Transcriptome assessment of the Pompe (Gaa-/-) mouse spinal cord indicates widespread neuropathology.

Pompe disease, caused by deficiency of acid alpha-glucosidase (GAA), leads to widespread glycogen accumulation and profound neuromuscular impairments. There has been controversy, however, regarding the role of central nervous system pathology in Pompe motor dysfunction. We hypothesized that absence of GAA protein causes progressive activation of neuropathological signaling, including pathways associated with cell death. To test this hypothesis, genomic data (Affymetrix Mouse Gene Array 2.0ST) from the midcervical spinal cord in 6 and 16 mo old Pompe (Gaa-/-) mice were evaluated (Broad Institute Molecular Signature Database), along with spinal cord histology. The midcervical cord was selected because it contains phrenic motoneurons, and phrenic-diaphragm dysfunction is prominent in Pompe disease. Several clinically important themes for the neurologic etiology of Pompe disease emerged from this unbiased genomic assessment. First, pathways associated with cell death were strongly upregulated as Gaa-/- mice aged, and motoneuron apoptosis was histologically verified. Second, proinflammatory signaling was dramatically upregulated in the Gaa-/- spinal cord. Third, many signal transduction pathways in the Gaa-/- cervical cord were altered in a manner suggestive of impaired synaptic function. Notably, glutamatergic signaling pathways were downregulated, as were "synaptic plasticity pathways" including genes related to neuroplasticity. Fourth, many genes and pathways related to cellular metabolism are dysregulated. Collectively, the data unequivocally confirm that systemic absence of GAA induces a complex neuropathological cascade in the spinal cord. Most importantly, the results indicate that Pompe is a neurodegenerative condition, and this underscores the need for early therapeutic intervention capable of targeting the central nervous system.

Ampakine CX717 potentiates intermittent hypoxia-induced hypoglossal long-term facilitation.

Glutamatergic currents play a fundamental role in regulating respiratory motor output and are partially mediated by α-amino-3-hydroxy-5-methyl-isoxazole-propionic acid (AMPA) receptors throughout the premotor and motor respiratory circuitry. Ampakines are pharmacological compounds that enhance glutamatergic transmission by altering AMPA receptor channel kinetics. Here, we examined if ampakines alter the expression of respiratory long-term facilitation (LTF), a form of neuroplasticity manifested as a persistent increase in inspiratory activity following brief periods of reduced O2 [intermittent hypoxia (IH)]. Current synaptic models indicate enhanced effectiveness of glutamatergic synapses after IH, and we hypothesized that ampakine pretreatment would potentiate IH-induced LTF of respiratory activity. Inspiratory bursting was recorded from the hypoglossal nerve of anesthetized and mechanically ventilated mice. During baseline (BL) recording conditions, burst amplitude was stable for at least 90 min (98 ± 5% BL). Exposure to IH (3 × 1 min, 15% O2) resulted in a sustained increase in burst amplitude (218 ± 44% BL at 90 min following final bout of hypoxia). Mice given an intraperitoneal injection of ampakine CX717 (15 mg/kg) 10 min before IH showed enhanced LTF (500 ± 110% BL at 90 min). Post hoc analyses indicated that CX717 potentiated LTF only when initial baseline burst amplitude was low. We conclude that under appropriate conditions ampakine pretreatment can potentiate IH-induced respiratory LTF. These data suggest that ampakines may have therapeutic value in the context of hypoxia-based neurorehabilitation strategies, particularly in disorders with blunted respiratory motor output such as spinal cord injury.

The effects of an oral preparation containing hyaluronic acid (Oralvisc®) on obese knee osteoarthritis patients determined by pain, function, bradykinin, leptin, inflammatory cytokines, and heavy water analyses.

