Essential function for the GTPase TC21 in homeostatic antigen receptor signaling.

T cell antigen receptors (TCRs) and B cell antigen receptors (BCRs) transmit low-grade signals necessary for the survival and maintenance of mature cell pools. We show here that TC21, a small GTPase encoded by Rras2, interacted constitutively with both kinds of receptors. Expression of a dominant negative TC21 mutant in T cells produced a rapid decrease in cell viability, and Rras2(-/-) mice were lymphopenic, possibly as a result of diminished homeostatic proliferation and impaired T cell and B cell survival. In contrast, TC21 was overexpressed in several human lymphoid malignancies. Finally, the p110delta catalytic subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recruited to the TCR and BCR in a TC21-dependent way. Consequently, we propose TC21 directly links antigen receptors to PI(3)K-mediated survival pathways.

Transjugular Intrahepatic Portosystemic Shunt for Very Severe Veno-Occlusive Disease/Sinusoidal Obstruction Syndrome (VOD/SOS) after Unmanipulated Haploidentical Hematopoietic Stem Cell Transplantation with Post-transplantation Cyclophosphamide.

Hepatic veno-occlusive disease or sinusoidal obstruction syndrome (VOD/SOS) is a threatening complication after both autologous and allogeneic hematopoietic stem cell transplantation (HSCT), with high mortality rates despite early medical treatment, including the use of defibrotide (DF). We retrospectively analyzed 185 unmanipulated haploidentical (haplo-) HSCT with post-transplantation cyclophosphamide as graft-versus-host disease prophylaxis performed consecutively between 2011 and June 2019 in a single center. Seventeen patients (9.2%) were diagnosed with VOD/SOS. Based on revised European Society for Blood and Marrow Transplantation severity criteria, the VOD/SOS cases were classified as mild in 2 patients (11.7%), moderate in 2 (11.7%), severe in 2 (11.7%), and very severe in 11 (64.9%). Thirteen patients (76%) were treated with DF, including all patients with severe or very severe VOD/SOS, except 1 patient with CNS hemorrhage. Sixteen patients (94%) were alive at day +100 after HSCT. Seven patients (41%) with very severe VOD/SOS were treated with transjugular intrahepatic portosystemic shunt (TIPS) owing to rapid clinical or analytical deterioration or refractory hepatorenal syndrome despite medical treatment, including DF. TIPS insertion was performed at a median time since VOD/SOS diagnosis of 4 days (range, 1 to 28 days) without technical complications in any case. The median hepatic venous pressure gradient before and after TIPS treatment was 24 mmHg (range, 14 to 29 mmHg) and 7 mmHg (range, 2 to 11 mmHg), respectively, with a median drop of 16 mmHg (range, 9 to 19 mmHg). Following TIPS insertion, all patients showed clinical improvement with hepatomegaly, ascites, and renal failure resolution, and all showed analytical improvement with reduced alanine aminotransferase (ALT), creatinine, and international normalized ratio values, except for patient 2, whose indication for TIPS was refractory hepatorenal syndrome with a normal ALT level. The 6 patients who had initiated DF before TIPS insertion completed 21 days of treatment. All patients met the criteria for complete remission (CR) at a median of 8 days after TIPS insertion (range, 2 to 82 days). The 100-day overall survival was 100%. For patients with rapid progressive VOD/SOS, early TIPS insertion allowed completion of DF therapy. The use of TIPS together with DF resulted in CR and no associated complications with no VOD/SOS-associated mortality despite high severity. In our experience, timely and individualized use of TIPS significantly improves outcomes of very severe VOD/SOS after haplo-HSCT. Therefore, TIPS should be promptly considered in rapidly progressive cases.

ß-Arrestin-1 mediates the TCR-triggered re-routing of distal receptors to the immunological synapse by a PKC-mediated mechanism.

