, 2011 and Wu et al., 2009). While light-induced photoactivation has been

in the cell biologists’ toolkit for decades, these methods have required chemical Vorinostat ic50 modification of molecules or proteins, which must then be somehow introduced to the intracellular environment and uncaged with UV light. It is only in recent times that the technique has been combined with the ease of genetically encoded expression and the use of visible light wavelengths, making the experiments very amenable to live neurons in culture or even in vivo. This can be attributed to exploitation of light sensitive domains isolated from plant proteins. These can be used to either allosterically block an active protein from interacting with its effectors (Wu et al., 2009), or to artificially dimerize two targets (Kennedy et al., 2010 and Levskaya et al., CDK inhibitor 2009). In the latter example, artificial interaction can be used to either anchor a target protein to a specific subcellular compartment or to cause an association

between two targets. In both cases, light absorption activates a synthetic signaling cascade that is both reversible and dose dependent. Like CALI, light-induced photoactivation is instantaneous and can be performed at the subcellular level. Overall, this type of manipulation is a better approximation of actual signaling events and an invaluable tool for deducing the true function of a protein in a specific cellular process. Together with the development and deployment of super resolution imaging techniques (Toomre and Bewersdorf, 2010), we might be closer to a better understanding of the full orchestra of the players that power the growth cone. Research in authors′ lab is supported in

part by grants from the National Institutes of Health to J.Q.Z. and a Ruth L. Kirschstein National Research Service Award to E.A.V. “
“The mammalian neocortex is characterized by its stereotyped laminar cytoarchitecture and regional variations in cellular architecture that differentiate cortical areas. As emphasized by Brodmann over a century ago through the creation of cytoarchitectonic cortical maps (Brodmann, 1909), cortical organization is conserved across species, particularly between humans and nonhuman primates (reviewed in Zilles Levetiracetam and Amunts, 2010). Gene expression is increasingly used as an empirical means of differentiating and delineating cortical areas, for example through identification of area-specific gene markers (Takahata et al., 2009) or boundary mapping based on differences in neurotransmitter receptor expression (Zilles et al., 2004). Whole-genome transcriptional profiling has particular potential to elucidate cortical areal specification and specialization through identification of differentially regulated genes and molecular pathways that underlie cytoarchitectural and functional areal identity (Johnson et al., 2009).

, 2012), and future work will determine which GTPases in vivo are regulated by plexin GAP activity. The Rho family of small GTPases, which includes Rho, Rac, and Cdc42, controls growth cone behavior through the regulation of actin dynamics ( Hall and Lalli, 2010). In Drosophila neuromuscular

development, overexpression of a dominant-negative (DN) Cdc42 causes motor neuron growth cone arrest. However, expression of DN Rac1 frequently results in parallel bypass phenotypes, www.selleckchem.com/products/bmn-673.html indicative of defects in target recognition and axonal defasciculation ( Kaufmann et al., 1998). Here, we observe that Rho1 plays a critical role in Sema-1a-mediated motor axon repulsion, and that its activity is modulated by opposing Pbl and p190 functions specified by Sema-1a-mediated repulsive,

and most likely attractive, signals ( Figure S6). We also observed highly penetrant motor axon pathfinding defects in Sema-1a, pbl, and p190 loss-of-function mutants (85%, 98%, and 66% of mutant hemisegments respectively). Furthermore, our finding that Sema-1a and Pbl collaborate to reduce cell size in cultured cells, and that this synergistic effect is reversed by inhibition of Rho1 activity, support an essential role for Rho1 in repulsive guidance at specific choice points. Previous studies show that pbl null alleles including pbl2, pbl3, and pbl5, which we find here show strong genetic interactions with Sema-1a, strongly suppress the Rho1-induced Rucaparib rough eye phenotypes in Drosophila, and that Pbl interacts with

