344 research outputs found
Nephrolithiasis, kidney failure and bone disorders in Dent disease patients with and without CLCN5 mutations
open9noDent disease (DD) is a rare X-linked recessive renal tubulopathy characterised by low-molecular-weight proteinuria (LMWP), hypercalciuria, nephrocalcinosis and/or nephrolithiasis. DD is caused by mutations in both the CLCN5 and OCRL genes. CLCN5 encodes the electrogenic chloride/proton exchanger ClC-5 which is involved in the tubular reabsorption of albumin and LMW proteins, OCRL encodes the inositol polyphosphate 5-phosphatase, and was initially associated with Lowe syndrome. In approximately 25 % of patients, no CLCN5 and OCRL mutations were detected. The aim of our study was to evaluate whether calcium phosphate metabolism disorders and their clinical complications are differently distributed among DD patients with and without CLCN5 mutations. Sixty-four male subjects were studied and classified into three groups: Group I (with CLCN5 mutations), Group II (without CLCN5 mutations) and Group III (family members with the same CLCN5 mutation). LMWP, hypercalciuria and phosphaturic tubulopathy and the consequent clinical complications nephrocalcinosis, nephrolithiasis, bone disorders, and chronic kidney disease (CKD) were considered present or absent in each patient. We found that the distribution of nephrolithiasis, bone disorders and CKD differs among patients with and without CLCN5 mutations. Only in patients harbouring CLCN5 mutations was age-independent nephrolithiasis associated with hypercalciuria, suggesting that nephrolithiasis is linked to altered proximal tubular function caused by a loss of ClC-5 function, in agreement with ClC-5 KO animal models. Similarly, only in patients harbouring CLCN5 mutations was age-independent kidney failure associated with nephrocalcinosis, suggesting that kidney failure is the consequence of a ClC-5 dysfunction, as in ClC-5 KO animal models. Bone disorders are a relevant feature of DD phenotype, as patients were mainly young males and this complication occurred independently of age. The triad of symptoms, LMWP, hypercalciuria, and nephrocalcinosis, was present in almost all patients with CLCN5 mutations but not in those without CLCN5 mutations. This lack of homogeneity of clinical manifestations suggests that the difference in phenotypes between the two groups might reflect different pathophysiological mechanisms, probably depending on the diverse genes involved. Overall, our results might suggest that in patients without CLCN5 mutations several genes instead of the prospected third DD underpin patients' phenotypes.openAnglani, Franca; D’Angelo, Angela; Bertizzolo, Luisa Maria; Tosetto, Enrica; Ceol, Monica; Cremasco, Daniela; Bonfante, Luciana; Addis, Maria Antonietta; Del Prete, DorellaAnglani, Franca; D'Angelo, Angela; Bertizzolo, Luisa Maria; Tosetto, Enrica; Ceol, Monica; Cremasco, Daniela; Bonfante, Luciana; Addis, Maria Antonietta; DEL PRETE, Dorell
Influence of Moraxella sp. colonization on the kidney proteome of farmed gilthead sea breams (Sparus aurata, L.)
Background: Currently, presence of Moraxella sp. in internal organs of fish is not considered detrimental for fish
farming. However, bacterial colonization of internal organs can affect fish wellness and decrease growth rate, stress
resistance, and immune response. Recently, there have been reports by farmers concerning slow growth, poor feed
conversion, and low average weight increase of fish farmed in offshore floating sea cages, often associated with
internal organ colonization by Moraxella sp. Therefore, presence of these opportunistic bacteria deserves further
investigations for elucidating incidence and impact on fish metabolism.
