169 research outputs found
Adjuvant Treatment in Pancreatic Cancer: Shaping the Future of the Curative Setting
Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease even in the early stages, despite progresses in surgical and pharmacological treatment in recent years. High potential for metastases is the main cause of therapeutic failure in localized disease, highlighting the current limited knowledge of underlying pathological processes. However, nowadays research is focusing on the search for personalized approaches also in the adjuvant setting for PDAC, by implementing the use of biomarkers and investigating new therapeutic targets. In this context, the aim of this narrative review is to summarize the current treatment scenario and new potential therapeutic approaches in early stage PDAC, from both a preclinical and clinical point of view. Additionally, the review examines the role of target therapies in localized PDAC and the influence of neoadjuvant treatments on survival outcomes
Pancreatic cancer molecular classifications: From bulk genomics to single cell analysis
Pancreatic cancer represents one of the most lethal disease worldwide but still orphan of a molecularly driven therapeutic approach, although many genomic and transcriptomic classifications have been proposed over the years. Clinical heterogeneity is a hallmark of this disease, as different patients show different responses to the same therapeutic regimens. However, genomic analyses revealed quite a homogeneous disease picture, with very common mutations in four genes only (KRAS, TP53, CDKN2A, and SMAD4) and a long tail of other mutated genes, with doubtful pathogenic meaning. Even bulk transcriptomic classifications could not resolve this great heterogeneity, as many informations related to small cell populations within cancer tissue could be lost. At the same time, single cell analysis has emerged as a powerful tool to dissect intratumoral heterogeneity like never before, with possibility of generating a new disease taxonomy at unprecedented molecular resolution. In this review, we summarize the most relevant genomic, bulk and single-cell transcriptomic classifications of pancreatic cancer, and try to understand how novel technologies, like single cell analysis, could lead to novel therapeutic strategies for this highly lethal disease
Serum gamma-glutamyltransferase fractions in Myotonic Dystrophy type I: Differences with healthy subjects and patients with liver disease.
Objectives: Elevation of serum gamma-glutamyltransferase (GGT), in absence of a clinically significant
liver damage, is often found in Myotonic Dystrophy type-1 (DM1).
In this study we investigated if a specific GGT fraction pattern is present in DM1.
Designs and methods: We compared total and fractional GGT values (b-, m-, s-, f-GGT) among patients
with DM1 or liver disease (LD) and healthy subjects (HS).
Results: The increase of GGT in DM1 and LD, vs HS, was mainly due to s-GGT (median: 32.7; 66.7; and
7.9 U/L, respectively), and b-GGT (8.5; 18.9; and 2.1 U/L). The subset of DM1 patients matched with HS with
corresponding serum GGT showed higher b-GGT (6.0 vs 4.2 U/L).
Conclusions: DM1 patients with normal total GGT values showed an alteration of the production and
release in the blood of GGT fractions. Since increased s-GGT is also found in LD, a sub-clinical liver damage
likely occurs in DM1 subjects apparently free of liver disease
Elevated levels of erythrocyte-conjugated dienes indicate increased lipid peroxidation in schistosomiasis mansoni patients
Optimizing expression and purification of an ATP-binding gene gsiA from Escherichia coli k-12 by using GFP fusion
The cloning, expression and purification of the glutathione (sulfur) import system ATP-binding protein (gsiA) was carried out. The coding sequence of Escherichia coli gsiA, which encodes the ATP-binding protein of a glutathione importer, was amplified by PCR, and then inserted into a prokaryotic expression vector pWaldo-GFPe harboring green fluorescent protein (GFP) reporter gene. The resulting recombinant plasmid pWaldo-GFP-GsiA was transformed into various E. coli strains, and expression conditions were optimized. The effect of five E. coli expression strains on the production of the recombinant gsiA protein was evaluated. E. coli BL21 (DE3) was found to be the most productive strain for GsiA-GFP fusion-protein expression, most of which was insoluble fraction. However, results from in-gel and Western blot analysis suggested that expression of recombinant GsiA in Rosetta (DE3) provides an efficient source in soluble form. By using GFP as reporter, the most suitable host strain was conveniently obtained, whereby optimizing conditions for overexpression and purification of the proteins for further functional and structural studies, became, not only less laborious, but also time-saving
Contribution by Polymorphonucleate Granulocytes to Elevated Gamma-Glutamyltransferase in Cystic Fibrosis Sputum
Background: Cystic fibrosis (CF) is an autosomal recessive disorder characterized by a chronic neutrophilic airways
inflammation, increasing levels of oxidative stress and reduced levels of antioxidants such as glutathione (GSH). Gammaglutamyltransferase
(GGT), an enzyme induced by oxidative stress and involved in the catabolism of GSH and its derivatives,
is increased in the airways of CF patients with inflammation, but the possible implications of its increase have not yet been
investigated in detail.
Principal Findings: The present study was aimed to evaluate the origin and the biochemical characteristics of the GGT
detectable in CF sputum. We found GGT activity both in neutrophils and in the fluid, the latter significantly correlating with
myeloperoxidase expression. In neutrophils, GGT was associated with intracellular granules. In the fluid, gel-filtration
chromatography showed the presence of two distinct GGT fractions, the first corresponding to the human plasma b-GGT
fraction, the other to the free enzyme. The same fractions were also observed in the supernatant of ionomycin and fMLPactivated
neutrophils. Western blot analysis confirmed the presence of a single band of GGT immunoreactive peptide in the
CF sputum samples and in isolated neutrophils.
Conclusions: In conclusion, our data indicate that neutrophils are able to transport and release GGT, thus increasing GGT
activity in CF sputum. The prompt release of GGT may have consequences on all GGT substrates, including major
inflammatory mediators such as S-nitrosoglutathione and leukotrienes, and could participate in early modulation of
inflammatory response
Rapid Dopaminergic Modulation of the Fish Hypothalamic Transcriptome and Proteome
Background - Dopamine (DA) is a major neurotransmitter playing an important role in the regulation of vertebrate reproduction. We developed a novel method for the comparison of transcriptomic and proteomic data obtained from in vivo experiments designed to study the neuroendocrine actions of DA. // Methods and Findings - Female goldfish were injected (i.p.) with DA agonists (D1-specific; SKF 38393, or D2-specific; LY 171555) and sacrificed after 5 h. Serum LH levels were reduced by 57% and 75% by SKF 38393 and LY 171555, respectively, indicating that the treatments produced physiologically relevant responses in vivo. Bioinformatic strategies and a ray-finned fish database were established for microarray and iTRAQ proteomic analysis of the hypothalamus, revealing a total of 3088 mRNAs and 42 proteins as being differentially regulated by the treatments. Twenty one proteins and mRNAs corresponding to these proteins appeared on both lists. Many of the mRNAs and proteins affected by the treatments were grouped into the Gene Ontology categorizations of protein complex, signal transduction, response to stimulus, and regulation of cellular processes. There was a 57% and 14% directional agreement between the differentially-regulated mRNAs and proteins for SKF 38393 and LY 171555, respectively. // Conclusions - The results demonstrate the applicability of advanced high-throughput genomic and proteomic analyses in an amendable well-studied teleost model species whose genome has yet to be sequenced. We demonstrate that DA rapidly regulates multiple hypothalamic pathways and processes that are also known to be involved in pathologies of the central nervous system
Transcriptomic analysis of maternally provisioned cues for phenotypic plasticity in the annual killifish, Austrofundulus limnaeus
Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology
- …