Clinical features and outcomes of Hodgkin's lymphoma in Korea: Consortium for Improving Survival of Lymphoma (CISL).
Ethnic and regional differences in the epidemiology and pathological aspects of Hodgkin's lymphoma (HL) between Western and Asian patients may be associated with differences in clinical features and prognosis. We retrospectively analyzed the clinical and histopathological characteristics, therapeutic outcomes, and prognostic factors of 539 HL patients treated at 16 centers in Korea. We found that the incidence of histological subtypes of HL in Korea was similar to that in Western and other Asian countries. However, the incidence peaked between 16 and 30 years of age, unlike the bimodal age distribution seen in Western countries. In patients with stage I-IIA non-bulky disease, the complete response (CR) rate was similar between combined modality therapy and chemotherapy alone (93% vs. 84%, P = 0.44), and there was no difference in relapse-free survival (RFS) and overall survival (OS). Patients with stage I-II disease plus unfavorable factors and those with advanced-stage disease treated with combination chemotherapy regimens had an overall CR rate of 77%, with no difference between doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) and non-ABVD regimens (77.2% vs. 76.8%, P = 0.95). Among those patients who achieved final CR, there was no significant difference in RFS or OS between those who achieved interim CR and PR. Only the presence of B symptoms was independently predictive of a shorter RFS. Age > 45 years, Eastern Cooperative Oncology Group 2-4, and B symptoms were independent risk factors for death. Although the incidence of HL was lower in Korea than in Western countries, the distribution of morphological subtypes, treatment outcomes, and patient prognosis were similar.


SHIV infection protects against heterologous pathogenic SHIV challenge in macaques: a gold-standard for HIV-1 vaccine development?
A current debate in the HIV-1 vaccine field concerns the ability of an immunodeficiency virus to elicit a protective response. One argument is that HIV-1 superinfections are frequent in healthy individuals, because virus evades conventional immune surveillance, a serious obstacle to vaccine design. The opposing argument is that protection from superinfection is significant, reflecting a robust immune response that might be harnessed by vaccination to prevent disease. In an experiment designed to address the debate, two macaques received an I.V. inoculation with SHIV KU-1-d (a derivative of SHIV KU-1) and were rested for >10 months. Infection elicited diverse neutralizing antibody activities in both animals. Animals were then exposed to SHIV 89.6P (I.V.), a virus carrying a heterologous envelope protein relative to the vaccine strain. Infection was monitored by viral load and CD4+ T-cell measurements. All control animals were infected and most succumbed to disease. In contrast, protection from superinfection was statistically significant in test monkeys; one animal showed no evidence of superinfection at any time point and the second showed evidence of virus at only one time point over a 6-month observation period. Neither animal showed signs of disease. Perhaps this protective state may serve as a 'gold-standard' for HIV-1 vaccine development, as a similar degree of protection against immunodeficiency virus infections in humans would be much desired.


Dietary flavonoids, antioxidant vitamins, and incidence of stroke: the Zutphen study.
Epidemiological studies suggested that consumption of fruit and vegetables may protect against stroke. The hypothesis that dietary antioxidant vitamins and flavonoids account for this observation is investigated in a prospective study. A cohort of 552 men aged 50 to 69 years was examined in 1970 and followed up for 15 years. Mean nutrient and food intake was calculated from cross-check dietary histories taken in 1960, 1965, and 1970. The association between antioxidants, selected foods, and stroke incidence was assessed by Cox proportional hazards regression analysis. Adjustment was made for confounding by age, systolic blood pressure, serum cholesterol, cigarette smoking, energy intake, and consumption of fish and alcohol. Forty-two cases of first fatal or nonfatal stroke were documented. Dietary flavonoids (mainly quercetin) were inversely associated with stroke incidence after adjustment for potential confounders, including antioxidant vitamins. The relative risk (RR) of the highest vs the lowest quartile of flavonoid intake ( > or = 28.6 mg/d vs <18.3 mg/d) was 0.27 (95% confidence interval [CI], 0.11 to 0.70). A lower stroke risk was also observed for the highest quartile of beta-carotene intake (RR, 0.54; 95% CI, 0.22 to 1.33). The intake of vitamin C and vitamin E was not associated with stroke risk. Black tea contributed about 70% to flavonoid intake. The RR for a daily consumption of 4.7 cups or more of tea vs less than 2.6 cups of tea was 0.31 (95% CI, 0.12 to 0.84). The habitual intake of flavonoids and their major source (tea) may protect against stroke.


Spectral signatures of L-DOPA-induced dyskinesia depend on L-DOPA dose and are suppressed by ketamine.
L-DOPA-induced dyskinesias (LID) are debilitating motor symptoms of dopamine-replacement therapy for Parkinson's disease (PD) that emerge after years of L-DOPA treatment. While there is an abundance of research into the cellular and synaptic origins of LID, less is known about how LID impacts systems-level circuits and neural synchrony, how synchrony is affected by the dose and duration of L-DOPA exposure, or how potential novel treatments for LID, such as sub-anesthetic ketamine, alter this activity. Sub-anesthetic ketamine treatments have recently been shown to reduce LID, and ketamine is known to affect neural synchrony. To investigate these questions, we measured movement and local-field potential (LFP) activity from the motor cortex (M1) and the striatum of preclinical rodent models of PD and LID. In the first experiment, we investigated the effect of the LID priming procedures and L-DOPA dose on neural signatures of LID. Two common priming procedures were compared: a high-dose procedure that exposed unilateral 6-hydroxydopamine-lesioned rats to 12 mg/kg L-DOPA for 7 days, and a low-dose procedure that exposed rats to 7 mg/kg L-DOPA for 21 days. Consistent with reports from other groups, 12 mg/kg L-DOPA triggered LID and 80-Hz oscillations; however, these 80-Hz oscillations were not observed after 7 mg/kg administration despite clear evidence of LID, indicating that 80-Hz oscillations are not an exclusive signature of LID. We also found that weeks-long low-dose priming resulted in the emergence of non-oscillatory broadband gamma activity (> 30 Hz) in the striatum and theta-to-high-gamma cross-frequency coupling (CFC) in M1. In a second set of experiments, we investigated how ketamine exposure affects spectral signatures of low-dose L-DOPA priming. During each neural recording session, ketamine was delivered through 5 injections (20 mg/kg, i.p.) administered every 2 h. We found that ketamine exposure suppressed striatal broadband gamma associated with LID but enhanced M1 broadband activity. We also found that M1 theta-to-high-gamma CFC associated with the LID on-state was suppressed by ketamine. These results suggest that ketamine's therapeutic effects are region specific. Our findings also have clinical implications, as we are the first to report novel oscillatory signatures of the common low-dose LID priming procedure that more closely models dopamine replacement therapy in individuals with PD. We also identify neural correlates of the anti-dyskinetic activity of sub-anesthetic ketamine treatment.


The new sequencer on the block: comparison of Life Technology's Proton sequencer to an Illumina HiSeq for whole-exome sequencing.
We assessed the performance of the new Life Technologies Proton sequencer by comparing whole-exome sequence data in a Centre d'Etude du Polymorphisme Humain trio (family 1463) to the Illumina HiSeq instrument. To simulate a typical user's results, we utilized the standard capture, alignment and variant calling methods specific to each platform. We restricted data analysis to include the capture region common to both methods. The Proton produced high quality data at a comparable average depth and read length, and the Ion Reporter variant caller identified 96 % of single nucleotide polymorphisms (SNPs) detected by the HiSeq and GATK pipeline. However, only 40 % of small insertion and deletion variants (indels) were identified by both methods. Usage of the trio structure and segregation of platform-specific alleles supported this result. Further comparison of the trio data with Complete Genomics sequence data and Illumina SNP microarray genotypes documented high concordance and accurate SNP genotyping of both Proton and Illumina platforms. However, our study underscored the problem of accurate detection of indels for both the Proton and HiSeq platforms.


A general method for the computer simulation of biological systems interacting with fluids.
At this Symposium on Biological Fluid Dynamics, it is appropriate to ask whether there is any common theme that unites the diverse problems that arise in the study of living systems interacting with fluids. The answer that immediately comes to mind is this: biological fluid dynamics invariably involves the interaction of elastic flexible tissue with viscous incompressible fluid. (In many cases the tissue is not only elastic, it is also active, i.e. capable of doing work on the fluid). This paper describes the immersed boundary method, which is a general framework for the computer simulation of biofluid dynamic systems. This method has already been applied to blood flow in the heart (including the computer-assisted design of prosthetic cardiac valves), platelet aggregation during blood clotting, aquatic animal locomotion, wave propagation along the basilar membrane of the inner ear, and flow in collapsible tubes. In the immersed boundary method, the elastic (and possibly active) biological tissue is treated as a part of the fluid in which additional forces (derived from the tissue stresses) are applied. Because the tissue is represented in terms of its force field, the method remains straightforward, even when the geometry of the biological tissue is complicated, dynamic and not known in advance.


