A meckelin-filamin A interaction mediates ciliogenesis

MKS3, encoding the transmembrane receptor meckelin, is mutated in Meckel-Gruber syndrome (MKS), an autosomal-recessive ciliopathy. Meckelin localizes to the primary cilium, basal body and elsewhere within the cell. Here, we found that the cytoplasmic domain of meckelin directly interacts with the actin-binding protein filamin A, potentially at the apical cell surface associated with the basal body. Mutations in FLNA, the gene for filamin A, cause periventricular heterotopias. We identified a single consanguineous patient with an MKS-like ciliopathy that presented with both MKS and cerebellar heterotopia, caused by an unusual in-frame deletion mutation in the meckelin C-terminus at the region of interaction with filamin A. We modelled this mutation and found it to abrogate the meckelin-filamin A interaction. Furthermore, we found that loss of filamin A by siRNA knockdown, in patient cells, and in tissues from Flna(Dilp2) null mouse embryos results in cellular phenotypes identical to those caused by meckelin loss, namely basal body positioning and ciliogenesis defects. In addition, morpholino knockdown of flna in zebrafish embryos significantly increases the frequency of dysmorphology and severity of ciliopathy developmental defects caused by mks3 knockdown. Our results suggest that meckelin forms a functional complex with filamin A that is disrupted in MKS and causes defects in neuronal migration and Wnt signalling. Furthermore, filamin A has a crucial role in the normal processes of ciliogenesis and basal body positioning. Concurrent with these processes, the meckelin-filamin A signalling axis may be a key regulator in maintaining correct, normal levels of Wnt signalling.     

Persistent efficacy of topical eprinomectin against nematode parasites in cattle

Six studies were conducted to evaluate the persistent efficacy of eprinomectin pour-on against experimental challenges with infective nematode larvae in calves. In each study, calves were randomly assigned to one untreated group and up to four test groups, which were treated with eprinomectin at 500 μg/kg body weight at weekly intervals before single bolus challenge. The calves were necropsied approximately 4 weeks after challenge infection for nematode recovery. Eprinomectin pour-on provided ≥90% efficacy against challenge with Haemonchus placei, Trichostrongylus axei and T. colubriformis at 21 days after treatment and against Cooperia oncophora, C. punctata, C. surnabada, Dictyocaulus viviparus, Nematodirus helvetianus, Oesophagostomum radiatum and Ostertagia ostertagi at 28 days after treatment. Received: 14 April 2000 / Accepted: 18 May 2000     

Clinical,biochemical, and genetic features of four patients with short‐chain enoyl‐CoA hydratase (ECHS1) deficiency

Short‐chain enoyl‐CoA hydratase (SCEH or ECHS1) deficiency is a rare inborn error of metabolism caused by biallelic mutations in the gene ECHS1 (OMIM 602292). Clinical presentation includes infantile‐onset severe developmental delay, regression, seizures, elevated lactate, and brain MRI abnormalities consistent with Leigh syndrome (LS). Characteristic abnormal biochemical findings are secondary to dysfunction of valine metabolism. We describe four patients from two consanguineous families (one Pakistani and one Irish Traveler), who presented in infancy with LS. Urine organic acid analysis by GC/MS showed increased levels of erythro‐2,3‐dihydroxy‐2‐methylbutyrate and 3‐methylglutaconate (3‐MGC). Increased urine excretion of methacrylyl‐CoA and acryloyl‐CoA related metabolites analyzed by LC‐MS/MS, were suggestive of SCEH deficiency; this was confirmed in patient fibroblasts. Both families were shown to harbor homozygous pathogenic variants in the ECHS1 gene; a c.476A > G (p.Gln159Arg) ECHS1variant in the Pakistani family and a c.538A > G, p.(Thr180Ala) ECHS1 variant in the Irish Traveler family. The c.538A > G, p.(Thr180Ala) ECHS1 variant was postulated to represent a Canadian founder mutation, but we present SNP genotyping data to support Irish ancestry of this variant with a haplotype common to the previously reported Canadian patients and our Irish Traveler family. The presence of detectable erythro‐2,3‐dihydroxy‐2‐methylbutyrate is a nonspecific marker on urine organic acid analysis but this finding, together with increased excretion of 3‐MGC, elevated plasma lactate, and normal acylcarnitine profile in patients with a Leigh‐like presentation should prompt consideration of a diagnosis of SCEH deficiency and genetic analysis of ECHS1. ECHS1 deficiency can be added to the list of conditions with 3‐MGA.     

