Structure-Function Analysis of the HrpB2-HrcU Interaction in the Xanthomonas citri Type III Secretion System

Bacterial type III secretion systems deliver protein virulence factors to host cells. Here we characterize the interaction between HrpB2, a small protein secreted by the Xanthomonas citri subsp. citri type III secretion system, and the cytosolic domain of the inner membrane protein HrcU, a paralog of the flagellar protein FlhB. We show that a recombinant fragment corresponding to the C-terminal cytosolic domain of HrcU produced in E. coli suffers cleavage within a conserved Asn264-Pro265-Thr266-His267 (NPTH) sequence. A recombinant HrcU cytosolic domain with N264A, P265A, T266A mutations at the cleavage site (HrcUAAAH) was not cleaved and interacted with HrpB2. Furthermore, a polypeptide corresponding to the sequence following the NPTH cleavage site also interacted with HrpB2 indicating that the site for interaction is located after the NPTH site. Non-polar deletion mutants of the hrcU and hrpB2 genes resulted in a total loss of pathogenicity in susceptible citrus plants and disease symptoms could be recovered by expression of HrpB2 and HrcU from extrachromossomal plasmids. Complementation of the ΔhrcU mutant with HrcUAAAH produced canker lesions similar to those observed when complemented with wild-type HrcU. HrpB2 secretion however, was significantly reduced in the ΔhrcU mutant complemented with HrcUAAAH, suggesting that an intact and cleavable NPTH site in HrcU is necessary for total functionally of T3SS in X. citri subsp. citri. Complementation of the ΔhrpB2 X. citri subsp. citri strain with a series of hrpB2 gene mutants revealed that the highly conserved HrpB2 C-terminus is essential for T3SS-dependent development of citrus canker symptoms in planta

Sugarcane (Saccharum X officinarum): A Reference Study for the Regulation of Genetically Modified Cultivars in Brazil

Global interest in sugarcane has increased significantly in recent years due to its economic impact on sustainable energy production. Sugarcane breeding and better agronomic practices have contributed to a huge increase in sugarcane yield in the last 30 years. Additional increases in sugarcane yield are expected to result from the use of biotechnology tools in the near future. Genetically modified (GM) sugarcane that incorporates genes to increase resistance to biotic and abiotic stresses could play a major role in achieving this goal. However, to bring GM sugarcane to the market, it is necessary to follow a regulatory process that will evaluate the environmental and health impacts of this crop. The regulatory review process is usually accomplished through a comparison of the biology and composition of the GM cultivar and a non-GM counterpart. This review intends to provide information on non-GM sugarcane biology, genetics, breeding, agronomic management, processing, products and byproducts, as well as the current technologies used to develop GM sugarcane, with the aim of assisting regulators in the decision-making process regarding the commercial release of GM sugarcane cultivars

Phase correlations in chaotic dynamics: a Shannon entropy measure

In the present work, we investigate phase correlations by recourse to the Shannon entropy. Using theoretical arguments, we show that the entropy provides an accurate measure of phase correlations in any dynamical system, in particular when dealing with a chaotic diffusion process. We apply this approach to different low-dimensional maps in order to show that indeed the entropy is very sensitive to the presence of correlations among the successive values of angular variables, even when it is weak. Later on, we apply this approach to unveil strong correlations in the time evolution of the phases involved in the Arnold’s Hamiltonian that lead to anomalous diffusion, particularly when the perturbation parameters are comparatively large. The obtained results allow us to discuss the validity of several approximations and assumptions usually introduced to derive a local diffusion coefficient in multidimensional near-integrable Hamiltonian systems, in particular the so-called reduced stochasticity approximation.Fil: Cincotta, Pablo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; ArgentinaFil: Giordano, Claudia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; Argentin

Metabolic reprogramming identifies the most aggressive lesions at early phases of hepatic cencerogenesis

Metabolic changes are associated with cancer, but whether they are just bystander effects of deregulated oncogenic signaling pathways or characterize early phases of tumorigenesis remains unclear. Here we show in a rat model of hepatocarcinogenesis that early preneoplastic foci and nodules that progress towards hepatocellular carcinoma (HCC) are characterized both by inhibition of oxidative phosphorylation (OXPHOS) and by enhanced glucose utilization to fuel the pentose phosphate pathway (PPP). These changes respectively require increased expression of the mitochondrial chaperone TRAP1 and of the transcription factor NRF2 that induces the expression of the rate-limiting PPP enzyme glucose-6-phosphate dehydrogenase (G6PD), following miR-1 inhibition. Such metabolic rewiring exclusively identifies a subset of aggressive cytokeratin-19 positive preneoplastic hepatocytes and not slowly growing lesions. No such metabolic changes were observed during non-neoplastic liver regeneration occurring after two/third partial hepatectomy. TRAP1 silencing inhibited the colony forming ability of HCC cells while NRF2 silencing decreased G6PD expression and concomitantly increased miR-1; conversely, transfection with miR-1 mimic abolished G6PD expression. Finally, in human HCC patients increased G6PD expression levels correlates with grading, metastasis and poor prognosis. Our results demonstrate that the metabolic deregulation orchestrated by TRAP1 and NRF2 is an early event restricted to the more aggressive preneoplastic lesions

Coming-Out to Family Members and Internalized Sexual Stigma in Bisexual, Lesbian and Gay People

Coming out has been described as an essential component in the lesbian, gay, and bisexual identity development process and in the mental health of sexual minority people. This study investigated the coming out to family members in lesbian, gay, and bisexual people and analyzed the potential predictors associated to the choice to come out. For this purpose, disclosure of sexual orientation to family members, internalized sexual stigma (evaluated through an adapted short version of the internalized sexual stigma for lesbians, gay men, and bisexual people, measure of the internalized sexual stigma for lesbians, gay, and bisexual), gender, age, sexual orientation, background information, current romantic relationship and the wealth of social network with sexual minority people were assessed on an Italian sample of 291 adolescents and young adults (206 lesbian/gay men and 85 bisexual people). Hierarchical multiple regression showed that higher scores of coming out to family were associated with gay/lesbian identity, liberal political orientation, higher education level, presence of a stable romantic relationship, higher number of lesbian, gay, and bisexual contacts, and lower levels of internalized sexual stigma. Results suggest that bisexual people encounter major difficulties regarding the coming out to family members, respect to lesbian/gay people and indicate that internalized sexual stigma is strongly associated with concealing one’s sexual orientation. Clinical implications for the present findings are discussed

Chapter 15: Dilated Cardiomyopathy at the Crossroad: Multidisciplinary Approach

Patients with dilated cardiomyopathy (DCM) can present for the first time with typical cardiac symptoms, as heart failure, arrhythmias, and syncope, with cardiomyopathy as the main presenting feature. However, DCM may be a common feature of systemic diseases or presenting in the contest of different clinical settings. Symptoms of multisystem disease may be described by patients themselves or be determined on routine examination. A cardiomyopathy-specific mindset, which combines conventional cardiologic assessment with non-cardiac clinical acumen and a systematic approach, is important, because it can be used to identify specific disorders, guide rational selection of diagnostic tests, and increase the chance that disorders with tailored management strategies can be identified. Moreover, in different clinical settings, phenotypic expression in other organs may precede cardiac manifestations, and the cardiologist\u2019s role is to search for cardiac involvement in a patient who already has a non-cardiac diagnosis. In the present chapter, we consider DCM and ventricular dysfunction in the contest of peculiar clinical settings ranging from inflammatory or autoimmune disorders to specific forms of systemic disease, infectious disease, as well as toxin-induced DCM