Endovenous laser treatment for varicose veins in patients with active ulcers: Measurement of intravenous and perivenous temperatures during the procedure

Background Conventional saphenous vein stripping is difficult to be indicated for the treatment of varicose veins in patients classified as CEAP C4, C5, or C6. Objective This study was developed to evaluate treatment results for varicose veins with active ulcers using endovenous laser (EVL), compared to a group undergoing clinical treatment, during 1 year. Patients and method Fifty-two patients presenting with varicose veins with active ulcers for more than 1 year were divided for treatment into two randomized groups: Group 1, clinical treatment, composed of 25 subjects, was submitted to elastic or inelastic compression therapy; Group 2, EVL treatment, composed of 27 subjects, was submitted to great and or small saphenous vein ablation with a 980-nm diode EVL, plus the clinical treatment. Intravenous and perivenous temperatures were measured continuously during the EVL treatment. All patients were followed for 12 months and studied with ultrasound at the beginning and end of the study. The ulcers' areas were evaluated initially and at every 3 months. Results In 12 months, 81.5% of the wounds in patients in Group 2 and only 24% in patients in Group 1 had healed. Ulcer recurrence rate was 44.4% in Group 1. The mean wound area in Group 1 decreased from 17.48 to 12.76 cm(2) at the end of the year. In Group 2, the wound area decreased from 22.26 to 2.7 cm(2) (p=.0037). Mean intravenous and perivenous temperatures of 79.3 and 43.0 degrees C were recorded. Conclusion The treatment for varicose veins with EVL is safe in patients with active ulcers. Wounds healed faster than in patients undergoing clinical treatment alone during a 1-year period. There was no ulcer recurrence in patients treated with EVL.33101234124

FEZ1 interacts with CLASP2 and NEK1 through coiled-coil regions and their cellular colocalization suggests centrosomal functions and regulation by PKC

FEZ1 was initially described as a neuronal protein that influences axonal development and cell polarization. CLASP2 and NEK1 proteins are present in a centrosomal complex and participate in cell cycle and cell division mechanisms, but their functions were always described individually. Here, we report that NEK1 and CLASP2 colocalize with FEZ1 in a perinuclear region in mammalian cells, and observed that coiled-coil interactions occur between FEZ1/CLASP2 and FEZ1/NEK1 in vitro. These three proteins colocalize and interact with endogenous gamma-tubulin. Furthermore, we found that CLASP2 is phosphorylated and interacts with active PKC isoforms, and that FEZ1/CLASP2 colocalization is inhibited by PMA treatment. Our results provide evidence that these three proteins cooperate in centrosomal functions and open new directions for future studies.338416713545Fundacao de Amparo a Pesquisa do Estado Sao PauloConselho Nacional de Pesquisa e DesenvolvimentoLNL

Characterization of hNek6 Interactome Reveals an Important Role for Its Short N-Terminal Domain and Colocalization with Proteins at the Centrosome

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Physical protein protein interactions are fundamental to all biological processes and are organized in complex networks One branch of the kinome network is the evolutionarily conserved NIMA-related serine/threonine kinases (Neks) Most of the 11 mammalian Neks studied so far are related to cell cycle regulation and due to association with diverse human pathologies, Neks are promising chemotherapeutic targets Human Nek6 was associated to carcinogenesis but its interacting partners and signaling pathways remain elusive Here we introduce hNek6 as a highly connected member in the human kinase interactome In a more global context, we performed a broad data bank comparison based on degree distribution analysis and found that the human kinome is enriched in hubs Our networks include a broad set of novel hNek6 interactors as identified by our yeast two hybrid screens classified into 18 functional categories All of the tested interactions were confirmed and the majority of tested substrates were phosphorylated in vitro by hNek6 Notably, we found that hNek6 N-terminal is important to mediate the interactions with its partners Some novel interactors also colocalized with hNek6 and gamma-tubulin in human cells, pointing to a possible centrosomal interaction The interacting proteins link hNek6 to novel pathways, for example, Notch signaling and actin cytoskeleton regulation, or give new insights on how hNek6 may regulate previously proposed pathways such as cell cycle regulation, DNA repair response, and NF-kappa B signaling Our findings open new perspectives in the study of hNek6 role in cancer by analyzing its novel interactions in specific pathways in tumor cells which may provide important implications for drug design and cancer therapy91262986316Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Associacao Brasileira de Tecnologia de Luz Sincrotron (ABTLuS)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Human Nek6 is a monomeric mostly globular kinase with an unfolded short N-terminal domain

Background: The NIMA-related kinases (Neks) are widespread among eukaryotes. In mammalians they represent an evolutionarily conserved family of 11 serine/threonine kinases, with 40 45% amino acid sequence identity to the Aspergillus nidulans mitotic regulator NIMA within their catalytic domains. Neks have cell cycle-related functions and were recently described as related to pathologies, particularly cancer, consisting in potential chemotherapeutic targets. Human Nek6, -7 and -9 are involved in the control of mitotic spindle formation, acting together in a mitotic kinase cascade, but their mechanism of regulation remain elusive. Results: In this study we performed a biophysical and structural characterization of human Nek6 with the aim of obtaining its low resolution and homology models. SAXS experiments showed that hNek6 is a monomer of a mostly globular, though slightly elongated shape. Comparative molecular modeling together with disorder prediction analysis also revealed a flexible disordered N-terminal domain for hNek6, which we found to be important to mediate interactions with diverse partners. SEC-MALS experiments showed that hNek6 conformation is dependent on its activation/phosphorylation status, a higher phosphorylation degree corresponding to a bigger Stokes radius. Circular dichroism spectroscopy confirmed our in silico predictions of secondary structure content and thermal stability shift assays revealed a slightly higher stability of wild-type hNek6 compared to the activation loop mutant hNek6(S206A). Conclusions: Our data present the first low resolution 3D structure of hNek6 protein in solution. SAXS, comparative modeling and SEC-MALS analysis revealed that hNek6 is a monomeric kinase of slightly elongated shape and a short unfolded N-terminal domain.11Fundacao de Amparo a Pesquisa do Estado Sao PauloConselho Nacional de Pesquisa e DesenvolvimentoLNL