Rho1, but not with Rac1 or Cdc42, in yeast two-hybrid assays ( Prokopenko et al., 1999). Taken together, these observations strongly suggest that Sema-1a regulates the GEF activity of Pbl directed toward the Rho1 GTPase. Phenotypic analysis of and pbl and p190 mutants demonstrates an additional role for Rho1 in regulating axon target recognition. In summary, our results suggest that Sema-1a-mediated reverse signaling pathways converge on Rho1 to control motor axon target recognition and guidance. It will be important to determine whether other transmembrane guidance cues best known as ligands, including vertebrate transmembrane semaphorins, also mediate receptor functions through direct modulation of Rho GTPase activities. We used the w1118 strain as a wild-type control. The following flies were obtained from the Bloomington Stock Center: pbl2, pbl3, pbl5, pblKG07669, pbl09645, UAS-pbl RNAi[t28343], UAS-p190 RNAi[8.2], UAS-p190 RNAi[5.2], UAS-mycp190, e16E-GAL4, and Rho172F. UAS-pbl RNAi[v35349] and UAS-pbl RNAi[v35350] were obtained from the Vienna Drosophila RNAi Center. All other strains were described previously: Sema-1aPI and UAS-Sema-1a ( Yu et al., 1998); PlexADf(4)C3 ( Winberg et al., 1998); PlexBKG00878 ( Ayoob et al., 2006); Elav(2)-GAL4, Elav(3E)-GAL4, and 24B-GAL4 ( Luo et al., 1994); Sca-GAL4 ( Klaes et al., 1994); GMR37C03GAL4 and GMR37D12GAL4 ( Pfeiffer et al., 2008). p1902 was obtained from J. Cho.

As a federal state, responsibility for health in Canada is shared by the national and

provincial-territorial governments. Numerous federal–provincial–territorial consultative processes enable coordination and collaboration among different levels of government while preserving local independence. The Public Health Agency of Canada (PHAC), created in 2004 Selleckchem Epigenetics Compound Library and led by Canada’s Chief Public Health Officer, is the main federal agency responsible for public health. PHAC reports to Parliament through the Minister of Health, and collaborates closely with all levels of government (provincial, territorial, municipal), as well as non-governmental organizations, Stem Cell Compound Library cell assay other countries, and international organizations like the WHO. NACI is an expert advisory committee of the PHAC and was established and mandated by the agency itself through its legislative ability to seek views about public health issues [2]. NACI is charged with providing medical and scientific advice on immunization for Canadians, focusing on scientific evidence to evaluate vaccine safety and efficacy. The planning and delivery of immunization programs in Canada falls under the jurisdiction of each province/territory. A federal/provincial/territorial committee, the Canadian

Immunization Committee, considers these programmatic issues, including economic considerations, in light of NACI statements, and produces recommendations to the Pan-Canadian Public Health Network. The overarching framework for the administration of these committees is the National Immunization 3-mercaptopyruvate sulfurtransferase Strategy (available at: http://www.phac-aspc.gc.ca/publicat/nis-sni-03/index-eng.php). Recommendations for the prevention of vaccine-preventable infections and other health hazards for

Canadians who travel outside Canada’s borders are made by a separate scientific committee, the Committee to Advise on Tropical Medicine and Travel. A broad range of stakeholders depend on NACI’s recommendations, including decision-makers in provinces and territories, public health practitioners, health care providers, individuals; as well as vaccine manufacturers, non-governmental organizations (e.g. professional societies and immunization advocacy groups), and federal departments (e.g. First Nations Inuit Health Branch, Citizenship & Immigration Canada, Department of National Defence). In fact, in a recent report from the national Advisor on Healthy Children and Youth, it was recommended that “the federal government continue to support the work of the National Advisory (Committee) on Immunization in getting valuable information to health care providers and parents” [3].

Here, we detail how synaptic signaling exclusively from glutamate spillover engages neural circuits not previously predicted by anatomical mapping. First, we show that CF-mediated glutamate spillover affects the excitability of closely and distantly located MLIs. MLIs excited by spillover inhibit MLIs outside the spillover

limit, resulting in segregated activity based on proximity to the active CF. Single CF stimulation recruits AMPARs and NMDARs on MLIs within the spillover limit to trigger spiking and thus mediate a long-lasting component of spillover-mediated FFI to MLIs outside the spillover limit. Concerted activity of MLIs within and outside the spillover limit converges on neighboring PCs to generate a biphasic change in http://www.selleckchem.com/products/i-bet151-gsk1210151a.html inhibitory synaptic tone that initially decreases and subsequently increases evoked spike probability. These results demonstrate a pathway for information transfer in the cerebellar Cabozantinib mouse cortex that extends the influence of CFs beyond the conventional one-to-one relationship with postsynaptic PCs. Synaptic transmission