The purpose of this study was to determine the effects of an oral preparation containing hyaluronic acid on osteoarthritic knee joint pain and function as well as changes in inflammatory cytokines, bradykinin, and leptin. We also used heavy water to determine the turnover rates of glycosaminoglycans in synovial fluid. This was a double-blind, randomized, placebo-controlled study of 40 subjects over a period of 3 months. Visual analog scale, Western Ontario McMaster pain, and WOMAC function scores were recorded. Serum and synovial fluid were measured by enzyme-linked immunosorbent assays for inflammatory cytokines, bradykinin, and leptin. In 20 subjects, terminal heavy water ingestion was used for spectral analyses of serum and joint fluid samples. There were statistically significant improvements in pain and function. Both serum and synovial fluid samples showed significant decreases for a majority of inflammatory cytokines, leptin, and bradykinin in the oral hyaluronic acid preparation group. Heavy water analyses revealed a significant decrease in hyaluronic acid turnover in the synovial fluid of the treatment group. A preparation containing hyaluronic acid and other glycosaminoglycans holds promise for a safe and effective agent for the treatment for patients with knee osteoarthritis and who are overweight. Further studies will be required to see whether this is a disease-modifying agent.

Effect of bile acid sequestrants on glucose metabolism, hepatic de novo lipogenesis, and cholesterol and bile acid kinetics in type 2 diabetes: a randomised controlled study.

AIMS/HYPOTHESIS: The primary aim of this completed multicentre randomised, parallel, double-blind placebo-controlled study was to elucidate the mechanisms of glucose-lowering with colesevelam and secondarily to investigate its effects on lipid metabolism (hepatic de novo lipogenesis, cholesterol and bile acid synthesis). METHODS: Participants with type 2 diabetes (HbA(1c) 6.7-10.0% [50-86 mmol/mol], fasting glucose <16.7 mmol/l, fasting triacylglycerols <3.9 mmol/l and LDL-cholesterol >1.55 mmol/l) treated with diet and exercise, sulfonylurea, metformin or a combination thereof, were randomised by a central coordinator to either 3.75 g/day colesevelam (n = 30) or placebo (n = 30) for 12 weeks at three clinical sites in the USA. The primary measure was the change from baseline in glucose kinetics with colesevelam compared to placebo treatment. Fasting and postprandial glucose, lipid and bile acid pathways were measured at baseline and post-treatment using stable isotope techniques. Plasma glucose, insulin, total glucagon-like peptide-1 (GLP-1), total glucose-dependent insulinotropic polypeptide (GIP), glucagon and fibroblast growth factor-19 (FGF-19) concentrations were measured during the fasting state and following a meal tolerance test. Data was collected by people blinded to treatment. RESULTS: Compared with placebo, colesevelam improved HbA(1c) (mean change from baseline of 0.3 [SD 1.1]% for placebo [n = 28] and -0.3 [1.1]% for colesevelam [n = 26]), glucose concentrations, fasting plasma glucose clearance and glycolytic disposal of oral glucose. Colesevelam did not affect gluconeogenesis or appearance rate (absorption) of oral glucose. Fasting endogenous glucose production and glycogenolysis significantly increased with placebo but were unchanged with colesevelam (treatment effect did not reach statistical significance). Compared with placebo, colesevelam increased total GLP-1 and GIP concentrations and improved HOMA-beta cell function while insulin, glucagon and HOMA-insulin resistance were unchanged. Colesevelam increased cholesterol and bile acid synthesis and decreased FGF-19 concentrations. However, no effect was seen on fractional hepatic de novo lipogenesis. CONCLUSIONS/INTERPRETATION: Colesevelam, a non-absorbed bile acid sequestrant, increased circulating incretins and improved tissue glucose metabolism in both the fasting and postprandial states in a manner different from other approved oral agents. TRIAL REGISTRATION: ClinicalTrials.gov NCT00596427 FUNDING: The study was funded by Daiichi Sankyo.

Evidence for a Diagenetic Origin of Vera Rubin Ridge, Gale Crater, Mars: Summary and Synthesis of Curiosity's Exploration Campaign.