The etiology of osteoarthritis of the hip: an integrated mechanical concept.
The etiology of osteoarthritis of the hip has long been considered secondary (eg, to congenital or developmental deformities) or primary (presuming some underlying abnormality of articular cartilage). Recent information supports a hypothesis that so-called primary osteoarthritis is also secondary to subtle developmental abnormalities and the mechanism in these cases is femoroacetabular impingement rather than excessive contact stress. The most frequent location for femoroacetabular impingement is the anterosuperior rim area and the most critical motion is internal rotation of the hip in 90 degrees flexion. Two types of femoroacetabular impingement have been identified. Cam-type femoroacetabular impingement, more prevalent in young male patients, is caused by an offset pathomorphology between head and neck and produces an outside-in delamination of the acetabulum. Pincer-type femoroacetabular impingement, more prevalent in middle-aged women, is produced by a more linear impact between a local (retroversion of the acetabulum) or general overcoverage (coxa profunda/protrusio) of the acetabulum. The damage pattern is more restricted to the rim and the process of joint degeneration is slower. Most hips, however, show a mixed femoroacetabular impingement pattern with cam predominance. Surgical attempts to restore normal anatomy to avoid femoroacetabular impingement should be performed in the early stage before major cartilage damage is present. Level V, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Pedagogical tools to explore Cartesian mind-body dualism in the classroom: philosophical arguments and neuroscience illusions.
A fundamental discussion in lower-level undergraduate neuroscience and psychology courses is Descartes's "radical" or "mind-body" dualism. According to Descartes, our thinking mind, the res cogitans, is separate from the body as physical matter or substance, the res extensa. Since the transmission of sensory stimuli from the body to the mind is a physical capacity shared with animals, it can be confused, misled, or uncertain (e.g., bodily senses imply that ice and water are different substances). True certainty thus arises from within the mind and its capacity to doubt physical stimuli. Since this doubting mind is a thinking thing that is distinct from bodily stimuli, truth and certainty are reached through the doubting mind as cogito ergo sum, or the certainty of itself as it thinks: hence Descartes's famous maxim, I think, therefore I am. However, in the last century of Western philosophy, with nervous system investigation, and with recent advances in neuroscience, the potential avenues to explore student's understanding of the epistemology and effects of Cartesian mind-body dualism has expanded. This article further explores this expansion, highlighting pedagogical practices and tools instructors can use to enhance a psychology student's understanding of Cartesian dualistic epistemology, in order to think more critically about its implicit assumptions and effects on learning. It does so in two ways: first, by offering instructors an alternative philosophical perspective to dualistic thinking: a mind-body holism that is antithetical to the assumed binaries of dualistic epistemology. Second, it supplements this philosophical argument with a practical component: simple mind-body illusions that instructors may use to demonstrate contrary epistemologies to students. Combining these short philosophical and neuroscience arguments thereby acts as a pedagogical tool to open new conceptual spaces within which learning may occur.


Gastrinomas demonstrate amplification of the HER-2/neu proto-oncogene.
This study determined whether genomic amplification of HER-2/neu or mutations of the p53 and ras genes were present in gastrinomas. Amplification of HER-2/neu, a proto-oncogene related to the epidermal growth factor receptor, and mutation of the ras proto-oncogene and p53 tumor suppressor gene appear to play a role in the pathogenesis of some human cancers. Little is known about possible molecular alterations in gastrinomas, tumors that may be particularly virulent because of gastrin overproduction, resulting in the severe ulcer diathesis, the Zollinger-Ellison syndrome. The differential polymerase chain reaction (PCR) procedure was used to detect amplification of the HER-2/neu gene in DNA samples from the novel human gastrinoma cell line (PT) and from paraffin-embedded samples of gastrinomas. Sequencing techniques were used to determine whether mutations of the p53 or ras (Ha-ras, N-ras, Ki-ras) genes were present. Amplification (> twofold) occurred in all gastrinoma tumor samples. Compared with normal pancreas or ileum, a 4- to 12-fold amplification of HER-2/neu was found in 3 gastrinomas, 3 to 3.3-fold in four samples and 2.1- to 2.4-fold in the remaining five tumors. A heterozygous point mutation in the p53 gene (codon 273) was found in a single sample; none of the gastrinomas contained a mutation of the ras genes. Amplification of the HER-2/neu gene, but not alterations of either p53 or ras, may be involved in the pathogenesis of gastrinomas. The unique PT cell line will be a useful model to further elucidate the molecular mechanisms that contribute to gastrinoma formation and growth.


When adolescents stop psychological therapy: Rupture-repair in the therapeutic alliance and association with therapy ending.
The therapeutic alliance predicts dropout from psychological therapy, and ruptures in the therapeutic alliance may also predict dropout, yet there is a dearth of research with adolescents. This study investigated whether markers of rupture-repair in the alliance were indicative of different types of treatment ending in adolescents receiving psychological treatment for depression. Data were from the IMPACT study, a trial investigating the effectiveness of therapies for adolescent depression. Participants were randomly allocated to receive a brief psychosocial intervention, cognitive-behavioral therapy, or short-term psychoanalytic psychotherapy. The sample (N = 35) comprised adolescents who had either completed their treatment (n = 14) or dropped out (n = 21) according to their therapist. Dropout cases were further classified as dissatisfied (n = 14) or got-what-they-needed (n = 7) based on posttherapy interviews with the adolescent and therapist. Selected recordings of therapy sessions were rated using the Rupture Resolution Rating System and Working Alliance Inventory (observer version). Therapeutic alliance and rupture-repair during therapy were similar for completers and got-what-they-needed dropouts, whereas dissatisfied dropouts had poorer therapeutic alliance, more ruptures, ruptures that were frequently unresolved, and greater therapist contribution to ruptures. Qualitative analysis of sessions led to the construction of three categories of therapist contribution to ruptures: therapist minimal response, persisting with a therapeutic activity, and focus on risk. Results suggest that ruptures, especially when unresolved, could be warning signs of disengagement and dropout from therapy. Future research should investigate how ruptures may be effectively identified and resolved in treatment with adolescents. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Dynamic properties of ankyrin in T lymphocytes: colocalization with spectrin and protein kinase C beta.
Ankyrin is a well characterized membrane skeletal protein which has been implicated in the anchorage of specific integral membrane proteins to the spectrin-based membrane skeleton in a number of systems. In this study, the organization of ankyrin was examined in lymphocytes in relation to T cell function. Light and electron microscope immunolocalization studies revealed extensive heterogeneity in the subcellular distribution of ankyrin in murine tissue-derived lymphocytes. While ankyrin can be localized at the lymphocyte plasma membrane, it can also be accumulated at some distance from the cell periphery, in small patches or in a single discrete, nonmembrane-bound structure. Double immunofluorescence studies demonstrated that ankyrin colocalizes with spectrin and with the signal transducing molecule protein kinase C beta (PKC beta) in tissue-derived lymphocytes, suggesting a functional association between these molecules in the lymphocyte cytoplasm. In addition, T lymphocyte activation-related signals and phorbol ester treatment, both of which lead to PKC activation, cause a rapid translocation of ankyrin, together with spectrin and PKC beta, to a single Triton X-100-insoluble aggregate in the cytoplasm. This finding suggests a mechanism for the reported appearance of PKC in the particulate fraction of cells after activation: activated lymphocyte PKC beta may interact with insoluble cytoskeletal elements like ankyrin and spectrin. Further evidence for a link between the subcellular organization of these proteins and PKC activity is provided by the observation that inhibitors of PKC activity cause their concomitant redistribution to the cell periphery. The dynamic nature of lymphocyte ankyrin and its ability to accumulate at sites distant from the plasma membrane are properties which may be unique to the lymphocyte form of the molecule. Its colocalization with PKC beta in the lymphocyte cytoplasm, together with its redistribution in response to physiological signals, suggests that structural protein(s) may play a role in signal transduction pathways in this cell type. Our data support the conclusion that ankyrin is not solely involved in anchorage of proteins at the plasma membrane in lymphoid cells.


Effectiveness of HypoAware, a Brief Partly Web-Based Psychoeducational Intervention for Adults With Type 1 and Insulin-Treated Type 2 Diabetes and Problematic Hypoglycemia: A Cluster Randomized Controlled Trial.
To establish the effectiveness of a brief, partly web-based group intervention, HypoAware, in patients with problematic hypoglycemia, in terms of the reduction of severe hypoglycemic episodes, impaired hypoglycemia awareness, and worries. A two-arm, cluster randomized controlled trial was conducted in insulin-treated patients with problematic hypoglycemia in eight Dutch clinics comparing HypoAware with care as usual. Primary outcomes included self-reported severe hypoglycemia, impaired hypoglycemia awareness (Gold score), and worries and distress about hypoglycemia (Hypoglycemia Fear Survey-II/Problem Areas In Diabetes scale hypo-item), assessed at baseline, and at 2, 4, and 6 months of follow-up. We used t tests, nonparametric tests, and intention-to-treat generalized estimation equation (GEE) analyses with linear, logistic, and Poisson or negative binomial models. We enrolled 137 participants. Adjusted GEE analyses over four time points showed a nonsignificant 33% fewer episodes of severe hypoglycemia in the HypoAware group compared with the control group (relative risk [RR] 0.67 [95% CI 0.39-1.16], P = 0.150); a significant reduced odds of impaired awareness (odds ratio 0.38 [95% CI 0.15-0.95], P = 0.038), a trend toward 20% fewer worries about hypoglycemia (RR 0.80 [95% CI 0.64-1.01], P = 0.059), and a significant 30% less hypo-distress (RR 0.70 [95% CI 0.56-0.88], P = 0.002). Over the 6-month study duration, participants experienced a median of 2.5 events of severe hypoglycemia (interquartile range [IQR] 1-10) in the control condition versus 1 event (IQR 0-6.5) in the HypoAware group (P = 0.030). There was no significant change in HbA1c level within and between both groups. HypoAware resulted in fewer severe hypoglycemic episodes, significantly improved hypoglycemia awareness, and less hypo-distress in comparison with usual care, and deserves further dissemination.


Testing Syndromes of Psychopathology in Parent and Youth Ratings Across Societies.
As societies become increasingly diverse, mental health professionals need instruments for assessing emotional, behavioral, and social problems in terms of constructs that are supported within and across societies. Building on decades of research findings, multisample alignment confirmatory factor analyses tested an empirically based 8-syndrome model on parent ratings across 30 societies and youth self-ratings across 19 societies. The Child Behavior Checklist for Ages 6-18 and Youth Self-Report for Ages 11-18 were used to measure syndromes descriptively designated as Anxious/Depressed, Withdrawn/Depressed, Somatic Complaints, Social Problems, Thought Problems, Attention Problems, Rule-Breaking Behavior, and Aggressive Behavior. For both parent ratings (N = 61,703) and self-ratings (N = 29,486), results supported aggregation of problem items into 8 first-order syndromes for all societies (configural invariance), plus the invariance of item loadings (metric invariance) across the majority of societies. Supported across many societies in both parent and self-ratings, the 8 syndromes offer a parsimonious phenotypic taxonomy with clearly operationalized assessment criteria. Mental health professionals in many societies can use the 8 syndromes to assess children and youths for clinical, training, and scientific purposes.


