Comparative expression profiling of wild type Drosophila Malpighian tubules and von Hippel-Lindau haploinsufficient mutant

The von-Hippel Lindau (VHL) disease is a hereditary genetic disorder that predisposes to the onset of several highly vascularized benign and malignant tumors, developing with elevate frequency in the central nervous system and kidneys. The most-aggressive VHL tumor is ccRCC, the clear-cell renal cell carcinoma, affecting the kidney. VHL disease etiology can be attributed to the inheritance of a VHL loss-of-function allele, typically a deletion (Gnarra et al., 1994; Herman et al., 1994); this facilitates the somatic inactivation of the other allele (through amorphic mutations or gene silencing through promoter methylation), leading to the onset of the tumorous phenotype (Latif et al., 1993). This reveals the haploinsufficient behavior of the VHL gene. The high vascularization of VHL tumors can be explained considering that human VHL protein is the substrate-binding subunit of an E3 ubiquitin ligase (Lonergan et al., 1998; Iwai et al., 1999; Kamura et al., 1999) involved in the poly-ubiquitination of HIF-1α transcription factor. This post-translational modification leads HIF-1α to proteosomal degradation (Maxwell et al., 1999). Loss of VHL function causes the stabilization of HIF-1α, triggering cellular response and adaptation to hypoxic conditions (expression of genes involved in glycolysis, angiogenesis and erythropoiesis) (Bader and Hsu, 2012). While this represents the canonical function of VHL, other HIF-1α-independent function of VHL have been identified, thanks to the contribution of model organisms (Hsu, 2012). Indeed, VHL gene function is conserved and also Drosophila has a VHL homolog, the dVHL gene (Adryan et al., 2000; Aso et al., 2000). dVHL is involved in the development of Drosophila vascular system (Adryan et al., 2000; Hsouna et al., 2010) and in morphogenesis of follicular epithelium of the egg chamber (Duchi et al., 2010). Interestingly, some VHL functions are mediated by Awd, an endocytic mediator whose human orthologs are NME1/2 metastasis suppressors (Rosengard et al., 1989). Awd is broadly required during Drosophila development since it is involved in epithelial morphogenesis (Nallamothu et al., 2008; Woolworth et al., 2009; Ignesti et al., 2014) and required for maintaining genomic stability (Romani et al., 2017). Moreover, Awd is also present into the extracellular fluids of Drosophila larvae (Romani et al., 2016, 2018). In Drosophila, two pairs of monolayered epithelial Malpighian tubules, each composed of 100-150 cells, absolve to osmoregulation and excretion functions (Denholm and Skaer, 2009). Transcriptomic analysis of Malpighian tubules revealed that among genes that are here enriched there are homologs of human genes implicated into renal pathologies (Wang et al., 2004). This justifies the use of Drosophila Malpighian tubules as model system to gain insights into pathophysiology of human kidneys (Dow and Romero, 2010; Miller et al., 2013). The dVHL1.1 allele is a loss of function mutation of the dVHL locus (Duchi et al., 2010; Hsouna et al., 2010). dVHL1.1/+ flies mimic the genetic condition of VHL patients. We carried out a genome-wide gene expression profiling of whole Malpighian tubules dissected from Drosophila females both heterozygous for the dVHL1.1 mutation and with two wild type copies of the dVHL gene. The comparison of differentially expressed genes in the two genetic backgrounds potentially allows to identify genes that are sensible to dVHL functional copy number. Quality control assessments of the data were performed and results obtained from the differential expression analysis were confirmed by qRT-PCR. With this approach we aimed to provide a well-controlled dataset for a better understanding of the VHL disease. Indeed, even if further molecular and functional characterization are needed, human homologs of the differentially expressed genes, if existing, could have a role in the somatic inactivation of the wild type copy of VHL and/or into the very first phase of cancer onset

Ward identity and optical-conductivity sum rule in the d-density wave state

We consider the role of the Ward identity in dealing with the transport properties of an interacting system forming a d-wave modulated charge-density wave or staggered flux phase. In particular, we address this issue from the point of view of the restricted optical-conductivity sum rule. Our aim is to provide a controlled approximation for the current-current correlation function which allows us also to determine analytically the corresponding sum rule. By analyzing the role of the vertex functions in both the microscopic interacting model and in the effective mean-field Hamiltonian, we propose a non-standard low-energy sum-rule for this system. We also discuss the possible applicability of these results for the description of cuprate superconductors in the pseudogap regime.Comment: Revised version, accepted for publication in Phys. Rev.

