Towards real-time 4D radiation dosimetry on an MRI-Linac.

4D radiation dosimetry using a highly radiation-sensitive polymer gel dosimeter with real-time quantitative magnetic resonance imaging (MRI) readout is presented as a technique to acquire the accumulated radiation dose distribution during image-guided radiotherapy on an MRI-Linac. Optimized T 2-weighted Turbo-Spin-Echo (TSE) scans are converted into quantitative ΔR 2 maps and subsequently to radiation dose maps. The concept of temporal uncertainty is introduced as a metric of effective temporal resolution. A mathematical framework is presented to optimize the echo time of the TSE sequence in terms of dose resolution, and the trade-off between temporal resolution and dose resolution is discussed. The current temporal uncertainty achieved with the MAGAT gel dosimeter on a 1 T MRI-Linac is 3.8 s which is an order of magnitude better than what has been achieved until now. The potential of real-time 4D radiation dosimetry in a theragnostic MRI-Linac is demonstrated for two scenarios: an irradiation with three coplanar beams on a head phantom and a dynamic arc treatment on a cylindrical gel phantom using a rotating couch. The dose maps acquired on the MRI-Linac are compared with a treatment plan and with dose maps acquired on a clinical 3 T MRI scanner. 3D gamma map evaluations for the different modalities are provided. While the presented method demonstrates the potential of gel dosimetry for tracking the dose delivery during radiotherapy in 4D, a shortcoming of the MAGAT gel dosimeter is a retarded dose response. The effect of non-ideal radiofrequency pulses resulting from limitations in the specific absorption rate or B1-field inhomogeneity on the TSE acquired ΔR 2 values is analysed experimentally and by use of computational modelling with a Bloch simulator.

Neurovascular Lesions in Pediatric Epilepsy.

INTRODUCTION: Neurovascular lesions are rare and understudied in the pediatric population. Their initial presentation can range from seizures to focal neurologic deficits, as well as headaches. The goal of this study was to examine the clinical presentation and natural history of neurovascular lesions in children with epilepsy. METHODS: We reviewed all pediatric epilepsy patients with neurovascular lesions diagnosed between 2006 and 2018 at the University of Alberta and the Stollery Children's Hospital, Edmonton, Canada. Initial clinical presentation and brain imaging, as well as long-term epilepsy and postsurgical outcome, were assessed. RESULTS: Of the 14 patients, 10 patients had an initial presentation of focal seizures with impaired awareness, whereas 2 patients presented with headache, 1 presented with visual field defects as well as chronic headaches, and 1 with decreased level of consciousness. Seven patients had cavernous angiomas, 6 had arteriovenous malformation, and 1 patient had an arteriovenous fistula. Notably, all patients with cavernous angiomas and 4 of 6 patients with arteriovenous malformations presented with seizures. Among 9 of the 14 who underwent neurovascular corrective surgery, all 9 patients required long-term antiepileptic treatment of at least 1 antiepileptic drug for seizure control after the operation. CONCLUSION: In this novel case series, we describe focal seizures as the initial presentation of pediatric neurovascular lesions. This clinical presentation appears to be independent of the type of neurovascular lesion. Furthermore, unlike our pediatric surgical patients with epilepsy due to other causes, seizure freedom following neurovascular surgery is limited, and patients require long-term antiepileptic treatment.

Physico-chemical properties and optimization of the deformable FlexyDos3D radiation dosimeter.

Deformable 3D radiation dosimetry is receiving growing interest for the validation of image-guided radiotherapy treatments (IGRT) of moving and deformable targets. Previously, a proof-of-concept of a flexible anthropomorphic 3D dosimeter called 'FlexyDos3D' has been demonstrated. One of the concerns with respect to the FlexyDos3D dosimeter is its dose-response instability. The effect of different formulations of the dosimeter on its stability were investigated. A stable formulation for the dosimeter was found by optimising the ratios of curing agent and base of the silicone matrix between 3% and 4.5% [w/w] curing agent. The effects of elevated curing temperatures and times upon the dosimetric properties were also investigated and the dose-response was found to be independent of curing times for curing times over an hour at 120 °C. 1H NMR spectra of the dosimeter chemical constituents and the effect of radiation dose were determined. The evaporation and diffusion rates of chloroform in the dosimeter were determined and are the likely cause of the dosimeters depth-dose profile uncertainties. A composition for a stable silicone dosimeter which can be cured quickly at elevated temperatures was found, demonstrating the potential for 3D printing of patient-specific dosimeters. However, it is suggested that another radical initiator be used in future formulations of the dosimeter.