can be divided into fast and slow forms based on the kinetics of the postsynaptic response (Isaacson et al., 1993). Slow synaptic transmission is mediated by transmitters that act at diffusely distributed receptors located outside synapses (Fuxe and Agnati, 1991; but see Beckstead et al., 2004), whereas fast transmission is typically confined to synapses. In this view, spillover can Linifanib (ABT-869) be considered as an intermediate form of transmission, in which traditional fast neurotransmitters act at receptors distant from release sites. Spillover is not only associated with indirect modulation of fast transmission through G protein-coupled receptors (i.e., Isaacson et al., 1993; Scanziani et al., 1997; Mitchell and Silver, 2000), but also with direct signaling through activation of ionotropic receptors (i.e., Isaacson, 1999; DiGregorio

et al., 2002; Rancz et al., 2007; Scimemi et al., 2009). Direct spillover-mediated transmission improves efficacy and reliability of point-to-point transmission at some specialized synapses (DiGregorio et al., 2002; Sargent et al., 2005; Rancz et al., 2007) and has recently been implicated in the ability of synaptic inputs to generate nonlinear responses mediated by NMDARs (NMDA spikes; Chalifoux and Carter, 2011). In a few cases, synaptic signaling between neurons occurs solely via spillover in the absence of morphologically identified synaptic contacts (Isaacson, 1999; Szapiro and Barbour, 2007; Szmajda and Devries, 2011). Although there is debate about the prevalence of spillover at typical small glutamatergic synapses, our demonstration that spillover-mediated signaling recruits local microcircuits supports its functional significance.

6 and 7 Heart rate variability (HRV) has been examined as a simple non-invasive indicator of cardiac control and a useful tool in assessing autonomic nervous system activity across a range of populations.8, 9, 10 and 11 Further, fluctuations in cardiac autonomic regulation and HRV have been shown to decrease with periods of intense training and competition9 and increase during taper in elite athletes.12, 13 and 14 Garet and colleagues13 reported

a negative correlation between cardiac parasympathetic indices of HRV and swimming performance during intensive training, coupled with an increase in HRV and performance during learn more taper, in seven regional level adolescent swimmers. Subsequently, HRV has been suggested as a simple, non-invasive method of gauging cardiac autonomic nervous system fluctuations. Although HRV has been examined within specific training phases, there has been minimal longitudinal

assessment of daily variations in HRV throughout a periodised training program.3 Recently, Plews and colleagues3 observed daily HRV responses over a 10-week period in two elite triathletes. While recent studies have highlighted the prospective use of HRV for able-bodied athletes, minimal research has focussed on elite athletes with a disability competing in the Paralympics. It has been shown that Olympic and Paralympic swimmers follow similar periodised training programs.15 However, Lapatinib ic50 despite the similar training characteristics, it is unknown whether Paralympic swimmers exhibit a similar cardiac autonomic profile comparable to athletes competing at the Olympic level. To our knowledge, no studies have examined the impact that neuromuscular disabilities, limb deficiency, or the loss of a limb(s) has on HRV. To further understand training

adaptations for elite athletes, the aim of this case study was to examine cardiac autonomic variations in Paralympic swimmers as they prepared for the London 2012 Paralympic games. These case studies were designed to explore the cardiac autonomic profiles of three elite (gold medallist) swimmers with a disability. Due to the unique nature of the study population a case study approach was employed to best analyse and compare each athlete’s individual HRV responses over the 17-week monitoring period. Three Paralympic swimmers selected for the London 2012 Paralympic games were recruited for this study. Each swimmer had competed at this level Phosphoprotein phosphatase previously and leading into the event were ranked in the top three in the world in their respective sprint distance events (<200 m). Each athlete was monitored daily for their resting HRV over 17 weeks immediately prior to the 2012 Paralympics games. The periodised training program prescribed by the head swimming coach was individualised for each athlete and incorporated periods of speed (decreased km’s and higher intensity), aerobic (higher km’s and a decreased intensity) and quality (a mix of speed and aerobic, focussing on race specific pace and drills) training phases.

