1,721,010 research outputs found
LIBRERIA FAGICA DI FRAMMENTI DELLA PROTEINA HER2
No full textLa presente invenzione riguarda una libreria fagica di frammenti della
proteina HER2, i suoi usi come strumento di ricerca o per la diagnosi e terapia
del tumore. La libreria si basa sull’espressione sulla superficie di un fago di
un gruppo di frammenti peptidici che rappresentano l’intera sequenza
amminoacidica della proteina HER2. Alcuni frammenti, posizionati in
successione, coprono l’intero dominio extracellulare della molecola, altri sono
sovrapposti ai precedenti, ricoprendo metà di un frammento e metà del
successivo. Questo consente l’esposizione sulla superficie dei fagi di
un repertorio di epitopi conformazionali riconosciuti dal sistema immunitario
Protein transport into the nucleus: characterization of nuclear localization signals in the protozoan ciliate Euplotes
No full textIn E. raikovi, a nuclear protein kinase, designated Er-MAPK1, appears to be phosphorylated in association with the mechanism of signal transduction which promotes cell proliferation through autocrine interactions between cell type-specific signaling protein pheromones and their membrane receptors. This kinase shows significant structural matching to mammalian kinases that are localized in the nucleus of specialized cell types, such as the "Male germ cell-Associated Kinases" and the "Intestinal Cell Kinases". Two Arg/Lys-rich motifs were identified in the Er-MAPK1 C-terminal domain as putative “nuclear localization signals” and their effective function in directing this protein into the nucleus was studied by expressing GFP-tagged protein constructs in mammalian fibroblasts. The results obtained provide evidence that distant related organisms such as ciliates and mammals use the same molecular language for the nuclear translocation and localization of proteins, suggesting that this language arose early in the evolution of the eukaryotic cell
Evolution of the intracellular transport mechanisms in eukaryotes: ciliates and mammals use the same translocation and nuclear localization signals
Full text linkIn the ciliate E. raikovi, self/non-self recognition phenomena are controlled by cell type-specific, water-borne signal proteins (pheromones) by their binding to target cell-surface receptors. The downstream signal transduction pathway activated by the pheromone-receptor interactions of self type (that promote the vegetative, mitogenic cell growth) involves the phosphorylation of a nuclear protein kinase, designated Er-MAPK1, which is structurally similar to the "intestinal-cell kinase" and "male germ cell-associated kinase" described in mammals. To identify the sequence segments responsible for Er-MAPK1 nuclear localization, mouse fibroblasts were transfected with plasmids containing the reporter gene for the "Green-Fluorescent Protein" (GFP) associated to different fragments of the Er-MAPK1 coding sequence. By expressing GFP-tagged protein constructs in mammalian cells, in the C-terminal domain of Er-MAPK1 it was effectively possible to identify an Arg/Lys-rich motif that is required for the nuclear entry of GFP-fused constructs. These results provide evidence that distant related organisms such as ciliates and mammals use the same molecular language for the nuclear translocation and localization of proteins, thus suggesting that this language arose early in the evolution of the eukaryotic cell
Early life permethrin treatment induces in striatum of older rats changes in alpha-synuclein content
No full textObjectives: Many reports have demonstrated the strong relationship between environmental exposure during the first stages of life and the development in adult phase of neurodegenerative disorders. Previous studies performed in rats exposed from postnatal day 6 to 21 to low dose of permethrin (PERM) induced Parkinson-like neurodegeneration characterized in the striatum by a decrease of Nurr1 gene and protein expression, reduced dopamine level, together with its accelerated turnover (1-3). α-Synuclein is a presynaptic neuronal protein that contributes to Parkinson pathogenesis through its aggregation in oligomeric conformations called protofibrils that mediate disruption of cellular homeostasis and neuronal death. The present work aims to analyze in the striatum of early PERM treated rats the -sinuclein content in the adolescent, adult and old rats.
Methods PERM was dissolved in corn oil and administered orally to Wistar rats for 15 days, once a day in the morning from PND6 to PND21. Control rats were treated with vehicle (corn oil 4 ml/kg) on a similar schedule. Rats were sacrificed after 90 days (adolescent age), 180 days (adult age) and 300 days (old age) from the treatment and the striatum collected and kept at -80°C. Tissue lysates were prepared using two different buffers that solubilize -sinuclein in the free and aggregated forms. 40 g of protein were used for western blot analysis, separated using SDS–PAGE (7,5%) and electrophoretically blotted on a nitrocellulose support (Hybond C, Amersham Bioscience, Little Chalfont, UK). -synuclein antibody was purchased by Santa Cruz.
Results -synuclein free resulted more expressed in striatum from adolescent treated rats with respect to control while in samples from adult and old rats we observed an opposite situation with decreased level of -sinuclein in treated respect to their controls. Different oligomeric conformations of the protein were also observed when we analyzed by western blotting the lysate with solubilised aggregated protein. Striatum from adolescent treated rats shows higher expression of protein respect to the control with the formations of oligomers
Conclusions The early life treatment with PERM induces more significant effect on adolescent rats with the increase of -synuclein in both form, free and aggregated. This behavior could be explained with a more effective impairment of dopaminergic system in the first important phase of the life such as adolescence.
