Fast diffusion kurtosis imaging (DKI) with Inherent COrrelation‐based Normalization (ICON) enhances automatic segmentation of heterogeneous diffusion MRI lesion in acute stroke

Diffusion kurtosis imaging (DKI) has been shown to augment diffusion‐weighted imaging (DWI) for the definition of irreversible ischemic injury. However, the complexity of cerebral structure/composition makes the kurtosis map heterogeneous, limiting the specificity of kurtosis hyperintensity to acute ischemia. We propose an Inherent COrrelation‐based Normalization (ICON) analysis to suppress the intrinsic kurtosis heterogeneity for improved characterization of heterogeneous ischemic tissue injury. Fast DKI and relaxation measurements were performed on normal (n = 10) and stroke rats following middle cerebral artery occlusion (MCAO) (n = 20). We evaluated the correlations between mean kurtosis (MK), mean diffusivity (MD) and fractional anisotropy (FA) derived from the fast DKI sequence and relaxation rates R₁ and R₂, and found a highly significant correlation between MK and R₁ (p < 0.001). We showed that ICON analysis suppressed the intrinsic kurtosis heterogeneity in normal cerebral tissue, enabling automated tissue segmentation in an animal stroke model. We found significantly different kurtosis and diffusivity lesion volumes: 147 ± 59 and 180 ± 66 mm³, respectively (p = 0.003, paired t‐test). The ratio of kurtosis to diffusivity lesion volume was 84% ± 19% (p < 0.001, one‐sample t‐test). We found that relaxation‐normalized MK (RNMK), but not MD, values were significantly different between kurtosis and diffusivity lesions (p < 0.001, analysis of variance). Our study showed that fast DKI with ICON analysis provides a promising means of demarcation of heterogeneous DWI stroke lesions.     

MRI quantification of non‐Gaussian water diffusion in normal human kidney: a diffusional kurtosis imaging study

Our aim was to prospectively evaluate the feasibility of diffusional kurtosis imaging (DKI) in normal human kidney and to report preliminary DKI measurements. Institutional review board approval and informed consent were obtained. Forty‐two healthy volunteers underwent diffusion‐weighted imaging (DWI) scans with a 3‐T MR scanner. b values of 0, 500 and 1000 s/mm² were adopted. Maps of fractional anisotropy (FA), mean diffusivity (MD), radial diffusivity (D_⊥), axial diffusivity (D_||), mean kurtosis (MK), radial kurtosis (K_⊥) and axial kurtosis (K_||) were produced. Three representative axial slices in the upper pole, mid‐zone and lower pole were selected in the left and right kidney. On each selected slice, three regions of interest were drawn on the renal cortex and another three on the medulla. Statistical comparison was performed with t‐test and analysis of variance. Thirty‐seven volunteers successfully completed the scans. No statistically significant differences were observed between the left and right kidney for all metrics (p values in the cortex: FA, 0.114; MD, 0.531; D_⊥, 0.576; D_||, 0.691; MK, 0.934; K_⊥, 0.722; K_||, 0.891; p values in the medulla: FA, 0.348; MD, 0.732; D_⊥, 0.470; D_||, 0.289; MK, 0.959; K_⊥, 0.780; K_||, 0.287). Kurtosis metrics (MK, K_||, K_⊥) obtained in the renal medulla were significantly (p <0.001) higher than those in the cortex (0.552 ± 0.04, 0.637 ± 0.07 and 0.530 ± 0.08 in the medulla and 0.373 ± 0.04, 0.492 ± 0.06 and 0.295 ± 0.06 in the cortex, respectively). For the diffusivity measures, FA of the medulla (0.356 ± 0.03) was higher than that of the cortex (0.179 ± 0.03), whereas MD, D_⊥ and D_|| (mm²/ms) were lower in the medulla than in the cortex (3.88 ± 0.09, 3.50 ± 0.23 and 4.65 ± 0.29 in the cortex and 2.88 ± 0.11, 2.32 ± 0.20 and 3.47 ± 0.31 in the medulla, respectively). Our results indicate that DKI is feasible in the human kidney. We have reported the preliminary DKI measurements of normal human kidney that demonstrate well the non‐Gaussian behavior of water diffusion, especially in the renal medulla. Copyright © 2014 John Wiley & Sons, Ltd.     