Gardner Transition in Physical Dimensions.

The Gardner transition is the transition that at mean-field level separates a stable glass phase from a marginally stable phase. This transition has similarities with the de Almeida-Thouless transition of spin glasses. We have studied a well-understood problem, that of disks moving in a narrow channel, which shows many features usually associated with the Gardner transition. We show that some of these features are artifacts that arise when a disk escapes its local cage during the quench to higher densities. There is evidence that the Gardner transition becomes an avoided transition, in that the correlation length becomes quite large, of order 15 particle diameters, even in our quasi-one-dimensional system.

Transitions of females from adolescent secure to adult secure services: a qualitative pilot project.

BACKGROUND: For young people, the transition from adolescent to adult services is particularly problematic. This may be particularly difficult for female service users. AIMS: The aim of this study is to gain a fuller account of the experience of young people during transition from adolescent services to adult services and to add to the knowledge around the transitional process. METHOD: This study used a qualitative analysis of female adolescents' experience of transition to adult secure services. A four-layer coding structure grouped responses by time frame. RESULTS: The results were consistent with previous research indicating that adolescents are sensitive to the behaviour of others throughout the transition process. Particularly, strong themes were the negative impact of aggression from other patients, the importance of relationships with staff and other patients, and the need for informed involvement in all aspects of the transition process. CONCLUSIONS: An increase in positive statements regarding the post-transition experience suggests that moves have been positive although this could be explained by admission to settings of lower security. The discussion highlights the importance of moving beyond procedural issues of transition to a focus on the social and culture gaps that appear to divide CAMHS and AMHS.

Lifting the lid on GPCRs: the role of extracellular loops.

GPCRs exhibit a common architecture of seven transmembrane helices (TMs) linked by intracellular loops and extracellular loops (ECLs). Given their peripheral location to the site of G-protein interaction, it might be assumed that ECL segments merely link the important TMs within the helical bundle of the receptor. However, compelling evidence has emerged in recent years revealing a critical role for ECLs in many fundamental aspects of GPCR function, which supported by recent GPCR crystal structures has provided mechanistic insights. This review will present current understanding of the key roles of ECLs in ligand binding, activation and regulation of both family A and family B GPCRs.

Enhanced Liver Fibrosis (ELF) test accurately identifies liver fibrosis in patients with chronic hepatitis C.

Assessment of liver fibrosis is important in determining prognosis and evaluating interventions. Due to limitations of accuracy and patient hazard of liver biopsy, non-invasive methods have been sought to provide information on liver fibrosis, including the European liver fibrosis (ELF) test, shown to have good diagnostic accuracy for the detection of moderate and severe fibrosis. Access to independent cohorts of patients has provided an opportunity to explore if this test could be simplified. This paper reports the simplification of the ELF test and its ability to identity severity of liver fibrosis in external validation studies in patients with chronic hepatitis C (CHC). Paired biopsy and serum samples from 347 naïve patients with CHC in three independent cohorts were analysed. Diagnostic performance characteristics were derived (AUROC, sensitivity and specificity, predictive values), and clinical utility modelling performed to determine the proportion of biopsies that could have been avoided if ELF test was used in this patient group. It was possible to simplify the original ELF test without loss of performance and the new algorithm is reported. The simplified ELF test was able to predict severe fibrosis [pooled AUROC of 0.85 (95% CI 0.81-0.89)] and using clinical utility modelling to predict severe fibrosis (Ishak stages 4-6; METAVIR stages 3 and 4) 81% of biopsies could have been avoided (65% correctly). Issues of spectrum effect in diagnostic test evaluations are discussed. In chronic hepatitis C a simplified ELF test can detect severe liver fibrosis with good accuracy.

Development and optimization of a doxorubicin loaded poly(lactic acid) contrast agent for ultrasound directed drug delivery.