AMPA-Kainate Receptor Inhibition Promotes Neurologic Recovery in Premature Rabbits with Intraventricular Hemorrhage.
Intraventricular hemorrhage (IVH) in preterm infants leads to cerebral inflammation, reduced myelination of the white matter, and neurological deficits. No therapeutic strategy exists against the IVH-induced white matter injury. AMPA-kainate receptor induced excitotoxicity contributes to oligodendrocyte precursor cell (OPC) damage and hypomyelination in both neonatal and adult models of brain injury. Here, we hypothesized that IVH damages white matter via AMPA receptor activation, and that AMPA-kainate receptor inhibition suppresses inflammation and restores OPC maturation, myelination, and neurologic recovery in preterm newborns with IVH. We tested these hypotheses in a rabbit model of glycerol-induced IVH and evaluated the expression of AMPA receptors in autopsy samples from human preterm infants. GluR1-GluR4 expressions were comparable between preterm humans and rabbits with and without IVH. However, GluR1 and GluR2 levels were significantly lower in the embryonic white matter and germinal matrix relative to the neocortex in both infants with and without IVH. Pharmacological blockade of AMPA-kainate receptors with systemic NBQX, or selective AMPA receptor inhibition by intramuscular perampanel restored myelination and neurologic recovery in rabbits with IVH. NBQX administration also reduced the population of apoptotic OPCs, levels of several cytokines (TNFα, IL-β, IL-6, LIF), and the density of Iba1(+) microglia in pups with IVH. Additionally, NBQX treatment inhibited STAT-3 phosphorylation, but not astrogliosis or transcription factors regulating gliosis. Our data suggest that AMPA-kainate receptor inhibition alleviates OPC loss and IVH-induced inflammation and restores myelination and neurologic recovery in preterm rabbits with IVH. Therapeutic use of FDA-approved perampanel treatment might enhance neurologic outcome in premature infants with IVH. Intraventricular hemorrhage (IVH) is a major complication of prematurity and a large number of survivors with IVH develop cerebral palsy and cognitive deficits. The development of IVH leads to inflammation of the periventricular white matter, apoptosis and arrested maturation of oligodendrocyte precursor cells, and hypomyelination. Here, we show that AMPA-kainate receptor inhibition by NBQX suppresses inflammation, attenuates apoptosis of oligodendrocyte precursor cells, and promotes myelination as well as clinical recovery in preterm rabbits with IVH. Importantly, AMPA-specific inhibition by the FDA-approved perampanel, which unlike NBQX has a low side-effect profile, also enhances myelination and neurological recovery in rabbits with IVH. Hence, the present study highlights the role of AMPA-kainate receptor in IVH-induced white matter injury and identifies a novel strategy of neuroprotection, which might improve the neurological outcome for premature infants with IVH.


Dynamic Stabilization with Active Locking Plates Delivers Faster, Stronger, and More Symmetric Fracture-Healing.
Axial dynamization of fractures can promote healing, and overly stiff fixation can suppress healing. A novel technology, termed active plating, provides controlled axial dynamization by the elastic suspension of locking holes within the plate. This prospective, controlled animal study evaluated the effect of active plates on fracture-healing in an established ovine osteotomy model. We hypothesized that symmetric axial dynamization with active plates stimulates circumferential callus and delivers faster and stronger healing relative to standard locking plates. Twelve sheep were randomly assigned to receive a standard locking plate or an active locking plate for stabilization of a 3-mm tibial osteotomy gap. The only difference between plates was that locking holes of active plates were elastically suspended, allowing up to 1.5 mm of axial motion at the fracture. Fracture-healing was analyzed weekly on radiographs. After sacrifice at nine weeks postoperatively, callus volume and distribution were assessed by computed tomography. Finally, to determine their strength, healed tibiae and contralateral tibiae were tested in torsion until failure. At each follow-up, the active locking plate group had more callus (p < 0.001) than the standard locking plate group. At postoperative week 6, all active locking plate group specimens had bridging callus at the three visible cortices. In standard locking plate group specimens, only 50% of these cortices had bridged. Computed tomography demonstrated that all active locking plate group specimens and one of the six standard locking plate group specimens had developed circumferential callus. Torsion tests after plate removal demonstrated that active locking plate group specimens recovered 81% of their native strength and were 399% stronger than standard locking plate group specimens (p < 0.001), which had recovered only 17% of their native strength. All active locking plate group specimens failed by spiral fracture outside the callus zone, but standard locking plate group specimens fractured through the osteotomy gap. Symmetric axial dynamization with active locking plates stimulates circumferential callus and yields faster and stronger healing than standard locking plates. The stimulatory effect of controlled motion on fracture-healing by active locking plates has the potential to reduce healing complications and to shorten the time to return to function.


Establishing and testing the "reuse potential" indicator for managing wastes as resources.
This study advances contemporary ideas promoting the importance of managing wastes as resources such as closed-loop or circular material economies, and sustainable materials management by reinforcing the notion of a resource-based paradigm rather than a waste-based one. It features the creation of a quantitative tool, the "reuse potential indicator" to specify how "resource-like" versus how "waste-like" specific materials are on a continuum. Even with increasing attention to waste reuse and resource conservation, constant changes in product composition and complexity have left material managers without adequate guidance to make decisions about what is technically feasible to recover from the discard stream even before markets can be considered. The reuse potential indicator is developed to aid management decision-making about waste based not on perception but more objectively on the technical ability of the materials to be reused in commerce. This new indicator is based on the extent of technological innovation and commercial application of actual reuse approaches identified and cataloged. Coal combustion by-products (CCBs) provide the test case for calculating the reuse potential indicator. While CCBs are often perceived as wastes and then isolated in landfills or surface impoundments, there is also a century-long history in the industry of developing technologies to reuse CCBs. The recent statistics show that most CCBs generated in Europe and Japan are reused (90-95%), but only 40-45% of CCBs are used in the United States. According to the reuse potential calculation, however, CCBs in the United States have high technical reusability. Of the four CCBs examined under three different regulatory schemes, reuse potential for boiler slag and flue-gas desulfurization gypsum maintains a value greater than 0.8 on a 0-1 scale, indicating they are at least 80% resource-like. Under current regulation in the United States, both fly ash and bottom ash are 80-90% resource-like. Very strict regulation would remove many reuse options decreasing potential for these two CCBs to 30% resource-like. A more holistic view of waste and broad application of the new indicator would make clear what technologies are available and assist public and private decision makers in setting quantitative material reuse targets from a new knowledge base that reinforces a resource-based paradigm.


Metabolic changes detected by in vivo magnetic resonance studies of HEPA-1 wild-type tumors and tumors deficient in hypoxia-inducible factor-1beta (HIF-1beta): evidence of an anabolic role for the HIF-1 pathway.
Hypoxia-inducible factor-1 (HIF-1) regulates many pathways potentially important for tumor growth, including angiogenesis and glycolysis. Most attention has focused on its role in the response to hypoxia, but HIF-1 is also constitutively expressed in many tumors. To analyze the role of this pathway in vivo, we used magnetic resonance (MR) methods and complementary techniques to monitor metabolic changes in tumors derived from HEPA-1 mouse hepatoma lines that were either wild type (WT) or deficient in hypoxia-inducible transcription factor HIF-1beta (c4). The c4 tumors grew significantly more slowly than the WT tumors (P < 0.05), but were examined at a similar size (0.4-0.6 g). At the tumor size used in these studies, no differences in vascularity were observed, and MR parameters measured that related to tumor blood flow, vascularity, and oxygenation demonstrated no significant differences between the two tumor types. Unexpectedly, the ATP content of the c4 tumor was approximately 5 times less than in the WT tumor [measured in tumor extracts (P < 0.001) and by metabolic imaging (P < 0.05)]. Noninvasive (31)P MR spectroscopy showed that the nucleoside triphosphate/P(i) ratio of the two tumor types was similar, so the low ATP content of the c4 tumors was not caused by (or a cause of) impaired cellular bioenergetics. Rather, glycine, an essential precursor for de novo purine formation, was significantly lower in the c4 tumors (P < 0.05), suggesting that ATP synthesis was impaired in the mutant tumor cells. Supporting evidence for this hypothesis came from the significantly lower concentrations of betaine, phosphocholine, and choline in the c4 tumors (P < 0.05); these are intermediates in an alternative pathway for glycine synthesis. No significant differences were seen in lactate or glucose content. MR resonances from phosphodiesters, which relate to the metabolic turnover of phospholipid membranes, were significantly lower in the WT tumors than in the c4 tumors, both in vivo (P < 0.05) and in extracts (P < 0.01). We propose that loss of up-regulation of expression of the genes for glucose transporters and glycolytic enzymes in the c4 tumors decreased formation of glycine, an essential precursor of ATP synthesis, and thus caused the low ATP content of the c4 tumors. In summary, these data suggest that disruption of the HIF-1 pathway in these tumor cells impairs the supply of anabolic precursors required for cell synthesis. They suggest potential biochemical targets that may be modified by therapy blocking HIF-1 function.


Improving access to affordable quality-assured inhaled medicines in low- and middle-income countries.
BACKGROUND: Access to affordable inhaled medicines for chronic respiratory diseases (CRDs) is severely limited in low- and middle-income countries (LMICs), causing avoidable morbidity and mortality. The International Union Against Tuberculosis and Lung Disease convened a stakeholder meeting on this topic in February 2022.METHODS: Focused group discussions were informed by literature and presentations summarising experiences of obtaining inhaled medicines in LMICs. The virtual meeting was moderated using a topic guide around barriers and solutions to improve access. The thematic framework approach was used for analysis.RESULTS: A total of 58 key stakeholders, including patients, healthcare practitioners, members of national and international organisations, industry and WHO representatives attended the meeting. There were 20 pre-meeting material submissions. The main barriers identified were 1) low awareness of CRDs; 2) limited data on CRD burden and treatments in LMICs; 3) ineffective procurement and distribution networks; and 4) poor communication of the needs of people with CRDs. Solutions discussed were 1) generation of data to inform policy and practice; 2) capacity building; 3) improved procurement mechanisms; 4) strengthened advocacy practices; and 5) a World Health Assembly Resolution.CONCLUSION: There are opportunities to achieve improved access to affordable, quality-assured inhaled medicines in LMICs through coordinated, multi-stakeholder, collaborative efforts.


