Does SOFA predict outcomes better than SIRS in Brazilian ICU patients with suspected infection? A retrospective cohort study

ABSTRACT We compared the discriminatory capacity of the sequential organ failure assessment (SOFA) versus the systemic inflammatory response syndrome (SIRS) score for predicting ICU mortality, need for and length of mechanical ventilation, ICU stay, and hospitalization in patients with suspected infection admitted to a mixed Brazilian ICU. We performed a retrospective analysis of a longitudinal ICU database from a tertiary hospital in Southern Brazil. Patients were categorized according to whether they met the criteria for sepsis according to SOFA (variation ≥2 points over the baseline clinical condition) and SIRS (SIRS score ≥2 points). From January 2008 to December 2014, 1487 patients were admitted to the ICU due to suspected infection. SOFA ≥2 identified more septic patients than SIRS ≥2 (79.0% [n = 1175] vs. 68.5% [n = 1020], p < 0.001). There was no difference between the two scores in predicting ICU mortality (area under the receiver operating characteristic curve (AUROC) = 0.64 vs. 0.64, p = 0.99). SOFA ≥2 was marginally better than SIRS ≥2 in predicting need for mechanical ventilation (AUROC = 0.64 vs. 0.62, p = 0.001), ICU stay > 7 days (AUROC = 0.65 vs. 0.63, p = 0.004), and length of hospitalization >10 days (AUROC = 0.61 vs. 0.59, p < 0.001). There was no difference between the two scores in predicting mechanical ventilation >7 days.

Does SOFA predict outcomes better than SIRS in Brazilian ICU patients with suspected infection? A retrospective cohort study.

We compared the discriminatory capacity of the sequential organ failure assessment (SOFA) versus the systemic inflammatory response syndrome (SIRS) score for predicting ICU mortality, need for and length of mechanical ventilation, ICU stay, and hospitalization in patients with suspected infection admitted to a mixed Brazilian ICU. We performed a retrospective analysis of a longitudinal ICU database from a tertiary hospital in Southern Brazil. Patients were categorized according to whether they met the criteria for sepsis according to SOFA (variation ≥2 points over the baseline clinical condition) and SIRS (SIRS score ≥2 points). From January 2008 to December 2014, 1487 patients were admitted to the ICU due to suspected infection. SOFA ≥2 identified more septic patients than SIRS ≥2 (79.0% [n=1175] vs. 68.5% [n=1020], p<0.001). There was no difference between the two scores in predicting ICU mortality (area under the receiver operating characteristic curve (AUROC)=0.64 vs. 0.64, p=0.99). SOFA ≥2 was marginally better than SIRS ≥2 in predicting need for mechanical ventilation (AUROC=0.64 vs. 0.62, p=0.001), ICU stay>7 days (AUROC=0.65 vs. 0.63, p=0.004), and length of hospitalization >10 days (AUROC=0.61 vs. 0.59, p<0.001). There was no difference between the two scores in predicting mechanical ventilation >7 days.

Glutamate promotes cell growth by EGFR signaling on U-87MG human glioblastoma cell line.

Accumulating evidences suggest that glutamate plays a key role in the proliferation and invasion of malignant glioblastoma (GBM) tumors. It has been shown that GBM cells release and exploit glutamate for proliferation and invasion through AMPA glutamate receptors. Additionally, amplification of the epidermal growth factor receptor (EGFR) gene occurs in 40-50% of GBM. Since, PI3K/Akt is considered one of the main intracellular pathways involved in EGFR activation, AKT functions could trigger EGFR signaling. Thus, we investigated whether EGFR-phospho-Akt pathway is involved on the glutamate inducing U-87MG human GBM cell line proliferation. For these purpose, we treated the U-87MG cell line with 5 to 200 mM of glutamate and assessed the number of viable cells by trypan blue dye exclusion test. An increase in cell number (50%) was found at 5 mM glutamate, while the addition of DNQX (500 microM), an antagonist of AMPA receptor, inhibited the effect of glutamate on the U87-MG cells proliferation. Also, at 5 mM glutamate we observed an increase on the EGFR and phospho-Akt contents evaluated by immunohistochemistry. Moreover, U-87MG cells treated with glutamate exhibited an increase about 2 times in the EGFR mRNA expression. While, in the presence of the anti-EGFR gefitinib (50 muM) or the PI3K inhibitor wortmannin (5 muM), the U-87MG proliferation was restored to control levels. Together, our data suggest that glutamate signaling mediated by AMPA receptor induces U-87MG human GBM cell line proliferation via EGFR-phospho-Akt pathway.