This paper provides an overview of the Curiosity rover's exploration at Vera Rubin ridge (VRR) and summarizes the science results. VRR is a distinct geomorphic feature on lower Aeolis Mons (informally known as Mount Sharp) that was identified in orbital data based on its distinct texture, topographic expression, and association with a hematite spectral signature. Curiosity conducted extensive remote sensing observations, acquired data on dozens of contact science targets, and drilled three outcrop samples from the ridge, as well as one outcrop sample immediately below the ridge. Our observations indicate that strata composing VRR were deposited in a predominantly lacustrine setting and are part of the Murray formation. The rocks within the ridge are chemically in family with underlying Murray formation strata. Red hematite is dispersed throughout much of the VRR bedrock, and this is the source of the orbital spectral detection. Gray hematite is also present in isolated, gray-colored patches concentrated toward the upper elevations of VRR, and these gray patches also contain small, dark Fe-rich nodules. We propose that VRR formed when diagenetic event(s) preferentially hardened rocks, which were subsequently eroded into a ridge by wind. Diagenesis also led to enhanced crystallization and/or cementation that deepened the ferric-related spectral absorptions on the ridge, which helped make them readily distinguishable from orbit. Results add to existing evidence of protracted aqueous environments at Gale crater and give new insight into how diagenesis shaped Mars' rock record.

A stable isotope method for the simultaneous measurement of matrix synthesis and cell proliferation in articular cartilage in vivo.

OBJECTIVE: Measurements of cell proliferation and matrix synthesis in cartilage explants have identified regulatory factors [e.g., interleukin-1 (IL-1)] that contribute to osteoarthritis and anabolic mediators [e.g., bone morphogenic protein-7 (BMP-7)] that may have therapeutic potential. The objective of this study was to develop a robust method for measuring cell proliferation and glycosaminoglycan synthesis in articular cartilage that could be applied in vivo. METHODS: A stable isotope-mass spectrometry approach was validated by measuring the metabolic effects of IL-1 and BMP-7 in cultures of mature and immature bovine cartilage explants. The method was also applied in vivo to quantify physiologic turnover rates of matrix and cells in the articular cartilage of normal rats. Heavy water was administered to explants in the culture medium and to rats via drinking water, and cartilage was analyzed for labeling of chondroitin sulfate (CS), hyaluronic acid (HA) and DNA. RESULTS: As expected, IL-1 inhibited the synthesis of DNA and CS in cartilage explants. However, IL-1 inhibited HA synthesis only in immature cartilage. Furthermore, BMP-7 was generally stimulatory, but immature cartilage was significantly more responsive than mature cartilage, particularly in terms of HA and DNA synthesis. In vivo, labeling of CS and DNA in normal rats for up to a year indicated half-lives of 22 and 862 days, respectively, in the joint. CONCLUSIONS: We describe a method by which deuterium from heavy water is traced into multiple metabolites from a single cartilage specimen to profile its metabolic activity. This method was demonstrated in tissue culture and rodents but may have significant clinical applications.

Catecholamine uptake, accumulation, and release in acute porphyria.

Hypertension and tachycardia are well known features of acute porphyria and have been shown to be related to increased circulating catecholamines. The mechanism by which circulating catecholamines are increased was studied using the isolated perfused rat heart and human platelets as a model of adrenergic neuronal function. It was found that neither delta-aminolevulinate (ALA) nor porphobilinogen (PBG) blocked uptake or caused release in the isolated perfused rat heart. Platelets from six patients with acute prophyria, three in remission and three latent, with matching normal controls were studied with regard to their uptake of [(3)H]norepinephrine in the presence of ALA or PBG. It was found that ALA and PBG significantly reduced uptake and accumulation of [(3)H]-norepinephrine in patients with acute porphyria; however, no similar reduction in uptake and accumulation was observed in the platelets of normal controls. Therefore, it appears that there is a latent defect in the catecholamine uptake and (or) accumulation of platelets of patients with acute prophyria which only manifests itself in the presence of ALA or PBG. If platelet uptake serves as a model of adrenergic neuron uptake, this suggests that elevated circulating catecholamine levels during acute attacks of acute porphyria are caused at least partially by blockade of re-uptake into the sympathetic neurons.

High-resolution X-ray diffraction with no sample preparation.