Mechanotransduction and fracture repair.
Fracture-healing is regulated in part by mechanical factors. Study of the processes by which the mechanical environment of a fracture modulates healing can yield new strategies for the treatment of bone injuries. This article focuses on several key unanswered questions in the study of mechanotransduction and fracture repair. These questions concern identifying the mechanical stimuli that promote bone-healing, defining the mechanisms that are involved in this process, and examining the potential for cross-talk between investigations of mechanotransduction in bone-healing and in healing of other mesenchymally derived tissues. Several approaches to obtain accurate estimates of the mechanical stimuli present within a fracture callus are proposed, and our current understanding of the mechanotransduction processes involved in bone-healing is reviewed. Further study of mechanotransduction mechanisms is needed in order to identify those that are most critical and active during the various phases of fracture repair. A better understanding of the effect of mechanical factors on bone-healing will also benefit the study of healing, regeneration, and engineering of other skeletal tissues.


Cephalometric comparison of vertical changes between Begg and preadjusted edgewise appliances.
The purpose of this retrospective study was to quantify and compare the vertical dimensional changes in bialveolar dental protrusion patients undergoing extraction of all four first premolars between the preadjusted edgewise appliance (PEA) and the Begg appliance. The cephalometric records of 55 patients (14 males and 41 females) with Class I bimaxillary protrusion were selected and divided into two groups based on the appliance used, i.e Begg or PEA mechanotherapy. To minimize the effects of growth, the subjects were at least in stage VI of skeletal maturation. The mean age was 18.25 ± 3.2 years for the Begg group and 18.03 ± 3.5 years for the PEA group. Skeletal and dental changes were analysed in both groups on lateral cephalograms taken pre- (T1) and post- (T2) treatment. A Student's t-test was used to analyse the treatment changes. Within group results showed a significant increase in face height and extrusion and mesial movement of the upper and lower molars for both groups. However, no significant differences were observed when the groups were compared. A significant correlation was found between the change in lower molar to mandibular plane angle and lower anterior face height for both groups. No significant difference was found when the Begg mechanotherapy was compared to the PEA technique on vertical dimensional changes. It can be speculated that mesial movement of the molars tended to keep pace with their extrusion and negated any bite opening effect with both mechanotherapies.


Tissue plasminogen activator induction in Purkinje neurons after cerebellar motor learning.
The cerebellar cortex is implicated in the learning of complex motor skills. This learning may require synaptic remodeling of Purkinje cell inputs. An extracellular serine protease, tissue plasminogen activator (tPA), is involved in remodeling various nonneural tissues and is associated with developing and regenerating neurons. In situ hybridization showed that expression of tPA messenger RNA was increased in the Purkinje neurons of rats within an hour of their being trained for a complex motor task. Antibody to tPA also showed the induction of tPA protein associated with cerebellar Purkinje cells. Thus, the induction of tPA during motor learning may play a role in activity-dependent synaptic plasticity.


Impact of Chronic Total Occlusion Revascularization Attempts on Subsequent Clinical Outcomes.
We examined a contemporary, unselected cohort of patients with coronary chronic total occlusions (CTOs) to determine the impact of CTO revascularization on long-term outcomes. We retrospectively assessed the impact of CTO revascularization on clinical outcomes of consecutive patients found to have a CTO during coronary angiography performed at our institution during 2011 and 2012. The primary endpoint was the incidence of a major adverse cardiac event (MACE, defined as a composite of death, myocardial infarction, stroke, and target-vessel revascularization [TVR]). Survival analysis was performed in the overall and propensity-matched retrospective cohorts of patients stratified by prior coronary artery bypass graft (CABG) surgery. Propensity-adjusted hazard ratio (HR) and 95% confidence interval (95% CI) were calculated with Cox proportional hazards analysis. All analyses were by intention to treat. Of 624 patients (319 without prior CABG and 305 with prior CABG) included in the present analysis, CTO revascularization (surgical or percutaneous) was attempted in 60% and 16% of patients without and with prior CABG, respectively. During a median follow-up of 26 months (range, 18-40 months), the incidence of MACE was 20.6%. CTO revascularization (achieved or attempted) was associated with lower incidence of MACE among patients without prior CABG (propensity-adjusted HR, 0.51; 95% CI, 0.27-0.94; P=.03), but not among prior CABG patients (propensity-adjusted HR, 1.38; 95% CI, 0.64-2.96; P=.41). In a large, unselected patient population with coronary CTOs, a CTO revascularization attempt was associated with lower incidence of subsequent MACE among patients without prior CABG, but not among prior CABG patients.


Macaque parvocellular mediodorsal thalamus: dissociable contributions to learning and adaptive decision-making.
Distributed brain networks govern adaptive decision-making, new learning and rapid updating of information. However, the functional contribution of the rhesus macaque monkey parvocellular nucleus of the mediodorsal thalamus (MDpc) in these key higher cognitive processes remains unknown. This study investigated the impact of MDpc damage in cognition. Preoperatively, animals were trained on an object-in-place scene discrimination task that assesses rapid learning of novel information within each session. Bilateral neurotoxic (NMDA and ibotenic acid) MDpc lesions did not impair new learning unless the monkey had also sustained damage to the magnocellular division of the MD (MDmc). Contralateral unilateral MDpc and MDmc damage also impaired new learning, while selective unilateral MDmc damage produced new learning deficits that eventually resolved with repeated testing. In contrast, during food reward (satiety) devaluation, monkeys with either bilateral MDpc damage or combined MDpc and MDmc damage showed attenuated food reward preferences compared to unoperated control monkeys; the selective unilateral MDmc damage left performance intact. Our preliminary results demonstrate selective dissociable roles for the two adjacent nuclei of the primate MD, namely, MDpc, as part of a frontal cortical network, and the MDmc, as part of a frontal-temporal cortical network, in learning, memory and the cognitive control of behavioural choices after changes in reward value. Moreover, the functional cognitive deficits produced after differing MD damage show that the different subdivisions of the MD thalamus support distributed neural networks to rapidly and fluidly incorporate task-relevant information, in order to optimise the animals' ability to receive rewards.


Diagnostic strategies in CADASIL.
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited autosomal dominant condition characterized by migraine, recurrent stroke, and dementia. It results from mutations in the notch3 gene but mutations may occur at multiple sites making molecular diagnosis time consuming. It has been suggested that the presence of granular osmiophilic material (GOM) on skin biopsy and involvement of the anterior temporal lobe and external capsule on MRI may help in diagnosis. The authors identified 83 potential index cases from the British population and screened exons 2 to 23 of notch3. MRI scans were scored using a modified Scheltens scale. Skin biopsy was performed in a subgroup. Fifteen different point mutations were identified in 48 families, 73% of which were in exon 4, 8% in exon 3, and 6% in each of exons 5 and 6. Moderate or severe involvement of the anterior temporal pole on MRI had a sensitivity of 89% and specificity of 86% for diagnosis of CADASIL, whereas external capsule involvement had a high sensitivity of 93% but a low specificity of 45%. Skin biopsy, performed in 18 cases, had a sensitivity of 45% and specificity of 100%. The spectrum of mutations in this study can be used to plan appropriate screening protocols; a suggested protocol is to screen exon 4, and proceed to exons 3, 5, and 6 where indicated. GOM on skin biopsy is diagnostic but can be negative. Anterior temporal pole involvement on MRI is a useful diagnostic marker.


Knee range of motion during the first two years after use of posterior cruciate-stabilizing or posterior cruciate-retaining total knee prostheses. A randomized clinical trial.
The impact of posterior cruciate ligament-substituting and posterior cruciate ligament-retaining devices on the range of motion of the knee following primary total knee arthroplasty is unclear. The primary objective of our study was to compare the range of motion of the knee over the first two postoperative years between subjects who had received the ligament-substituting design and those who had received the ligament-retaining design. Secondarily, pain, function, and health-related quality of life were compared between the two groups. We undertook a prospective randomized study of 100 patients with noninflammatory osteoarthritis who were undergoing primary total knee arthroplasty. Patients were randomized at the time of surgery to receive one of the two study prosthesis designs. They were evaluated preoperatively, at the time of hospital discharge, at three months postoperatively, and at up to two years postoperatively by a physical therapist who was blinded to the group allocation. Active knee flexion and extension, measured with a goniometer, were the primary outcome measures. Self-reported pain, function, and health-related quality of life were measured with the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the RAND 36-Item Health Survey (RAND-36). Complications and revision rates were determined through hospital record review and at each patient evaluation. Eighty percent (seventy-eight) of the ninety-eight available patients returned for the final assessment. At two years postoperatively, the mean difference between the groups with regard to knee flexion was 0.03 degrees (95% confidence interval, -5.9 degrees to 6.0 degrees ) and the mean difference in knee extension was 1.0 degrees (95% confidence interval, -0.36 degrees to 2.4 degrees ). Ninety-one (93%) of the ninety-eight available patients completed questionnaires at the time of the final assessment. The two treatment groups had similar pain, function, and quality-of-life scores and complication rates. One subject in the cruciate-retaining group required revision, secondary to a deep joint infection, and one subject in the cruciate-substituting group required manipulation to address poor knee flexion. Overall, the two treatment groups had a similar range of motion of the knee over the initial two-year postoperative time period. A satisfactory range of motion was achieved by three months postoperatively and was maintained at the final assessment.


Where in the cell is SIRT3?--functional localization of an NAD+-dependent protein deacetylase.
Sirtuins are NAD+-dependent enzymes that have been implicated in a wide range of cellular processes, including pathways that affect diabetes, cancer, lifespan and Parkinson's disease. To understand their cellular function in these age-related diseases, identification of sirtuin targets and their subcellular localization is paramount. SIRT3 (sirtuin 3), a human homologue of Sir2 (silent information regulator 2), has been genetically linked to lifespan in the elderly. However, the function and localization of this enzyme has been keenly debated. A number of reports have indicated that SIRT3, upon proteolytic cleavage in the mitochondria, is an active protein deacetylase against a number of mitochondrial targets. In stark contrast, some reports have suggested that full-length SIRT3 exhibits nuclear localization and histone deacetylase activity. Recently, a report comparing SIRT3-/- and SIRT+/+ mice have provided compelling evidence that endogenous SIRT3 is mitochondrial and appears to be responsible for the majority of protein deacetylation in this organelle. In this issue of the Biochemical Journal, Cooper et al. present additional results that address the mitochondrial and nuclear localization of SIRT3. Utilizing fluorescence microscopy and cellular fractionation studies, Cooper et al. have shown that SIRT3 localizes to the mitochondria and is absent in the nucleus. Thus this study provides additional evidence to establish SIRT3 as a proteolytically modified, mitochondrial deacetylase.


