. The HorRat values were calculated and found to be in the 0.26-0.49 range. CONCLUSIONS: The method for determining the lycopene content in tomato was evaluated by means of a collaborative study, and the reproducibility of this method was found to be acceptable. HIGHLIGHTS: Intended for standardization in Japan, a method to determine lycopene content in tomato has been developed and shown to have acceptable precision in a collaborative study.

Sign of Hard-X-Ray Pulsation from the γ-Ray Binary System LS 5039.

To understand the nature of the brightest γ-ray binary system LS 5039, hard x-ray data of the object, taken with the Suzaku and NuSTAR observatories in 2007 and 2016, respectively, were analyzed. The two data sets jointly gave tentative evidence for a hard x-ray periodicity, with a period of ∼9 s and a period increase rate by ∼3×10^{-10} s s^{-1}. Therefore, the compact object in LS 5039 is inferred to be a rotating neutron star, rather than a black hole. Furthermore, several lines of arguments suggest that this object has a magnetic field of several times ∼10^{10} T, two orders of magnitude higher than those of typical neutron stars. The object is hence suggested to be a magnetar, which would be the first to be found in a binary. The results also suggest that the highly efficient particle acceleration process, known to be operating in LS 5039, emerges through interactions between dense stellar winds from the massive primary star, and ultrastrong magnetic fields of the magnetar.

C-reactive protein : albumin ratio in patients with resectable intrahepatic cholangiocarcinoma.

BACKGROUND: The C-reactive protein : albumin ratio (CAR) has been reported as a novel prognostic marker in several cancers. The aim of this study was to investigate the prognostic value of CAR in patients with intrahepatic cholangiocarcinoma (ICC). METHODS: This was a single-centre retrospective study of patients who underwent surgery for ICC in a university hospital in Japan between 1998 and 2018. CAR, Glasgow Prognostic Score (GPS) and modified GPS (mGPS) were calculated. Their correlation with recurrence-free survival (RFS) and overall survival (OS) was analysed with Cox proportional hazards models. RESULTS: Seventy-two patients were included in the study. Patients were divided into two groups according to the optimal CAR cut-off value of 0·02. CAR above 0·02 was associated with higher carbohydrate antigen 19-9 levels (20·5 versus 66·1 units/ml for CAR of 0·02 or less; P = 0·002), larger tumour size (3·2 versus 4·4 cm respectively; P = 0·031) and a higher rate of microvascular invasion (9 of 28 versus 25 of 44; P = 0·041). RFS and OS were shorter in patients with CAR above 0·02: hazard ratio (HR) 4·31 (95 per cent c.i. 2·02 to 10·63) and HR 4·80 (1·85 to 16·40) respectively. In multivariable analysis CAR above 0·02 was an independent prognostic factor of RFS (HR 3·29 (1·33 to 8·12); P < 0·001), but not OS. CONCLUSIONS: CAR was associated with prognosis in patients who had hepatic resection for ICC.

ANTECEDENTES: La relación proteína C reactiva/albumina (C-reactive protein/albumin ratio, CAR) ha sido descrita como un marcador pronóstico novedoso en varios tipos de cáncer. El objetivo de este estudio fue investigar el valor pronóstico de CAR en pacientes con colangiocarcinoma intrahepático (intrahepatic cholangiocarcinoma, ICC). MÉTODOS: Se trata de un estudio retrospectivo y unicéntrico de pacientes sometidos a cirugía por ICC en un hospital universitario de Japón entre 1998 y 2018. Se calcularon CAR, puntuación pronóstica de Glasgow (Glasgow prognostic score, GPS), y GPS modificada (mGPS). Se analizó su correlación con la supervivencia libre de recidiva (recurrence-free survival, RFS) y con la supervivencia global (overall survival, OS) mediante modelos de riesgos proporcionales de Cox. RESULTADOS: Se incluyeron un total de 72 pacientes. El valor de corte óptimo de CAR fue de 0,02. Los pacientes se dividieron en dos grupos de acuerdo a este valor de corte. La presencia de CAR > 0,02 se asoció con niveles más elevados de antígeno carbohidrato 19-9 (20,5 U/ml versus 66,1 U/ml; P = 0,002), mayor tamaño tumoral (3,2 cm versus 4,4 cm; P = 0,031) y una tasa más elevada de invasión microvascular (32,1% versus 56,8%; P = 0,041). La RFS y OS fueron más cortas en pacientes con CAR > 0,02 (cociente de riesgos instantáneos, hazard ratio, HR 4,305; i.c. del 95% 2,016-10,63 y HR 4,803; i.c. del 95% 1,846-16,40, respectivamente). En los análisis multivariables, CAR de > 0,02 fue un factor pronóstico independiente para RFS (HR 3,286; i.c. del 95% 1,330-8,118; P < 0,001), pero no para la OS. CONCLUSIÓN: CAR se asoció con el pronóstico en pacientes sometidos a resección hepática por ICC.

Direct observation of imploded core heating via fast electrons with super-penetration scheme.

Fast ignition (FI) is a promising approach for high-energy-gain inertial confinement fusion in the laboratory. To achieve ignition, the energy of a short-pulse laser is required to be delivered efficiently to the pre-compressed fuel core via a high-energy electron beam. Therefore, understanding the transport and energy deposition of this electron beam inside the pre-compressed core is the key for FI. Here we report on the direct observation of the electron beam transport and deposition in a compressed core through the stimulated Cu Kα emission in the super-penetration scheme. Simulations reproducing the experimental measurements indicate that, at the time of peak compression, about 1% of the short-pulse energy is coupled to a relatively low-density core with a radius of 70 µm. Analysis with the support of 2D particle-in-cell simulations uncovers the key factors improving this coupling efficiency. Our findings are of critical importance for optimizing FI experiments in a super-penetration scheme.