Notational Analysis of Wheelchair Paralympic Table Tennis Matches

Paralympic table tennis is the third largest paralympic sport for the number of players. Performance analysis was conducted for the rally duration and interval and impact of serve, whilst none investigated the shots distribution among classes of physical impairment. Therefore, the purpose of this study was to conduct a notational analysis of international competitions in relation to the wheelchair classes. Five matches for each wheelchair class (C1-to-C5) were evaluated from 20 elite male right-handed players. Both players for each match were analyzed for the following performance indicators: strokes type, the area of ball bouncing, and the shots outcome. Backhand shots were the most used technique for all classes. The most used strokes for C1 players were backhand and forehand drive and backhand lob, while for C5 players they were backhand and forehand push and backhand topspin. Similar shots distribution was registered for C2-to-C5 players. The central and far-from-the-net zone was mainly reached by the serve for all classes. Errors shots were similar in all classes, whilst winning shots were more frequent in C1. The current notational analysis provided a meaningful performance modelling of indicators for coaches and athletes that can be used to design training programs for each class

Immunohistochemical analysis of axillary skin biopsies for the detection of adrenergic innervation of sweat glands in normal subjects and Parkinson’s disease patients

Beside the typical motor symptoms Parkinson’s disease (PD) is characterized, with varying severity, by autonomic dysfunction. Several studies have shed light on the anatomical and molecular changes that underlie the peripheral neuronal degeneration associated with PD and other Lewy body (LB) diseases (LBDs). By using skin biopsies from LBDs patients it was possible to detect misfolded phospho-α-synuclein (p-syn) deposits within dermal nerve fibers and correlate them with a reduced density of small nerve fibers. (1, 2). The skin biopsy approach is an inexpensive and minimally invasive technique. To date, there is not a standardized procedure for sampling site, tissue processing and nerve fibre assessment, so the goal of a diagnostic instrument for an early diagnosis of (LBDs) still remains a challenge. We have carried out a retrospective study setting up a novel protocol based on 10 µm thick serial sections from FFPE axillary skin biopsies. This choice take advantage from the presence of apocrine glands in the axillary region, as they receive a dense sympathetic adrenergic innervation, exploitable for a clear nervous fibers tracking. The biopsies were taken from 14 individuals who had been, in the first instance, diagnosed with various traits of motor and neurological dysfunction and two control subjects. Serial tissue sections were analysed by IHC (DAB chromogen) and by immunofluorescent labelling, using anti-p-α-synuclein (S129), anti- α -synuclein, anti-PGP9.5 and anti-tyrosine hydroxylase antibodies. This particular setting has proven useful to well highlight the adrenergic fibers surrounding the apocrine sweat glands and to visualize the fibers α -synuclein deposition. Our results enabled us to support the first diagnosis in various cases with probable PD but gave a negative p-Syn-S129 immunoreactivity results for samples from vascular Parkinson, multiple system atrophy, essential tremor and frontotemporal dementia. Our methodological setting is able to detect the adrenergic innervation of sweat apocrine glands and both the presence of Lewy bodies and Lewy neurites in axillary skin biopsies

Effects of culture system and hypoxia on long-term expansion and differentiation of mesenchymal stem cells derived from periodontal ligament

Periodontal ligament stem cells (PDLSCs), located in the perivascular space of the periodontium were able to differentiate into periodontal cell types in vitro [1]. In this study, we investigated the effect of three different culture media and of low oxygen tension (1%) on the immunophenotype, proliferation rate and osteogenic potential of PDLSCs. This study was the first report to compare the PDLSCs from the same person in different culture systems. PDLSCs were harvested from three healthy third molars and the single-cells suspensions were cultured in the culture media a-MEM, DMEM and a new medium formulation (Enriched Ham’s F12 Medium, EHFM), respectively. PDLSCs were subcultured (4 x 103/cm2) until passage 7. The characterization of PDLSCs included FACS, immunofluorescence analysis and cell proliferation assay in both normoxia and hypoxia (1%). After culture in osteogenic medium for 7, 14 and 21 days, osteoblastic differentiation was evaluated by alkaline phosphatase activity, mineralization (alizarin red staining) and gene expression of osteogenic markers. Osteoblastic differentiation was also evaluated under hypoxic conditions. PDLSCs cultured in EHFM showed increased proliferation rate and CD73 overexpression compared to cells maintained in a-MEM and DMEM. On the other hand, PDLSCs grown in a-MEM and DMEM showed higher osteogenic differentiation potential compared to EHFM. Hypoxia affected both proliferation rate and osteogenic potential. On the basis of these results, we propose a two stages protocol for the osteogenic induction of PDLSCs, in which the early expansion stage could be performed in EHFM without loss of cell stemness. Furthermore, the results obtained in the different conditions (normoxia and hypoxia) suggest that oxygen tension plays a critical role in PDLSCs physiology