References:
1) Carloni M et al. 2012, Exp Gerontol. 47(1):60-66 2) Nasuti C et al.2013, Toxicology 303:162-168 3) Nasuti C et al. 2007, Toxicology 229(3):194-20
α-synuclein and DNMT imbalance, following early life permethrin treatment, leads to Nurr1 down regulation
No full textEnvironmental factors interface the genotype-phenotype correlation and result conferring a stable memory of activity status of genes. Early-life exposure (postnatal day 6-21) to low doses of the pesticide permethrin leads to down regulation of Nurr1 expression and also dopamine levels. This is correlated with hyper methylation of CpG in the promoter region of Nurr1 in adolescent rats exposed to permethrin while no alteration in methylation was seen in the age-matched control group. In order to define the mechanisms associated with the decreased Nurr1 gene expression, DNMTs and -synuclein levels were measured in adolescent, adult and old rats. The results obtained confirm their imbalance depending on the treatment and the age of rats
Structural stability and increase in size rationalize the efficiency of lipoplexes in serum.
Full text linkWe have investigated the effect of serum on nanometric structure, size, surface potential, DNA-hinding capacity, and transfection efficiency of DDAB-DOPE/DNA and DC-Chol-DOPE/DNA lipoplexes as a function of membrane charge density and cationic lipid/DNA charge ratio. In the absence of serum, the nanometric structure and DNA binding capacity of lipoplexes determined the transfection efficiency. When serum was added, the transfection efficiency of all lipoplex formulations was found to increase. We identified structural stability and an increase in size in serum as major parameters regulating the efficiency of lipofection. By extrapolation, we propose that serum, regulating the size of resistant lipid - DNA complexes, can control the mechanism of internalization of lipoplexes and, in turn, their efficiency
Effect of Electrolyzed Reduced Water in an Animal Model of Parkinson-Like Disease
No full textEffect of Electrolyzed Reduced Water in an Animal Model of
Parkinson-Like Disease
Cinzia Nasuti1, Donatella Fedeli1, Laura Bordoni2, Maura Montani3,
Ivan Dus4, Rosita Gabbianelli1
1School of Pharmacy, University of Camerino, Italy; 2School of Advanced
Studies, University of Camerino, Italy; 3School of Biosciences and
Veterinary Medicine, University of Camerino, Italy; 4Head of Research for
Chanson Water Company Taiwan
Objectives: Early life environmental factors, life style and diet have a
profound impact on the organism’s later development and subsequent onset of
age-related diseases such as neurodegenerative diseases. In previous studies, we
developed an animal model where the early life exposure (from postnatal day 6
to 21) to low dose of pesticide permethrin (PERM, 34 mg/kg) induced Parkinsonlike
neurodegeneration in rats characterized by decreased dopamine, Nurr1 and
glutathione levels in the basal ganglia, altered immune responses and gut
disbiosis. The animal model was exposed to a neurotoxin PERM that is an insecticide
widely used for indoor and outdoor applications (i.e. carpets, kitchen
worktops other treated wood furniture, lawn, mosquito control). The presence
of its metabolites in the urine of 98% of population makes this insecticide a
reliable environmental risk factor to health.
The present study aims to test the effect of electrolyzed reduced water
(ERW) on this animal model developing neurodegenerative disease after
exposure to PERM pesticide in neonatal age.
Methods: The effect of ERW is determined by cognitive tests, dopamine and
Nurr1 levels measured in basal ganglia and Tyrosine Hydroxylase (TH) immunostaining
in the Substantia Nigra pars compacta (SNc) in 60-day-old rats.
Results: When working memory is assessed in a T-maze, PERM group has a
worst performance compared with healthy controls, whereas the performance
of PERM+ERW is similar to control group. The same trend is observed for the
average of perseverative errors.
With regard to dopamine levels measured by HPLC in basal ganglia,
decreased levels are observed in PERM group compared to control group,
whereas the co-treatment with PERM+ERW protects against the damage induced
by the pesticide even increasing the levels of dopamine. Similar results are
obtained with regard to Nurr1 levels measured by western blot in basal ganglia.
Immunohistochemical analysis with anti-TH antibody marking dopamine
neurons in SNc shows a reduced number of TH-positive neurons in PERM rats
compared to control rats, whereas a slight increase (not statistically significant)
was observed in the PERM+ERW group with respect to PERM group.
Conclusion: The use of ERW as functional water could be helpful as a therapeutic
tool in the prevention of neurodegenerative diseases
Self-assembled ternary complexes of neutral liposomes, DNA and bivalent metal cations. Promising vectors for gene transfer?
No full textBy means of synchrotron x-ray diffraction we demonstrate the self-assembled formation of the neutral ternary dioleoyl-phosphatidylcholine-deoxyribonucleic acid (plasmid) - Me2+ (Me=Ca and Mn) complexes in the liquid-crystalline Lα phase. We also report an attempt of an in vitro transfection on mouse fibroplast NIH 3T3 cell lines, which shows the capability of these complexes to transfect DNA. Based on the reported results, efficient encapsulation of DNA plasmids in these ternary neutral complexes may represent an important alternative to current systemic gene approaches
Sviluppo di in anticorpo monoclonale anti-Asaia (anti-Asaia monoclonal antibody)
No full textLa presente invenzione riguarda lo sviluppo di un anticorpo monoclonale prodotto contro batteri del genere Asaia. Il succitato anticorpo è caratterizzato da elevata specificità ed è stato ottenuto mediante la fusione di cellule mielomatose murine con splenociti ottenuti da topi Balb/c immunizzati con il batterio di nostro interesse, ovvero Asaia, secondo la tecnica messa a punto da Kohler e Milstein nel 1974 (Nature 256: 495-497)
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