Genetic Variants of Human Granzyme B Predict Transplant Outcomes after HLA Matched Unrelated Bone Marrow Transplantation for Myeloid Malignancies

Serine protease granzyme B plays important roles in infections, autoimmunity, transplant rejection, and antitumor immunity. A triple-mutated granzyme B variant that encodes three amino substitutions (Q48R, P88A, and Y245H) has been reported to have altered biological functions. In the polymorphism rs8192917 (2364A>G), the A and G alleles represent wild type QPY and RAH mutant variants, respectively. In this study, we analyzed the impact of granzyme B polymorphisms on transplant outcomes in recipients undergoing unrelated HLA-fully matched T-cell-replete bone marrow transplantation (BMT) through the Japan Donor Marrow Program. The granzyme B genotypes were retrospectively analyzed in a cohort of 613 pairs of recipients with hematological malignancies and their unrelated donors. In patients with myeloid malignancies consisting of acute myeloid leukemia and myelodysplastic syndrome, the donor G/G or A/G genotype was associated with improved overall survival (OS; adjusted hazard ratio [HR], 0.60; 95% confidence interval [CI], 0.41–0.89; P = 0.01) as well as transplant related mortality (TRM; adjusted HR, 0.48; 95% CI, 0.27–0.86, P = 0.01). The recipient G/G or A/G genotype was associated with a better OS (adjusted HR, 0.68; 95% CI, 0.47–0.99; P = 0.05) and a trend toward a reduced TRM (adjusted HR, 0.61; 95% CI, 0.35–1.06; P = 0.08). Granzyme B polymorphism did not have any effect on the transplant outcomes in patients with lymphoid malignancies consisting of acute lymphoid leukemia and malignant lymphoma. These data suggest that there is an association between the granzyme B genotype and better clinical outcomes in patients with myeloid malignancies after unrelated BMT

Search for gravitational-lensing signatures in the full third observing run of the LIGO-Virgo network

Gravitational lensing by massive objects along the line of sight to the source causes distortions of gravitational wave-signals; such distortions may reveal information about fundamental physics, cosmology and astrophysics. In this work, we have extended the search for lensing signatures to all binary black hole events from the third observing run of the LIGO--Virgo network. We search for repeated signals from strong lensing by 1) performing targeted searches for subthreshold signals, 2) calculating the degree of overlap amongst the intrinsic parameters and sky location of pairs of signals, 3) comparing the similarities of the spectrograms amongst pairs of signals, and 4) performing dual-signal Bayesian analysis that takes into account selection effects and astrophysical knowledge. We also search for distortions to the gravitational waveform caused by 1) frequency-independent phase shifts in strongly lensed images, and 2) frequency-dependent modulation of the amplitude and phase due to point masses. None of these searches yields significant evidence for lensing. Finally, we use the non-detection of gravitational-wave lensing to constrain the lensing rate based on the latest merger-rate estimates and the fraction of dark matter composed of compact objects

Electronic structure of self-doped layered Eu 3 F 4 Bi 2 S 4 material revealed by x-ray absorption spectroscopy and photoelectron spectromicroscopy

International audienceWe have studied the electronic structure of Eu 3 F 4 Bi 2 S 4 using a combination of Eu L 3-edge x-ray absorption spectroscopy (XAS) and space-resolved angle-resolved photoemission spectroscopy (ARPES). From the Eu L 3-edge XAS, we have found that the Eu in this system is in mixed valence state with coexistence of Eu 2+ /Eu 3+. The bulk charge doping was estimated to be ∼0.3 per Bi site in Eu 3 F 4 Bi 2 S 4 , which corresponds to the nominal x in a typical REO 1−x F x BiS 2 system (RE: rare-earth elements). From the space-resolved ARPES, we have ruled out the possibility of any microscale phase separation of Eu valence in the system. Using a microfocused beam we have observed the band structure as well as the Fermi surface that appeared similar to other compounds of this family with disconnected rectangular electronlike pockets around the X point. The Luttinger volume analysis gives the effective carrier to be 0.23 electrons per Bi site in Eu 3 F 4 Bi 2 S 4 , indicating that the system is likely to be in the underdoped region of its superconducting phase diagram. The BiS 2-based superconductors are composed of a layered structure with active BiS 2 bilayers intercalated by insulating spacer layers [1]. The typical BiS 2-based system REOBiS 2 (RE: rare-earth elements) is usually a band insulator, and the band filling can be controlled by electron doping; by substituting O by F [2] or tetravalent ions by trivalent La at RE site [3]. As the doping level increases, superconductivity develops within the BiS 2 layers with the maximum T c of 10.5 K found in LaO 0.5 F 0.5 BiS 2 [4]. The conduction bands are composed of Bi 6p x/y orbitals giving rise to rectangular electron pockets around the X point of the Brillouin zone [5,6]. With the same crystal structure, EuFBiS 2 shows super-conductivity without any doping. It is thought to be caused by the self-doped electrons coming from Eu that is in a mixed valence state with coexisting Eu 2+ and Eu 3+ [7]. A similar situation seems to occur in a new class of the BiS 2-based family Eu 3 F 4 Bi 2 S 4 system that is found to show bulk superconductivity with T c = 1.5 K induced by self-doping [8]. The structure is rather complex with an additional EuF 2 layer intercalated into the EuFBiS 2 structure as shown in Fig. 1(a). In this work we have studied the electronic structure of Eu 3 F 4 Bi 2 S 4 by means of Eu L 3-edge x-ray absorption spectroscopy (XAS) and space-resolved angle-resolved pho-toemission spectroscopy (ARPES); the Eu L 3-edge XAS is a bulk sensitive probe on the valence state while the space-resolved ARPES can reveal the space-dependent electronic structure, which is especially important on the present system with a mixed valence of Eu and highly disordered local symmetry [9-11]. High-quality single crystals of Eu 3 F 4 Bi 2 S 4 were grown by the CsCl-flux method with powders of EuS, Bi 2 S 3 , and BiF 3. The excess CsCl flux was removed using H 2 O. The crystals were well characterized for their crystal structures and transport properties prior to the spectroscopy measurements [12]. The Eu L 3-edge XAS measurements have been performed at BM30B beamline of the European Synchrotron Radiation Facility. At the BM30B, the synchrotron radiation was monochromatized by a double crystal Si(220) monochromator, and the energy resolution is close to the intrinsic resolution of the Si crystals, i.e., around 0.35 eV at the Eu L 3 edge [13]. Two Si mirrors covered by a Rh layer allow us to avoid high energy photons (harmonics). The Eu 3 F 4 Bi 2 S 4 crystals were mounted in a continuous flow He cryostat and XAS measurements were carried out in a partial fluorescence yield mode at a temperature of 21 K. The experimental geometry in sketched in the inset of Fig. 1(b) with the linearly polarized light falling at an angle of 33 • with respect to the normal direction of the sample. This angle avoids the polarization effects and provides the angle-independent bulk spectrum. The Eu L 3-edge absorption spectrum was collected by detecting the Eu L α1 fluorescence photons over a large solid angle using a multielement Ge detector. The obtained XAS spectrum was corrected for the self-absorption using the FLUO algorithm embedded in Athena software [14]. Space-resolved ARPES measurements have been performed at the Spectromicroscopy beamline, Elettra synchrotron facility, Trieste [15]. Incident photons of 74 eV were focused using a Schwarzschild optics down to a 500 × 500 nm 2 beam spot. For the present measurements the total energy resolution is about ∼100 meV while the angular resolution is 0.5 •. The single crystals of Eu 3 F 4 Bi 2 S 4 were cleaved and oriented in situ at 40 K under ultrahigh vacuum condition (<10 −10 mbar). All the measurements were carried 2469-9950/2017/95(3)/035152(5) 035152-