Do You Know What I Know? The Impact of Participant Role in Children's Referential Communication.
For successful language use, interlocutors must be able to accurately assess their shared knowledge ("common ground"). Such knowledge can be accumulated through linguistic and non-linguistic context, but the same context can be associated with different patterns of knowledge, depending on the interlocutor's participant role (Wilkes-Gibbs and Clark, 1992). Although there is substantial evidence that children's ability to model partners' knowledge develops gradually, most such evidence focuses on non-linguistic context. We investigated the extent to which 8- to 10-year-old children can assess common ground developed through prior linguistic context, and whether this is sensitive to variations in participant role. Children repeatedly described tangram figures to another child, and then described the same figures to a third child who had been a side-participant, an overhearer, or absent during the initial conversation. Children showed evidence of partner modeling, producing shorter referential expressions with repeated mention to the same partner. Moreover, they demonstrated sensitivity to differences in common ground with the third child based on participant role on some but not all measures (e.g., description length, but not definiteness). Our results suggest that by ten, children make distinctions about common ground accumulated through prior linguistic context but do not yet consistently deploy this knowledge in an adult-like way.


Protocatechuic acid promotes cell proliferation and reduces basal apoptosis in cultured neural stem cells.
Protocatechuic acid (PCA), a phenolic compound isolated from the kernels of Alpinia oxyphylla, showed anti-oxidant neuroprotective property in our previous study. However, it is still unknown whether PCA have effects on the cultured neural stem cells (NSCs). In this study, we investigated the roles of PCA in the survival and apoptosis of rat NSCs under normal conditions. NSCs obtained from 13.5-day-old rat embryos were propagated as neurospheres and cultured under normal conditions with or without PCA for 4 and 7 days. The cell viability was determined by the cell counting kit-8 (CCK-8) test, while cell proliferation was assayed by bromodeoxyuridine (BrdU) labeling. PCA increased the cellular viability of NSCs and stimulated cell proliferation in a dose- and time-dependent manner. Apoptotic cells were detected after 4 days by observing the nuclear morphological changes and flow cytometric analysis. Compared with the control on both culture days, treatment with PCA effectively reduced the levels of apoptosis of NSCs. At the same time, the reactive oxygen species (ROS) level in NSCs was depressed. In addition, PCA also significantly decreased the activity of elevated caspase-3, indicating that PCA may inhibit apoptosis of NSCs via suppression of the caspase cascade. These results suggest that PCA may be a potential growth inducer and apoptosis inhibitor for NSCs.


Expression and function of junctional adhesion molecule-C in human and experimental arthritis.
Junctional adhesion molecule-C (JAM-C) is an adhesion molecule involved in transendothelial migration of leukocytes. In this study, we examined JAM-C expression in the synovium and investigated the role of this molecule in two experimental mouse models of arthritis. JAM-C expression was investigated by reverse transcriptase-polymerase chain reaction and immunohistochemistry. The effects of a monoclonal anti-JAM-C antibody were assessed in antigen-induced arthritis (AIA) and K/BxN serum transfer-induced arthritis. JAM-C was expressed by synovial fibroblasts in the lining layer and associated with vessels in the sublining layer in human and mouse arthritic synovial tissue. In human tissue, JAM-C expression was increased in rheumatoid arthritis (RA) as compared to osteoarthritis synovial samples (12.7 +/- 1.3 arbitrary units in RA versus 3.3 +/- 1.1 in OA; p < 0.05). Treatment of mice with a monoclonal anti-JAM-C antibody decreased the severity of AIA. Neutrophil infiltration into inflamed joints was selectively reduced as compared to T-lymphocyte and macrophage infiltration (0.8 +/- 0.3 arbitrary units in anti-JAM-C-treated versus 2.3 +/- 0.6 in isotype-matched control antibody-treated mice; p < 0.05). Circulating levels of the acute-phase protein serum amyloid A as well as antigen-specific and concanavalin A-induced spleen T-cell responses were significantly decreased in anti-JAM-C antibody-treated mice. In the serum transfer-induced arthritis model, treatment with the anti-JAM-C antibody delayed the onset of arthritis. JAM-C is highly expressed by synovial fibroblasts in RA. Treatment of mice with an anti-JAM-C antibody significantly reduced the severity of AIA and delayed the onset of serum transfer-induced arthritis, suggesting a role for JAM-C in the pathogenesis of arthritis.


Isolation breeds naivety: island living robs Australian varanid lizards of toad-toxin immunity via four-base-pair mutation.
Since their introduction to the toad-free Australian continent cane toads (Bufo marinus) have caused a dramatic increase in naïve varanid mortality when these large lizards attempt to feed on this toxic amphibian. In contrast Asian-African varanids, which have coevolved with toads, are resistant to toad toxin. Toad toxins, such as Bufalin target the H1-H2 domain of the α(1) subunit of the sodium-potassium-ATPase enzyme. Sequencing of this domain revealed identical nucleotide sequences in four Asian as well as in three African varanids, and identical sequences in all 11 Australian varanids. However, compared to the Asian-African varanids, the Australian varanids showed four-base-pair substitutions, resulting in the alteration in three of the 12 amino acids representing the H1-H2 domain. The phenotypic effect of the substitutions was investigated in human embryonic kidney (HEK) 293 cells stably transfected with the Australian and the Asian-African H1-H2 domains. The transfections resulted in an approximate 3000-fold reduction in resistance to Bufalin in the Australian HEK293 cells compared to the Asian-African HEK293 cells, demonstrating the critical role of this minor mutation in providing Bufalin resistance. Our study hence presents a clear link between genotype and phenotype, a critical step in understanding the evolution of phenotypic diversity.


Weighted multiple testing procedures for genomic studies.
With the rapid development of biological technology, measurement of thousands of genes or SNPs can be carried out simultaneously. Improved procedures for multiple hypothesis testing when the number of tests is very large are critical for interpreting genomic data. In this paper, we review recent developments on three distinct but closely related methods involving p-value weighting to improve statistical power while also controlling for the false discovery rate or the family wise error rate.


Surrogate consent for critical care research: exploratory study on public perception and influences on recruitment.
Critical Care research involves an increasing level of technical and clinical interventions for the unconscious patient. If the general public has a negative (unfavourable) view of surrogate consent, low recruitment rates are likely. Results bias will be introduced if study populations are small, hindering knowledge generation and transfer through research. In the rapidly expanding healthcare industry of South East Asia, opportunities for critical care research will grow given a positive willingness (favourability) by the general public to act as a surrogate in the consent process when the (unconscious) patient cannot consent for him/herself. To determine public willingness for surrogate consent, a quantitative cross-sectional study was undertaken at a University Teaching Hospital in South East Asia during a three month interval. Four hypothetical critical care research scenarios were presented and responses from the public were analysed using a three-part questionnaire. Three hundred and five members of the public were recruited. In general, participants had a positive view of research. The level of education was significantly associated with a person's views about research especially in studies regarded as high risk. For low risk studies, a person's perception of research and willingness to be recruited to a study in the event that they were the (unconscious) patient, was the same whether they were the study subject or the person (legally acceptable representative) giving surrogate consent' on behalf of another (spouse, parent, child). Across all study scenarios, 60-80% of the public preferred to be approached by doctors to discuss the surrogate consent process. Given the hypothetical scenarios presented in this study, the odds of a person having a positive view and willingness to be consented to a critical care research study on the advice of another (surrogate consent) was greater than for those who had a negative or unfavourable view. Nurses may be disadvantaged in leading on the recruitment process due to a preference for information to be delivered by medically qualified clinicians. In the setting of South East Asia, cultural attitudes to nurse-led research in critical care must be taken in to consideration in the multidisciplinary approaches to building the research team.


IGF-2 coated porous collagen microwells for the culture of pancreatic islets.
Islet transplantation, the only curative therapy for type I diabetes, requires isolation of the graft in highly specialized facilities for its later dispatch to remote transplantation centres. During transport and culture, many valuable cells are lost due to several factors such as mechanical stress, islet aggregation and dissociation. Here, we evaluate a porous microwell array sheet made of natural collagen type I extracellular matrix (ECM) protein as a novel islet culture substrate. This culture platform can be coated with IGF-2, a growth factor favorable for islet survival, and allows segregation of the islets within the porous microwell sheet, preventing aggregation. This design shows promising results for improving human pancreatic islets viability and function during culture and could form a novel paradigm for the transport of islets between isolation and transplantation centres.


The positioning of Aboriginal students and their languages within Australia's education system: A human rights perspective.
This paper is a critical review of past and present languages policies in Australian schooling. We highlight the One Literacy movement that contravenes the human rights of Australia's Aboriginal students. This in turn impacts students' right to freedom of opinion and expression as stated in Article 19 of the Universal Declaration of Human Rights. The One Literacy movement operates by equating Standard Australian English literacy acquisition with Australia's global competitiveness and economic success. There is only one pathway through the Australian English curriculum with common assessments and standards. However, the Australian Curriculum provides three distinctive pathways when students from an English-speaking background learn languages other than English. We reveal this double standard, where current educational policies prioritise the languages of trade (e.g. Chinese) and accommodate speakers of these languages. Meanwhile Aboriginal-language-speaking students are not provided with the same accommodations. For educational equity, there should be a distinctive English language learner pathway that recognises that the majority of remote Aboriginal students from the Northern Territory are learning English as an additional language. We advocate for these changes because all children have a right to an appropriate education that will enable them to flourish as learners and citizens.