It is shown that energy-dispersive X-ray diffraction (EDXRD) implemented in a back-reflection geometry is extremely insensitive to sample morphology and positioning even in a high-resolution configuration. This technique allows high-quality X-ray diffraction analysis of samples that have not been prepared and is therefore completely non-destructive. The experimental technique was implemented on beamline B18 at the Diamond Light Source synchrotron in Oxfordshire, UK. The majority of the experiments in this study were performed with pre-characterized geological materials in order to elucidate the characteristics of this novel technique and to develop the analysis methods. Results are presented that demonstrate phase identification, the derivation of precise unit-cell parameters and extraction of microstructural information on unprepared rock samples and other sample types. A particular highlight was the identification of a specific polytype of a muscovite in an unprepared mica schist sample, avoiding the time-consuming and difficult preparation steps normally required to make this type of identification. The technique was also demonstrated in application to a small number of fossil and archaeological samples. Back-reflection EDXRD implemented in a high-resolution configuration shows great potential in the crystallographic analysis of cultural heritage artefacts for the purposes of scientific research such as provenancing, as well as contributing to the formulation of conservation strategies. Possibilities for moving the technique from the synchrotron into museums are discussed. The avoidance of the need to extract samples from high-value and rare objects is a highly significant advantage, applicable also in other potential research areas such as palaeontology, and the study of meteorites and planetary materials brought to Earth by sample-return missions.

Identification of the Beagle 2 lander on Mars.

The 2003 Beagle 2 Mars lander has been identified in Isidis Planitia at 90.43° E, 11.53° N, close to the predicted target of 90.50° E, 11.53° N. Beagle 2 was an exobiology lander designed to look for isotopic and compositional signs of life on Mars, as part of the European Space Agency Mars Express (MEX) mission. The 2004 recalculation of the original landing ellipse from a 3-sigma major axis from 174 km to 57 km, and the acquisition of Mars Reconnaissance Orbiter High Resolution Imaging Science Experiment (HiRISE) imagery at 30 cm per pixel across the target region, led to the initial identification of the lander in 2014. Following this, more HiRISE images, giving a total of 15, including red and blue-green colours, were obtained over the area of interest and searched, which allowed sub-pixel imaging using super high-resolution techniques. The size (approx. 1.5 m), distinctive multilobed shape, high reflectivity relative to the local terrain, specular reflections, and location close to the centre of the planned landing ellipse led to the identification of the Beagle 2 lander. The shape of the imaged lander, although to some extent masked by the specular reflections in the various images, is consistent with deployment of the lander lid and then some or all solar panels. Failure to fully deploy the panels-which may have been caused by damage during landing-would have prohibited communication between the lander and MEX and commencement of science operations. This implies that the main part of the entry, descent and landing sequence, the ejection from MEX, atmospheric entry and parachute deployment, and landing worked as planned with perhaps only the final full panel deployment failing.

Sources of plasma glucose and liver glycogen in fasted ob/ob mice.

Alterations in intrahepatic carbohydrate fluxes in ob/ob mice and the effects of acute leptin administration were studied in vivo by use of a dual-isotope tracer infusion. Metabolic sources of plasma glucose (gluconeogenesis (GNG) and glycogenolysis) and hepatic glycogen (GNG, direct synthesis and pre-existing) were determined in 20-h-fasted mice infused with [2-13C1]glycerol and [U13C6]glucose for 3 h. Total glucose output (TGO) and the rate of appearance (Ra) of plasma glycerol were measured by isotope dilution. GNG, the direct pathway of hepatic glycogen synthesis and hepatic triose-phosphate flux were determined by mass isotopomer distribution analysis (MIDA). Serum glucose, insulin, leptin and liver glycogen concentrations were also measured. After a 24-h fast, ob/ob mice had 2-fold higher TGO, 2.5-fold elevated liver glycogen content and markedly higher glycogenolytic flux to glucose, absolute GNG and direct glycogen synthesis rates (10-fold increased) compared to the control group. Ob/ob mice also had elevated triose-phosphate flux compared to controls (40 vs. 22 mg/kg lean body mass/min). A model of intrahepatic flux distributions in control and ob/ob mice is presented. In summary, elevated fasting plasma glucose concentrations are due to increased TGO in ob/ob mice, which is maintained by both increased GNG and increased glycogenolysis. Furthermore, the ob/ob mice have major alterations in fasting hepatic carbohydrate fluxes into triose-phosphate pools and glycogen. We support the model that actions of leptin on hepatic glucose metabolism require insulin or other factors.

