Mutant p53 reprograms TNF signaling in cancer cells through interaction with the tumor suppressor DAB2IP.

Inflammation is a significant factor in cancer development, and a molecular understanding of the parameters dictating the impact of inflammation on cancers could significantly improve treatment. The tumor suppressor p53 is frequently mutated in cancer, and p53 missense mutants (mutp53) can acquire oncogenic properties. We report that cancer cells with mutp53 respond to inflammatory cytokines increasing their invasive behavior. Notably, this action is coupled to expression of chemokines that can expose the tumor to host immunity, potentially affecting response to therapy. Mechanistically, mutp53 fuels NF-κB activation while it dampens activation of ASK1/JNK by TNFα, and this action depends on mutp53 binding and inhibiting the tumor suppressor DAB2IP in the cytoplasm. Interfering with such interaction reduced aggressiveness of cancer cells in xenografts. This interaction is an unexplored mechanism by which mutant p53 can influence tumor evolution, with implications for our understanding of the complex role of inflammation in cancer.

A genome-scale protein interaction profile of Drosophila p53 uncovers additional nodes of the human p53 network.

The genome of the fruitfly Drosophila melanogaster contains a single p53-like protein, phylogenetically related to the ancestor of the mammalian p53 family of tumor suppressors. We reasoned that a comprehensive map of the protein interaction profile of Drosophila p53 (Dmp53) might help identify conserved interactions of the entire p53 family in man. Using a genome-scale in vitro expression cloning approach, we identified 91 previously unreported Dmp53 interactors, considerably expanding the current Drosophila p53 interactome. Looking for evolutionary conservation of these interactions, we tested 41 mammalian orthologs and found that 37 bound to one or more p53-family members when overexpressed in human cells. An RNAi-based functional assay for modulation of the p53 pathway returned five positive hits, validating the biological relevance of these interactions. One p53 interactor is GTPBP4, a nucleolar protein involved in 60S ribosome biogenesis. We demonstrate that GTPBP4 knockdown induces p53 accumulation and activation in the absence of nucleolar disruption. In breast tumors with wild-type p53, increased expression of GTPBP4 correlates with reduced patient survival, emphasizing a potential relevance of this regulatory axis in cancer.

Deconstructing Haptic Feedback Information in Robot-Assisted Needle Insertion in Soft Tissues.

This paper evaluates the role and effectiveness of different types of haptic feedback in presenting relevant feedback information during needle insertion in soft tissues through a remotely operated robot. We carried out three experiments with human subjects to analyze the effect of grounded kinesthetic feedback, cutaneous vibrotactile feedback, and cutaneous pressure feedback for rendering the elastic and the viscous force components of a simplified needle-tissue interaction model in a simulated environment. Results showed that providing the two pieces of feedback information through different channels, i.e., kinesthetic and cutaneous, led to the best performance, yielding an improvement in detecting a different tissue layer with respect to providing both information through the same commercial grounded kinesthetic interface. Moreover, results indicate that cutaneous pressure feedback is more suited for rendering the elastic component of the interaction with respect to vibrotactile cutaneous sensations. Finally, results suggest that rendering this elastic component where the user holds the input interface is not so important, confirming that delocalized cutaneous sensations can be an effective solution.

A Wearable Haptic Device for the Hand with Interchangeable End-Effectors.

This paper presents a 4-degrees-of-freedom (4-DoF) hand wearable haptic device for Virtual Reality (VR). It is designed to support different end-effectors, that can be easily exchanged so as to provide a wide range of haptic sensations. The device is composed of a static upper body, secured to the back of the hand, and the (changeable) end-effector, placed in contact with the palm. The two parts of the device are connected by two articulated arms, actuated by four servo motors housed on the upper body and along the arms. The paper summarizes the design and kinematics of the wearable haptic device and presents a position control scheme able to actuate a broad range of end-effectors. As a proof of concept, we present and evaluate three representative end-effectors during interactions in VR, rendering the sensation of interacting (E1) with rigid slanted surfaces and sharp edges having different orientations, (E2) with curved surfaces having different curvatures, and (E3) with soft surfaces having different stiffness characteristics. A few additional end-effector designs are discussed. A human-subjects evaluation in immersive VR shows the broad applicability of the device, able to render rich interactions with a diverse set of virtual objects.

