Production of intracellular superoxide mediates dithiothreitol-dependent inhibition of apoptotic cell death

Apoptotic cell death proceeds from the activation of cysteine proteinases called caspases. As full enzymatic activity of caspases requires reduction of cysteine residues in and around the catalytic site of the proteases, cysteine- reducing agents such as dithiothreitol (DTT) are expected to facilitate caspase activity upon induction of apoptosis. However, DTT has been shown to efficiently protect cells from apoptosis. The mechanism involved in DTT-mediated inhibition of apoptosis has been attributed to its antioxidant activity. Interestingly, under physiological conditions, thiol-mediated antioxidant reaction has also been shown to result in intracellular generation of superoxide (O(2) (.-)). In line with our earlier findings implicating a slight prooxidant state in resistance to apoptosis, we set out to investigate if the death-inhibitory activity of DTT could be mediated by intracellular O2 (.-). Our results show that incubation of human melanoma cell line M14TF or human bladder carcinoma cell line T24 with DTT induced an increase in intracellular O2 (.-) with concomitant inhibition of apoptosis triggered by CD95 signaling, staurosporine, or hydrogen peroxide. Moreover, preincubation of either cells with Tiron, a specific O2 (.-) scavenger, reverted DTT-induced inhibition of apoptosis. These results show that the apoptosis-inhibitory activity of DTT may not be due to its antioxidant property, but instead linked to its ability to induce an increase in intracellular O2 (.-) level.     

The prognostic value of the stem-like group in colorectal cancer using a panel of immunohistochemistry markers

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the Western world. It is becoming increasingly clear that CRC is a diverse disease, as exemplified by the identification of subgroups of CRC tumours that are driven by distinct biology. Recently, a number of studies have begun to define panels of diagnostically relevant markers to align patients into individual subgroups in an attempt to give information on prognosis and treatment response. We examined the immunohistochemical expression profile of 18 markers, each representing a putative role in cancer development, in 493 primary colorectal carcinomas using tissue microarrays. Through unsupervised clustering in stage II cancers, we identified two cluster groups that are broadly defined by inflammatory or immune-related factors (CD3, CD8, COX-2 and FOXP3) and stem-like factors (CD44, LGR5, SOX2, OCT4). The expression of the stem-like group markers was associated with a significantly worse prognosis compared to cases with lower expression. In addition, patients classified in the stem-like subgroup displayed a trend towards a benefit from adjuvant treatment. The biologically relevant and poor prognostic stem-like group could also be identified in early stage I cancers, suggesting a potential opportunity for the identification of aggressive tumors at a very early stage of the disease.     

Predicting GFR in children and adults: a comparison of the Cockcroft-Gault,Schwartz, and modification of diet in renal disease formulas

BACKGROUND: A rapid prediction of glomerular filtration rate (GFR) is often needed in clinics. Formulas based on plasma creatinine level are being increasingly used, Schwartz for children, supposed to give GFR; Cockcroft-Gault for adults, supposed to indicate the creatinine clearance; and a recent formula introduced by the Modification of Diet in Renal Disease (MDRD) group. Our objective was to test whether one single formula could suffice and which one gives the best estimation of GFR. METHODS: In 198 children (with two kidneys, single kidney, or renal transplant) and 116 adults (single kidney and transplanted), we measured inulin clearance and creatinine clearance and calculated Cockcroft-Gault, MDRD and, in children only, Schwartz. Data were compared with analysis of variance (ANOVA), regression statistics, and concordance studies. RESULTS: In patients over 12 years of age, Cockcroft-Gault was almost similar to GFR corrected for body surface and creatinine clearance exceeded GFR by more than 20%; Schwartz was above creatinine clearance excepted for transplanted children. In younger children, no prediction approached GFR. Predictions were well correlated with GFR, but concordance studies showed Schwartz with dispersed results and GFR overestimated (20 mL/min/1.73 m2); Cockcroft-Gault was close to GFR and results were dispersed; MDRD in children gave a large overestimation and badly dispersed results; in transplanted adults its prediction was good. CONCLUSION: Cockcroft-Gault prediction could be used for children over 12 years of age and adults; it should not be considered as creatinine clearance but as GFR corrected for body surface, it is merely a prediction, 95% of the results are between +/-40 mL/min/1.73 m(2) in children and +/-30 mL/min/1.72 m(2) in adults. In younger children no formula is satisfying.     

The prevention of pain during botulinum toxin injections in children

According to the recommendations of the French health products safety agency (AFSSAPS) in 2009, botulinum toxin injections are recommended to reduce spasticity in children with cerebral palsy. Apprehension of the session can increase the pain. Therefore, programmes have been put in place which help to win over the child and gain their trust. Since 2008 a nurse consultation which informs the child through play has been set up in the centre for physical medicine and rehabilitation of the Association for the social and professional integration of disabled people.