Pseudogap and spectral function from superconducting fluctuations to the bosonic limit

The crossover from weak to strong coupling for a three dimensional continuum model of fermions interacting via an attractive contact potential is studied above the superconducting critical temperature. The pair-fluctuation propagator, the one-loop self-energy, and the spectral function are investigated in a systematic way from the superconducting fluctuation regime (weak coupling) to the bosonic regime (strong coupling). Analytic and numerical results are reported. In the strong-coupling regime, where the pair fluctuation propagator has bosonic character, two quite different peaks appear in the spectral function, a broad one at negative frequencies and a narrow one at positive frequencies. By decreasing coupling, the two-peak structure evolves smoothly. In the weak-coupling regime, where the fluctuation propagator has diffusive Ginzburg-Landau character, the overall line-shape of the spectral function is more symmetric. The systematic analysis of the spectral function identifies specific features which allow one to distinguish by ARPES whether a system is in the weak- or strong-coupling regime. Connection of the results of our analysis with the phenomenology of cuprate superconductors is also attempted and rests on the recently introduced two-gap model.Comment: 19 pages, 18 figure

Density-induced BCS to Bose-Einstein crossover

We investigate the zero-temperature BCS to Bose-Einstein crossover at the mean-field level, by driving it with the attractive potential and the particle density.We emphasize specifically the role played by the particle density in this crossover.Three different interparticle potentials are considered for the continuum model in three spatial dimensions, while both s- and d-wave solutions are analyzed for the attractive (extended) Hubbard model on a two-dimensional square lattice. For this model the peculiar behavior of the crossover for the d-wave solution is discussed.In particular, in the strong-coupling limit when approaching half filling we evidence the occurrence of strong correlations among antiparallel-spin fermions belonging to different composite bosons, which give rise to a quasi-long-range antiferromagnetic order in this limit.Comment: 10 pages, 5 enclosed figure

Temperature-doping phase diagram of layered superconductors

The superconducting properties of a layered system are analyzed for the cases of zero- and non-zero angular momentum of the pairs. The effective thermodynamic potential for the quasi-2D XY-model for the gradients of the phase of the order parameter is derived from the microscopic superconducting Hamiltonian. The dependence of the superconducting critical temperature T_c on doping, or carrier density, is studied at different values of coupling and inter-layer hopping. It is shown that the critical temperature T_c of the layered system can be lower than the critical temperature of the two-dimensional Berezinskii-Kosterlitz-Thouless transition T_BKT at some values of the model parameters, contrary to the case when the parameters of the XY-model do not depend on the microscopic Hamiltonian parameters.Comment: To be published in Phys. Rev.

Interleukin-6 neutralization ameliorates symptoms in prematurely aged mice

Hutchinson\u2013Gilford progeria syndrome (HGPS) causes premature aging in children, with adipose tissue, skin and bone deterioration, and cardiovascular impairment. In HGPS cells and mouse models, high levels of interleukin-6, an inflammatory cytokine linked to aging processes, have been detected. Here, we show that inhibition of interleukin-6 activity by tocilizumab, a neutralizing antibody raised against interleukin-6 receptors, counteracts progeroid features in both HGPS fibroblasts and LmnaG609G/G609G progeroid mice. Tocilizumab treatment limits the accumulation of progerin, the toxic protein produced in HGPS cells, rescues nuclear envelope and chromatin abnormalities, and attenuates the hyperactivated DNA damage response. In vivo administration of tocilizumab reduces aortic lesions and adipose tissue dystrophy, delays the onset of lipodystrophy and kyphosis, avoids motor impairment, and preserves a good quality of life in progeroid mice. This work identifies tocilizumab as a valuable tool in HGPS therapy and, speculatively, in the treatment of a variety of aging-related disorders