The evolutionary conserved gene C16orf35 encodes a nucleo-cytoplasmic protein that interacts with p73.

C16orf35 is a highly conserved gene positioned upstream of the alpha-globins in humans and other vertebrates. The deduced protein is also highly conserved, it has no defined structural features or domains, and its function is currently unknown. Here we show that the C16orf35 protein has nuclear and cytosolic distribution, and can localize to PML nuclear bodies. The C16orf35 protein was detected in several human transformed cells lines, and studies of transient and stable overexpression indicate that increased levels of C16orf35 inhibit cell proliferation. We also find that C16orf35 interacts with human p73, and represses transcription by TAp73gamma but not by TAp73alpha. This selectivity is not due to differential interaction, since C16orf35 binds both p73 variants. Our data suggest that C16orf35 can modulate differentially the specific activities of selected p73 isoforms.

Phage Display Technology for Human Monoclonal Antibodies.

During the last 20 years in vitro technologies opened powerful routes to combine the generation of large libraries together with fast selection and screening procedures to identify lead candidates. One of the most successful methods is based on the use of filamentous phages. Functional Antibodies (Abs) fragments can be displayed on the surface of phages by fusing the coding sequence of the antibody variable (V) regions to the phage minor coat protein pIII. By creating large libraries, antibodies with affinities comparable to those obtained using traditional hybridoma technology can be isolated by a series of cycles of selection on the antigen of interest. In this system, antibody genes can be recovered simultaneously with selection and can be easily further engineered, for example by increasing their affinity to levels unobtainable in the immune system, or by modulating their specificity and their effector functions (by recloning into a full-length immunoglobulin scaffold). This chapter describes the basic protocols for antibody library construction and selection of binder with desired specificity.

Mutant p53 improves cancer cells' resistance to endoplasmic reticulum stress by sustaining activation of the UPR regulator ATF6.

Missense mutations in the TP53 gene are frequent in human cancers, giving rise to mutant p53 proteins that can acquire oncogenic properties. Gain of function mutant p53 proteins can enhance tumour aggressiveness by promoting cell invasion, metastasis and chemoresistance. Accumulating evidences indicate that mutant p53 proteins can also modulate cell homeostatic processes, suggesting that missense p53 mutation may increase resistance of tumour cells to intrinsic and extrinsic cancer-related stress conditions, thus offering a selective advantage. Here we provide evidence that mutant p53 proteins can modulate the Unfolded Protein Response (UPR) to increase cell survival upon Endoplasmic Reticulum (ER) stress, a condition to which cancer cells are exposed during tumour formation and progression, as well as during therapy. Mechanistically, this action of mutant p53 is due to enhanced activation of the pro-survival UPR effector ATF6, coordinated with inhibition of the pro-apoptotic UPR effectors JNK and CHOP. In a triple-negative breast cancer cell model with missense TP53 mutation, we found that ATF6 activity is necessary for viability and invasion phenotypes. Together, these findings suggest that ATF6 inhibitors might be combined with mutant p53-targeting drugs to specifically sensitise cancer cells to endogenous or chemotherapy-induced ER stress.

On the Triggering Mechanisms of Upward Lightning.

Upward lightning studies took place in Rapid City, South Dakota, USA and S. Paulo, Brazil during the summer thunderstorm seasons from 2011 to 2016. One of the main objectives of these campaigns was to evaluate and characterize the triggering of upward positive leaders from tall objects due to preceding nearby flash activity. 110 upward flashes were observed with a combination of high- and standard-speed video and digital still cameras, electric field meters, fast electric-field antenna systems, and for two seasons, a Lightning Mapping Array. These data were analyzed, along with correlated lightning location system data, to determine the triggering flash type responsible for the initiation of upward leaders from towers. In this paper, we describe the various processes during flash activity that can trigger upward leaders from tall objects in the USA and in Brazil. We conclude that the most effective triggering component is the propagation of the in-cloud negative leader during the continuing current that follows a positive return stroke.

