In Vitro Characterization of Human CD24hiCD38hi Regulatory B Cells Shows CD9 Is Not a Stable Breg Cell Marker

Regulatory B (Breg) cells are endowed with immune suppressive functions. Various human and murine Breg subtypes have been reported. While interleukin (IL)-10 intracellular staining remains the most reliable way to identify Breg cells, this technique hinders further essential functional studies. Recent findings suggest that CD9 is an effective surface marker of murine IL-10 competent Breg cells. However, the stability of CD9 and its relevance as a unique marker for human Breg cells, which have been widely characterized as CD24^hiCD38^hi, have not been investigated. Here, we demonstrate that CD9 expression is sensitive to in vitro B cell stimulations. CD9 expression could either be re-expressed or downregulated in purified CD9-negative B cells and CD9-positive B cells, respectively. We found no significant differences in the Breg differentiation capacity of the CD9-negative and CD9-positive B cells. Furthermore, CD9-positive B cells co-express CD40 and CD86, suggesting their nature as B cell activation or co-stimulatory molecules, rather than regulatory ones. Therefore, we report the relatively unstable CD9 as a distinct surface molecule, indicating the need for further research for a more reliable marker to purify human Breg cells.     

Aligned texture map creation for pose invariant face recognition

In last years, Face recognition based on 3D techniques is an emergent technology which has demonstrated better results than conventional 2D approaches. Using texture (180° multi-view image) and depth maps is supposed to increase the robustness towards the two main challenges in Face Recognition: Pose and illumination. Nevertheless, 3D data should be acquired under highly controlled conditions and in most cases depends on the collaboration of the subject to be recognized. Thus, in applications such as surveillance or control access points, this kind of 3D data may not be available during the recognition process. This leads to a new paradigm using some mixed 2D-3D face recognition systems where 3D data is used in the training but either 2D or 3D information can be used in the recognition depending on the scenario. Following this concept, where only part of the information (partial concept) is used in the recognition, a novel method is presented in this work. This has been called Partial Principal Component Analysis (P²CA) since they fuse the Partial concept with the fundamentals of the well known PCA algorithm. This strategy has been proven to be very robust in pose variation scenarios showing that the 3D training process retains all the spatial information of the face while the 2D picture effectively recovers the face information from the available data. Furthermore, in this work, a novel approach for the automatic creation of 180° aligned cylindrical projected face images using nine different views is presented. These face images are created by using a cylindrical approximation for the real object surface. The alignment is done by applying first a global 2D affine transformation of the image, and afterward a local transformation of the desired face features using a triangle mesh. This local alignment allows a closer look to the feature properties and not the differences. Finally, these aligned face images are used for training a pose invariant face recognition approach (P²CA).     

A formal analysis of database replication protocols with SI replicas and crash failures

This paper provides a formal specification and proof of correctness of a basic Generalized Snapshot Isolation certification-based data replication protocol for database middleware architectures. It has been modeled using a state transition system, as well as the main system components, allowing a perfect match with the usual deployment in a middleware system. The proof encompasses both safety and liveness properties, as it is commonly done for a distributed algorithm. Furthermore, a crash failure model has been assumed for the correctness proof, although recovery analysis is not the aim of this paper. This allows an easy extension toward a crash-recovery model support in future works. The liveness proof focuses in the uniform commit: if a site has committed a transaction, the rest of sites will either commit it or it would have crashed.     

Microfluidic device with dual-channel fluorescence acquisition for quantification/identification of cancer cells

This paper presents an optofluidic device for cell discrimination with two independent interrogation regions. Pumping light is coupled to the device, and cell fluorescence is extracted from the two interrogation zones by using optical fibers embedded in the optofluidic chip. To test the reliability of this device, AU-565 cells—expressing EpCAM and HER2 receptors—and RAMOS cells were mixed in a controlled manner, confined inside a hydrodynamic focused flow in the microfluidic chip and detected individually so that they could be discriminated as positive (signal reception from fluorescently labeled antibodies from the AU-565 cells) or negative events (RAMOS cells). A correlation analysis of the two signals reduces the influence of noise on the overall data.     