An echogenic, intravenous drug delivery platform is proposed in which an encapsulated chemotherapeutic can travel to a desired location and drug delivery can be triggered using external, focused ultrasound at the area of interest. Three methods of loading poly(lactic acid) (PLA) shelled ultrasound contrast agents (UCA) with doxorubicin are presented. Effects on encapsulation efficiency, in vitro enhancement, stability, particle size, morphology and release during UCA rupture are compared by loading method and drug concentration. An agent containing doxorubicin within the shell was selected as an ideal candidate for future hepatocellular carcinoma studies. The agent achieved a maximal drug load of 6.2 mg Dox/g PLA with an encapsulation efficiency of 20.5%, showed a smooth surface morphology and tight size distribution (poly dispersity index=0.309) with a peak size of 1865 nm. Acoustically, the agent provided 19 dB of enhancement in vitro at a dosage of 10 microg/ml, with a half life of over 15 min. In vivo, the agent provided ultrasound enhancement of 13.4+/-1.6 dB within the ascending aorta of New Zealand rabbits at a dose of 0.15 ml/kg. While the drug-incorporated agent is thought to be well suited for future drug delivery experiments, this study has shown that agent properties can be tailored for specific applications based on choice of drug loading method.

Delivery of encapsulated Doxorubicin by ultrasound-mediated size reduction of drug-loaded polymer contrast agents.

Low delivery efficiency combined with systemic toxicity of traditional chemotherapy provides a need for improved chemotherapeutic delivery. Within our laboratory, we have developed polymer ultrasound contrast agents (1.2-1.8 mum in diameter) containing doxorubicin (Dox) within the shell (100-150 nm). In vivo this platform is expected to circulate through the vasculature until activated at the tumor site with external focused ultrasound (US). In vitro, the agent is responsive to US and when insonated at peak positive pressure amplitudes of 0.69 MPa and above, shows dramatic size reduction, eventually reaching a mean particle size of 350 nm, presumably due to fragmentation of, or gas release from the agent. The resulting Dox-polymer particles retain the drug and are small enough to pass through the leaky pores (350-400 nm) within the tumor vasculature, providing a sustained intratumoral release of chemotherapeutic as the polymer degrades. In vivo studies using a VX2 liver tumor model have shown that the combination of the agent and US results in nearly 50% less drug delivered to the nontargeted, healthy liver ( p = 0.009) and a 110% increase ( p = 0.004) in Dox delivery to the viable peripheral tissue of the tumor, relative to the uninsonated controls. This study shows how US-mediated destruction of drug-loaded polymer contrast agent can be used to deliver encapsulated drug for potential sustained release. Penetration mechanisms of these resulting particles and their ability to provide a sustained release from the tumor interstia will be explored in the future.

Selective amygdalohippocampectomy: surgical outcome in children versus adults.

INTRODUCTION: The objective of the study was to review our experience with selective amygdalohippocampectomy (SAH) in children and adults with intractable temporal lobe epilepsy. METHODS: A retrospective case series was used in the setting of a tertiary care hospital which provides epilepsy care to both children and adults. All patients underwent a selective amygdalohippocampectomy procedure and had at least one year of follow-up. Adults and children were divided into two groups and the data was compared between children and adults. RESULTS: Twenty three patients, 9 children and 14 adults were studied. Age of surgery varied from 6 to 58 years. Surgical outcome was variable between the two groups. Amongst the children, three patients (33%) were seizure-free (Engel Class I), two patients (22%) had rare seizures (Engel Class II), one patient (11%) had a worthwhile decrease in seizures (Engel class III) and three patients (32%) had refractory seizures that required re-operation with an anterior temporal lobectomy. This differed from the adults, who all had a good outcome. Ten patients (71%) were seizure-free (Engel Class I) and the remainder (29%) had rare seizures (Engel Class II). CONCLUSION: Selective amygdalohippocampectomy can lead to excellent seizure surgical outcome in adults with refractory temporal lobe epilepsy. However, preliminary results show less favorable results in children. The difference is probably related to the different pathology between the two groups. Anterior temporal lobe resection may prove to be a more successful operation than SAH in children with intractable temporal lobe epilepsy.

Ultrasound triggered cell death in vitro with doxorubicin loaded poly lactic-acid contrast agents.

