Co-chaperones are limiting in a depleted chaperone network

To probe the limiting nodes in the chaperoning network which maintains cellular proteostasis, we expressed a dominant negative mutant of heat shock factor 1 (dnHSF1), the regulator of the cytoplasmic proteotoxic stress response. Microarray analysis of non-stressed dnHSF1 cells showed a two- or more fold decrease in the transcript level of 10 genes, amongst which are the (co-)chaperone genes HSP90AA1, HSPA6, DNAJB1 and HSPB1. Glucocorticoid signaling, which requires the Hsp70 and the Hsp90 folding machines, was severely impaired by dnHSF1, but fully rescued by expression of DNAJA1 or DNAJB1, and partially by ST13. Expression of DNAJB6, DNAJB8, HSPA1A, HSPB1, HSPB8, or STIP1 had no effect while HSP90AA1 even inhibited. PTGES3 (p23) inhibited only in control cells. Our results suggest that the DNAJ co-chaperones in particular become limiting in a depleted chaperoning network. Our results also suggest a difference between the transcriptomes of cells lacking HSF1 and cells expressing dnHSF1

Defining the Sister Rat Mammary Tumor Cell Lines HH-16 cl.2/1 and HH-16.cl.4 as an In Vitro Cell Model for Erbb2

Cancer cell lines have been shown to be reliable tools in genetic studies of breast cancer, and the characterization of these lines indicates that they are good models for studying the biological mechanisms underlying this disease. Here, we describe the molecular cytogenetic/genetic characterization of two sister rat mammary tumor cell lines, HH-16 cl.2/1 and HH-16.cl.4, for the first time. Molecular cytogenetic analysis using rat and mouse chromosome paint probes and BAC/PAC clones allowed the characterization of clonal chromosome rearrangements; moreover, this strategy assisted in revealing detected breakpoint regions and complex chromosome rearrangements. This comprehensive cytogenetic analysis revealed an increase in the number of copies of the Mycn and Erbb2 genes in the investigated cell lines. To analyze its possible correlation with expression changes, relative RNA expression was assessed by real-time reverse transcription quantitative PCR and RNA FISH. Erbb2 was found to be overexpressed in HH-16.cl.4, but not in the sister cell line HH-16 cl.2/1, even though these lines share the same initial genetic environment. Moreover, the relative expression of Erbb2 decreased after global genome demethylation in the HH-16.cl.4 cell line. As these cell lines are commercially available and have been used in previous studies, the present detailed characterization improves their value as an in vitro cell model. We believe that the development of appropriate in vitro cell models for breast cancer is of crucial importance for revealing the genetic and cellular pathways underlying this neoplasy and for employing them as experimental tools to assist in the generation of new biotherapies

DNA repair: the culprit for tumor-initiating cell survival?

The existence of “tumor-initiating cells” (TICs) has been a topic of heated debate for the last few years within the field of cancer biology. Their continuous characterization in a variety of solid tumors has led to an abundance of evidence supporting their existence. TICs are believed to be responsible for resistance against conventional treatment regimes of chemotherapy and radiation, ultimately leading to metastasis and patient demise. This review summarizes DNA repair mechanism(s) and their role in the maintenance and regulation of stem cells. There is evidence supporting the hypothesis that TICs, similar to embryonic stem (ES) cells and hematopoietic stem cells (HSCs), display an increase in their ability to survive genotoxic stress and injury. Mechanistically, the ability of ES cells, HSCs and TICs to survive under stressful conditions can be attributed to an increase in the efficiency at which these cells undergo DNA repair. Furthermore, the data presented in this review summarize the results found by our lab and others demonstrating that TICs have an increase in their genomic stability, which can allow for TIC survival under conditions such as anticancer treatments, while the bulk population of tumor cells dies. We believe that these data will greatly impact the development and design of future therapies being engineered to target and eradicate this highly aggressive cancer cell population

Processamento auditivo em indivíduos com epilepsia de lobo temporal Auditory processing in patients with temporal lobe epilepsy

A epilepsia do lobo temporal ocasiona descargas elétricas excessivas onde a via auditiva tem sua estação final. É uma das formas mais comuns e de mais difícil controle da doença. O correto processamento dos estímulos auditivos necessita da integridade anatômica e funcional de todas as estruturas envolvidas na via auditiva. OBJETIVO: Verificar o Processamento Auditivo de pacientes portadores de epilepsia do lobo temporal quanto aos mecanismos de discriminação de sons em seqüência e de padrões tonais, discriminação da direção da fonte sonora e atenção seletiva para sons verbais e não-verbais. MÉTODO: Foram avaliados oito indivíduos com epilepsia do lobo temporal confirmada e com foco restrito a essa região, através dos testes auditivos especiais: Teste de Localização Sonora, Teste de Padrão de Duração, Teste Dicótico de Dígitos e Teste Dicótico Não-Verbal. O seu desempenho foi comparado ao de indivíduos sem alteração neurológica (estudo caso-controle). RESULTADO: Os sujeitos com epilepsia do lobo temporal apresentaram desempenho semelhante aos do grupo controle quanto ao mecanismo de discriminação da direção da fonte sonora e desempenho inferior quanto aos demais mecanismos avaliados. CONCLUSÃO: Indivíduos com epilepsia do lobo temporal apresentaram maior prejuízo no processamento auditivo que os sem danos corticais, de idades semelhantes.<br>Temporal epilepsy, one of the most common presentation of this pathology, causes excessive electrical discharges in the area where we have the final station of the auditory pathway. Both the anatomical and functional integrity of the auditory pathway structures are essential for the correct processing of auditory stimuli. AIM: to check the Auditory Processing in patients with temporal lobe epilepsy regarding the auditory mechanisms of discrimination from sequential sounds and tone patterns, discrimination of the sound source direction and selective attention to verbal and nonverbal sounds. METHOD: eight individuals with temporal lobe epilepsy were assessed, after excluding those with non-confirmed diagnosis or with the focus of discharges not limited to this lobe. The evaluation was carried out through special auditory tests: Sound Localization Test, Duration Pattern Test, Digits Dichotic Test and Non-Verbal Dichotic Test. Their performances were compared to the performances of individuals without neurological diseases (case-control study). RESULTS: similar performances were observed between patients with temporal lobe epilepsy and the control group regarding the auditory mechanism of sound source direction discrimination. Comparing the other auditory mechanisms assessed, the patients with temporal lobe epilepsy presented worse results. CONCLUSION: individuals with temporal lobe epilepsy had more deficits in auditory processing than those without cortical damage