Wall proteins of Vitis vinifera pollen II. Influence of environment and rootstock on the electrophoretic pattern

Soluble wall proteins from Vitis vinifera pollen are genotypically determined and their expression, if not the extent of their expression, is independent from external factors. After we had previously observed that the time of sample collection does not influence their electrophoretic pattern, we demonstrate in the present work that this is true also for environment where the vine is growing and for the rootstock on which the scion is grafted

SLC6A14, a Pivotal Actor on Cancer Stage: When Function Meets Structure

SLC6A14 (ATB0,+) is a sodium- and chloride-dependent neutral and dibasic amino acid transporter that regulates the distribution of amino acids across cell membranes. The transporter is overexpressed in many human cancers characterized by an increased demand for amino acids; as such, it was recently acknowledged as a novel target for cancer therapy. The knowledge on the molecular mechanism of SLC6A14 transport is still limited, but some elegant studies on related transporters report the involvement of the 12 transmembrane \u3b1-helices in the transport mechanism, and describe structural rearrangements mediated by electrostatic interactions with some pivotal gating residues. In the present work, we constructed a SLC6A14 model in outward-facing conformation via homology modeling and used molecular dynamics simulations to predict amino acid residues critical for substrate recognition and translocation. We docked the proteinogenic amino acids and other known substrates in the SLC6A14 binding site to study both gating regions and the exposed residues involved in transport. Interestingly, some of these residues correspond to those previously identified in other LeuT-fold transporters; however, we could also identify a novel relevant residue with such function. For the first time, by combined approaches of molecular docking and molecular dynamics simulations, we highlight the potential role of these residues in neutral amino acid transport. This novel information unravels new aspects of the human SLC6A14 structure-function relationship and may have important outcomes for cancer treatment through the design of novel inhibitors of SLC6A14-mediated transport

Vitis vinifera - a chemotaxonomic approach: Seed storage proteins

The IEF pattern of the constituent peptides for the storage protein from Viris vinifera endosperm is used for the construction of a dendrogram relating 74 seed specimens

Peyronie's disease development and management in diabetic men

Background: Peyronie's disease (PD) is a fibrosing disorder of the penis resulting in plaque formation and penile deformity that negatively affect sexual and psychosocial function of patients. A multifactorial etiology of PD is assumed with diabetes mellitus (DM) being a potential risk factor. Objectives: The aim of this narrative review was to investigate diabetes role in PD pathophysiology, diagnosis, and treatment. Materials and methods: A non-systematic narrative review of original articles, meta-analyses, and randomized trials was conducted, including articles in the pre-clinical setting to support relevant findings. Results: Diabetes is one of the most common comorbidity observed in PD patients, with a prevalence of about 11% and a strong association with erectile dysfunction (ED). DM is associated with both a higher risk of developing PD and has also an impact on the outcomes of PD's treatments. Discussion: Evidence from literature underlines that metabolic alterations typical of DM are pivotal factors in the development of PD and resistance to its medical treatment. Conclusion: The role of DM in development of PD is still debated, while its role in PD development is not completely clear, there is a clear impact of DM on PD treatment outcomes