34 and 2.05 log CFU/ml, Ivacaftor respectively) obtained through single treatments of ozone or 50 °C. This indicates that the combination of ozone and heat treatments at 50 °C can produce a synergistic effect in pathogen inactivation of apple juice. Achen and Yousef (2001) confirmed that there was

no significant difference among various temperatures (4, 22, 45 °C) with bubbling ozone treatment in the inactivation of E. coli from apple surfaces. They explained that conflict between increasing solubility and decreasing stability and reaction rate reduced the efficacy of ozone at low temperatures. The current study also shows that there was no significant difference between surviving populations after treatments at 25 °C and 45 °C except for S. Typhimurium treated for 60 s as shown in (a) of Fig. 2, Fig. 3 and Fig. 4. In Fig. 2, Fig. 3 and Fig. 4(b), it is evident that microbial reduction following a 50 °C heat treatment alone was much greater than that of ozone treatment at 25 °C, but the combination treatment at 50 °C was more effective than the 50 °C heat treatment alone, selleck kinase inhibitor and this trend was observed for all pathogens. In Fig. 2, Fig. 3 and Fig. 4(c),

pathogens were greatly inactivated by the 55 °C heat treatment alone. Therefore, there were no significant differences in pathogen reduction between the 55 °C heat treatment alone and the combination treatment for 60 s except in E. coli O157:H7. In other words, both S. Typhimurium and L. monocytogenes were reduced to below the detection limit (1.0 log CFU/ml) after both heat only and combination treatments for 60 s. Some researchers reported that ozonation of fruit juices resulted in color change. When apple juice was treated with ozone (1–4.8% w/w) for 10 min, the color of the

juice was changed significantly (Torres et al., 2011). Patil et al. (2010a) reported that the color of apple juice samples lightened after ozone treatment (0.048 mg O3 at a flow rate of 0.12 l/min) for 0 to 10 min. While L- and b-values showed significant increases, the a-value of apple juice samples decreased as treatment time and concentration of ozone increased. In the case of ozone treated freshly squeezed orange juice ( Tiwari et al., 2008) and blackberry juice ( Tiwari Resminostat et al., 2009a), an increase in L-value and decreases in a- and b-values resulted. However, in our study, no significant changes of L-, a-, and b-values were found in apple juice treated with ozone and/or heat. The differences in color changes of fruit juices among the various studies may be caused by different systems consisting of various control parameters such as concentration, gas flow of ozone, and treatment time. Especially, treatment time in the current study was shorter than that of the cited studies. According to the Code of Federal Regulations, the maximum residual ozone level is 0.4 mg/l when water is bottled (FDA, 2012).

Fifty-five male and female subjects volunteered to participate in the DLW assessments to estimate their TEE. Six subjects were excluded from this report because of anomalies in their 2H and 18O decay curves. An additional three subjects were excluded because of missing HR logs. The final sample of 46 subjects included 16 middle-aged men (mean age 51.4 years), 14 middle-aged women (mean Selleckchem MG 132 age 49.9 years) and 16 young women (mean age 19.1 years). This study was conducted in accordance with the Helsinki Declaration, and was approved by the ethics committee of the Central Finland Health Care District. Written informed consent was obtained from all subjects

prior to the assessments. All measurements were performed after an overnight fast. The participants were weighed while wearing light clothes and without shoes. The weight was determined within 0.1 kg for each subject using an electronic scale calibrated before each measurement session. The height was determined using a fixed wall-scale measuring device to the nearest 0.1 cm. The body mass index (BMI, kg/m2) was calculated as weight/height2.

The TEE (kcal/day) was measured by the DLW method as previously described.23 Briefly, on day 1, the first urine sample was collected from each participant in the morning before the intake of a body weight-dependent (1 g per kg of body weight) dose of DLW (pre-mixed aliquots of 100 g of 10% 18O enriched water with 8 g of 99.9% 2H enriched Selleckchem Ribociclib water). Four to six hours after until consuming the DLW, the subjects provided a second urine sample. On day 14, the first sample was obtained in the morning after an overnight fasting, and the second sample was collected 4–6 h afterwards. The urine samples were stored at −80 °C until analysis. The samples were analyzed in triplicate for the 2H and 18O isotope ratio by mass spectrometry (Metabolic Solutions

Inc., Merrimack, NH, USA) at the University of Alabama, Birmingham, USA. The TEE was calculated using the validated described by Schoeller et al.24 and 25 The subjects wore an HR monitor (Body Guard, FirstBeat Oy, Jyväskylä, Finland) for 24 h before the REE measurement. The device was attached directly to the skin at three points, eliminating the need to wear a belt around the body and thus minimizing skin contact. The recorded 24-h HR data were then downloaded to a computer. The TEE was estimated using newly-developed software (MoveSense HRAnalyzer 2011a, RC1) provided by Suunto Company (Vantaa, Finland). The algorithm used in this software for the TEE estimation is based on HR reserve, taking into account age, gender and BMI. The REE (kcal/day) was assessed by respiratory gas exchange analysis (GEA) using a ventilated-hood system (VIASYS Healthcare, Yorba Linda, CA, USA). Calibration of the GEA was carried out before each measurement according to the manufacturer’s instructions.