Shear-induced Interleukin-6 Synthesis in Chondrocytes: ROLES OF E PROSTANOID (EP) 2 AND EP3 IN cAMP/PROTEIN KINASE A- AND PI3-K/Akt-DEPENDENT NF-κB ACTIVATION*

Mechanical overloading of cartilage producing hydrostatic stress, tensile strain, and fluid flow can adversely affect chondrocyte function and precipitate osteoarthritis (OA). Application of high fluid shear stress to chondrocytes recapitulates the earmarks of OA, as evidenced by the release of pro-inflammatory mediators, matrix degradation, and chondrocyte apoptosis. Elevated levels of cyclooxygenase-2 (COX-2), prostaglandin (PG) E2, and interleukin (IL)-6 have been reported in OA cartilage in vivo, and in shear-activated chondrocytes in vitro. Although PGE2 positively regulates IL-6 synthesis in chondrocytes, the underlying signaling pathway of shear-induced IL-6 expression remains unknown. Using the human T/C-28a2 chondrocyte cell line as a model system, we demonstrate that COX-2-derived PGE2 signals via up-regulation of E prostanoid (EP) 2 and down-regulation of EP3 receptors to raise intracellular cAMP, and activate protein kinase A (PKA) and phosphatidylinositol 3-kinase (PI3-K)/Akt pathways. PKA and PI3-K/Akt transactivate the NF-κB p65 subunit via phosphorylation at Ser-276 and Ser-536, respectively. Binding of p65 to the IL-6 promoter elicits IL-6 synthesis in sheared chondrocytes. Selective knockdown of EP2 or ectopic expression of EP3 blocks PKA- and PI3-K/Akt-dependent p65 activation and markedly diminishes shear-induced IL-6 expression. Similar inhibitory effects on IL-6 synthesis were observed by inhibiting PKA, PI3-K, or NF-κB using pharmacological and/or genetic interventions. Reconstructing the signaling network regulating shear-induced IL-6 expression in chondrocytes may provide insights for developing therapeutic strategies for arthritic disorders and for culturing artificial cartilage in bioreactors

Current best estimates for the average lifespans of mouse and human leukocytes : reviewing two decades of deuterium-labeling experiments

Deuterium is a non-toxic, stable isotope that can safely be administered to humans and mice to study their cellular turnover rates in vivo. It is incorporated into newly synthesized DNA strands during cell division, without interference with the kinetics of cells, and the accumulation and loss of deuterium in the DNA of sorted (sub-)populations of leukocytes can be used to estimate their cellular production rates and lifespans. In the past two decades, this powerful technology has been used to estimate the turnover rates of various types of leukocytes. Although it is the most reliable technique currently available to study leukocyte turnover, there are remarkable differences between the cellular turnover rates estimated by some of these studies. We have recently established that part of this variation is due to (a) difficulties in estimating deuterium availability in some deuterium-labeling studies, and (b) assumptions made by the mathematical models employed to fit the data. Being aware of these two problems, we here aim to approach a consensus on the life expectancies of different types of T cells, B cells, monocytes, and neutrophils in mice and men. We address remaining outstanding problems whenever appropriate and discuss for which immune subpopulations we currently have too little information to draw firm conclusions about their turnover