Parkinson's disease DJ-1 L166P alters rRNA biogenesis by exclusion of TTRAP from the nucleolus and sequestration into cytoplasmic aggregates via TRAF6.

Mutations in PARK7/DJ-1 gene are associated to autosomal recessive early onset forms of Parkinson's disease (PD). Although large gene deletions have been linked to a loss-of-function phenotype, the pathogenic mechanism of missense mutations is less clear. The L166P mutation causes misfolding of DJ-1 protein and its degradation. L166P protein may also accumulate into insoluble cytoplasmic aggregates with a mechanism facilitated by the E3 ligase TNF receptor associated factor 6 (TRAF6). Upon proteasome impairment L166P activates the JNK/p38 MAPK apoptotic pathway by its interaction with TRAF and TNF Receptor Associated Protein (TTRAP). When proteasome activity is blocked in the presence of wild-type DJ-1, TTRAP forms aggregates that are localized to the cytoplasm or associated to nucleolar cavities, where it is required for a correct rRNA biogenesis. In this study we show that in post-mortem brains of sporadic PD patients TTRAP is associated to the nucleolus and to Lewy Bodies, cytoplasmic aggregates considered the hallmark of the disease. In SH-SY5Y neuroblastoma cells, misfolded mutant DJ-1 L166P alters rRNA biogenesis inhibiting TTRAP localization to the nucleolus and enhancing its recruitment into cytoplasmic aggregates with a mechanism that depends in part on TRAF6 activity. This work suggests that TTRAP plays a role in the molecular mechanisms of both sporadic and familial PD. Furthermore, it unveils the existence of an interplay between cytoplasmic and nucleolar aggregates that impacts rRNA biogenesis and involves TRAF6.

Nefrectomia para doação de rim: insuficiência coronariana aguda peroperatório

Introdução: a isquemia miocárdica peroperatória está associada à alta morbidade e mortalidade. Seu diagnóstico precoce e a pronta abordagem terapêutica são de fundamental importância. Relato de caso: paciente masculino, 44 anos, 85 kg, ASA II, hipertenso leve, sem tratamento, tabagista (30 cigarros/dia), submetido à nefrectomia para doação de rim. Após punção venesa e monitorização com oximetria de pulso, ECG, capnografia e PANI, foi feita sedação com midazolan (3mg) e realizado bloqueio peridural com passagem de cateter e injeção fracionada de 20mçl de ropivacaína (0,33%). Apó0s pré-oxigenação, foi realizada indução da anestesia com injeção sequencial de propofol (150mg), atracúrio (40mg), afentanil realizada (2,5mg) e manutenção com isoflurano. Aproximadamente 50 minutos após o início do procedimento cirúrgico observamos no ECG supradesnivelamento amplo e progressivo do segmento ST acompanhado de instabilidade hemodinâmica (PA=90x70mmHg) e taquicardia sinusal (FC maior130bat/min). O isoflurano foi desligado e iniciou-se infusão de nitroglicerina (0,7 ug.kg-1. min-1). Houve melhora do padrão hemodinâmico, mas sem alterações na morfologia do segmento ST. Optamos pelo uso de 5 mg de metropolol EV injetados por um período de 5 minutos, sendo esta dose repetida 10 minutos após. Na metade da segunda dose observamos retorno do ST ao padrão normal e estabilidade hemodinâmica plena. Os eletrocardiogramas seriados realizados no pós-operatório não mostraram qualquer anormalidade. A dosagem de troponina e da CPKMB não pôde ser realizada por falta de reagentes no hospital. O paciente teve boa evolução clínica e alta hospitalar sem outras intercorrências. Discussão: o presente relato nos alerta para a possibilidade de isquemia miocárdia em pacientes com fatores de risco, como hipertensão e tabagismo. No período per e pós-operatório os pacientes estão especialmente susceptíveis devido ao estresse da resposta neuro-humoral gerada pela cirurgia. Apesar de a nitroglicerina ser fundamental no tratamento da isquemia ela não é recomendada na vigência de estados hipovolêmicos ou hipotensivos. O uso do beta bloqueador se mostrou efetivo na mudança da relação suprimento-demanda de oxigênio para o miocárdio.