T-cell receptors (TCR) recognize their antigen ligand at the interface between T cells and antigen-presenting cells, known as the immunological synapse (IS). The IS provides a means of sustaining the TCR signal which requires the continual supply of new TCRs. These are endocytosed and redirected from distal membrane locations to the IS. In our search for novel cytoplasmic effectors, we have identified ß-arrestin-1 as a ligand of non-phosphorylated resting TCRs. Using dominant-negative and knockdown approaches we demonstrate that ß-arrestin-1 is required for the internalization and downregulation of non-engaged bystander TCRs. Furthermore, TCR triggering provokes the ß-arrestin-1-mediated downregulation of the G-protein coupled chemokine receptor CXCR4, but not of other control receptors. We demonstrate that ß-arrestin-1 recruitment to the TCR, and bystander TCR and CXCR4 downregulation, are mechanistically mediated by the TCR-triggered PKC-mediated phosphorylation of ß-arrestin-1 at Ser163. This mechanism allows the first triggered TCRs to deliver a stop migration signal, and to promote the internalization of distal TCRs and CXCR4 and their translocation to the IS. This receptor crosstalk mechanism is critical to sustain the TCR signal.

Quantitative parameters for the examination of InGaN QW multilayers by low-loss EELS.

We present a detailed examination of a multiple InxGa1-xN quantum well (QW) structure for optoelectronic applications. The characterization is carried out using scanning transmission electron microscopy (STEM), combining high-angle annular dark field (HAADF) imaging and electron energy loss spectroscopy (EELS). Fluctuations in the QW thickness and composition are observed in atomic resolution images. The impact of these small changes on the electronic properties of the semiconductor material is measured through spatially localized low-loss EELS, obtaining band gap and plasmon energy values. Because of the small size of the InGaN QW layers additional effects hinder the analysis. Hence, additional parameters were explored, which can be assessed using the same EELS data and give further information. For instance, plasmon width was studied using a model-based fit approach to the plasmon peak; observing a broadening of this peak can be related to the chemical and structural inhomogeneity in the InGaN QW layers. Additionally, Kramers-Kronig analysis (KKA) was used to calculate the complex dielectric function (CDF) from the EELS spectrum images (SIs). After this analysis, the electron effective mass and the sample absolute thickness were obtained, and an alternative method for the assessment of plasmon energy was demonstrated. Also after KKA, the normalization of the energy-loss spectrum allows us to analyze the Ga 3d transition, which provides additional chemical information at great spatial resolution. Each one of these methods is presented in this work together with a critical discussion of their advantages and drawbacks.

Formation mechanisms of GaN nanowires grown by selective area growth homoepitaxy.

This work provides experimental evidence and theoretical explanations regarding the formation mechanisms of GaN nanowires grown by selective area growth on GaN-on-sapphire templates. The first growth stage, driven by selective area growth kinetics, consists of initial nucleation (along the nanohole inner periphery), coalescence onset and full coalescence, producing a single nanocrystal within each nanohole. In the second growth stage, driven by free-surface-energy minimization, the formed nanocrystal undergoes morphological evolution, exhibiting initial cylindrical-like shape, intermediate dodecagonal shape and a final, thermodynamically stable hexagonal shape. From this point on, the nanowire vertical growth proceeds while keeping the stable hexagonal form.

Investigation of III-V nanowires by plan-view transmission electron microscopy: InN case study.

We discuss observations of InN nanowires (NWs) by plan-view high-resolution transmission electron microscopy (TEM). The main difficulties arise from suitable methods available for plan-view specimen preparation. We explore different approaches and find that the best results are obtained using a refined preparation method based on the conventional procedure for plan-view TEM of thin films, specifically modified for the NW morphology. The fundamental aspects of such a preparation are the initial mechanical stabilization of the NWs and the minimization of the ion-milling process after dimpling the samples until perforation. The combined analysis by plan-view and cross-sectional TEM of the NWs allows determination of the degree of strain relaxation and reveals the formation of an unintentional shell layer (2-3-nm thick) around the InN NWs. The shell layer is composed of bcc In2O3 nanocrystals with a preferred orientation with respect to the wurtzite InN: In2O3 [111] || InN [0001] and In2O3<110>||InN<1120>.