Recovery of NIS expression in thyroid cancer cells by overexpression of Pax8 gene

BACKGROUND: Recovery of iodide uptake in thyroid cancer cells by means of obtaining the functional expression of the sodium/iodide symporter (NIS) represents an innovative strategy for the treatment of poorly differentiated thyroid cancer. However, the NIS gene expression alone is not always sufficient to restore radioiodine concentration ability in these tumour cells. METHODS: In this study, the anaplastic thyroid carcinoma ARO cells were stably transfected with a Pax8 gene expression vector. A quantitative RT-PCR was performed to assess the thyroid specific gene expression in selected clones. The presence of NIS protein was detected by Western blot and localized by immunofluorescence. A iodide uptake assay was also performed to verify the functional effect of NIS induction and differentiation switch. RESULTS: The clones overexpressing Pax8 showed the re-activation of several thyroid specific genes including NIS, Pendrin, Thyroglobulin, TPO and TTF1. In ARO-Pax8 clones NIS protein was also localized both in cell cytoplasm and membrane. Thus, the ability to uptake the radioiodine was partially restored, associated to a high rate of efflux. In addition, ARO cells expressing Pax8 presented a lower rate of cell growth. CONCLUSION: These finding demonstrate that induction of Pax8 expression may determine a re-differentiation of thyroid cancer cells, including a partial recovery of iodide uptake, fundamental requisite for a radioiodine-based therapeutic approach for thyroid tumours

A Functional Variant of the Dimethylarginine Dimethylaminohydrolase-2 Gene Is Associated with Insulin Sensitivity

Background: Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide synthase, which was associated with insulin resistance. Dimethylarginine dimethylaminohydrolase (DDAH) is the major determinant of plasma ADMA. Examining data from the DIAGRAM+ (Diabetes Genetics Replication And Meta-analysis), we identified a variant (rs9267551) in the DDAH2 gene nominally associated with type 2 diabetes (P =3610 25). Methodology/Principal Findings: initially, we assessed the functional impact of rs9267551 in human endothelial cells (HUVECs), observing that the G allele had a lower transcriptional activity resulting in reduced expression of DDAH2 and decreased NO production in primary HUVECs naturally carrying it. We then proceeded to investigate whether this variant is associated with insulin sensitivity in vivo. To this end, two cohorts of nondiabetic subjects of European ancestry were studied. In sample 1 (n = 958) insulin sensitivity was determined by the insulin sensitivity index (ISI), while in sample 2 (n = 527) it was measured with a euglycemic-hyperinsulinemic clamp. In sample 1, carriers of the GG genotype had lower ISI than carriers of the C allele (67633 vs.79644; P = 0.003 after adjusting for age, gender, and BMI). ADMA levels were higher in subjects carrying the GG genotype than in carriers of the C allele (0.6860.14 vs. 0.5760.14 mmol/l; P = 0.04). In sample 2, glucose disposal was lower in GG carriers as compared with C carriers (9.364.1 vs. 11.064.2 mg6Kg 21 free fat mass6min 21; P = 0.009)

Natural Form of Noncytolytic Flexible Human Fc as a Long-Acting Carrier of Agonistic Ligand, Erythropoietin

Human IgG1 Fc has been widely used as a bioconjugate, but exhibits shortcomings, such as antibody- and complement-mediated cytotoxicity as well as decreased bioactivity, when applied to agonistic proteins. Here, we constructed a nonimmunogenic, noncytolytic and flexible hybrid Fc (hyFc) consisting of IgD and IgG4, and tested its function using erythropoietin (EPO) conjugate, EPO-hyFc. Despite low amino acid homology (20.5%) between IgD Fc and IgG4 Fc, EPO-hyFc retained “Y-shaped” structure and repeated intravenous administrations of EPO-hyFc into monkeys did not generate EPO-hyFc-specific antibody responses. Furthermore, EPO-hyFc could not bind to FcγR I and C1q in contrast to EPO-IgG1 Fc. In addition, EPO-hyFc exhibited better in vitro bioactivity and in vivo bioactivity in rats than EPO-IgG1 Fc, presumably due to the high flexibility of IgD. Moreover, the mean serum half-life of EPO-hyFc(H), a high sialic acid content form of EPO-hyFc, was approximately 2-fold longer than that of the heavily glycosylated EPO, darbepoetin alfa, in rats. More importantly, subcutaneous injection of EPO-hyFc(H) not only induced a significantly greater elevation of serum hemoglobin levels than darbepoetin alfa in both normal rats and cisplatin-induced anemic rats, but also displayed a delayed time to maximal serum level and twice final area-under-the-curve (AUClast). Taken together, hyFc might be a more attractive Fc conjugate for agonistic proteins/peptides than IgG1 Fc due to its capability to elongate their half-lives without inducing host effector functions and hindering bioactivity of fused molecules. Additionally, a head-to-head comparison demonstrated that hyFc-fusion strategy more effectively improved the in vivo bioactivity of EPO than the hyperglycosylation approach