Direct assessment of mental health and metabolic syndrome amongst Indonesian adolescents: a study design for a mixed-methods study sampled from school and community settings.
Non-communicable diseases (NCDs) are the leading cause of morbidity and mortality globally, with the burden largely borne by people living in low- and middle-income countries. Adolescents are central to NCD control through the potential to modify risks and alter the trajectory of these diseases across the life-course. However, an absence of epidemiological data has contributed to the relative exclusion of adolescents from policies and responses. This paper documents the design of a study to measure the burden of metabolic syndrome (a key risk for NCDs) and poor mental health (a key outcome) amongst Indonesian adolescents. Using a mixed-method design, we sampled 16-18-year-old adolescents from schools and community-based settings across Jakarta and South Sulawesi. Initial formative qualitative enquiry used focus group discussions to understand how young people conceptualise mental health and body weight (separately); what they perceive as determinants of these NCDs; and what responses to these NCDs should involve. These findings informed the design of a quantitative survey that adolescents self-completed electronically. Mental health was measured using the Centre for Epidemiologic Studies Depression Scale-Revised (CESD-R) and Kessler-10 (both validated against formal psychiatric interview in a subsample), with the metabolic syndrome measured using biomarkers and anthropometry. The survey also included scales relating to victimisation, connectedness, self-efficacy, body image and quality of life. Adolescents were sampled from schools using a multistage cluster design, and from the community using respondent-driven sampling (RDS). This study will substantially advance the field of NCD measurement amongst adolescents, especially in settings like Indonesia. It demonstrates that high quality, objective measurement is acceptable and feasible, including the collection of biomarkers in a school-based setting. It demonstrates how comparable data can be collected across both in-school and out of school adolescents, allowing a more comprehensive measure of NCD burden, risk and correlates.


Myosin light-chain expression during avian muscle development.
Monoclonal antibodies to adult chicken myosin light chains were generated and used to quantitate the types of myosin light-chain (MLC) isoforms expressed during development of the pectoralis major (PM), anterior latissimus dorsi (ALD), and medial adductor (MA) muscles of the chicken. These are muscles which, in the adult, are composed predominantly of fast, slow, and a mixture of fiber types, respectively. Three distinct phases of MLC expression characterized the development of the PM and MA muscles. The first identifiable pase occurred during the period of 5-7 d of incubation in ovo. Extracts of muscles from the pectoral region (which included the presumptive PM muscle) contained only fast MLC isoforms. This period of exclusive fast light-chain synthesis was followed by a phase (8- 12 d of incubation in ovo) in which coexpression of both fast and slow MLC isoforms was apparent in both PM and MA muscles. During the period, the composition of both fast and slow MLC isoforms in the PM and MA muscles was identical. Beginning at day 12 in ovo, the ALD was also subjected to immunochemical analyses. The proportion of fast and slow MLCs in this muscle at day 12 was similar to that present in the other muscles studied. The third development phase of MLC expression began at approximately 12 d of incubation in ovo and encompassed the transition in MLC composition to the isoform patterns incubation in ovo and encompassed the transition in MLC composition to the isoform patterns typical of adult muscle. During this period, the relative proportion of slow MLC rose in both the MA and ALD and fell in the PM. By day 16, the third fast light chain, LC(3f), was apparent in extracts of both the PM and MA. These results show that there is a developmental progression in the expression of MLC in the two avian muscles studied from day 5 in ovo; first, only fast MLCs are accumulated, then both fast and slow MLC isoforms are expressed. Only during the latter third of development in ovo is the final MLC isoform pattern characteristic of a particular muscle type expressed.


Socioeconomic Status, Vocational Aspirations, School Tracks, and Occupational Attainment in South Korea.
Adults' career choice is not an abrupt event, but an outcome of continuous development throughout childhood and adolescence. In the developmental process of one's career, personal characteristics and contextual resources come into play. The goal of the present study is to examine how family socioeconomic status, adolescents' vocational aspirations, and high school contexts affect their occupational attainment in young adulthood, using two cohorts of data from the Korean Education and Employment Panel. Cohort 1 consisted of 1535 individuals (49.3% female), and cohort 2 consisted of 1473 individuals (53.5% female). Both cohorts were surveyed during their senior year of high school (Time 1; Mage = 17.8) and followed up until young adulthood (Time 2; Mage = 25.8). The results reveal that having high vocational aspirations and attending academic high school predict attaining higher-status occupations for both cohorts. Family background has positive direct and indirect effects on occupational attainment for cohort 2, while it only has an indirect effect on occupational attainment via types of high school for cohort 1. Implications in the context of constructing social systems to support adolescents' career development are discussed.


Antisense PNA effects in Escherichia coli are limited by the outer-membrane LPS layer.
Antisense peptide nucleic acids (PNAs) can inhibit Escherichia coli gene expression and cell growth through sequence-specific RNA binding, and this opens possibilities for novel anti-infective agents and tools for microbial functional genomics. However, the cellular effects of PNAs are limited relative to effects in cell extracts, presumably because of cell barrier components such as the outer-membrane lipopolysaccharide (LPS) layer or drug efflux pumps, both of which function to exclude antibiotics and other foreign molecules. To evaluate the importance of such cellular factors on PNA effects, the authors developed a positive assay for antisense inhibition by targeting the lac operon repressor and compared PNA susceptibilities in mutant and wild-type E. coli by assessing lacZ induction. Strains with defective LPS (AS19 and D22) were more permeable to the antibiotic nitrocefin and more susceptible to PNA than the wild-type. Also, PNA potency was improved in wild-type cells grown in the presence of certain cell-wall-permeabilizing agents. In contrast, the activities of the Acr and Emr drug efflux pumps were not found to affect PNA susceptibility. The results show that the LPS layer is a major barrier against cell entry, but PNAs that can enter E. coli are likely to remain active inside cells.


Model-Based Reconstruction of Large Three-Dimensional Optoacoustic Datasets.
Iterative model-based algorithms are known to enable more accurate and quantitative optoacoustic (photoacoustic) tomographic reconstructions than standard back-projection methods. However, three-dimensional (3D) model-based inversion is often hampered by high computational complexity and memory overhead. Parallel implementations on a graphics processing unit (GPU) have been shown to efficiently reduce the memory requirements by on-the-fly calculation of the actions of the optoacoustic model matrix, but the high complexity still makes these approaches impractical for large 3D optoacoustic datasets. Herein, we show that the computational complexity of 3D model-based iterative inversion can be significantly reduced by splitting the model matrix into two parts: one maximally sparse matrix containing only one entry per voxel-transducer pair and a second matrix corresponding to cyclic convolution. We further suggest reconstructing the images by multiplying the transpose of the model matrix calculated in this manner with the acquired signals, which is equivalent to using a very large regularization parameter in the iterative inversion method. The performance of these two approaches is compared to that of standard back-projection and a recently introduced GPU-based model-based method using datasets from in vivo experiments. The reconstruction time was accelerated by approximately an order of magnitude with the new iterative method, while multiplication with the transpose of the matrix is shown to be as fast as standard back-projection.


Clinical hints to diagnosis of attenuated forms of Mucopolysaccharidoses.
The mucopolysaccharidoses (MPS) are clinically similar but also heterogeneous in terms of major or minor involvement of different organs/systems, burden of disease, and rate of progression. The attenuated forms of MPS, due to their less severe presentations, are more difficult to diagnose and often receive a significantly delayed diagnosis. On the other hand, the diagnosis is very important since the attenuated forms may benefit from earlier treatments. The aim of this paper is to describe the natural history and the clinical signs useful to arise a suspicion of an attenuated form of MPS. MPS patients usually show a cluster of signs and symptoms, one of which may be the trigger for an evaluation by a specialist. Individuals with attenuated MPS are mostly cognitively normal, and dysmorphisms of the facies may be mild or absent. The most frequently involved organs/systems are the osteoarticular system, heart, and eyes. These patients may also have hepatosplenomegaly, hearing loss, and respiratory problems. When they are referred to a specialist (rheumatologist, cardiologist, ophthalmologist, surgeon, orthopedist, etc.) for their main complaint, the other signs and symptoms are likely to be missed in the medical history. To avoid missing data and to save time, we propose a semistructured medical history form to be filled in by the patients or their caregivers while waiting for evaluation by a specialist.


Integrin-receptor-mediated differentiation and growth inhibition are enhanced by transforming growth factor-beta in colorectal tumour cells grown in collagen gel.
We have studied the role of cell-matrix interactions and the modulating effect on these of transforming growth factor-beta s (TGF-beta s) in controlling differentiation and proliferation in a series of human colorectal carcinoma cell lines. Two (SW1222 and SW480) out of 7 cell lines specifically bound type-I collagen, via integrin-like (SW1222) and non-integrin-like (SW480) collagen receptors. Binding of these receptors may be responsible for regulating the degree of epithelial differentiation of the cells when grown in a 3-dimensional (3D) collagen gel. We have also shown that TGF-beta s enhance the binding of SW1222 cells to collagen and that this is accompanied by greatly increased crypt-like glandular differentiation and inhibition of cell proliferation. Inhibition of cell proliferation was only seen when cells were grown in 3D collagen gel and were thus expressing a fully differentiated phenotype. The enhanced collagen binding induced by TGF-beta s was partially inhibited by an Arg-Gly-Asp (RGD)-containing peptide which is a cell recognition signal for collagen binding. This suggests that TGF-beta s mediate their effects on differentiation of SW1222 cells specifically by modulating the expression of the integrin-like collagen receptor. The other colorectal carcinoma cell lines which lack this integrin-like receptor either failed to bind collagen or, in the case of SW480 binding, exhibited differentiation and proliferation which were not affected by TGF-beta s. This suggests that cell responsiveness to TGF-beta s may depend, at least in part, upon the cell-matrix interaction.


Postoperative Complications in the Ahmed Baerveldt Comparison Study During Five Years of Follow-up.
To compare the late complications in the Ahmed Baerveldt Comparison Study during 5 years of follow-up. Multicenter, prospective randomized clinical trial. setting: Sixteen international clinical centers. Two hundred seventy-six subjects aged 18-85 years with previous intraocular surgery or refractory glaucoma with intraocular pressure of >18 mm Hg. Ahmed Glaucoma Valve FP7 or Baerveldt Glaucoma Implant BG 101-350. Late postoperative complications (beyond 3 months), reoperations for complications, and decreased vision from complications. Late complications developed in 56 subjects (46.8 ± 4.8 5-year cumulative % ± SE) in the Ahmed Glaucoma Valve group and 67 (56.3 ± 4.7 5-year cumulative % ± SE) in the Baerveldt Glaucoma Implant group (P = .082). The cumulative rates of serious complications were 15.9% and 24.7% in the Ahmed Glaucoma Valve and Baerveldt Glaucoma Implant groups, respectively (P = .034), although this was largely driven by subjects who had tube occlusions in the 2 groups (0.8% in the Ahmed Glaucoma Valve group and 5.7% in the Baerveldt Glaucoma Implant group, P = .037). Both groups had a relatively high incidence of persistent diplopia (12%) and corneal edema (20%), although half of the corneal edema cases were likely due to pre-existing causes other than the aqueous shunt. The incidence of tube erosion was 1% and 3% in the Ahmed Glaucoma Valve and Baerveldt Glaucoma Implant groups, respectively (P = .04). Long-term rates of vision-threatening complications and complications resulting in reoperation were higher in the Baerveldt Glaucoma Implant than in the Ahmed Glaucoma Valve group over 5 years of follow-up.