Validation of the Social Phobia and Anxiety Inventory for Children (SPAI-C) in a sample of Brazilian children.

The purpose of the present study was to examine the factor structure and psychometric properties of the Social Phobia and Anxiety Inventory for Children (SPAI-C), an instrument developed in the United States and applied to a sample of Brazilian schoolchildren. The process included the translation of the original material from English into Portuguese by two bilingual psychiatrists and a back translation by a bilingual physician. Both the front and back translations were revised by a bilingual child psychiatrist. The study was performed using a cross-sectional design and the Portuguese version of the SPAI-C was applied to a sample of 1954 children enrolled in 3rd to 8th grade attending 2 private and 11 public schools. Eighty-one subjects were excluded due to an incomplete questionnaire and 2 children refused to participate. The final sample consisted of 1871 children, 938 girls (50.1%) and 933 boys (49.8%), ranging in age from 9 to 14 years. The majority of the students were Caucasian (89.0%) and the remainder were African-Brazilian (11.0%). The Pearson product-moment correlation showed that the two-week test-retest reliability coefficient was r = 0.780 and Cronbach's alpha was 0.946. The factor structure was almost similar to that reported in previous studies. The results regarding the internal consistency, the test-retest reliability and the factor structure were similar to the findings obtained in studies performed on English speaking children. The present study showed that the Portuguese language version of SPAI-C is a reliable and valid measure of social anxiety for Brazilian children.

Avasucular necrosis of pheochromocytoma followed by spontaneous remission.

Following an acute spontaneous hypertensive crisis and shock a patient with pheochromocytoma was found to have an exceedingly high catecholamine excretion rate. After this episode, the patient remained normotensive and urinary excretion of catecholamines returned to normal. During surgery, a large pheochromocytoma was found and removed that showed avascualr necrosis. In phenochromocytoma, a sudden and exceedingly high rate of catecholamine release may cause intense vasoconstriction both generally and within the tumor itself. In this patient, avascular tumor necrosis led to a spontaneous remission of clinical symptoms.

Tricyclic antidepressants in obsessive-compulsive disorder: antiobsessional or antidepressant agents? II.

The authors explored the relationship between the antiobsessional and antidepressant effects of tricyclic drugs in primary obsessive-compulsive disorders. Study 1 consisted of a controlled 12-week trial with clomipramine (N = 7) and placebo (N = 5); study 2 analyzed the pooled data from 15 patients uniformly selected and treated with either clomipramine or imipramine. Although the antiobsessional and antidepressant effects of the drugs covaried, their antidepressant action was not a prerequisite for their antiobsessional effect. The findings suggest that clomipramine and probably imipramine possess specific antiobsessive effects that are at least partially independent of their antidepressant effects.

Otoneurological examination in panic disorder and agoraphobia with panic attacks: a pilot study.

A battery of vestibular and audiological tests was administered to eight patients with panic disorder and 13 patients with agoraphobia and panic attacks, all of whom experienced dizziness during their panic attacks. Positional or spontaneous nystagmus was present in 67% of the subjects. Abnormal responses were found in caloric testing (56%), rotational testing (35%), and posturography (32%). Pure tone audiograms were abnormal in 26% of the subjects and acoustic reflexes were abnormal in 44% of the subjects. Six of eight patients tested had an abnormal brainstem auditory evoked potential. The possible importance of the findings and their implications for further research are discussed.

Panic, agoraphobia, and vestibular dysfunction.