Self-regulated radius of spontaneously formed GaN nanowires in molecular beam epitaxy.

We investigate the axial and radial growth of GaN nanowires upon a variation of the Ga flux during molecular beam epitaxial growth. An increase in the Ga flux promotes radial growth without affecting the axial growth rate. In contrast, a decrease in the Ga flux reduces the axial growth rate without any change in the radius. These results are explained by a kinetic growth model that accounts for both the diffusion of Ga adatoms along the side facets toward the nanowire tip and the finite amount of active N available for the growth. The model explains the formation of a new equilibrium nanowire radius after increasing the Ga flux and provides an explanation for two well-known but so far not understood experimental facts: the necessity of effectively N-rich conditions for the spontaneous growth of GaN nanowires and the increase in nanowire radius with increasing III/V flux ratio.

Insight into the compositional and structural nano features of AlN/GaN DBRs by EELS-HAADF.

III-V nitride (AlGa)N distributed Bragg reflector devices are characterized by combined high-angle annular dark-field (HAADF) and electron energy loss spectroscopy (EELS) in the scanning transmission electron microscope. Besides the complete structural characterization of the AlN and GaN layers, the formation of AlGaN transient layers is revealed using Vegard law on profiles of the position of the bulk plasmon peak maximum. This result is confirmed by comparison of experimental and simulated HAADF intensities. In addition, we present an advantageous method for the characterization of nano-feature structures using low-loss EELS spectrum image (EEL-SI) analysis. Information from the materials in the sample is extracted from these EEL-SI at high spatial resolution.The log-ratio formula is used to calculate the relative thickness, related to the electron inelastic mean free path. Fitting of the bulk plasmon is performed using a damped plasmon model (DPM) equation. The maximum of this peak is related to the chemical composition variation using the previous Vegard law analysis. In addition, within the context of the DPM, information regarding the structural properties of the material can be obtained from the lifetime of the oscillation. Three anomalous segregation regions are characterized, revealing formation of metallic Al islands.

III-Nitrides Resonant Cavity Photodetector Devices.

III-nitride resonant cavity-enhanced Schottky barrier photodetectors were fabricated on 2 µm thick GaN templates by radio frequency plasma-assisted molecular beam epitaxy. The optical cavity was formed by a bottom distributed Bragg reflector based on 10 periods of Al0.3Ga0.7N/GaN, an Au-based Schottky contact as top mirror, and an active zone of 40 nm-thick GaN layer. The devices were fabricated with planar geometry. To evaluate the main benefits allowed by the optical cavity, conventional Schottky photodetectors were also processed. The results revealed a planar spectral response for the conventional photodetector, unlike the resonant devices that showed two raised peaks at 330 and 358 nm with responsivities of 0.34 and 0.39 mA/W, respectively. Both values were 80 times higher than the planar response of the conventional device. These results demonstrate the strong effect of the optical cavity to achieve the desired wavelength selectivity and to enhance the optical field thanks to the light resonance into the optical cavity. The research of such a combination of nitride-based Bragg mirror and thin active layer is the kernel of the present paper.

Electron Tomography of Pencil-Shaped GaN/(In,Ga)N Core-Shell Nanowires.

The three-dimensional structure of GaN/(In,Ga)N core-shell nanowires with multi-faceted pencil-shaped apex is analyzed by electron tomography using high-angle annular dark-field mode in a scanning transmission electron microscope. Selective area growth on GaN-on-sapphire templates using a patterned mask is performed by molecular beam epitaxy to obtain ordered arrays of uniform nanowires. Our results of the tomographic reconstruction allow the detailed determination of the complex morphology of the inner (In,Ga)N multi-faceted shell structure and its deviation from the perfect hexagonal symmetry. The tomogram unambiguously identifies a dot-in-a-wire configuration at the nanowire apex including the exact shape and size, as well as the spatial distribution of its chemical composition.

Studying the Dynamics of TCR Internalization at the Immune Synapse.