Traditional chemotherapy generally results in systemic toxicity, which also limits drug levels at the area of need. Two ultrasound contrast agents (UCA), with diameters between 1-2 microm in diameter and shell thicknesses of 100-200 nm, composed of poly lactic-acid (PLA), one loaded by surface adsorption and the other loaded by drug incorporation in the shell, were compared in vitro for potential use in cancer therapy. These poly lactic-acid (PLA) UCA platforms contain a gas core that in an ultrasound (US) field can cause the UCA to oscillate or rupture. Following a systemic injection of drug loaded UCA with external application of US focused at the area of interest, this platform could potentially increase drug toxicity at the area of need, while protecting healthy tissue through microencapsulation of the drug. In vitro toxicity in MDA-MB-231 breast cancer cells of the surface-adsorbed and shell-incorporated doxorubicin (Dox) loaded UCA were examined at 5 MHz insonation using a pulse repetition frequency of 100 Hz at varying pressure amplitudes. Both platforms resulted in equivalent cell death compared to free Dox and US when insonated at peak positive pressure amplitudes of 1.26 MPa and above. While no significant changes in cell death were seen for surface adsorbed Dox-UCA with or without insonation, cell death using the platform with Dox incorporated within the shell increased from 16.12% to 25.78% (p=0.0272), approaching double the potency of the platform when insonated at peak positive pressure amplitudes of 1.26 MPa and above. This mechanism is believed to be the result of UCA rupture at higher insonation pressure amplitudes, resulting in more exposed drug and shell surface area as well as increased cellular uptake of Dox containing polymer shell fragments. This study has shown that a polymer UCA with drug housed within the shell may be used for US-triggered cell death. US activation can be used to make a carrier significantly more potent once in the area of need.

Family resemblances? Ligand binding and activation of family A and B G-protein-coupled receptors.

In April 2007, the Biochemical Society held a meeting to compare and contrast ligand binding and activation of Family A and B GPCRs (G-protein-coupled receptors). Being the largest class, Family A GPCRs usually receive the most attention, although a previous Biochemical Society meeting has focused on Family B GPCRs. The aim of the present meeting was to bring researchers of both families together in order to identify commonalities between the two. The present article introduces the proceedings of the meeting, briefly commenting on the focus of each of the following articles.

Extracellular loops and ligand binding to a subfamily of Family A G-protein-coupled receptors.

GPCRs (G-protein-coupled receptors) are a large family of structurally related proteins which mediate their effects by coupling to G-proteins. The V(1a)R (V(1a) vasopressin receptor) is a member of a family of related GPCRs that are activated by vasopressin {AVP ([Arg(8)]vasopressin)}, OT (oxytocin) and related peptides. These receptors are members of a subfamily of Family A GPCRs called the neurohypophysial peptide hormone receptor family. GPCRs exhibit a conserved tertiary structure comprising a bundle of seven TM (transmembrane) helices linked by alternating ECLs (extracellular loops) and ICLs (intracellular loops). The cluster of TM helices is functionally important for ligand binding, and, furthermore, activation of GPCRs involves movement of these TM helices. Consequently, it might be assumed that the extracellular face of GPCRs is composed of peptide linkers that merely connect important TM helices. However, using a systematic mutagenesis approach and focusing on the N-terminus and the second ECL of the V(1a)R, we have established that these extracellular domains fulfil a range of important roles with respect to GPCR signalling, including agonist binding, ligand selectivity and receptor activation.

Ligand binding and activation of the CGRP receptor.

The receptor for CGRP (calcitonin gene-related peptide) is a heterodimer between a GPCR (G-protein-coupled receptor), CLR (calcitonin receptor-like receptor) and an accessory protein, RAMP1 (receptor activity-modifying protein 1). Models have been produced of RAMP1 and CLR. It is likely that the C-terminus of CGRP interacts with the extracellular N-termini of CLR and RAMP1; the extreme N-terminus of CLR is particularly important and may interact directly with CGRP and also with RAMP1. The N-terminus of CGRP interacts with the TM (transmembrane) portion of the receptor; the second ECL (extracellular loop) is especially important. Receptor activation is likely to involve the relative movements of TMs 3 and 6 to create a G-protein-binding pocket, as in Family A GPCRs. Pro(321) in TM6 appears to act as a pivot. At the base of TMs 2 and 3, Arg(151), His(155) and Glu(211) may form a loose equivalent of the Family A DRY (Asp-Arg-Tyr) motif. Although the details of this proposed activation mechanism clearly do not apply to all Family B GPCRs, the broad outlines may be conserved.