OBJECTIVE: Otoneurological abnormalities have been reported in panic disorder. The purpose of this investigation was to determine the prevalence of such findings in panic disorder with and without agoraphobia and to discern whether vestibular dysfunction was associated with specific symptoms. METHOD: Clinical audiological and vestibular tests were administered to 30 patients with uncomplicated panic disorder (without agoraphobia or with only mild agoraphobia), 29 patients with panic disorder with moderate to severe agoraphobia, 27 patients with anxiety but no history of panic attacks, 13 patients with depressive disorders but no history of anxiety or panic attacks, and 45 normal comparison subjects. Evaluators were blind to subjects' diagnostic group. Quantitative measures of subjects' discomfort with space and motion and of the frequency of certain symptoms between and during panic attacks were obtained. Anxiety state levels were measured during the vestibular tests. RESULTS: Vestibular abnormalities were common in all the groups but most prevalent in the patients with panic disorder with moderate to severe agoraphobia. Vestibular dysfunction was associated with space and motion discomfort and with frequency of vestibular symptoms between, but not during, panic attacks. There were no major differences between the two panic groups in anxiety levels during vestibular testing. There were no significant differences between groups on the audiological component of the test battery. Exploratory data analysis indicated that the constellation of vestibular tests most specific for agoraphobia was one indicating compensated peripheral vestibular dysfunction. CONCLUSIONS: Subclinical vestibular dysfunction, as identified by clinical tests, may contribute to the phenomenology of panic disorder, particularly to the development of agoraphobia in panic disorder patients.

Anxiety disorders research with African Americans: current status.

Anxiety disorders are the most common psychiatric disorders in the general population of the United States. Recent findings indicate that the prevalence of certain anxiety disorders may be higher in African Americans. In addition, anxiety disorders in African Americans may be manifested differently. Despite the high prevalence and the suggestion of different patterns of manifestation, a paucity of research exists on anxiety disorders in this population. This may be due in part to the pattern of help-seeking behavior among African Americans, a negative perception of research by members of this community, the small number of African American researchers, and perhaps to some extent a general disinterest on the part of researchers. The available research on anxiety disorders in African Americans is critically reviewed, the need for research in the area is highlighted, and ways to facilitate such research are discussed.

The effect of body temperature and cell-mediated immunity on the growth of Mycobacterium marinum and Mycobacterium leprae in mice.

Evidence is presented that the high susceptibility of armadillos to infection with Mycobacterium leprae cannot be explained solely in terms of body temperature because mutant mice maintained with a body temperature similar to that of armadillos do not become heavily infected with M. leprae. The depression of cell-mediated immunity accompanying the low body temperature is not sufficient to produce an overwhelming infection. The results obtained with M. marinum suggest that whereas lack of cell-mediated immunity or a low body temperature result in a moderately enhanced infection in the mouse a combination of both of these factors is required to produce an overwhelming infection involving the internal organs.

At risk for anxiety: I. Psychopathology in the offspring of anxious parents.

OBJECTIVE AND METHOD: Children of parents with anxiety disorders, depressive disorders, mixed anxiety/depressive disorders, and no psychiatric disorder were assessed with semistructured interviews to determine rates of overall psychopathology and to determine specifically the presence of anxiety disorders. RESULTS: Children of the three "high-risk" groups were significantly more likely to have a diagnosable disorder (including anxiety disorders) than offspring of normal parents, but there were no differences among the children from the three parental diagnostic groups. However, when examined specifically for anxiety disorders, offspring of anxious parents were significantly more likely to have only anxiety disorders. Offspring of depressed or mixed anxious/depressed parents had a broader range of disorders and more comorbid disorders. Family socioeconomic status was related to the probability that a child would have a disorder. CONCLUSIONS: Anxiety disorders are common among offspring of anxious and depressed parents. However, when a parent has depression, children exhibit a broader range of psychopathology than when a parent has an anxiety disorder alone.

Assessing community violence: the children's report of exposure to violence.

OBJECTIVE: To describe the development and initial psychometric features of a brief self-report instrument for assessing exposure to community violence, the Children's Report of Exposure to Violence (CREV). METHOD: The CREV was administered to 228 schoolchildren between the ages of 9 and 15 years. A subset (n = 42) was retested 2 weeks later. RESULTS: The CREV has good test-retest reliability, internal consistency, and construct validity. Analyses performed by race and gender revealed no differences in frequency of exposure to community violence as assessed by the CREV. CONCLUSIONS: This initial report suggests that the CREV has promise as a self-report questionnaire, and its construction represents a significant advance in the development of psychometrically sound instruments for use in community violence research.