Innate and Adaptive Immunity Linked to Recognition of Antigens Shared by Neural Crest-Derived Tumors

In the adult, many embryologic processes can be co-opted by during cancer progression. The mechanisms of divisions, migration, and the ability to escape immunity recognition linked to specific embryo antigens are also expressed by malignant cells. In particular, cells derived from neural crests (NC) contribute to the development of multiple cell types including melanocytes, craniofacial cartilage, glia, neurons, peripheral and enteric nervous systems, and the adrenal medulla. This plastic performance is due to an accurate program of gene expression orchestrated with cellular/extracellular signals finalized to regulate long-distance migration, proliferation, differentiation, apoptosis, and survival. During neurulation, prior to initiating their migration, NC cells must undergo an epithelial–mesenchymal transition (EMT) in which they alter their actin cytoskeleton, lose their cell–cell junctions, apicobasal polarity, and acquire a motile phenotype. Similarly, during the development of the tumors derived from neural crests, comprising a heterogeneous group of neoplasms (Neural crest-derived tumors (NCDTs)), a group of genes responsible for the EMT pathway is activated. Here, retracing the molecular pathways performed by pluripotent cells at the boundary between neural and non-neural ectoderm in relation to the natural history of NCDT, points of contact or interposition are highlighted to better explain the intricate interplay between cancer cells and the innate and adaptive immune response

UbcH10 a Major Actor in Cancerogenesis and a Potential Tool for Diagnosis and Therapy

Malignant transformation is a multistep process in which several molecular entities become dysregulated and result in dysfunction in the regulation of cell proliferation. In past years, scientists have gradually dissected the pathways involved in the regulation of the cell cycle. The mitotic ubiquitin-conjugating enzymes UbcH10, has been extensively studied since its cloning and characterization and it has been identified as a constantly overexpressed factor in many types of cancer. In this paper, we have reviewed the literature about UbcH10 in human cancer, pointing out the association between its overexpression and exacerbation of cancer phenotype. Moreover, many recalled studied demonstrated how immunohistochemistry or RT-PCR analysis can distinguish normal tissues and benign lesions from malignant neoplasms. In other experimental studies, many of the consequences of UbcH10 overexpression, such as increased proliferation, metastasizing, cancer progression and resistance to anticancer drugs are reversed through gene silencing techniques. In recent years, many authors have defined UbcH10 evaluation in cancer patients as a useful tool for diagnosis and therapy. This opinion is shared by the authors who advertise how it would be useful to start using in clinical practice the notions acquired about this important moleculein the carcinogenesis of many human malignancies

Regulation by human chorionic gonadotropin of sodium/iodide symporter gene expression in the JAr human choriocarcinoma cell line

Sodium/iodide symporter (NIS) gene and protein expressions have been recently described in human cytotrophoblasts, emphasizing its potential function in the active transport of iodide from the mother to the fetus. In this study we analyzed NIS expression and function in the human JAr placental choriocarcinoma cell line. Using real-time quantitative RT-PCR, we first demonstrated that NIS transcripts are expressed at a high level in JAr cells compared with other cell lines, including thyroid cancer cells. Functional analysis clearly showed that Jar cells are able to concentrate iodide in presence of hCG. Iodide accumulation increased after 2-h exposure to 5 IU/ml hCG, to 6-fold over the basal level after 8 h. This effect was reproduced using forskolin, the cAMP analog (BU)(2)-cAMP, and phorbol acetate. Moreover, hCG increased both NIS mRNA after 2 h and NIS protein levels after 4 h, reaching a maximum after 8 h in both cases. In conclusion, our data demonstrate that 1) NIS is expressed in JAr cells; 2) iodide transport in JAr cells is regulated by hCG and by cAMP-dependent and -independent mechanisms; 3) the stimulation of iodide uptake is due to an increase in both NIS mRNA and protein levels; and 4) JAr cells may represent an excellent in vitro model suitable to analyze the molecular mechanisms involved in iodide transport from mother to fetus