Temporal resolution and SNR requirements for accurate DCE‐MRI data analysis using the AATH model

Dynamic contrast‐enhanced MRI has been used in conjunction with tracer kinetics modeling in a wide range of tissues for treatment monitoring, oncology drug development, and investigation of disease processes. Accurate measurement of model parameters relies on acquiring data with high temporal resolution and low noise, particularly for models with large numbers of free parameters, such as the adiabatic approximation to the tissue homogeneity model for separate measurements of blood flow and vessel permeability. In this simulation study, accuracy of the adiabatic approximation to the tissue homogeneity model was investigated, examining the effects of temporal resolution, noise levels, and error in the measured arterial input function. A temporal resolution of 1.5 s and high SNR (noise sd = 0.05) were found to ensure minimal bias (<5%) in all four model parameters (extraction fraction, blood flow, mean transit time, and extravascular extracellular volume), and the sampling interval can be relaxed to 6 s, if the transit time need not be measured accurately (bias becomes >10%). A 10% error in the measured height of the arterial input function first pass peak resulted in an error of at most 10% in each model parameter. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.     

EGCG induces apoptosis in human laryngeal epidermoid carcinoma Hep2 cells via mitochondria with the release of apoptosis-inducing factor and endonuclease G

(−)-Epigallocatechin-3-gallate (EGCG), a major green tea polyphenol, was tested for in vitro cytotoxicity against human laryngeal epidermoid carcinoma of the larynx Hep2 cells. EGCG-induced apoptotic cell death accompanied by a change in the cell cycle. However, EGCG did not result in caspase activation, nor did a caspase inhibitor block cell death. Furthermore, EGCG caused no change in the intracellular levels of reactive oxygen species (ROS). The levels of p53 were increased in the EGCG-treated cells, with a corresponding decrease in Bcl-2 and Bid protein levels as well as an increase in the Bax level. In addition, EGCG induced the cytoplasmic release of cytochrome c from the mitochondria accompanied by a decreased mitochondrial membrane potential, and subsequently upregulated translocation of apoptosis-inducing factor (AIF) and endonuclease G (EndoG) into the nucleus during the apoptotic process. Taken together, these findings indicate that the p53-mediated mitochondrial pathway and the nuclear translocation of AIF and EndoG play a crucial role in EGCG-induced apoptosis of human laryngeal epidermoid carcinoma Hep2 cells, which proceeds through a caspase-independent pathway.     

Aloe-emodin induces apoptosis of human nasopharyngeal carcinoma cells via caspase-8-mediated activation of the mitochondrial death pathway

Aloe-emodin (AE), a natural, biologically active compound from the rhizome of Rheum palmatum, has been shown to induce apoptosis in several cancer cell lines in vitro. However, its molecular mechanism of action in the apoptosis induction of human nasopharyngeal carcinoma (NPC) cells has not been explored. This study shows that AE induced G₂/M phase arrest by increasing levels of cyclin B1 bound to Cdc2, and also caused an increase in apoptosis of NPC cells, which was characterized by morphological changes, nuclear condensation, DNA fragmentation, caspase-3 activation, cleavage of poly (ADP-ribose) polymerase (PARP) and increased sub-G₁ population. Treatment of NPC cells with AE also resulted in a decrease in Bcl-X_L and an increase in Bax expression. Ectopic expression of Bcl-X_L but not Bcl-2 or small interfering RNA (siRNA)-mediated attenuation of Bax suppressed AE-induced apoptotic cell death. AE-induced loss of mitochondrial membrane potential (MMP) and increase in cellular Ca⁺⁺ content, reactive oxygen species (ROS) and apoptotic cell death were suppressed by the treatment of cyclosporin A (CsA) or caspase-8 inhibitor Z-IETD-FMK. Co-treatment with caspase-9 inhibitor Z-LEHD-FMK could inhibit AE-induced cell death and the activation of caspase-3 and -9. In addition, suppression of caspase-8 with the specific inhibitor Z-IETD-FMK inhibited AE-induced the activation of Bax, the cleavage of Bid, the translocation of tBid to the mitochondria and the release of cytochrome c, apoptosis-inducing factor (AIF) and Endo G from the mitochondria and subsequent apoptosis. Taken together, these results indicate that the caspase-8-mediated activation of the mitochondrial death pathway plays a critical role in AE-induced apoptosis of NPC cells.     