Adolescent alcohol use is linked to disruptions in age-appropriate cortical thinning: an unsupervised machine learning approach.
Cortical thickness changes dramatically during development and is associated with adolescent drinking. However, previous findings have been inconsistent and limited by region-of-interest approaches that are underpowered because they do not conform to the underlying spatially heterogeneous effects of alcohol. In this study, adolescents (n = 657; 12-22 years at baseline) from the National Consortium on Alcohol and Neurodevelopment in Adolescence (NCANDA) study who endorsed little to no alcohol use at baseline were assessed with structural magnetic resonance imaging and followed longitudinally at four yearly intervals. Seven unique spatial patterns of covarying cortical thickness were obtained from the baseline scans by applying an unsupervised machine learning method called non-negative matrix factorization (NMF). The cortical thickness maps of all participants' longitudinal scans were projected onto vertex-level cortical patterns to obtain participant-specific coefficients for each pattern. Linear mixed-effects models were fit to each pattern to investigate longitudinal effects of alcohol consumption on cortical thickness. We found in six NMF-derived cortical thickness patterns, the longitudinal rate of decline in no/low drinkers was similar for all age cohorts. Among moderate drinkers the decline was faster in the younger adolescent cohort and slower in the older cohort. Among heavy drinkers the decline was fastest in the younger cohort and slowest in the older cohort. The findings suggested that unsupervised machine learning successfully delineated spatially coordinated patterns of vertex-level cortical thickness variation that are unconstrained by neuroanatomical features. Age-appropriate cortical thinning is more rapid in younger adolescent drinkers and slower in older adolescent drinkers, an effect that is strongest among heavy drinkers.


Camellia oleifera Abel shells as a new biosorbent to remove methylene blue from aqueous solutions.
This study aimed at investigating the feasibility of using Camellia oleifera Abel shell (COAS), an agriculture product in middle-west region in China, for the adsorption of methylene blue (MB), a cationic dye. Batch adsorption experiments were carried out to evaluate the effects of contact time, pH, adsorbent dosage, temperature and initial concentration on the adsorption of MB. Adsorption equilibrium studies showed that MB adsorption followed Langmuir model with a maximum adsorption capacity of 454.54 mg/g. The results demonstrated that the COAS is a promising adsorbent in the removal of MB from aqueous solutions.


Wire transfer function analysis for castellated dual-energy x-ray detectors.
An investigation into the spatial resolving power of a castellated linear dual-energy x-ray detector array is reported. The detector was developed for use in aviation security screening applications. Experiments employing different gauges of lead wire are used to plot a wire transfer function. A numerical simulation is developed to predict and underpin the empirical results. The suitable processing of the castellated detector signals helps to maintain spatial resolving power while affording a 50% reduction in x-ray sensing elements. This encouraging result has formed the basis for an ongoing investigation into materials discrimination capability of the castellated detector array.


Hemodynamics of blood pressure responses during active and passive coping.
This laboratory study was designed to address a number of interrelated issues regarding cardiovascular reactivity to psychological stress. One objective was to extend the previous research comparing cardiovascular responses during active versus passive coping, by comparing responses to two task conditions designed to be similar in all ways except the opportunity to make a response influencing the task's outcome. A second objective was to compare responses to two different passive film tasks, which differed in outcome uncertainty and the degree of vicarious active coping achieved through identification with the role portrayed by the actors. A third objective was to evaluate whether individuals are predisposed to exhibit a particular hemodynamic pattern underlying their blood pressure adjustments, independently of the task demands imposed. Ninety healthy young adult male subjects were tested in pairs on a series of tasks that included a competitive reaction-time task, an active as well as a passive phase of a team reaction-time task, and passive viewing of two film segments. The tasks demanding active coping responses tended to raise blood pressure due primarily to an increase in cardiac output, while vascular resistance fell. During passive coping demands cardiac output increased to a lesser extent, but vascular resistance also tended to increase, thereby raising blood pressure by their synergistic effects. However, these patterns were not typical of all participating subjects. On the basis of their cardiac output and vascular resistance responses to the competitive reaction-time task, one third of the subjects were categorized as being high myocardial reactors (n = 30) and another third high vascular reactors (n = 31). Post-hoc analyses of responses to the other tasks, based on these categorizations, indicated that the hemodynamic basis of reactivity is an individual characteristic only partially modified by coping demands. The active/passive coping dimension is discussed both conceptually and in relation to the role of stress in the etiology of hypertension.


Astrocytic involvement in the maturation phenomenon after temporary cerebral ischemia.
Astrocytes support neuronal functions by regulating the extracellular ion homeostasis and levels of neurotransmitters, and by providing fuel such as lactate to the neurons via their processes (APs). After two 10-min unilateral carotid occlusions with a 5-h interval in gerbils, we investigated maturing disseminated selective neuronal necrosis (DSNN) on the coronal surface sectioned at the infundibular level. We chronologically counted the normal appearing, degenerated, and dead neurons and astrocytes in the cerebral cortex; observed the ultrastructure of APs, and counted the number of their cut-ends and mitochondria in the neuropil; determined the percentage volume of APs according to Weibel's point-counting method; compared the number of cut-ends and mitochondria and percentage volume of APs around the astrocytes and around the normal-appearing, degenerated, and dead -neurons. Heterogeneous degeneration of APs was concluded to be closely associated with the maturation of DSNN.Using the same model, at the coronally sectioned surface on the chiasmatic level, we investigated the mechanism of development of focal infarction in the maturing DSNN. Same as in the above study, we chronologically counted various neurons and astrocytes; observed and measured the area of the ultrastructure of astrocytic end-feet; counted the number of carbon-black-suspension-perfused microvessels. We concluded that after temporary cerebral ischemia, secondary focal ischemia was induced by microvascular obstruction compressed by swollen astrocytic end-feet, resulting in delayed focal infarction.


Carboxymethyl Chitosan-Functionalized Polyaniline/Polyacrylonitrile Nano-Fibers for Neural Differentiation of Mesenchymal Stem Cells.
Electroconductive scaffolds based on polyaniline (PANi)/polyacrylonitrile (PAN) were fabricated and surface-functionalized by carboxymethyl chitosan (CMC) as efficient scaffolds for nerve tissue regeneration. The results of scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and water contact angle measurement approved the successful fabrication of CMC-functionalized PANi/PAN-based scaffolds. Human adipose-derived mesenchymal stem cells (hADMSCs) were cultured on the scaffolds for 10 d in the presence or absence of β-carotene (βC, 20 µM) as a natural neural differentiation agent. The MTT and SEM results confirmed the attachment and proliferation of hADMSCs on the scaffolds. The expression of MAP2 at the mRNA and protein levels showed the synergic neurogenic induction effect of CMC-functionalization and βC for hADMSCs on the scaffolds. The CMC-functionalized nanofibrous PANi/PAN-based scaffolds are potential candidates for nerve tissue engineering.


The phytopathogenic bacteria Erwinia carotovora infects Drosophila and activates an immune response.
Although Drosophila possesses potent immune responses, little is known about the microbial pathogens that infect Drosophila. We have identified members of the bacterial genus Erwinia that induce the systemic expression of genes encoding antimicrobial peptides in Drosophila larvae after ingestion. These Erwinia strains are phytopathogens and use flies as vectors; our data suggest that these strains have also evolved mechanisms for exploiting their insect vectors as hosts. Erwinia infections induce an antimicrobial response in Drosophila larvae with a preferential expression of antibacterial versus antifungal peptide-encoding genes. Antibacterial peptide gene expression after Erwinia infection is reduced in two Drosophila mutants that have reduced numbers of hemocytes, suggesting that blood cells play a role in regulating Drosophila antimicrobial responses and also illustrating that this Drosophila-Erwinia interaction provides a powerful model for dissecting host-pathogen relationships.


Four Garlic Viruses Identified by Reverse Transcription-Polymerase Chain Reaction and Their Regional Distribution in Northern Japan.
A specific and highly efficient indexing method for four major garlic viruses, GV1 carlavirus, GV2 potyvirus, onion yellow dwarf virus (OYDV), and mite-borne mosaic virus, was developed using the reverse transcription-polymerase chain reaction method (RT-PCR). When specific primers were synthesized to amplify the coat protein genes of these viruses, the amplified PCR bands had the same expected sizes. This indexing method can be performed using an extremely small amount of leaf tissue (50 mg) to obtain reproducible data. Diseased garlic plants collected from five fields in northern Japan were tested using this method. As GV2 was detected in all the fields, and high frequency of disease symptoms was obtained in the case of heavy infection, GV2 may well be a major garlic virus in this region. By contrast, GV1 was detected in only one of the five fields. OYDV and mite-borne mosaic viruses were also detected in various fields and at different frequencies. Judging from the virus indexing data and the disease symptoms, these two viruses appeared to cause severe symptoms in the case of a mixed infection with GV2. The frequency of virus re-infection to virus-free clones was low in isolated fields, and GV2 was the major virus that re-infected virus-free clones.


Efficient transduction of non-human primate motor neurons after intramuscular delivery of recombinant AAV serotype 6.
Retrograde transport of viral vectors in the rodent spinal cord provides a powerful means to administer a therapeutic transgene from the innervated musculature. With the aim of scaling up this approach to non-human primates, we have injected recombinant adeno-associated vectors (rAAV) serotype 6 expressing enhanced green fluorescent protein (eGFP) into the gastrocnemius muscle of African green monkeys to determine whether this results in efficient transgene delivery to lumbar motor neurons. Cells expressing eGFP were observed across more than 1 cm of the spinal cord 4 weeks after intramuscular injection, reaching more than half of motor neurons in some cross-sections. Furthermore, quantitative PCR on the spinal cord tissue confirmed that eGFP expression within motor neurons was due to bona fide retrograde transport of the vector genome from the muscle. Although infiltrations of macrophages and lymphocytes were observed in the rAAV2/6-injected muscle, there was no detectable immune response within the transduced region of the spinal cord. These findings imply that retrograde delivery of rAAV serotype 6 in a primate species constitutes a non-invasive and robust approach to transduce motor neurons, a crucial target cell population in neurodegenerative disorders, such as amyotrophic lateral sclerosis and spinal muscular atrophy.