Results: A total of 960 gilthead sea breams (Sparus aurata, L.), collected along 17 months from four offshore sea
cage plants and two natural lagoons in Sardinia, were subjected to routine microbiological examination of internal
organs throughout the production cycle. Thirteen subjects (1.35%) were found positive for Moraxella sp. in the
kidney (7), brain (3), eye (1), spleen (1), and perivisceral fat (1). In order to investigate the influence of Moraxella sp.
colonization, positive and negative kidney samples were subjected to a differential proteomics study by means of
2-D PAGE and mass spectrometry. Interestingly, Moraxella sp. infected kidneys displayed a concerted upregulation
of several mitochondrial enzymes compared to negative tissues, reinforcing previous observations following
lipopolysaccharide (LPS) challenge in fish.
Conclusions: Presence of Moraxella sp. in farmed sea bream kidney is able to induce proteome alterations similar
to those described following LPS challenge in other fish species. This study revealed that Moraxella sp. might be
causing metabolic alterations in fish, and provided indications on proteins that could be investigated as markers of
infection by Gram-negative bacteria within farming plants.Pubblicat
L’ATTO AMMINISTRATIVO PRESUPPOSTO
My thesis is about the supposed administrative act. I deal with the subject in three
chapters speaking about three essential points. In the first chapter I deal with the way
in which the supposed act influences the validity, the efficacy and the essential being
of the acts supposing it. At the first I make a survey of the doctrinal reconstruction of
the phenomenon, then I deal with the subject in a general theory by the light of
juridical dynamics. In the second chapter I analyze the phenomenon of derivative
invalidity, lingering over the invalidity cancelling of which I deny the theoretical
foundations. At the last, in the third chapter, I dwell upon the legal spin-off of
practical reconstructions. In this chapter, by the light of new set of rules introduced in
legal administrative law, I question the practical foundations too, and the real utility of
invalidity cancelling
The impact of sequence database choice on metaproteomic results in gut microbiota studies
Background: Elucidating the role of gut microbiota in physiological and pathological processes has recently emerged as a key research aim in life sciences. In this respect, metaproteomics, the study of the whole protein complement of a microbial community, can provide a unique contribution by revealing which functions are actually being expressed by specific microbial taxa. However, its wide application to gut microbiota research has been hindered by challenges in data analysis, especially related to the choice of the proper sequence databases for protein identification.
Results: Here, we present a systematic investigation of variables concerning database construction and annotation and evaluate their impact on human and mouse gut metaproteomic results. We found that both publicly available and experimental metagenomic databases lead to the identification of unique peptide assortments, suggesting parallel database searches as a mean to gain more complete information. In particular, the contribution of experimental metagenomic databases was revealed to be mandatory when dealing with mouse samples. Moreover, the use of a "merged" database, containing all metagenomic sequences from the population under study, was found to be generally preferable over the use of sample-matched databases. We also observed that taxonomic and functional results are strongly database-dependent, in particular when analyzing the mouse gut microbiota. As a striking example, the Firmicutes/Bacteroidetes ratio varied up to tenfold depending on the database used. Finally, assembling reads into longer contigs provided significant advantages in terms of functional annotation yields.
Conclusions: This study contributes to identify host- and database-specific biases which need to be taken into account in a metaproteomic experiment, providing meaningful insights on how to design gut microbiota studies and to perform metaproteomic data analysis. In particular, the use of multiple databases and annotation tools has to be encouraged, even though this requires appropriate bioinformatic resources
The Wolf of Wall Street. Sopravvivere nello stato di natura
El cine contemporáneo ha profundizado en el retrato de la violencia del
capitalismo financiero y The Wolf of Wall Street (2013) de Martin Scorsese
denuncia el intento de establecer una representaciĂłn plausible de las formas
de vida y relaciones de poder que emergen de la globalizaciĂłn y de la
volatilizaciĂłn de los mercados y de los bienes. El retorno a una dimensiĂłn
salvaje y primordial de la violencia pone en escena un estado de naturaleza
ajeno a las leyes que regulan el comportamiento colectivo, y que se convierten
en el fundamento de un contrato social opuesto al del estado de derecho.