低剂量电离辐射诱导小鼠睾丸NO含量、NOS活性及AIF基因的变化

目的 研究低剂量电离辐射诱导小鼠睾丸一氧化氮(NO)含量、一氧化氮合酶(NOS)活性及凋亡诱导因子(apoptosis inducing factor,AIF)基因的变化。方法 酶法检测不同剂量(0、0.025、0.05、0.075、0.1及0.2Gy)及0.075Gy照射后不同时间(0、3、6、12、18和24h)小鼠睾丸组织中NO含量及NOS活性的变化;Real-time PCR和Western blot法检测不同剂量(0、0.05、0.075、0.1及0.2Gy)及0.075Gy照射后不同时间(0、6、12和24h)小鼠睾丸组织中AIF mRNA表达及蛋白的表达。结果 不同剂量X射线照射后12h,小鼠睾丸组织中NO含量及NOS活性随照射剂量增加而增加,0.075Gy增至最大(P<0.05),0.1~0.2Gy一直维持较高水平(P<0.05);AIF mRNA随剂量增加而增加,0.1Gy时达到最大(P<0.05);而AIF蛋白在0.075Gy时达到最大。0.075Gy X射线照射后,NO含量与NOS活性随时间延长而增加,NO含量在24h时增到峰值(P<0.05),而NOS活性在12h时达到峰值(P<0.05);AIFmRNA随时间延长而增加,在24h时达到峰值(P<0.05);其蛋白表达在12h时达到峰值。结论 低剂量电离辐射照射后,小鼠睾丸生精细胞中NO含量、NOS活性及AIF基因表达增加,具有时程和剂量效应规律性。     

Neuroprotective actions of ovarian hormones without insult in the raphe region of rhesus macaques

Using a nonhuman primate model of surgical menopause, our laboratory has shown that ovarian hormone treatment (HT) improves 5-HT neural function in the dorsal raphe nucleus (DRN). We further hypothesize that HT may increase 5-HT neuronal resilience. Recent data from microarray analysis indicated that HT regulates gene expression in pathways that lead to apoptosis. In this study, we questioned whether HT alters protein expression in caspase-dependent and independent pathways. Ovariectomized monkeys received Silastic implants containing placebo (empty), estrogen (E) or E+ progesterone (P). A small block of the midbrain containing the DRN was dissected and subjected to subcellular fractionation, yielding cytosolic, nuclear and mitochondrial fractions (n=4/group). The pro-apoptotic protein, c-jun n-terminal kinase (JNK1) and its phosphorylation were decreased by E+P treatment in the cytosolic fraction. Downstream of JNK are proteins in the caspase-dependent and -independent pathways. First, in the caspase-dependent pathway, cytoplasmic and mitochondrial fractions were immunoblotted for Bcl-2 family members, cytochrome c, Apaf1 and XIAP. However, the expression of these proteins did not differ among treatments. Pro-caspase 3 was decreased by E+P, but there was no evidence of active caspase in any group. Then, we examined the involvement of a protein in the caspase-independent pathway, called apoptosis-inducing factor (AIF). AIF mRNA (n=3/group) and AIF mitochondrial protein tended to decrease with hormone treatment. However, AIF protein in the nuclear fraction in E+P treated monkeys was significantly reduced. This indicates that HT is reducing the translocation of AIF from mitochondria to nucleus, thus inhibiting AIF-mediated apoptosis. AIF was immunocytochemically localized to large 5-HT-like neurons of the dorsal raphe. These data suggest that in the absence of global trauma or ischemia, HT may act through the caspase-independent pathway to promote neuroprotection in the 5-HT system.     

Consistency of normalized cerebral blood volume values in glioblastoma using different leakage correction algorithms on dynamic susceptibility contrast magnetic resonance imaging data without and with preload

Background and purpose

Several leakage correction algorithms for dynamic susceptibility contrast (DSC) magnetic resonance imaging (MRI)-based cerebral blood volume (CBV) measurement have been proposed, and combination with a preload of contrast agent is generally recommended. A single bolus application scheme would largely simplify and facilitate standardized clinical applications, while reducing contrast agent (CA) dose. The aim of this study was, therefore, to investigate whether appropriate leakage correction redundantizes prebolus application by comparing normalized DSC-based CBV (nCBV) measures of two consecutive CA boli.