Reduced risk of peanut sensitization following exposure through breast-feeding and early peanut introduction

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Different strains of Mycobacterium tuberculosis cause various spectrums of disease in the rabbit model of tuberculosis

The rabbit model of tuberculosis has been used historically to differentiate between Mycobacterium tuberculosis and Mycobacterium bovis based on their relative virulence in this animal host. M. tuberculosis infection in market rabbits is cleared over time, whereas infection with M. bovis results in chronic, progressive, cavitary disease leading to death. Because of the innate resistance of commercial rabbits to M. tuberculosis, 320 to 1,890 log-phase, actively growing inhaled bacilli were required to form one grossly visible pulmonary tubercle at 5 weeks. The range of inhaled doses required to make one tubercle allows us to determine the relative pathogenicities of different strains. Fewer inhaled organisms of the M. tuberculosis Erdman strain were required than of M. tuberculosis H37Rv to produce a visible lesion at 5 weeks. Furthermore, with the Erdman strain, only 7 of 15 rabbits had healed lesions at 16 to 18 weeks; among the other animals, two had chronic, progressive cavitary disease, a phenotype usually seen only with M. bovis infection. Genotypic investigation of the Erdman strain with an H37Rv-based microarray identified gene differences in the RD6 region. Southern blot and PCR structural genetic analysis showed significant differences between M. tuberculosis strains in this region. Correlation of the relative pathogenicity, including disease severity, in the rabbit model with the strain genotype may help identify stage-specific M. tuberculosis genes important in human disease.     

Child functional independence and maternal psychosocial stress as risk factors threatening adaptation in mothers of physically or sensorially handicapped children

This study investigated the contribution of child functional independence and maternal psychosocial stress to the adaptation of 119 mothers. Each mother had a child, aged 2-18 years, with a physical or sensory disability. Multiple dimensions of each construct were measured through self-report. Child functional independence did not uniquely explain variation in mothers' adaptation. However, maternal stress was uniquely associated with maternal mental health, but not physical health or social functioning, even when controlling for demographic status, disability type, and functional independence. Daily hassles and handicap-related psychosocial stress in particular put mothers at risk for reporting mental health problems.     

A randomized pilot trial of brief versus prolonged heparin after successful reperfusion in acute myocardial infarction

Controversy exists as to whether and how long heparin treatment is necessary after infarct vessel recanalization. To determine the role of heparin, patients with suitable angiographic features after reperfusion therapy were randomly allocated to receive a brief infusion of intravenous heparin for less than or equal to 24 hours (group 1), adjusted to a partial thromboplastin time of 2 times control or a prolonged infusion for greater than or equal to 72 hours (group 2), using the same titration mechanism. Patients were excluded for complex intimal dissections, large residual filling defects, less than Thrombolysis in Myocardial Infarction grade 3 flow pattern or greater than 50% residual stenosis. Heparin was sustained except for discontinuation 2 to 4 hours before periaccess sheath removal, or if significant bleeding (greater than or equal to 2 units blood transfusion) occurred. The primary endpoints were 1-week patency determined by repeat catheterization or recurrent ischemia, or both, and the incidence of bleeding complications. Fifty patients were randomized, 25 in both groups. Baseline variables were similar; 14 group 1 and 15 group 2 patients received thrombolytic treatment; 20 patients in each group had coronary angioplasty. Two documented reocclusions occurred in both groups. Significant bleeding complications occurred in 0 of 25 (0%) group 1 versus 6 of 25 (24%) group 2 patients (p less than 0.05). Thus, in low-risk patients after successful reperfusion, prolonged heparin therapy does not protect against rethrombosis and is associated with a significantly higher rate of bleeding complications. Therefore, prolonged heparin therapy for greater than 24 hours does not appear to be justified in low-risk patients with successful reperfusion.     

Myocardial perfusion imaging and gated cardiac blood pool scanning: clinical application.

Myocardial perfusion imaging with thallium-201 and gated cardiac blood pool scanning are finding increasing use in clinical cardiology. These noninvasive techniques have been found useful in detecting myocardial infarction independent of the electrocardiogram and determining the site and extent of the infarct as well as its effect on left ventricular function. These studies provide important prognostic data and are proving to be of value in evaluating patients with cardiogenic shock. Neither the thallium-201 myocardial perfusion image nor the gated cardiac blood pool scan can distinguish between acute and chronic myocardial damage. In clinical situations where this is important, infarct avid imaging with technetium-99m pyrophosphate allows determination of whether a given perfusion defect or wall motion abnormality is acute. Myocardial perfusion imaging with thallium-201 at rest and after exercise is also proving to be of value in evaluating patients with suspected ischemic heart disease. Initial studies suggest that the technique may be more sensitive than exercise electrocardiography and is of special value in minimizing the occurrence of false positive exercise tests for the diagnosis of ischemic heart disease. The combined tracers technique is also of value in the evaluation of patients undergoing coronary bypass graft surgery and those with cardiomyopathy.