Esta forma de violencia se desdobla en la enunciación con imágenes que
agreden al espectador, imponiéndole un único punto de vista que fagocita
el relato. Desde una perspectiva que combina la metodologĂa semiĂłtica y la
teorĂa crĂtica, se analiza aquĂ el modo en que el film aborda las formas de
subjetivaciĂłn especĂficas del capitalismo financiero.Contemporary cinema has widely inquired the regime of violence deployed
by financial capitalism and The Wolf of Wall Street (2013) by Martin Scorsese
elaborates an appropriate representation of power relationships arisen from
market globalization and the dematerialization of goods. The narration of a
return to a savage and primordial dimension of violence thus stages a state
of nature which is foreign to the laws ruling social behavior and nevertheless
becomes the ground for a new contractuality opposed to the existing state of
right. In turn, this thematic dimension seems to be constantly doubled by an
enunciational level, since it is the image itself which uses violence against the
spectatorial gaze, imposing the main character’s unique point of view, which
seems to expunge any form of otherness and difference. This paper aims
at proposing a reflexion through images about the forms of subjectivation
specific of the animal competition transposed to the virtual world of financial
trade, intertwining the theory in action inside the film with Aesthetics and
Critical Theory positions
Camera con ananas: Immagini di cibo, immaginari di luogo nella piattaforma Airbnb
Il presente contributo verte sulle fotografe che illustrano gli alloggi offerti su Airbnb a Siena e nei suoi dintorni, concentran dosi in particolare sulle fgure alimentari e gastronomiche al fne di rifettere sui processi di turistifcazione del territorio innescati
e accelerati dalle piattaforme
Merging the “Morphology–Performance–Fitness” Paradigm and Life-History Theory in the Eagle Lake Garter Snake Research Project
The morphology-performance-fitness paradigm for testing selection on morphological traits has seen decades of successful application. At the same time, life-history approaches using matrix methods and perturbation studies have also allowed the direct estimate of selection acting on vital rates and the traits that comprise them. Both methodologies have been successfully applied to the garter snakes of the long-term Eagle Lake research project to reveal selection on morphology, such as color pattern, number of vertebrae, and gape size; and life-history traits such as birth size, growth rates, and juvenile survival. Here we conduct a reciprocal transplant study in a common laboratory environment to study selection on morphology and life-history. To place our results in the ecomorphology paradigm, we measure performance outcomes (feeding rates, growth, insulin-like growth factor 1 titers) of morphological variation (body size, condition) and their fitness consequences for juvenile survival?a trait that has large fitness sensitivities in these garter snake populations, and therefore is thought to be subject to strong selection. To better merge these two complementary theories, we end by discussing our findings in a nexus of morphology-performance-fitness-life history to highlight what these approaches, when combined, can reveal about selection in the wild.Fil: Addis, Elizabeth A.. University of Iowa; Estados UnidosFil: Gangloff, Eric J.. University of Iowa; Estados UnidosFil: Palacios, MarĂa Gabriela. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Centro Nacional PatagĂłnico; Argentina. University of Iowa; Estados UnidosFil: Carr, Katherine E.. Gonzaga University; Estados UnidosFil: Bronikowski, Anne M.. University of Iowa; Estados Unido
Critical comparison of sample preparation strategies for shotgun proteomic analysis of formalin-fixed, paraffin-embedded samples: insights from liver tissue
Background: The growing field of formalin-fixed paraffin-embedded (FFPE) tissue proteomics holds promise for improving translational research. Direct tissue trypsinization (DT) and protein extraction followed by in solution digestion (ISD) or filter-aided sample preparation (FASP) are the most common workflows for shotgun analysis of FFPE samples, but a critical comparison of the different methods is currently lacking.
Experimental design: DT, FASP and ISD workflows were compared by subjecting to the same label-free quantitative approach three independent technical replicates of each method applied to FFPE liver tissue. Data were evaluated in terms of method reproducibility and protein/peptide distribution according to localization, MW, pI and hydrophobicity.
