WNK2 inhibits autophagic flux in human glioblastoma cell line

The following are available online at http://www.mdpi.com/2073-4409/9/2/485/s1, Figure S1: Validation of WNK2 overexpression by RT-PCR.Autophagy is a cell-survival pathway with dual role in tumorigenesis, promoting either tumor survival or tumor death. WNK2 gene, a member of the WNK (with no lysine (K)) subfamily, acts as a tumor suppressor gene in gliomas, regulating cell migration and invasion; however, its role in autophagy process is poorly explored. The WNK2-methylated human glioblastoma cell line A172 WT (wild type) was compared to transfected clones A172 EV (empty vector), and A172 WNK2 (WNK2 overexpression) for the evaluation of autophagy using an inhibitor (bafilomycin A1—baf A1) and an inducer (everolimus) of autophagic flux. Western blot and immunofluorescence approaches were used to monitor autophagic markers, LC3A/B and SQSTM1/p62. A172 WNK2 cells presented a significant decrease in LC3B and p62 protein levels, and in LC3A/B ratio when compared with control cells, after treatment with baf A1 + everolimus, suggesting that WNK2 overexpression inhibits the autophagic flux in gliomas. The mTOR pathway was also evaluated under the same conditions, and the observed results suggest that the inhibition of autophagy mediated by WNK2 occurs through a mTOR-independent pathway. In conclusion, the evaluation of the autophagic process demonstrated that WNK2 inhibits the autophagic flux in glioblastoma cell line.This project was supported by the Barretos Cancer Hospital Internal Research Funds (PAIP) to Rui Manuel Reis and by the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer Project), Campinas, Brazil. Ana Laura Vieira Alves is the recipient of a FAPESP master fellowship (2016/18907-0)

Estudo comparativo da neoformação óssea utilizando-se o enxerto autógeno e três substitutos: defeitos ósseos em ratos Comparative study of bone neoformation using autologous grafting and three replacements: bone defects in rats

OBJETIVO: Comparar a percentagem de neoformação óssea promovida pelo enxerto ósseo autógeno e três tipos de materiais de substituição de características distintas em cavidades em fêmures de ratos. MÉTODOS: Foram realizadas duas cavidades de 5,4 x 2,7mm, em cada fêmur (direito e esquerdo), de 14 ratos Wistar isogênicos. Cada um dos quatro defeitos criados foi preenchido com o osso autógeno ou com um dos três materiais testados - hidroxiapatita (HA), Genphos® (HA+ &#946;-TCP) e GenMix® (um enxerto ósseo bovino composto). Ao final dos períodos de seis semanas (n = 6) e 12 semanas (n = 8), os animais foram sacrificados. As lâminas (coradas com Picro-Sirius) foram analisadas por microscopia óptica normal e software específico. RESULTADOS: Os grupos com o osso autógeno se mostraram muito superiores aos demais nos dois tempos analisados, tendo média de neoformação óssea ± DP de 90,6 ± 10,8% em seis semanas, e 98 ± 9,2% em 12 semanas (p > 0,0001 em ambos os tempos analisados). Em seis semanas, os resultados para os demais grupos foram os seguintes: Genphos®, 46 ± 7,1%; HA, 43,1 ± 8,4%; e GenMix®, 57,3 ± 4,5%. Em 12 semanas: Genphos®, 47,8 ± 11,1%; HA, 39,9 ± 5,4%; GenMix®, 59,7 ± 4,8%, significativa (p = 0,007). CONCLUSÕES: Em ambos os tempos analisados, os três materiais de substituição óssea testados se mostraram inferiores ao osso autógeno quanto à percentagem de neoformação óssea.<br>OBJECTIVE: Compare the percentage of bone neoformation promoted by autologous bone grafting and three kinds of replacement materials with different characteristics in rats' femoral holes. METHODS: Two holes measuring 5.4 x 2.7mm, were produced on each femur (right and left) of 14 isogenic Wistar rats. Each of the four defects produced was filled by autologous bone or by one of three tested materials - hydroxyapatite (HA), Genphos® (HA+ &#946;-TCP) and GenMix® (a combined bovine bone graft). In the end of the 6-week (n = 6) and 12-week (n = 8) periods, the animals were sacrificed. The sections (stained with Picro-Sirius) were assessed by optical microscopy and specific software. RESULTS: The groups with autologous bone were shown to be significantly superior to the others at both assessed times, showing a mean bone formation rate ± SD of 90.6 ± 10.8% in six weeks, and 98 ± 9.2% in 12 weeks (p > 0.0001 for both assessed times). In six weeks, the results for the other groups were the following: Genphos®, 46 ± 7.1%; HA, 43.1 ± 8.4%; and GenMix®, 57.3 ± 4.5%. In 12 weeks: Genphos®, 47.8 ± 11.1%; HA, 39.9 ± 5.4%; GenMix®, 59.7 ± 4.8%, significant (p = 0.007). CONCLUSIONS: In both assessed times, the three bone replacement materials tested in the study showed to be inferior to autologous bone graft for bone neoformation percentage

COMPARATIVE STUDY OF BONE NEOFORMATION USING AUTOLOGOUS GRAFTING AND THREE REPLACEMENTS: BONE DEFECTS IN RATS

ABSTRACTObjective: Compare the percentage of bone neoformation promoted by autologous bone grafting and three kinds of replacement materials with different characteristics in rats' femoral holes. Methods: Two holes measuring 5.4×2.7mm, were produced on each femur (right and left) of 14 isogenic Wistar rats. Each of the four defects produced was filled by autologous bone or by one of three tested materials-hydroxyapatite (HA), Genphos® (HA+ β-TCP) and GenMix® (a combined bovine bone graft). In the end of the 6-week (n = 6) and 12-week (n = 8) periods, the animals were sacrificed. The sections (stained with Picro-Sirius) were assessed by optical microscopy and specific software. Results: The groups with autologous bone were shown to be significantly superior to the others at both assessed times, showing a mean bone formation rate ± SD of 90.6 ± 10.8% in six weeks, and 98 ± 9.2% in 12 weeks (p > 0.0001 for both assessed times). In six weeks, the results for the other groups were the following: Genphos®, 46 ± 7.1%; HA, 43.1 ± 8.4%; and GenMix®, 57.3 ± 4.5%. In 12 weeks: Genphos®, 47.8 ± 11.1%; HA, 39.9 ± 5.4%; GenMix®, 59.7 ± 4.8%, significant (p = 0.007). Conclusions: In both assessed times, the three bone replacement materials tested in the study showed to be inferior to autologous bone graft for bone neoformation percentage

Estrongiloidíase Diagnosticada por Lavado Broncoalveolar

Paciente masculino, branco, 76 anos de idade, com diabete melito, tabagista e portador de doença pulmonar obstrutiva crônica grave em uso crônico de prednisona na dose de 20 mg/dia nos últimos 7 anos, interna no hospital por piora da dispnéia e diarréia. Radiograma simples de tórax mostrou hiperinsuflação pulmonar bilateral e opacidade em lobo inferior esquerdo. Tomografia computadorizada de tórax revelou enfisema bilateral, bronquiectasias em lobo médio e opacidade de contornos indefinidos em lobo inferior esquerdo. Hemograma completo e bioquímica de rotina sem alterações