A proteomic approach to identify novel disease biomarkers in LCAT deficiency

Genetic LCAT deficiency is a rare recessive autosomal disease due to loss-of-function mutations in the gene coding for the enzyme lecithin:cholesterol acyltransferase (LCAT). Homozygous carriers are characterized by corneal opacity, haemolytic anaemia and renal disease, which represent the first cause of morbidity and mortality in these subjects. Diagnostic and prognostic markers capable of early detecting declining kidney function in these subjects are not available, and the specific serum or urine proteomic signature of LCAT deficient carriers has never been assessed. Taking advantage of a proteomic approach, we performed 2-DE analysis of carriers' plasma and identified proteins present at different concentration in samples from homozygous carriers. Our data confirm the well-known alterations in the concentration of circulating apolipoproteins, with a statistically significant decrease of both apoA-I and apoA-II and a statistically significant increase of apoC-III. Furthermore, we observed increased level of alpha-1-antitrypsin, zinc-alpha-2-glycoprotein and retinol-binding protein 4, and reduced level of clusterin and haptoglobin. Interestingly, only beta but not alpha subunit of haptoglobin is significant reduced in homozygous subjects. Despite the limited sample size, our findings set the basis for assessing the identified protein in a larger population and for correlating their levels with clinical markers of renal function and anaemia. Significance: This investigation defines the effects of LCAT deficiency on the level of the major plasma proteins in homozygous and heterozygous carriers. Increase for some proteins, with different function, together with a drop for haptoglobin, and specifically for haptoglobin beta chains, are reported for the first time as part of a coherent signature. We are glad to have the opportunity to report our findings on this subject, which is one of the main interests for our research group, when Journal of Proteomics celebrates its 10th anniversary. With its various sections devoted to different areas of research, this journal is a privileged forum for publishing proteomic investigations without restrictions either in sample type or in technical approach. It is as well a privileged forum for reviewing literature data on various topics related to proteomics investigation, as colleagues in our research group have done over the years; by the way, a good share of the reviewed papers were as well reports published in Journal of Proteomics itself. The journal also offers opportunities for focused surveys through thematic issues devoted to a variety of subjects, timely selected for their current relevance in research; it was an honour for colleagues in our group to recently act as editors of one of those. Out of this diverse experience, we express our appreciation for the endeavour of Journal of Proteomics in its first 10 years of life \u2013 and wish identical and possibly greater success for the time to come

Serum metal evaluation in a small cohort of Amyotrophic Lateral Sclerosis patients reveals high levels of thiophylic species

Amyotrophic Lateral Sclerosis (ALS) has often been associated with improper/altered metal metabolism. Analysis of thiophylic metals in serum from a small and geographically restricted cohort of ALS patients indicates contents of Pb and Ni much higher in patients than in controls (Ni, 5-fold; Pb, 2-fold). Se levels are also higher in the patients\u2019 group, which has instead lower As levels than controls. Thiophylic metals may impair biogenesis of FeS clusters or substitute for iron, even in folded proteins; Se may non-functionally replace S. Thus, improper assembly/function of FeS proteins could represent another possible issue to be considered in ALS pathogenesis

Metal and proteomic analysis of sporadic ALS patients with common geographical origin

Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder characterized by selective degeneration of both upper and lower motor neurons in the brain, brainstem, and spinal cord. This results in paralysis due to muscle weakness and atrophy, leading to death in 3-5 years. Genetic and environmental factors are involved in the pathogenesis of the disease and metals metabolism have been linked to ALS. This study enrolled seven patients and five controls (age matched, living in the same geographical area). For metal quantitation, samples of serum were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range. Statistical analyses were carried out with Student's t-test and Artificial Neural Networks. Among the metals analyzed, As concentration resulted significantly lower in patients than in controls (p=0.007); Hg too was found in lower concentration in patients, but with a lower statistical significance (p=0.13). Higher concentration of Al in patients was detected (p=0.08). In this study, we were not able to confirm the higher concentrations of Ni and Pb in patients previously described in a smaller cohort. Our proteomics data show that APOA2 is decreased by 30% in patients with respect to controls. Furthermore, AHSG and SAP showed a significant decrease in patients with a story of more than 10 years of disease. Impaired metal homeostasis, attributable to environmental exposure, could lead to mineral overload. Besides promoting oxidative stress, metals can compete for the binding sites of metal-containing proteins, such as those containing iron-sulfur clusters. At present, no literature data link APOA2 to ALS, but the fact that its mRNA is processed by TDP43, provides a possible connection with the disease. The proteins differentially expressed belong to the group of Acute Phase Reaction proteins, possibly linking ALS to a chronic inflammation status. Further experiments are still ongoing