Diffusion abnormalities in temporal lobes of children with temporal lobe epilepsy: a preliminary diffusional kurtosis imaging study and comparison with diffusion tensor imaging

In this preliminary study, we aimed to investigate the abnormalities of water diffusion in children with temporal lobe epilepsy (TLE). Eight children with unilateral TLE (according to electroencephalography, EEG) and eight age‐ and sex‐matched controls were recruited. Diffusion tensor imaging (DTI)/diffusional kurtosis imaging (DKI) acquisitions were performed. Radial diffusivity (λ_⊥), axial diffusivity (λ_∥), mean diffusivity (MD) and fractional anisotropy (FA) maps were calculated for both DTI and DKI, and radial kurtosis (K_⊥), axial kurtosis (K_∥) and mean kurtosis (MK) maps were calculated for DKI only. Mann–Whitney test showed that, for white matter in the temporal lobe, DKI‐derived λ_∥, MD and K_∥ were significantly different in bilateral temporal lobes and EEG‐abnormal and EEG‐normal sides of the temporal lobe between patients and controls, whereas DTI showed no abnormalities. For gray matter, DKI detected significantly higher MD and MK in the same three comparisons, whereas DTI detected abnormalities only in the comparison between bilateral temporal lobes and between EEG‐normal sides in cases and left–right matched sides in controls. No significant difference was observed between EEG‐abnormal and EEG‐normal sides in cases. These preliminary results indicate that DKI is more sensitive than DTI for the detection of diffusion abnormalities in the temporal lobes of children with TLE, even when EEG signals are normal. These findings pave the way for the application of DKI for in‐depth studies on TLE in children. Copyright © 2012 John Wiley & Sons, Ltd.     

Measuring healthy female nulliparous pubovisceral muscle from diffusion kurtosis imaging

This study explores the feasibility of using diffusion kurtosis imaging (DKI) in the pelvic floor region and assesses the water diffusivity of the pubovisceral muscle. Twenty-seven healthy young nulliparous females underwent DKI at 3.0 T that included 15 gradient directions and three b values (0, 750, and 1500 s/mm²). The diffusion tensor and diffusion kurtosis metrics values of the pubovisceral muscle were measured after image processing. Two independent sample t-tests, a paired-samples t-test, and a nonparametric hypothesis test were performed as appropriate to compare the differences among different metrics. Twenty-six subjects (mean ± standard deviation age, 25 ± 2 years) were successfully analyzed by measuring the diffusion tensor and diffusion kurtosis metrics of the bilateral pubovisceral muscles. The metrics included mean kurtosis, axial kurtosis, radial kurtosis, fractional anisotropy, mean diffusivity, axial diffusivity, and radial diffusivity. We found no statistically significant differences for these measurement values between the left and right pubovisceral muscles (p = 0.271–0.931). However, radial kurtosis was greater than axial kurtosis in both pubovisceral muscles (p < 0.001) and axial diffusivity was lower than radial diffusivity in both pubovisceral muscles (p < 0.001). We deem the application of DKI technology to the pelvic floor region to be feasible.     

Characterisation of brain microstructural alterations in children with obstructive sleep apnea syndrome using diffusion kurtosis imaging