Atopy and asthma in rural Poland: a paradigm for the emergence of childhood respiratory allergies in Europe.

BACKGROUND: We hypothesized that, in south-west Poland, a 'rural' protective effect on atopy and respiratory allergies would be most pronounced among children but that at all ages would be stronger among those with a rural background. METHODS: A cross-sectional survey of the inhabitants (age >5 years, n = 1657) of Sobotka, a town of 4000 people in south-west Poland: and seven neighbouring villages. We measured and analysed responses to skin prick tests (atopy) and to a standard questionnaire (asthma and hayfever). RESULTS: Atopy was very uncommon (7%) among villagers at all ages but not among townspeople (20%, P < 0.001); the differences were most marked among those aged under 40 years. Asthma and hayfever were similarly distributed, both being very rare among villagers. The differences appear to be explained by the cohort effect of a communal move away from rural life. This interpretation is supported by an ecological correlation (rho = -0.59) between rural populations and childhood wheeze in 22 European countries. CONCLUSION: The very striking differences in the prevalence of allergy between these two neighbouring communities of central Europe reflect the pan-continental population movements that may have been responsible for the emergence of childhood allergies in Europe.

Prognostic significance of serial magnetic resonance spectroscopies over the course of radiation therapy for patients with malignant glioma.

BACKGROUND: The standard treatment of high grade gliomas (HGG) involves maximal neuro-surgical debulking, followed by post-operative radiotherapy, with or without concurrent chemotherapy, depending on histologic grade. Despite this aggressive strategy, there are few long-term survivors. Proton magnetic resonance spectroscopy (MRS) is a non-invasive imaging method that can monitor metabolic changes in brain tumours. To date there is little data concerning the prognostic significance of the evolving spectral alterations during a course of radiotherapy. MATERIALS: We report herein a prospective study of patients with HGGs undergoing post-operative radiotherapy. Fourteen consecutively eligible patients with a confirmed histologic diagnosis of malignant glioma and completion of all required MRS imaging were included in this study. All patients had MRS imaging prior to radiotherapy, at week 4 of radiotherapy, and 2 months post-treatment. T1 and T2 weighted images as well as post-gadolinium multi-voxel proton MRS images were obtained. Normalized (tumour metabolite/normal brain metabolite) levels of choline, NAA, creatine, lipid and lactate were calculated. Kaplan-Meier (KM) curves of progression-free and overall survival were constructed based on the evolving patterns of metabolite changes over the course of the images. RESULTS: The mean tumour choline/NAA ratio decreased over the course of therapy, with a reduction observed between the baseline and post-radiotherapy studies (1.91 vs. 1.29, P=0.049). A similar decrease was identified with the mean normalized choline ratio, with a highly significant difference observed between the baseline and post-radiation images (1.61 vs. 0.96, P=0.001). Patients who exhibited more than 40% decrease in normalized choline between the week 4 and post-radiotherapy studies were associated with unfavourable survival (logrank test, P=0.003) and disease progression (logrank test, P=0.012). The Lactate/NAA ratio at the 4th week of radiotherapy and the change in normalized choline/creatine between baseline and week 4 of radiotherapy were also predictive of outcome suggesting the possibility of adaptive, response-based radiation treatment. Patients with two or more poor prognostic MRS indices had a significantly shorter progression-free survival compared with those with zero or one poor indices, with 15% and 68% at 1 year, respectively (logrank test, P=0.045). CONCLUSION: The evolving pattern of spectral changes over the course of radiotherapy, in particular those associated with choline-containing compounds, appears to be prognostic of tumour response and outcome. Based on our data, a decision point may exist in the mid course of radical radiotherapy, at which time consideration of the choline levels could indicate the extent of radiotherapeutic response, thus allowing for individualized treatment modification.

The third extracellular loop of G-protein-coupled receptors: more than just a linker between two important transmembrane helices.