Development of the first in vivo GPR17 ligand through an iterative drug discovery pipeline: A novel disease-modifying strategy for multiple sclerosis

The GPR17 receptor, expressed on oligodendroglial precursors (OPCs, the myelin producing cells), has emerged as an attractive target for a pro-myelinating strategy in multiple sclerosis (MS). However, the proof-of-concept that selective GPR17 ligands actually exert protective activity in vivo is still missing. Here, we exploited an iterative drug discovery pipeline to prioritize novel and selective GPR17 pro-myelinating agents out of more than 1,000,000 compounds. We first performed an in silico high-throughput screening on GPR17 structural model to identify three chemically-diverse ligand families that were then combinatorially exploded and refined. Top-scoring compounds were sequentially tested on reference pharmacological in vitro assays with increasing complexity, ending with myelinating OPC-neuron co-cultures. Successful ligands were filtered through in silico simulations of metabolism and pharmacokinetics, to select the most promising hits, whose dose and ability to target the central nervous system were then determined in vivo. Finally, we show that, when administered according to a preventive protocol, one of them (named by us as galinex) is able to significantly delay the onset of experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. This outcome validates the predictivity of our pipeline to identify novel MS-modifying agents

Dissecting Sporadic ALS in a geographical cluster of patients: a multidisciplinary study

Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder that results in paralysis and leading to death in 3-5 years. Genetic and environmental factors are involved in the pathogenesis of this disease and metals metabolism has been linked to ALS. Proteomic studies are currently being performed to search for possible biomarkers. Here we present a study aimed at investigating different aspects of the disease, based on a multidisciplinary approach. The cohort of ALS patients that we analyzed includes seven patients, all originating from a common, restricted, geographical area and five matched controls. Environmental exposure is the same for all these subjects. SOD1, FUS, TDP43, C9ORF72 and APOE genotypes were evaluated. For metal quantitation, samples of serum and whole blood were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range both in reducing and nonreducing conditions. Levels of DNA oxidation were evaluated by a comet assay. Statistical analyses were carried out with Student\u2019s t-test and Artificial Neural Networks. As concentration resulted significantly lower in patients than in controls ( Auto-CM analysis linked closely high concentrations of Al and Se to the ALS group. Levels of metals in whole blood have been correlated with levels in serum. Proteomics data show that some proteins related to Acute Phase Response (APR) and lipid homeostasis are decreased in patients. Apo\u3b54 allele is more represented in the patient\u2019s group than in controls\u2019. Impaired metal homeostasis, attributable to environmental exposure, could lead to mineral overload. Metals can compete for the binding sites of metal-containing proteins. The different expression of the APR proteins reported could be a reflection of the disease status of the subjects analyzed. Enrichment in Apo\u3b54 allele frequency in patients may provide a link between neurodegeneration and lipid metabolism disturbances

A multidisciplinary approach to study Sporadic Amyotrophic Lateral Sclerosis in patients with common geographical origin

Amyotrophic Lateral Sclerosis (ALS), a fatal neurodegenerative disorder, is object of intensive research as the causes are still unknown and a treatment not available yet. This project is aimed to study, with a multidisciplinary approach, a small cohort of ALS subjects with a common environmental exposure. For metal quantitation, samples of serum and whole blood were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range. Arsenic concentration resulted significantly lower in patients than in controls. Also, Mn and Hg showed lower levels in patients. Levels of plasma APOA2 protein resulted decreased in patients with respect to controls, whereas SAMP showed a significant decrease only in the late onset group. APOA1 and TTHY also were decreased, the latter in late-onset patients. RET4 was decreased only in the early-onset group. When evaluating APOE genotype we found a 3-fold increase in the frequency of E3/E4 genotype in the patient\u2019s group. DNA oxidative stress has been evaluated through a Comet Assay. The multidisciplinary approach applied in this study allowed to dissect different aspects of ALS, often are evaluated separately and in heterogeneous cohorts of patients