Analysis of the HLA class I associated peptide repertoire in a hepatocellular carcinoma cell line reveals tumor-specific peptides as putative targets for immunotherapy

HLA class I molecules present peptides on the cell surface to CD8(+) T cells. The repertoire of peptides that associate to class I molecules represents the cellular proteome. Therefore, cells expressing different proteomes could generate different class I-associated peptide repertoires. A large number of peptides have been sequenced from HLA class I alleles, mostly from lymphoid cells. On the other hand, T cell immunotherapy is a goal in the fight against cancer, but the identification of T cell epitopes is a laborious task. Proteomic techniques allow the definition of putative T cell epitopes by the identification of HLA natural ligands in tumor cells. In this study, we have compared the HLA class I-associated peptide repertoire from the hepatocellular carcinoma (HCC) cell line SK-Hep-1 with that previously described from lymphoid cells. The analysis of the peptide pool confirmed that, as expected, the peptides from SK-Hep-1 derive from proteins localized in the same compartments as in lymphoid cells. Within this pool, we have identified 12 HLA class I peptides derived from HCC-related proteins. This confirms that tumor cell lines could be a good source of tumor associated antigens to be used, together with MS, to define putative epitopes for cytotoxic T cells from cancer patients.     

The interpretation of X-ray photoelectron spectra of pyrolized S-containing carbonaceous materials

Theoretical models and ab initio Hartree-Fock wavefunctions have been used to investigate the S(2p) core level binding energies (BE), of pyrolized S-containing, carbonaceous materials. Comparison between experimental and calculated data for thiophene permits the accuracy of the present approach to be established. A systematic study of different situations demonstrates that, in these materials, non-oxidized S atoms can show peaks at very high BE relative to the C(1s) peak at 285.0 eV. This study confirms that the peak at 164.1 eV corresponds to ‘thiophenic’ S atoms. On the other hand, it shows that the peaks at higher BE could correspond to S atoms replacing C atoms in three-coordinated structures of graphene layers, in agreement with the arguments suggested in the experimental works. The energetic similarity between the two peaks at very high BE makes it difficult to differentiate between them, although the results of the present study seem to suggest that the peak at experimental BE ≈ 166 eV could correspond to S atoms coordinated to two C and one H atoms at the edge of graphene layers, while the peak at ≈ 169 eV would correspond to S atoms replacing C atoms in inner positions of the graphene layers, and it is bonded to three C atoms.     

A probabilistic integrated object recognition and tracking framework

This paper describes a probabilistic integrated object recognition and tracking framework called PIORT, together with two specific methods derived from it, which are evaluated experimentally in several test video sequences. The first step in the proposed framework is a static recognition module that provides class probabilities for each pixel of the image from a set of local features. These probabilities are updated dynamically and supplied to a tracking decision module capable of handling full and partial occlusions. The two specific methods presented use RGB color features and differ in the classifier implemented: one is a Bayesian method based on maximum likelihood and the other one is based on a neural network. The experimental results obtained have shown that, on one hand, the neural net based approach performs similarly and sometimes better than the Bayesian approach when they are integrated within the tracking framework. And on the other hand, our PIORT methods have achieved better results when compared to other published tracking methods in video sequences taken with a moving camera and including full and partial occlusions of the tracked object.     

Growth of La2Mo2O9 films on porous Al2O3 substrates by radio frequency magnetron sputtering

Synthesis conditions of La₂Mo₂O₉ thin film by radio frequency (RF) sputtering technique on Al₂O₃ ceramic substrates are studied. It is found that the deposition temperature and oxygen partial pressure are the most important factors for obtaining pure La₂Mo₂O₉ films. Varying both parameters, Mo-rich, stoichiometric, and Mo-deficient films are obtained. With increasing the La:Mo ratio, films become denser. A crust layer is observed on top of the Mo-rich and the Mo-deficient films. The formation of the La₂Mo₂O₉ phase is discussed with respect to the sputtering mechanism.     

The Deubiquitinating Enzyme USP48 Interacts with the Retinal Degeneration-Associated Proteins UNC119a and ARL3

Proteins related to the ubiquitin-proteasome system play an important role during the differentiation and ciliogenesis of photoreceptor cells. Mutations in several genes involved in ubiquitination and proteostasis have been identified as causative of inherited retinal dystrophies (IRDs) and ciliopathies. USP48 is a deubiquitinating enzyme whose role in the retina is still unexplored although previous studies indicate its relevance for neurosensory organs. In this work, we describe that a pool of endogenous USP48 localises to the basal body in retinal cells and provide data that supports the function of USP48 in the photoreceptor cilium. We also demonstrate that USP48 interacts with the IRD-associated proteins ARL3 and UNC119a, and stabilise their protein levels using different mechanisms. Our results suggest that USP48 may act in the regulation/stabilisation of key ciliary proteins for photoreceptor function, in the modulation of intracellular protein transport, and in ciliary trafficking to the photoreceptor outer segment.