We made whole-cell recordings from layer 2/3 pyramidal cells in the somatosensory and visual

cortex to monitor somatic voltage changes, and activated an increasing number of synapses with two-photon learn more glutamate uncaging (Gasparini and Magee, 2006, Losonczy and Magee, 2006 and Matsuzaki et al., 2001). We selected seven spines distributed over a region of ∼20–30 μm (Figure 1A), and recorded somatic EPSPs in response to the activation of one to all seven synapses (with a 1 ms interval between stimulation of each synapse). We found that the EPSP peak increased with the number of activated synapses, closely following a sigmoidal function that greatly deviated from the linear summation of each individual synapse (Figure 1B). We then tested different regions between the tip and the branch point of single branches, and analyzed how this function varied with location. Distal synapses had a much steeper function than proximal synapses (fraction of maximum per input

for tip: 0.48 ± 0.09, middle: 0.24 ± 0.06, LGK-974 in vivo base: 0.11 ± 0.01; n = 9, p = 0.004, ANOVA), which was also shifted to the left (number of inputs at half of the maximum for tip: 2.1 ± 0.4; middle: 3.4 ± 0.6; base: 6.1 ± 0.6, p = 0.006, ANOVA; Figures 1C and Figure S1A, available online). The gain of the input-output function increased more than 3-fold from the branch point to the dendritic tip, and was shifted by approximately five inputs (Figure 1D). Furthermore, the EPSP supralinearity increased from 128% ± 6% to 209% ± 16% between the base and the tip of the dendrite (p = 0.015, ANOVA; Figures 1E and S1B). These results were also observed with small unitary gluEPSPs (∼0.25 mV, see Figures S2A–S2D) and show that there is a gradient of nonlinear synaptic integration along individual dendritic branches, in which distal inputs are amplified more Megestrol Acetate strongly than proximal ones. To understand the biophysical

mechanism underlying supralinear integration, we used pharmacology to probe the role of specific dendritic active conductances (Johnston and Narayanan, 2008, Magee, 2000 and Spruston, 2008). Blocking L-type voltage-gated calcium channels (VGCCs) shifted the input-output curve to the right (Figure 2A; number of inputs at half maximum = 215% ± 39% of control; p = 0.032; supralinearity at three inputs = 118% ± 10%; p = 0.0078; n = 6) without significantly affecting the gain (fraction of maximum per input: 115% ± 13% of control; p = 0.33; Figures 2A and 2C). A similar effect was produced by blocking voltage-gated sodium channels (number of inputs at half maximum = 159% ± 15% of control; p = 0.030; supralinearity at three inputs = 127% ± 15%; p = 0.024; n = 4; Figure 2C), as well as by simultaneous VGCC and voltage-gated sodium channel (VGSC) block (Figure S3).

We conclude that HR analysis using

Suunto’s software (MoveSense HRAnalyzer 2011a, RC1) needs further development for use in estimations of the daily TEE in free-living individuals. The authors have no conflicts to disclose. This work was funded by the Academy of Finland, the Finnish Ministry of Education, Suunto Oy, the Shanghai overseas distinguish professor award program 2011, the Shanghai Key Lab of click here Human Performance (No. 11DZ2261100), and 2012 National Science and Technology Infrastructure Program (Grant No. 2012BAK21B00). “
“Obesity is a risk factor for several chronic diseases, including type 2 diabetes and cardiovascular disease.1 and 2 Lifestyle interventions, such as dietary weight loss and increasing physical activity (PA), are advocated for the treatment selleck chemical of obesity and prevention of future chronic diseases.3 and 4 The mechanisms through which dietary weight loss and exercise training alter adipose tissue lipid metabolism and lower adiposity need to be investigated. Lipolysis is the process by which triglycerides stored in adipocytes are broken down and free fatty acids and glycerol are released. One of the important enzymes to regulate adipocyte lipolysis is hormone sensitive lipase (HSL).5