Results: DT showed lower reproducibility, good preservation of high-MW proteins, a general bias towards hydrophilic and acidic proteins, much lower keratin contamination, as well as higher abundance of non-tryptic peptides. Conversely, FASP and ISD proteomes were depleted in high-MW proteins and enriched in hydrophobic and membrane proteins; FASP provided higher identification yields, while ISD exhibited higher reproducibility.
Conclusions: These results highlight that diverse sample preparation strategies provide significantly different proteomic information, and present typical biases that should be taken into account when dealing with FFPE samples. When a sufficient amount of tissue is available, the complementary use of different methods is suggested to increase proteome coverage and depth.Pubblicat
An Easy and efficient method for native and immunoreactive <i>Echinococcus granulosus</i> antigen 5 enrichment from hydatid cyst fluid
Background:Currently, the serodiagnosis of cystic echinococcosis relies mostly on crude Echinococcus granulosus hydatid cyst fluid as the antigen. Consequently, available immunodiagnostic tests lack standardization of the target antigen and, in turn, this is reflected on poor sensitivity and specificity of the serological diagnosis.
Methodology/Principal Findings: Here, a chromatographic method enabling the generation of highly enriched Antigen 5 (Ag5) is described. The procedure is very easy, efficient and reproducible, since different hydatid cyst fluid (HCF) sources produced very similar chromatograms, notwithstanding the clearly evident and extreme heterogeneity of the starting material. In addition, the performance of the antigen preparation in immunological assays was preliminarily assessed by western immunoblotting and ELISA on a limited panel of cystic echinococcosis patients and healthy controls. Following western immunoblotting and ELISA experiments, a high reactivity of patient sera was seen, with unambiguous and highly specific results.
Conclusions/Significance: The methods and results reported open interesting perspectives for the development of sensitive diagnostic tools to enable the timely and unambiguous detection of cystic echinococcosis antibodies in patient sera
Critical comparison of sample preparation strategies for shotgun proteomic analysis of formalin-fixed, paraffin-embedded samples
Introduction and objectives. The growing field of formalin-fixed paraffin-embedded (FFPE) tissue proteomics holds promise for improving translational research. Worldwide archival tissue banks hold a significant number and variety of tissue samples, as well as a wealth of retrospective information regarding diagnosis, prognosis, and response to therapy. This makes them an important resource for protein biomarker discovery and validation. Direct tissue trypsinization (DT) and protein extraction followed by in solution digestion (ISD) or filter-aided sample preparation (FASP) are the most common workflows for shotgun LC-MS/MS analysis of FFPE samples, but a critical comparison of the different methods is currently lacking. Methods DT was preceded by homogenization in ammonium bicarbonate, while ISD and FASP comprised protein extraction in SDS based-buffer, followed by SDS depletion with Detergent Removal Spin Columns and Microcon Ultracel YM-30 filtration devices, respectively. The three workflows were applied to consecutive tissue sections cut from an FFPE liver tissue block, and peptide mixtures were finally analyzed according to a label-free quantitative MS approach. Data were evaluated in terms of method reproducibility and protein/peptide distribution according to localization, MW, pI and hydrophobicity. Results and Discussion. DT showed lower reproducibility, good preservation of high-MW proteins, a general bias towards hydrophilic and acidic proteins, much lower keratin contamination, as well as higher abundance of non tryptic peptides. Conversely, FASP and ISD proteomes were depleted in high-MW proteins and enriched in hydrophobic and membrane proteins; FASP provided higher identification yields, while ISD exhibited higher reproducibility. Conclusion. These results highlight that diverse sample preparation strategies provide significantly different proteomic information, and present typical biases that should be taken into account when dealing with FFPE samples. When a sufficient amount of tissue is available, the complementary use of different methods is suggested to increase proteome coverage and depth.2014-10-06Madrid, Spain13th Human Proteome Organization World Congres
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