Establishing a stable interaction between a T cell and an antigen presenting cell (APC) involves the formation of an immune synapse (IS). It is through this structure that the T cell can integrate all the signals provided by the APC. The IS also serves as a mechanism for TCR downregulation through internalization. Here, we describe methods for visualizing MHC-engaged T cell receptor (TCR) internalization from the IS in human cell lines and mouse primary T cells by confocal fluorescence microscopy techniques.

First-in-class inhibitor of the T cell receptor for the treatment of autoimmune diseases.

Modulating T cell activation is critical for treating autoimmune diseases but requires avoiding concomitant opportunistic infections. Antigen binding to the T cell receptor (TCR) triggers the recruitment of the cytosolic adaptor protein Nck to a proline-rich sequence in the cytoplasmic tail of the TCR's CD3ε subunit. Through virtual screening and using combinatorial chemistry, we have generated an orally available, low-molecular weight inhibitor of the TCR-Nck interaction that selectively inhibits TCR-triggered T cell activation with an IC50 (median inhibitory concentration) ~1 nM. By modulating TCR signaling, the inhibitor prevented the development of psoriasis and asthma and, furthermore, exerted a long-lasting therapeutic effect in a model of autoimmune encephalomyelitis. However, it did not prevent the generation of a protective memory response against a mouse pathogen, suggesting that the compound might not exert its effects through immunosuppression. These results suggest that inhibiting an immediate TCR signal has promise for treating a broad spectrum of human T cell-mediated autoimmune and inflammatory diseases.

Critical aspects of substrate nanopatterning for the ordered growth of GaN nanocolumns.

Precise and reproducible surface nanopatterning is the key for a successful ordered growth of GaN nanocolumns. In this work, we point out the main technological issues related to the patterning process, mainly surface roughness and cleaning, and mask adhesion to the substrate. We found that each of these factors, process-related, has a dramatic impact on the subsequent selective growth of the columns inside the patterned holes. We compare the performance of e-beam lithography, colloidal lithography, and focused ion beam in the fabrication of hole-patterned masks for ordered columnar growth. These results are applicable to the ordered growth of nanocolumns of different materials.

[Cavernous haemangioma as the cause of ureteral pyelic junction obstruction]. / Hemangioma cavernoso como causa de obstrucción ureteropiélica.

OBJECTIVES: To report one case of cavernous haemangioma in the upper urinary tract. METHODS/RESULTS: 29-year-old female presenting with right renal colic whose IVP showed UPJ obstruction. Dismembered pyeloplasty was performed and pathology reported a cavernous haemangioma as the cause of stenosis. CONCLUSIONS: Cavernous haemangioma, a rare upper urinary tract pathology, should be considered in the differential diagnosis of causes of UPJ obstruction.

Columnar AlGaN/GaN nanocavities with AlN/GaN Bragg reflectors grown by molecular beam epitaxy on Si(111).

Self-assembled columnar AlGaN/GaN nanocavities, with an active region of GaN quantum disks embedded in an AlGaN nanocolumn and cladded by top and bottom AlN/GaN Bragg mirrors, were grown. The nanocavity has no cracks or extended defects, due to the relaxation at the Si interface and to the nanocolumn free-surface to volume ratio. The emission from the active region matched the peak reflectivity by tuning the Al content and the GaN disks thickness. Quantum confinement effects that depend on both the disk thickness and the inhomogeneous strain distribution within the disks are clearly observed.

[Bladder leiomyoma. Report of one case]. / Leiomioma vesical. Presentación de un caso clínico.

OBJECTIVES: To report a new case of bladder leiomyoma. METHODS: We describe the case of a 56-year-old female patient presenting with renal colic who was diognosed of a bladder filling defect after work up with intravenous urography ultrasound and urine cytology. Complete transurethral resection of the lesion was performed. RESULTS: Pathology reported bladder leiomyoma. CONCLUSIONS: Every pathological possibility should be considered in the differential diagnosis and treatment of a filling defect. Bladder leiomyoma is usually an incidental diagnosis on the pathologic report after resection of a bladder tumor.