Obstructive sleep apnea (OSA) is a common chronic sleep-related breathing disorder in children. Previous studies showed widespread alterations in white matter (WM) in children with OSA mainly by using diffusion tensor imaging (DTI), while diffusional kurtosis imaging (DKI) extended DTI and exhibited improved sensitivity in detecting developmental and pathological changes in neural tissues. Therefore, we conducted whole-brain DTI and DKI analyses and compared the differences in kurtosis and diffusion parameters within the skeleton between 41 children with OSA and 32 healthy children. Between-group differences were evaluated by tract-based spatial statistics (TBSS) analysis (p < 0.05, TFCE corrected), and partial correlations between DKI metrics and sleep parameters were assessed considering age and gender as covariates. Compared with the controls, children with OSA showed significantly decreased kurtosis fractional anisotropy (KFA) mainly in white matter regions with a complex fibre arrangement including the posterior corona radiate (PCR), superior longitudinal fasciculus (SLF), and inferior fronto-occipital fasciculus (IFOF), while decreased FA in white matter regions with a coherent fibre arrangement including the posterior limb of internal capsule (PLIC), anterior thalamic radiation (ATR), and corpus callosum (CC). Notably, the receiver operating characteristic (ROC) curve analysis demonstrated the KFA value in complex tissue regions significantly (p < 0.001) differentiated children with OSA from the controls. In addition, the KFA value in the left PCR, SLF, and IFOF showed significant partial correlations to the sleep parameters for children with OSA. Combining DKI derived kurtosis and diffusion parameters can provide complementary neuroimaging biomarkers for assessing white matter alterations, and reveal pathological changes and monitor disease progression in paediatric OSA.     

Validation of fast diffusion kurtosis MRI for imaging acute ischemia in a rodent model of stroke

Diffusion‐weighted imaging (DWI) captures ischemic tissue that is likely to infarct, and has become one of the most widely used acute stroke imaging techniques. Diffusion kurtosis imaging (DKI) has lately been postulated as a complementary MRI method to stratify the heterogeneously damaged DWI lesion. However, the conventional DKI acquisition time is relatively long, limiting its use in the acute stroke setting. Recently, a fast kurtosis mapping method has been demonstrated in fixed brains and control subjects. The fast DKI approach provides mean diffusion and kurtosis measurements under substantially reduced scan time, making it amenable to acute stroke imaging. Because it is not practical to obtain and compare different means of DKI to test whether the fast DKI method can reliably detect diffusion and kurtosis lesions in acute stroke patients, our study investigated its diagnostic value using an animal model of acute stroke, a critical step before fast DKI acquisition can be routinely applied in the acute stroke setting. We found significant correlation, per voxel, between the diffusion and kurtosis coefficients measured using the fast and conventional DKI protocols. In acute stroke rats, the two DKI methods yielded diffusion and kurtosis lesions that were in good agreement. Importantly, substantial kurtosis–diffusion lesion mismatch was observed using the conventional (26 ± 13%, P < 0.01) and fast DKI methods (23 ± 8%, P < 0.01). In addition, regression analysis showed that the kurtosis–diffusion lesion mismatches obtained using conventional and fast DKI methods were substantially correlated (R² = 0.57, P = 0.02). Our results confirmed that the recently proposed fast DKI method is capable of capturing heterogeneous diffusion and kurtosis lesions in acute ischemic stroke, and thus is suitable for translational applications in the acute stroke clinical setting. Copyright © 2014 John Wiley & Sons, Ltd.     

弥散峰度成像与直肠癌临床病理预后因素的相关性研究

目的 探讨弥散峰度成像(DKI)定量参数和表观扩散系数(ADC)与直肠癌临床病理预后因素之间的潜在关系,为临床预测评估直肠癌的恶性程度提供一定的参考依据。方法 对2016年11月—2017年4月山西省肿瘤医院122例术前行MRI检查的直肠腺癌患者的影像和临床资料进行回顾性分析,其中女48例(39.3%)、男74例 (60.7%),年龄42~81岁。利用相关软件测得平均表观扩散系数(MD)、平均峰度(MK)和ADC值,通过独立样本t检验或Mann-Whitney U检验、ROC曲线和Spearman相关性分析进行统计学分析。结果 直肠癌高T分期及淋巴结转移与低MD值(r=-0.367、-0.240)和低ADC值(r=-0.391、-0.254)相关,不同分组间差异均有统计学意义(P值均＜0.05);同样,不同组织病理学分级的MD值之间差异亦有统计学意义(P＜0.05)且呈负相关(r=-0.210, P＜0.05);随着组织病理学分级的升高、淋巴结受累、瘤周血管浸润(LVI)或神经侵犯及环周切缘(CRM)受侵,MK值相应增大,差异均有统计学意义(P值均＜0.05)呈正相关(r=0.478、0.206、0.237、0.228, P值均＜0.05)。MD、MK和ADC值均与淋巴结转移有相关(P值均＜0.01)性;ROC曲线显示MK值较其他参数在诊断淋巴结转移与否的曲线下面积高为0.784(95%CI 0.703~0.865),当其阈值取0.984时,具有较高的敏感度及特异度,分别为65.9%和88.7%。结论直肠癌DKI定量参数值和ADC值,尤其MK值,与其重要的临床病理预后因素均有明显相关性,对于预测直肠癌患者预后有一定意义。     