The primer extension assay.
In the primer extension assay, the transcription start site for a gene is determined experimentally by identifying the 5' end of the encoded messenger RNA (mRNA). The protocol begins with a primer, usually a synthetic oligonucleotide of about 20 residues, that is complementary to an mRNA sequence ∼50-150 nucleotides downstream of the anticipated 5' end. The primer is 5'-end-labeled using [γ-(32)P]ATP and T4 polynucleotide kinase and is annealed to the specific mRNA molecules within an RNA sample. Reverse transcriptase (RT), deoxyribonucleoside triphosphates, and appropriate buffer components are added to the primer-mRNA hybrids to catalyze elongation of the primer to the 5' end of the mRNA. The resulting radiolabeled complementary DNA (cDNA) products are analyzed by denaturing polyacrylamide gel electrophoresis, followed by autoradiography. The sizes of the bands detected on the gel, as compared to an adjacent sequencing ladder or molecular weight standards, provide a measure of the distance from the 5' end of the synthetic oligonucleotide to the beginning of the mRNA transcripts. In theory, the 3' end of the cDNA will coincide with the 5' end of the mRNA. Thus, the size of the radiolabeled cDNAs should represent the distance from the labeled 5' end of the primer to the 5' end of the mRNA (i.e., the 3' end of the cDNA). If the labeled cDNA products are within the resolution range of the gel, the transcription start site can be determined with an accuracy of plus or minus one nucleotide.


MRI-based inverse finite element approach for the mechanical assessment of patellar articular cartilage from static compression test.
The mechanical property of articular cartilage determines to a great extent the functionality of diarthrodial joints. Consequently, the early detection of mechanical and, thus, functional changes of cartilage is crucial for preventive measures to maintain the mobility and the quality of life of individuals. An alternative to conventional mechanical testing is the inverse finite element approach, enabling non-destructive testing of the tissue. We evaluated a method for the assessment of the equilibrium material properties of the patellar cartilage based on magnetic resonance imaging during patellofemoral compression. We performed ex vivo testing of two equine patellas with healthy cartilage, one with superficial defects, and one with synthetically degenerated cartilage to simulate a pre-osteoarthritic stage. Static compression with 400 N for 2 h resulted in morphological changes comparable to physiological in vivo deformations in humans. We observed a decrease of the equilibrium Young's modulus of the degenerated cartilage by -59%, which was in the range of the results from indentation (-74%) and confined compression tests (-58%). With the reported accuracy of magnetic resonance imaging and its reproducibility, the results indicate the potential to measure differences in Young's modulus with regard to cartilage degeneration and consequently to distinguish between healthy and pre-osteoarthritic cartilage.


Computed tomography of the tarsal joint in clinically normal dogs.
To use computed tomography to provide a detailed description of tarsal joint structures in clinically normal dogs. 6 clinically normal adult mixed-breed dogs weighing 25 to 35 kg and one 12-month-old Bullmastiff weighing 65 kg. To perform computed tomography (CT) of both tarsal regions, dogs were anesthetized and placed in ventral recumbency. One- and 2-mm contiguous slices were obtained, using a third generation CT scanner Individual images were reviewed, using bone (window width = 3,500 Hounsfield units; window level = 500 Hounsfield units) and soft-tissue (window width = 400 Hounsfield units; window level = 66 Hounsfield units) settings. After euthanasia, the hind limbs from the Bullmastiff were removed and frozen at -18 C. Tarsal joints were sectioned into approximately 1-mm-thick slab sections, using a cryomicrotome. Anatomic sections were photographed and compared with the corresponding CT images. Computed tomographic reconstructions of the tarsocrural joint were created in sagittal and dorsal planes. Structures on the CT images were matched with structures in the corresponding anatomic sections. The entire tarsocrural joint surface could be evaluated on the reconstructed images in the sagittal and dorsal planes. CT images provide full anatomic detail of the bony structures of the tarsal joint in dogs. Tendons and large blood vessels can also be evaluated. These results could be used as a basis for evaluation of CT images of the hind limbs of dogs with tarsal joint injuries.


Clinical and public health research using methylated DNA immunoprecipitation (MeDIP): a comparison of commercially available kits to examine differential DNA methylation across the genome.
The methylated DNA immunoprecipitation method (MeDIP) is a genome-wide, high-resolution approach that detects DNA methylation with oligonucleotide tiling arrays or high throughput sequencing platforms. A simplified high-throughput MeDIP assay will enable translational research studies in clinics and populations, which will greatly enhance our understanding of the human methylome. We compared three commercial kits, MagMeDIP Kit TM (Diagenode), Methylated-DNA IP Kit (Zymo Research) and Methylamp™ Methylated DNA Capture Kit (Epigentek), in order to identify which one has better reliability and sensitivity for genomic DNA enrichment. Each kit was used to enrich two samples, one from fresh tissue and one from a cell line, with two different DNA amounts. The enrichment efficiency of each kit was evaluated by agarose gel band intensity after Nco I digestion and by reaction yield of methylated DNA. A successful enrichment is expected to have a 1:4 to 10:1 conversion ratio and a yield of 80% or higher. We also evaluated the hybridization efficiency to genome-wide methylation arrays in a separate cohort of tissue samples. We observed that the MagMeDIP kit had the highest yield for the two DNA amounts and for both the tissue and cell line samples, as well as for the positive control. In addition, the DNA was successfully enriched from a 1:4 to 10:1 ratio. Therefore, the MagMeDIP kit is a useful research tool that will enable clinical and public health genome-wide DNA methylation studies.


A community change in the algal endosymbionts of a scleractinian coral following a natural bleaching event: field evidence of acclimatization.
The symbiosis between reef-building corals and their algal endosymbionts (zooxanthellae of the genus Symbiodinium) is highly sensitive to temperature stress, which makes coral reefs vulnerable to climate change. Thermal tolerance in corals is known to be substantially linked to the type of zooxanthellae they harbour and, when multiple types are present, the relative abundance of types can be experimentally manipulated to increase the thermal limits of individual corals. Although the potential exists for this to translate into substantial thermal acclimatization of coral communities, to date there is no evidence to show that this takes place under natural conditions. In this study, we show field evidence of a dramatic change in the symbiont community of Acropora millepora, a common and widespread Indo-Pacific hard coral species, after a natural bleaching event in early 2006 in the Keppel Islands (Great Barrier Reef). Before bleaching, 93.5% (n=460) of the randomly sampled and tagged colonies predominantly harboured the thermally sensitive Symbiodinium type C2, while the remainder harboured a tolerant Symbiodinium type belonging to clade D or mixtures of C2 and D. After bleaching, 71% of the surviving tagged colonies that were initially C2 predominant changed to D or C1 predominance. Colonies that were originally C2 predominant suffered high mortality (37%) compared with D-predominant colonies (8%). We estimate that just over 18% of the original A. millepora population survived unchanged leaving 29% of the population C2 and 71% D or C1 predominant six months after the bleaching event. This change in the symbiont community structure, while it persists, is likely to have substantially increased the thermal tolerance of this coral population. Understanding the processes that underpin the temporal changes in symbiont communities is key to assessing the acclimatization potential of reef corals.


Water transport and functional dynamics of aquaporins in osmoregulatory organs of fishes.
Aquaporins play distinct roles for water transport in fishes as they do in mammals-both at the cellular, organ, and organismal levels. However, with over 32,000 known species of fishes inhabiting almost every aquatic environment, from tidal pools, small mountain streams, to the oceans and extreme salty desert lakes, the challenge to obtain consensus as well as specific knowledge about aquaporin physiology in these vertebrate clades is overwhelming. Because the integumental surfaces of these animals are in intimate contact with the surrounding milieu, passive water loss and uptake represent two of the major osmoregulatory challenges that need compensation. However, neither obligatory nor regulatory water transport nor their mechanisms have been elucidated to the same degree as, for example, ion transport in fishes. Currently fewer than 60 papers address fish aquaporins. Most of these papers identify "what is present" and describe tissue expression patterns in various teleosts. The agnathans, chondrichthyans, and functionality of fish aquaporins generally have received little attention. This review emphasizes the functional physiology of aquaporins in fishes, focusing on transepithelial water transport in osmoregulatory organs in euryhaline species - primarily teleosts, but covering other taxonomic groups as well. Most current knowledge comes from teleosts, and there is a strong need for related information on older fish clades. Our survey aims to stimulate new, original research in this area and to bring together new collaborations across disciplines.


Change in public perspective on cholesterol and heart disease. Results from two national surveys.
The National Heart, Lung and Blood Institute, Bethesda, Md, and the Food and Drug Administration, Washington, DC, sponsored two national probability telephone surveys (N = 4000) of adults to assess attitudes and knowledge about heart disease risk from high blood cholesterol levels and the public's efforts to lower blood cholesterol levels. The first survey was conducted in 1983, before release of the results from the Lipid Research Clinics Coronary Primary Prevention Trial, which showed that a reduction in the blood cholesterol level reduced coronary heart disease; the second survey was conducted in 1986. The percentage of adults who believed that reducing high blood cholesterol levels would have a large effect on heart disease increased from 64% in 1983 to 72% in 1986, so that the importance attached to reducing high blood cholesterol levels approached that attributed to reducing smoking and high blood pressure. In 1983, 35% of adults reported that they had their cholesterol level checked vs 46% in 1986. In both years, diet changes were most frequently chosen (greater than 60%) as ways to control the blood cholesterol level; reducing dietary fat was believed to be as important as reducing dietary cholesterol. By 1986, 23% of adults reported that they made dietary changes specifically to lower their blood cholesterol level, up from 14% in 1983. These comparative data show gains in public awareness and action relating to high blood cholesterol level risk. The data can be used to develop education programs.