Investigação do efeito do glutamato sobre o conteúdo de rEGF em linhagens celulares derivadas de glioblastomas humanos estabelecidas e caracterizadas

Os gliomas são as neoplasias cerebrais primárias que mais freqüentemente acometem o Sistema Nervoso Central (SNC). Cerca de 50% dos casos de gliomas são neoplasias com fenótipo maligno, incluindo os glioblastomas. Os glioblastomas possuem grande capacidade de expansão e invasão do tecido cerebral adjacente, o que contribui para o seu prognóstico sombrio. O glutamato é o principal neurotransmissor excitatório do SNC, mas sob condições patológicas a ativação excessiva do sistema glutamatérgico provoca danos citotóxicos às células neurais. A toxicidade glutamatérgica está associada a uma série de doenças agudas e crônico-degenerativas que acometem o SNC. Os glioblastomas humanos são capazes de liberar quantidades tóxicas de glutamato sobre os tecidos adjacentes e esta liberação parece favorecer o crescimento e a expansão tumoral. A fim de avaliarmos se a progressão tumoral mediada pelo glutamato é desencadeada por elementos das vias de sinalização, investigamos o efeito do glutamato sobre o conteúdo e expressão do receptor do fator de crescimento epitelial (rEGF). Desta forma, os cultivos primários Gli1, Gli2 e Gli3, assim como a linhagem estabelecida U87MG, foram tratados com doses crescentes de glutamato (5-200 mM) por 48 horas e após o tratamento a viabilidade celular, os conteúdos de rEGF e de Akt foram avaliados. Os cultivos analisados apresentaram suscetibilidades distintas aos efeitos citotóxicos do glutamato, porém em todos os casos as doses efetivas foram muito superiores (valores de ICso de 45mM a 100mM) às concentrações tóxicas descritas para células neurais (valores de ICso de100uM a 1mM). A expressão protéica de rEGF e o conteúdo de Akt fosforilado aumentaram após o tratamento com 5mM de glutamato, sugerindo que a ativação dos receptores de EGF possam exercer função na via de sinalização desencadeada pelo glutamato. Após o término dos experimentos com glutamato dois dos três cultivos primários utilizados (Gli1 e Gli2) se mantiveram viáveis em cultivo por mais de 50 passagens, o que sugere estabilidade das alterações moleculares e a seleção de uma subpopulação especifica de células que pode ser denominada linhagem celular. Assim, em os à caracterização destas duas novas linhagens celulares derivadas de glioblastomas - Gli1 e Gli2. Para tal avaliamos as suas taxas de proliferação, morfologia celular, o perfil de cariótipo de ambas e os conteúdos de rEGF, Hsp70, 6, p53 e MMP2, marcadores relacionados à progressão tumoral. Em suma, os cultivos Gli1 e Gli2 demonstraram alterações cromossômicas e expressão de marcadores compatíveis com as anomalias descritas para a manifestação do fenótipo de glioblastomas. Igualmente, comparamos o crescimento destas linhagens glioblastomas cultivadas como monocamada com o cultivo tridimensional em esferóides multicelulares. Os resultados obtidos revelaram padrões de crescimento e conteúdos dos marcadores diferentes entre as condições de cultivo