Histological correlation of diffusional kurtosis and white matter modeling metrics in cuprizone‐induced corpus callosum demyelination

The cuprizone mouse model is well established for studying the processes of both demyelination and remyelination in the corpus callosum, and it has been utilized together with diffusion tensor imaging (DTI) to investigate myelin and axonal pathology. Although some underlying morphological mechanisms contributing to the changes in diffusion tensor (DT) metrics have been identified, the understanding of specific associations between histology and diffusion measures remains limited. Diffusional kurtosis imaging (DKI) is an extension of DTI that provides metrics of diffusional non‐Gaussianity, for which an associated white matter modeling (WMM) method has been developed. The main goal of the present study was to quantitatively assess the relationships between diffusion measures and histological measures in the mouse model of cuprizone‐induced corpus callosum demyelination. The diffusional kurtosis (DK) and WMM metrics were found to provide additional information that enhances the sensitivity to detect the morphological heterogeneity in the chronic phase of the disease process in the rostral segment of the corpus callosum. Specifically, in the rostral segment, axonal water fraction (d = 2.6; p < 0.0001), radial kurtosis (d = 2.0; p = 0.001) and mean kurtosis (d = 1.5; p = 0.005) showed the most sensitivity between groups with respect to yielding statistically significant p values and high Cohen's d values. These results demonstrate the ability of DK and WMM metrics to detect white mater changes and inflammatory processes associated with cuprizone‐induced demyelination. They also validate, in part, the application of these new WMM metrics for studying neurological diseases, as well as helping to elucidate their biophysical meaning. Copyright © 2014 John Wiley & Sons, Ltd.     

扩散峰度成像在预测直肠癌KRAS基因突变中的应用价值

目的 探讨扩散峰度成像(DKI)定量参数在预测直肠癌KRAS基因突变中的应用价值。方法 回顾性分析山西省肿瘤医院2016年11月-2018年6月经病理证实的152例直肠腺癌患者的资料,其中男92例,女60例;年龄33～86岁,平均61岁。患者均于术前行常规MRI和功能DKI序列检查,由2名放射科医师双盲勾画感兴趣区,应用MatLab软件计算两组患者DKI定量参数平均表观扩散系数(MD)、平均峰度(MK)以及表观扩散系数(ADC),并采用组内相关系数进行一致性分析。术后均进行KRAS基因检测,依据检测结果将患者分为野生组和突变组两组,采用独立样本t检验对比两组患者的MD、MK、ADC。绘制DKI定量参数诊断KRAS基因突变的受试者工作特征曲线(ROC) ,根据约登指数确定各定量参数的最佳诊断阈值,以及相应的灵敏度、特异度,并采用DeLong检验比较各定量参数的ROC的曲线下面积(AUC)。结果 152例直肠癌患者中,KRAS基因突变组74例,野生组78例,基因突变率为48.6%(74/152)。突变组患者的ADC、MD、MK值分别为(1.18±0.18)×10^-3 mm²/s、(1.28±0.18)×10^-3 mm²/s、0.97±0.11,野生组分别为(1.33±0.18)×10^-3 mm²/s、(1.42±0.17)×10^-3 mm²/s、0.82±0.09;突变组ADC、MD值均小于野生组患者,而MK值则大于野生组患者,差异均有统计学意义(t=5.424、4.882、-8.809, P值均＜0.01)。ROC曲线显示,ADC、MD、和MK值预测KRAS基因的AUC分别为0.758、0.740、0.845,灵敏度分别为75.7%、82.4%、77.0%,特异度分别为68.0%、57.8%、84.6%。DeLong检验结果显示,MK值的AUC明显高于ADC和MD值(P值均＜0.01),而ADC值和MD值间AUC差异无统计学意义(P＞0.05)。结论 DKI定量参数MD、MK和ADC值,在预测直肠癌的KRAS基因突变方面均有一定的价值,其中MK值有更高的AUC和特异度,有更高的诊断价值。     

