Peripheric facial paralysis as initial manifestation of occult metastatic neoplasia Paralisia facial periférica como manifestação inicial de neoplasia oculta metastática

Universidade Federal de São Paulo (UNIFESP) Escola Paulista de MedicinaUNIFESP ENT DepartmentUNIFESP, EPMUNIFESP, ENT DepartmentSciEL

A Reliable Stem Cell Carrier: An Experimental Study in Wistar Rats

Introduction Treatments based on cell biology need reliable and precise carriers for reaching the desired targets. For that reason, a PDO-based cell carrier was idealized, with the purpose of carrying stem cells to distant sites at room temperature. Materials and Methods Three modalities of the same carrier were evaluated: one containing undifferentiated human dental pulp stem cells (DPSCs); one loaded with stem cells induced to neurogenic differentiation (DPSCNs); and one without cells (Blank). The carriers were implanted in sciatic nerve gaps in 48 Wistar rats that were divided in three groups. Two other rats were included in a SHAM control group. Immunohistochemical, histological and clinical analyses were performed in two, four, six and eight weeks of time. Results Efficacy of human stem cell transportation at room temperature to rats was attested. Moreover, it was possible to confirm that those cells show tropism for inflamed environments and are also prone to induction of neurogenesis in the first two weeks, vanishing after that period. Conclusion Clinical evaluation of the animals' gait recovery shows a promising perspective of success with the inclusion of stem cell-loaded PDO tubes in nerve gaps, which may be positively compared to previously published studies. No level assigned This journal requires that authors assign a level of evidence to each article.4351353136

The Profile of Heparanase Expression Distinguishes Differentiated Thyroid Carcinoma from Benign Neoplasms

<div><p>Introduction</p><p>The search for a specific marker that could help to distinguish between differentiated thyroid carcinoma and benign lesions remains elusive in clinical practice. Heparanase (HPSE) is an endo-beta-glucoronidase implicated in the process of tumor invasion, and the heparanase-2 (HPSE2) modulates HPSE activity. The aim of this study was to evaluate the role of heparanases in the development and differential diagnosis of follicular pattern thyroid lesions.</p><p>Methods</p><p>HPSE and HPSE2 expression by qRT-PCR, immunohistochemistry evaluation, western blot analysis and HPSE enzymatic activity were evaluated.</p><p>Results</p><p>The expression of heparanases by qRT-PCR showed an increase of HPSE2 in thyroid carcinoma (P = 0.001). HPSE activity was found to be higher in the malignant neoplasms than in the benign tumors (P<0.0001). On Western blot analysis, HPSE2 isoforms were detected only in malignant tumors. The immunohistochemical assay allowed us to establish a distinct pattern for malignant and benign tumors. Carcinomas showed a typical combination of positive labeling for neoplastic cells and negative immunostaining in colloid, when compared to benign tumors (P<0.0001). The proposed diagnostic test presents sensitivity and negative predictive value of around 100%, showing itself to be an accurate test for distinguishing between malignant and benign lesions.</p><p>Conclusions</p><p>This study shows, for the first time, a distinct profile of HPSE expression in thyroid carcinoma suggesting its role in carcinogenesis.</p></div

HPSE and HPSE2 quantitative RT-PCR analysis.

<p>The values are represented by box plots and demonstrate the high expression of HPSE2 in DTC (**P = 0.001, Mann-Whitney test) and also of HPSE but with just a statistical trend (*P = 0.067, Mann-Whitney test); (N = 27).</p

Proteins and heparan sulfate expression in thyroid tissues.

<p>(A), Expression of heparanase-2 isoforms (HPSE2a, HPSE2b and HPSE2c); heparanase active enzyme (50 kDa) and pro-enzyme (65 kDa); heparan sulfate (HS). The samples were analyzed on a single sample of normal thyroid sample (“Normal”), and papillary thyroid carcinoma (“PTC”). (B), The values indicate relative expression obtained by the ratio of proteins and HS corrected by beta-actin expression.</p

Heparanase enzymatic assay in DTC and benign thyroid lesions (N = 27).

<p>* Mann-Whitney test.</p><p>** Relative fluorescence expresses the ratio between intensity of fluorescence from degraded heparan sulfate and non-degraded substrate.</p><p>*** Mean ± standard error</p><p>Legend: PTC, papillary thyroid carcinoma.</p><p>Heparanase enzymatic assay in DTC and benign thyroid lesions (N = 27).</p

Hypothetical scheme to explain herapanase physiology based on data of this study and also from Gengis-Velitski et al. [24], Levy-Adam et al. 2010 [25], Linke et al. 2002 [26], Zetzer et al. 2004 [27] and Nadav et al 2002 [17].

<p>(I) In the normal thyroid follicle or benign lesions HPSE and HPSE2 are first targeted to the endoplasmic reticulum lumen (ER), shuttled to the Golgi apparatus (<i>step 1</i>), partially processed and activated in lysosomes (<i>step 2</i>), and both heparanases (HPSE and HPSE2) are highly secreted to colloid (<i>step 3</i>); HPSE is therefore partially internalized (<i>step 4</i>) and also activated in the lysosomes (<i>step 5</i>) and directed to the basolateral surface of follicular thyroid cells where it presents low activity (<i>step 6</i>). (II) In the differentiated thyroid carcinoma there is overexpression of both heparanases (<i>step 1</i>), increased processing and activation of HPSE in lysosomes (<i>step 2</i>) and low secretion to colloid (<i>step 3</i>) and follows that described in steps 4 and 5 below. An increased secretion of HPSE to basolateral surface and extracellular matrix occurs to degrade HSPG (<i>step 6</i>, large arrows). * Additional information: (Ia) low presence of latent HPSE and no presence of HPSE2 in the cytoplasm of normal / benign lesion follicular cell (Western blotting); (Ib) immunoexpression of both heparanases by immunohistochemistry: low positivity in follicular cells for HPSE and positive staining in colloid for HPSE2; (IIa) high immunostaining in neoplastic cells for HPSE2; (IIb) high immunostaining in neoplastic cells for HPSE and Western blotting demonstrating the presence of the active form (50 kDa). Legend: HPSE 65 kDa, ο; HPSE 50 kDa, οο; HPSE2, Δ.</p

Immunostaining for HPSE2 in malignant and benign FNAB samples.

<p>Left panel represents malignant thyroid tissues (two cases of PTC) and the right panel represents benign thyroid tissues. Immunocytochemistry obtained using light microscopy, X400, showed a different pattern of HPSE2 staining: high expression on follicular cells of PTC and none expression on benign lesions and also a negative colloid expression on PTC (*) and positive on nodular goiter.</p