HSL and adipose triglyceride lipase (ATGL) work hierarchically to regulate complete lipolysis.6 Currently, HSL and ATGL have been considered to be the major regulators of lipolysis under catecholamine-stimulated and basal lipolysis, respectively.7 In the absence of adipose tissue HSL or ATGL, energy metabolism was altered Dichloromethane dehalogenase and exercise performance was impaired in mice.8 and 9 However, fasting, but not exercise, up-regulated ATGL expression in human adipose tissue,10 suggesting that exercise may be more effective in regulating HSL, but not ATGL in adipose tissue. The role of exercise training intensity on adipose tissue metabolism has been reported by several studies. In exercise-only studies, vigorous-intensity, but not moderate-intensity exercise,

tended to increase adipose lipolysis.11 and 12 However, it is unclear if this is due to an exercise training effect on adipose tissue HSL expression. In an animal study, exercise training increased adipose tissue HSL amount and activity.13 It is well known that an acute exercise session increases catecholamine levels and the release of catecholamines is directly related to exercise intensity.14 It is highly possible that acute and chronic exercise intensity also influences HSL, which is the key enzyme to regulate catecholamine-stimulate lipolysis. However, the effect of exercise training intensity on adipose tissue HSL has not been studied, especially in obese individuals during dietary weight loss. Identification of effective lifestyle interventions is needed for the treatment of obesity. Changes in adipose tissue metabolism by lifestyle interventions may be reflected in current or future changes in adiposity.

The first training session took place on day 1 (i.e., Monday), approximately 1.5 hr after the pretraining imaging session. During fMRI we used the same temporal discrimination task as during behavioral testing. Unlike training and psychophysics, in fMRI we used three different standard durations: i.e., the 200 ms “trained”

duration, plus two “untrained” durations (100 ms and 400 ms). Moreover, the duration of the comparison interval (T + ΔT) was not changed adaptively; instead, two fixed durations were used: T + ΔT1 and T + ΔT2 (see Results for more details). The ΔT1 obtained with the adaptive procedure outside the scanner was measured for Birinapant clinical trial the 200 ms standard duration only. This was done because of two reasons. First, by assessing the ΔT1 threshold for the trained duration only PD0332991 in vivo (i.e., 200 ms), we minimized the presentation of the nontrained stimuli (i.e., 100 and 400 ms) thus reducing any possible learning effects on these control durations. Second, previous literature on the scalar property of temporal judgment (Church et al., 1994; Gibbon, 1977) indicates that one should be able, for any duration (T), to

estimate the ΔT leading to equivalent performance discrimination using the Weber fraction (i.e., ΔT/T). Accordingly, we used the Weber fraction to generate ΔT1s for the 100 and 400 ms control durations. The visual and the auditory tasks were tested in separate imaging runs (two runs for each sensory modality). The order of the task

(visual versus auditory) was counterbalanced across participants. The three standard durations (100, 200, or 400 ms) were presented in different blocks, while ΔT1 and ΔT2 were presented pseudorandomly within each block. Each imaging run included 12 blocks (four blocks per standard duration) with eight trials per block. The total trial Astemizole duration was on average 6.48 s ranging from 5.65 to 7.41 s, the intertrial interval was a variable value randomly chosen from a uniform distribution ranging from 2.5 to 3.5 s. A 3T system (Siemens Magnetom Allegra, Siemens Medical Solutions, Erlangen, Germany) was used to acquire T2∗-weighted echoplanar image (EPI) volumes sensitized to blood oxygenation level-dependent (BOLD) contrast (TE = 30 ms). Each EPI volume comprised thirty-two 2.5 mm axial slices with an in-plane resolution of 3 × 3 mm positioned to cover the entire cortex (50% gap between slices). Each run consisted of 324 volumes. The first four volumes of each run were discarded to allow for T1 equilibration effects. Volumes were acquired continuously with a TR of 2.08 s per volume. A T1-weighted anatomical image was acquired for each participant using 3D modified driven equilibrium Fourier transform (MDEFT) sequence (TR = 1338 ms, TE = 2.4 ms, matrix = 256 × 224 × 176, in-plane FOV = 250 × 250 mm2, slice thickness = 1 mm). Diffusion weighted twice-refocused spin-echo EPI (TR = 170 ms, TE = 85 ms, maximum b factor = 1000 smm−2, isotropic resolution 2.