The gliomas are the primary brain tumors that more frequntIy accomplished the Central Nervous System (CNS). About 50% of the glioma cases presented malignant phenotype, including glioblastomas. Glioblastoma are tumors with high capacity of proliferation and invasion through the adjacent healthy nervous tissue, which largely contributes for its poor prognosis. The glutamate is the main excitatory neurotransmitter of the CNS, but under pathological conditions the extreme activation of the glutamatergic system induces cytotoxic damages to the neural cells. The glutamatergic citotoxicity is associated with severaI acute and chronic-degenerative diseases of the CNS. Human glioblastomas are capable of release toxic amounts of glutamate on adjacent brain tissue and this release seems to favor tumoral growth and expansion. In order to evaluate if the tumoral progression mediated by glutamato is a product of the activation of elements 6f1he signaling pathways, we investigated the glutamate effect on the content and expression of EGFi . For this reason, the primary culture cells Gli1, Gli2 and Gli3, as well as the established cell line U-87MG, were treated with increasing doses of glutamate (5-200 mM) for 48 hours and after the treatment the cell viability, EGFr and Akt contents were evaluated. The studied cells presented distinct susceptibilities to the cytotoxic effects of glutamate, however in all the cell cultures the toxicity was revealed only with very high glutamate doses (values of ICso range from 45mM to 100mM) when compared to the described toxic concentrations for neural cells (ICso values range from 100uM to 1 mM). The EGFr protein expression and the phosphor-Akt content increased after the treatment with 5mM of glutamate, suggesting that the activation of the EGF receptors can have a role in the glutamate signaling pathways. After the conclusion of these set of experiments, twoof out three tested primary cultures remained viable in cell culture for more than 50 passages, which suggests stability of the molecular profile and the selection of a specific ce11 subpopulation that can be named by 0011 line. Thus, in order to characterize these two new glioblastoma cell lines (Gli1 and Gli2), we carry out a series of experiments. The experiments had included the evaluation of the cellular growth rate, the cellular morphology, the kariotype profile and the identification of molecular markers related to tumor progression (EGF, Hsp70, Ki76, p53 e MMP2). 80th glioblastoma cell lines had demonstrated chromosomic alterations and expression of molecular markers similar to those described for glioblastoma phenotype. Equally, we compared the growth of these cells in monolayer cultures to three-dimensional multicellular spheroids cultures. Accordingly the results demonstrated different standarts of growth and molecular markers between the culture conditions

Effects of toxic doses of glutamate on Cu-Zn and Mn/superoxide dismutases activities in human glioma cell lines.

Recent research has implicated glutamate in the growth and invasive migration of gliomas. Superoxide dismutase (SOD) is involved in excitotoxicity and may influence cellular proliferative status. Thus, this study investigated the effects of gliotoxic doses of glutamate on Cu-Zn and Mn/SODs activities in human glioma cell lines. To this end, glioma cell lines (U87MG, U138MG and U251MG) were treated with glutamate (5-200 mM) during 48 h. Then, cell viability assays, clonogenic assay and Cu-Zn and Mn/SODs activities of the cell lines were performed. IC50values of glutamate were similar for both U87MG and U138MG cells (56 and 69 mM, respectively), while a higher value was detected for U251MG cells (110 mM). In the long term, 14 days after glutamate was removed from the culture media, cells showed partial or complete recovery. The effects of glutamate treatment on Cu-Zn and Mn/SODs activities varied among the distinct cell lines. While acute treatment with toxic doses of glutamate caused a significant decrease in the Cu-Zn/SOD activity of U138MG and U251MG cells, it did not affect Cu-Zn/SOD activity in U87MG cells. Only in U251MG cells, acute glutamate treatment decreased significantly Mn/SOD activity. In the long term (14 days after the 48 h treatment), glutamate did not affect either Cu-Zn or Mn/SODs activities. Thus, it may be suggested that SOD vulnerability to glutamate-mediated effects may be related to distinct tumoral cell behavior.