弥散峰度成像预测直肠腺癌UGT1A1*28基因突变的价值

目的:评估弥散峰度成像（DKI）对直肠腺癌尿苷二磷酸葡萄糖醛酸基转移酶1A1（UGT1A1）*28基因突变的预测价值。方法:回顾性研究。纳入山西省肿瘤医院2016年11月—2020年8月167例直肠腺癌患者的临床资料,其中男98例、女69例,年龄29~89岁、中位年龄为62岁。患者术前均行MR常规序列和DKI序列检查,...     

Fast diffusion kurtosis imaging of fibrotic mouse kidneys

Diffusion kurtosis imaging (DKI) is sensitive to tissue microstructure and may therefore be useful in the diagnosis and monitoring of disease in brain and body organs. Generally, diffusion magnetic resonance imaging (dMRI) in the body is challenging because of the heterogeneous body composition, which can cause image artefacts as a result of chemical shifts and susceptibility differences. In addition, the abdomen possesses physiological factors (e.g. breathing, heartbeat, blood flow) which may severely reduce image quality, especially when echo planar imaging is employed, as is typical in dMRI. Collectively, these challenging measurement conditions impede the use and exploration of DKI in the body. This impediment is further exacerbated by the traditionally large amount of data required for DKI and the low signal‐to‐noise ratio at the b‐values needed to effectively probe the kurtosis regime. Recently introduced fast DKI techniques reduce the challenge of DKI in the body by decreasing the data requirement substantially, so that, for example, triggering and breath‐hold techniques may be applied for the entire DKI acquisition without causing unfeasible scan times. One common pathological condition for which body DKI may be of immediate clinical value is kidney fibrosis, which causes progressive changes in organ microstructure. With its sensitivity to microstructure, DKI is an obvious candidate for a non‐invasive evaluation method. We present preclinical evidence indicating that the rapidly obtainable tensor‐derived mean kurtosis ( ) distinguishes moderately fibrotic kidneys from healthy controls. The presence and degree of fibrosis are confirmed by histology, which also indicates fibrosis as the main driver behind the DKI differences observed between groups. We therefore conclude that fast kurtosis is a likely candidate for an MRI‐based method for the detection and monitoring of renal fibrosis. We provide protocol recommendations for fast renal DKI in humans based on a b‐value optimisation performed using data acquired at 3 T in normal human kidney.     

Diffusional kurtosis MRI of the lower leg: changes caused by passive muscle elongation and shortening

Diffusional kurtosis MRI (DKI) quantifies the deviation of water diffusion from a Gaussian distribution. We investigated the influence of passive elongation and shortening of the lower leg muscles on the DKI parameters D (diffusion coefficient) and K (kurtosis). After approval by the local ethics committee, eight healthy volunteers (age, 29.1 ± 2.9 years) underwent MRI of the lower leg at 3 T. Diffusion‐weighted images were acquired with 10 different b values at three ankle positions (passive dorsiflexion 10°, neutral position 0°, passive plantar flexion 40°). Parametrical maps of D and K were obtained by voxel‐wise fitting of the signal intensities using a non‐linear Levenberg–Marquardt algorithm. D and K were measured in the tibialis anterior, medial and lateral gastrocnemius, and soleus muscles. In the neutral position, D and K values were in the range between 1.66–1.79 × 10^–3 mm²/s and 0.21–0.39, respectively. D and K increased with passive shortening, and decreased with passive elongation, which could also be illustrated on the parametrical maps. In dorsiflexion, D (p < 0.01) and K (p = 0.036) were higher in the tibialis anterior than in the medial gastrocnemius. In plantar flexion, the opposite was found for K (p = 0.035). DKI parameters in the lower leg muscles are significantly influenced by the ankle joint position, indicating that the diffusion of water molecules in skeletal muscle deviates from a Gaussian distribution depending on muscle tonus. Copyright © 2016 John Wiley & Sons, Ltd.     