Stability of condensate in superconductors

According to the BCS theory the superconducting condensate develops in a single quantum mode and no Cooper pairs out of the condensate are assumed. Here we discuss a mechanism by which the successful mode inhibits condensation in neighboring modes and suppresses a creation of noncondensed Cooper pairs. It is shown that condensed and noncondensed Cooper pairs are separated by an energy gap which is smaller than the superconducting gap but large enough to prevent nucleation in all other modes and to eliminate effects of noncondensed Cooper pairs on properties of superconductors. Our result thus justifies basic assumptions of the BCS theory and confirms that the BCS condensate is stable with respect to two-particle excitations

Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1

BACKGROUND: Anosmin-1, the protein implicated in the X-linked Kallmann's syndrome, plays a role in axon outgrowth and branching but also in epithelial morphogenesis. The molecular mechanism of its action is, however, widely unknown. Anosmin-1 is an extracellular protein which contains a cysteine-rich region, a whey acidic protein (WAP) domain homologous to some serine protease inhibitors, and four fibronectin-like type III (FnIII) repeats. Drosophila melanogaster Kal-1 (DmKal-1) has the same protein structure with minor differences, the most important of which is the presence of only two FnIII repeats and a C-terminal region showing a low similarity with the third and the fourth human FnIII repeats. We present a structure-function analysis of the different DmKal-1 domains, including a predicted heparan-sulfate binding site. RESULTS: This study was performed overexpressing wild type DmKal-1 and a series of deletion and point mutation proteins in two different tissues: the cephalopharyngeal skeleton of the embryo and the wing disc. The overexpression of DmKal-1 in the cephalopharyngeal skeleton induced dosage-sensitive structural defects, and we used these phenotypes to perform a structure-function dissection of the protein domains. The reproduction of two deletions found in Kallmann's Syndrome patients determined a complete loss of function, whereas point mutations induced only minor alterations in the activity of the protein. Overexpression of the mutant proteins in the wing disc reveals that the functional relevance of the different DmKal-1 domains is dependent on the extracellular context. CONCLUSION: We suggest that the role played by the various protein domains differs in different extracellular contexts. This might explain why the same mutation analyzed in different tissues or in different cell culture lines often gives opposite phenotypes. These analyses also suggest that the FnIII repeats have a main and specific role, while the WAP domain might have only a modulator role, strictly connected to that of the fibronectins

Cooper pairing with finite angular momentum: BCS vs Bose limits

We revisit the old problem of exotic superconductivity as Cooper pairing with finite angular momentum emerging from a central potential. Based on some general considerations, we suggest that the phenomenonn is associated with interactions that keep electrons at some particular, finite distance (r_{0}), and occurs at a range of intermediate densities (n\sim 1/r_{0}^{3}). We discuss the ground state and the critical temperature in the framework of a standard functional-integral theory of the BCS to Bose crossover. We find that, due to the lower energy of two-body bound states with (l=0), the rotational symmetry of the ground state is always restored on approaching the Bose limit. Moreover in that limit the critical temperature is always higher for pairs with (l=0.) The breaking of the rotational symmetry of the continuum by the superfluid state thus seems to be a property of weakly-attractive, non-monotonic interaction potentials, at intermediate densities.Comment: Proceedings of SCENM02 (to appear in J. Phys. A

Crustins: enigmatic WAP domain-containing antibacterial proteins from crustaceans

Crustins are antibacterial proteins of ca. 7�14 kDa with a characteristic four-disulphide core-containing whey acidic protein (WAP) domain, expressed by the circulating haemocytes of crustaceans. Over 50 crustin sequences have been now reported from a variety of decapods, including crabs, lobsters, shrimp and crayfish. Three main types seem to occur but all possess a signal sequence at the amino terminus and a WAP domain at the carboxyl end. Differences between types lie in the structure of the central region. Those crustins purified as the native protein or expressed recombinantly all kill Gram-positive bacteria, and gene studies have shown that they are constitutively expressed, often at high levels, but show no consistent patterns of change in expression following injection ofbacteria. This variable response to infection is enigmatic but indicates that these proteins could perform additional functions, perhaps as immune regulators in recovery from wounding, trauma or physiological stress