GPCRs (G-protein-coupled receptors) are a large family of structurally related proteins, which mediate their effects by coupling with G-proteins. Despite responding to a range of very diverse stimuli, these receptors exhibit a conserved tertiary structure comprising a bundle of seven TM (transmembrane) helices linked by alternating ECLs (extracellular loops) and ICLs (intracellular loops). The hydrophobic environment formed by the cluster of TM helices is functionally important. For example, the 11-cis retinal chromophore of rhodopsin forms a protonated Schiff base linkage to a lysine in TM7, deep within the helical bundle, and small ligands, such as amine neurotransmitters and non-peptide analogues of peptide hormones, also bind within the corresponding region of their cognate receptors. In addition, activation of GPCRs involves relative movement of TM helices to present G-protein interaction sites across the intracellular face of the receptor. Consequently, it might be assumed that the ECLs of the GPCR are inert peptide linkers that merely connect important TM helices. Focusing on ECL3 (third ECL), it is becoming increasingly apparent that this extracellular domain can fulfil a range of important roles with respect to GPCR signalling, including agonist binding, ligand selectivity and receptor activation.

Heterodimers and family-B GPCRs: RAMPs, CGRP and adrenomedullin.

RAMPs (receptor activity-modifying proteins) are single-pass transmembrane proteins that associate with certain family-B GPCRs (G-protein-coupled receptors). Specifically for the CT (calcitonin) receptor-like receptor and the CT receptor, this results in profound changes in ligand binding and receptor pharmacology, allowing the generation of six distinct receptors with preferences for CGRP (CT gene-related peptide), adrenomedullin, amylin and CT. There are three RAMPs: RAMP1-RAMP3. The N-terminus appears to be the main determinant of receptor pharmacology, whereas the transmembrane domain contributes to association of the RAMP with the GPCR. The N-terminus of all members of the RAMP family probably contains two disulphide bonds; a potential third disulphide is found in RAMP1 and RAMP3. The N-terminus appears to be in close proximity to the ligand and plays a key role in its binding, either directly or indirectly. BIBN4096BS, a CGRP antagonist, targets RAMP1 and this gives the compound very high selectivity for the human CGRP(1) receptor.

Pediatric temporal lobectomy for epilepsy.

BACKGROUND: Temporal lobectomy in adults is an accepted form of treatment for patients with intractable complex partial seizures. There have been few long-term studies of children undergoing temporal lobectomy for epilepsy. METHODS: We reviewed the pediatric cases of temporal lobectomy for intractable epilepsy performed by the Comprehensive Epilepsy Program at the University of Alberta Hospitals between 1988 and 2000. All patients had preoperative and postoperative clinical evaluations, seizure charts, drug levels, EEG, CT/MRI, long-term video EEG monitoring and neuropsychological testing. The patients were reassessed at 6 weeks, 6 months and 1 year postoperatively, then yearly. The duration of follow up was 1-10 years (mean 5 years). RESULTS: Forty-two patients were studied (25 males and 17 females). Age at surgery ranged from 18 months to 16 years. The interictal EEG was abnormal in 38 of the 42 patients. Twenty-two patients had focal epileptic discharge and 1 had generalized epileptic discharge. Focal slowing was seen in 9 patients and diffuse slowing in 5 patients. CT scan was abnormal in 17 of 39 patients and normal in 22 of 39. MRI was abnormal in 34 of 42 patients and normal in 8 of 42. Pathology included brain tumors in 14 patients, mesial temporal sclerosis in 8, focal cortical dysplasia in 4, tuberous sclerosis in 4, dual pathology in 4, porencephalic cyst in 1 and normal pathology or gliosis in 6. Thirty-three of 42 patients (78%) were seizure-free following surgery and an additional 5 (12%) had a decrease in seizure frequency. Three patients had complications, but there were no deaths. CONCLUSION: Temporal lobectomy is a safe and effective treatment for children with intractable complex partial seizures. Seventy-eight percent of patients are seizure-free following the surgery and there are few complications. MRI is superior to CT scan for detection of temporal lobe pathology yet failed to detect abnormalities in some patients. The most common pathologies found were brain tumors, mesial temporal sclerosis and developmental lesions. In addition to seizure control, many patients experienced improvement in cognitive and psychosocial function following surgery.