Comparison of image sensitivity between conventional tensor‐based and fast diffusion kurtosis imaging protocols in a rodent model of acute ischemic stroke

Diffusion kurtosis imaging (DKI) can offer a useful complementary tool to routine diffusion MRI for improved stratification of heterogeneous tissue damage in acute ischemic stroke. However, its relatively long imaging time has hampered its clinical application in the emergency setting. A recently proposed fast DKI approach substantially shortens the imaging time, which may help to overcome the scan time limitation. However, to date, the sensitivity of the fast DKI protocol for the imaging of acute stroke has not been fully described. In this study, we performed routine and fast DKI scans in a rodent model of acute stroke, and compared the sensitivity of diffusivity and kurtosis indices (i.e. axial, radial and mean) in depicting acute ischemic lesions. In addition, we analyzed the contrast‐to‐noise ratio (CNR) between the ipsilateral ischemic and contralateral normal regions using both conventional and fast DKI methods. We found that the mean kurtosis shows a relative change of 47.1 ± 7.3% between the ischemic and contralateral normal regions, being the most sensitive parameter in revealing acute ischemic injury. The two DKI methods yielded highly correlated diffusivity and kurtosis measures and lesion volumes (R² ? 0.90, p < 0.01). Importantly, the fast DKI method exhibited significantly higher CNR of mean kurtosis (1.6 ± 0.2) compared with the routine tensor protocol (1.3 ± 0.2, p < 0.05), with its CNR per unit time (CNR efficiency) approximately doubled when the scan time was taken into account. In conclusion, the fast DKI method provides excellent sensitivity and efficiency to image acute ischemic tissue damage, which is essential for image‐guided and individualized stroke treatment. Copyright © 2016 John Wiley & Sons, Ltd.     

PARK2 and PACRG are commonly downregulated in clear‐cell renal cell carcinoma and are associated with aggressive disease and poor clinical outcome

PARK2 is an E3 ligase, known to be involved in ubiquitination of several proteins and to play a role in neuronal protection. The gene PARK2 and its potentially co‐regulated gene PACRG have been previously found to be deleted in clear‐cell renal cell carcinomas (ccRCCs). The aim of our study was to evaluate the mRNA and protein expression of PARK2 and PACRG in a large cohort of ccRCC, and to investigate their association with outcome. The expression of both genes was measured by quantitative PCR in 94 primary ccRCCs and autologous nonmalignant kidney tissues. PACRG and PARK2 protein expression was determined immunohistochemically using tissue microarrays comprising 133 ccRCCs. The mRNA and protein expression of PARK2 and PACRG was significantly downregulated in ccRCCs compared with nonmalignant tissues. Low levels of PARK2 mRNA were associated with high‐grade ccRCC and lymph node metastasis. Patients with low PARK2 mRNA levels showed a higher tumor‐specific mortality rate and a shorter overall survival (OS) than those with high PARK2 expression. Patients without PACRG mRNA expression in the tumor had a shorter disease‐free survival and OS than those with tumors expressing PACRG. In multivariate analyses, neither PARK2 nor PACRG expression were independent prognostic factors. The protein expression of PARK2 and PACRG was significantly downregulated in ccRCCs (82.8, and 96.9%, respectively), but no association with clinical outcome was noticed. © 2012 Wiley Periodicals, Inc.     

The protein tyrosine phosphatase receptor type J is regulated by the pVHL–HIF axis in clear cell renal cell carcinoma

Mass spectrometry analysis of renal cancer cell lines recently suggested that the protein‐tyrosine phosphatase receptor type J (PTPRJ), an important regulator of tyrosine kinase receptors, is tightly linked to the von Hippel–Lindau protein (pVHL). Therefore, we aimed to characterize the biological relevance of PTPRJ for clear cell renal cell carcinoma (ccRCC). In pVHL‐negative ccRCC cell lines, both RNA and protein expression levels of PTPRJ were lower than those in the corresponding pVHL reconstituted cells. Quantitative RT‐PCR and western blot analysis of ccRCC with known VHL mutation status and normal matched tissues as well as RNA in situ hybridization on a tissue microarray (TMA) confirmed a decrease of PTPRJ expression in more than 80% of ccRCCs, but in only 12% of papillary RCCs. ccRCC patients with no or reduced PTPRJ mRNA expression had a less favourable outcome than those with a normal expression status (p = 0.05). Sequence analysis of 32 PTPRJ mRNA‐negative ccRCC samples showed five known polymorphisms but no mutations, implying other mechanisms leading to PTPRJ's down‐regulation. Selective silencing of HIF‐α by siRNA and reporter gene assays demonstrated that pVHL inactivation reduces PTPRJ expression through a HIF‐dependent mechanism, which is mainly driven by HIF‐2α stabilization. Our results suggest PTPRJ as a member of a pVHL‐controlled pathway whose suppression by HIF is critical for ccRCC development.     

White matter organization in relation to upper limb motor control in healthy subjects: exploring the added value of diffusion kurtosis imaging

Diffusion tensor imaging (DTI) characterizes white matter (WM) microstructure. In many brain regions, however, the assumption that the diffusion probability distribution is Gaussian may be invalid, even at low b values. Recently, diffusion kurtosis imaging (DKI) was suggested to more accurately estimate this distribution. We explored the added value of DKI in studying the relation between WM microstructure and upper limb coordination in healthy controls (N = 24). Performance on a complex bimanual tracking task was studied with respect to the conventional DTI measures (DKI or DTI derived) and kurtosis metrics of WM tracts/regions carrying efferent (motor) output from the brain, corpus callosum (CC) substructures and whole brain WM. For both estimation models, motor performance was associated with fractional anisotropy (FA) of the CC-genu, CC-body, the anterior limb of the internal capsule, and whole brain WM (r _s range 0.42–0.63). Although DKI revealed higher mean, radial and axial diffusivity and lower FA than DTI (p < 0.001), the correlation coefficients were comparable. Finally, better motor performance was associated with increased mean and radial kurtosis and kurtosis anisotropy (r _s range 0.43–0.55). In conclusion, DKI provided additional information, but did not show increased sensitivity to detect relations between WM microstructure and bimanual performance in healthy controls.     

Cyclin D1 expression in astrocytomas is associated with cell proliferation activity and patient prognosis

An important positive regulator of the cell cycle, cyclin D1, is often amplified and overexpressed in malignancies. Cyclin D1 aberrations were analysed in grade II–IV astrocytomas by fluorescence in situ hybridization (FISH), mRNA in situ hybridization and immunohistochemistry. Proliferation activity was determined by Ki-67^MIB-1 immunolabelling and mitotic counting. High cyclin D1 expression was observed in grade IV astrocytomas (grades II–III versus grade IV; mRNA expression: p < 0·001; immunoexpression: p = 0·013), and correlated with poor patient survival (p < 0·001, n = 46). Upregulated cyclin D1 expression was also closely associated with poor patient prognosis in grade II–III astrocytomas (p < 0·001, n = 30). Cyclin D1 gene was not found to be amplified (n = 7). Cell proliferation activity was significantly increased in tumours exhibiting high cyclin D1 mRNA levels (Ki-67^MIB-1: p < 0·001; mitotic count: p < 0·001) and high cyclin D1 protein expression (Ki-67^MIB-1: p = 0·002; mitotic count: p = 0·012). These results indicate that increased production of cyclin D1 is closely associated with high cell proliferation activity and aggressive behaviour in diffusely infiltrating astrocytomas. Copyright © 1999 John Wiley & Sons, Ltd.     

Precision and accuracy of diffusion kurtosis estimation and the influence of b‐value selection

Diffusion kurtosis imaging (DKI) is an extension of diffusion tensor imaging that accounts for leading non‐Gaussian diffusion effects. In DKI studies, a wide range of different gradient strengths (b‐values) is used, which is known to affect the estimated diffusivity and kurtosis parameters. Hence there is a need to assess the accuracy and precision of the estimated parameters as a function of b‐value. This work examines the error in the estimation of mean of the kurtosis tensor (MKT) with respect to the ground truth, using simulations based on a biophysical model for both gray (GM) and white (WM) matter. Model parameters are derived from densely sampled experimental data acquired in ex vivo rat brain and in vivo human brain. Additionally, the variability of MKT is studied using the experimental data. Prevalent fitting protocols are implemented and investigated. The results show strong dependence on the maximum b‐value of both net relative error and standard deviation of error for all of the employed fitting protocols. The choice of b‐values with minimum MKT estimation error and standard deviation of error was found to depend on the protocol type and the tissue. Protocols that utilize two terms of the cumulant expansion (DKI) were found to achieve minimum error in GM at b‐values less than 1 ms/μm², whereas maximal b‐values of about 2.5 ms/μm² were found to be optimal in WM. Protocols including additional higher order terms of the cumulant expansion were found to provide higher accuracy for the more commonly used b‐value regime in GM, but were associated with higher error in WM. Averaged over multiple voxels, a net average error of around 15% for both WM and GM was observed for the optimal b‐value choice. These results suggest caution when using DKI generated metrics for microstructural modeling and when comparing results obtained using different fitting techniques and b‐values.     

Double‐pulsed diffusional kurtosis imaging

Diffusional kurtosis imaging (DKI) is extended to double‐pulsed‐field‐gradient (d‐PFG) diffusion MRI sequences. This gives a practical approach for acquiring and analyzing d‐PFG data. In particular, the leading d‐PFG effects, beyond what conventional single‐pulsed field gradient (s‐PFG) provides, are interpreted in terms of the kurtosis for a diffusion displacement probability density function (dPDF) in a six‐dimensional (6D) space. The 6D diffusional kurtosis encodes the unique information provided by d‐PFG sequences up to second order in the b‐value. This observation leads to a compact expression for the signal magnitude, and it suggests novel data acquisition and analysis methods. Double‐pulsed DKI (DP‐DKI) is demonstrated for in vivo mouse brain with d‐PFG data obtained at 7 T. Copyright © 2014 John Wiley & Sons, Ltd.     

Ultra‐deep T cell receptor sequencing reveals the complexity and intratumour heterogeneity of T cell clones in renal cell carcinomas

The recognition of cancer cells by T cells can impact upon prognosis and be exploited for immunotherapeutic approaches. This recognition depends on the specific interaction between antigens displayed on the surface of cancer cells and the T cell receptor (TCR), which is generated by somatic rearrangements of TCR α‐ and β‐chains (TCRb). Our aim was to assess whether ultra‐deep sequencing of the rearranged TCRb in DNA extracted from unfractionated clear cell renal cell carcinoma (ccRCC) samples can provide insights into the clonality and heterogeneity of intratumoural T cells in ccRCCs, a tumour type that can display extensive genetic intratumour heterogeneity (ITH). For this purpose, DNA was extracted from two to four tumour regions from each of four primary ccRCCs and was analysed by ultra‐deep TCR sequencing. In parallel, tumour infiltration by CD4, CD8 and Foxp3 regulatory T cells was evaluated by immunohistochemistry and correlated with TCR‐sequencing data. A polyclonal T cell repertoire with 367–16 289 (median 2394) unique TCRb sequences was identified per tumour region. The frequencies of the 100 most abundant T cell clones/tumour were poorly correlated between most regions (Pearson correlation coefficient, –0.218 to 0.465). 3–93% of these T cell clones were not detectable across all regions. Thus, the clonal composition of T cell populations can be heterogeneous across different regions of the same ccRCC. T cell ITH was higher in tumours pretreated with an mTOR inhibitor, which could suggest that therapy can influence adaptive tumour immunity. These data show that ultra‐deep TCR‐sequencing technology can be applied directly to DNA extracted from unfractionated tumour samples, allowing novel insights into the clonality of T cell populations in cancers. These were polyclonal and displayed ITH in ccRCC. TCRb sequencing may shed light on mechanisms of cancer immunity and the efficacy of immunotherapy approaches. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.