Absence of alpha-sarcoglycan and novel missense mutations in the alpha-sarcoglycan gene in a young British girl with muscular dystrophy

An 11-year-old white female presented with progressive proximal muscle weakness and marked calf hypertrophy. Muscle biopsy showed severe dystrophy with normal expression of dystrophin. There was complete absence of the 50kDa dystrophin-associated glycoprotein (alpha-sarcoglycan). DNA analysis showed novel point mutations (one missense and one splicing) in the alpha-sarcoglycan gene at chromosomal location 17q21, confirming the diagnosis of limb-girdle muscular dystrophy type 2D (LGMD-2D). We believe this is one of the first confirmed white cases of primary alpha-sarcoglycanopathy identified in the UK. This case supports the assumption of a wide geographic prevalence of severe childhood onset autosomal recessive muscular dystrophy and genetic heterogeneity. In the future, with improved diagnostic accuracy it is likely that more cases demonstrating primary or secondary deficiency of alpha-sarcoglycan will be identified. We would recommend staining for dystrophin-associated glycoproteins (sarcoglycans) in all new cases of muscular dystrophy with normal dystrophin, and confirmation with DNA analysis where possible.     

大鼠海马注射胍基琥珀酸引起的惊厥及对神经细胞的损害作用

大鼠海马注射微量胍基琥珀酸,可引起典型的阵挛性惊厥和癫痫样放电,用高倍显微镜进行组织学分析发现,胍基琥珀酸能广泛损伤大鼠注射侧海马CA₁部位的锥体细胞。典型的抗癫痫药苯巴比妥和苯妥英钠不能对抗上述作用,而非竞争性NMDA受体的拮抗剂氯胺酮却有对抗作用,且能预防其对海马神经细胞的损害。结果提示,胍基琥珀酸在诱发惊厥和对神经细胞的损伤方面与兴奋性氨基酸 谷氨酸的作用非常相似。同时也说明,其作用与NMDA受体有一定的关系。     

Subcellular localization of the Huntington's disease gene product in cell lines by immunofluorescence and biochemical subcellular fractionation

Huntington's disease is a progressive neurodegenerative disorder, which is caused by expansion of a polymorphic (CAG)n repeat in the coding region of the Huntington's disease gene. The function of huntingtin has not been elucidated so far. Accordingly, detailed subcellular localization studies remain useful. In an immunohistochemical study, we have reported huntingtin to be present in the cytoplasm of cells in the majority of the tissues studied. In addition, we detected a signal in the nucleus of cells in some tissues, including neuronal cells. We have further extended these studies in various mammalian cell lines, using a panel of (affinity-purified) polyclonal huntingtin antibodies in immunofluorescence, confocal laser scanning microscopy and biochemical subcellular fractionation studies. In mouse embryonic fibroblasts, human skin fibroblasts and in mouse neuroblastoma cells huntingtin was present in the cytoplasm. All five antibodies, directed against different parts of huntingtin, also showed a signal in the nucleus. This signal could be competed by the original antigen. The localization of huntingtin in both cytoplasm and nucleus, was confirmed by biochemical subcellular fractionation studies. However, in most other studies, a nuclear location for huntingtin has not been found. Our results suggest, however, that besides its function(s) in the cytoplasm, a nuclear function of huntingtin at some stages of differentiation or in some phases of the cell cycle may not be excluded.      

Integrin function in chronic lymphocytic leukemia

Integrins are central to many aspects of the tissue localization of normal and malignant lymphocytes. We examined how integrin function, rather than simple expression, might determine disease behavior in chronic lymphocyte leukemia (CLL). Using fluorescence-activated cell sorting (FACS) and immunoprecipitation, we first established the precise integrin heterodimer expression of a representative group of CLL patients (beta1 consistently present, with variable alpha 3, alpha 4, and alpha 5; alpha 4 beta 7 often expressed; alpha L beta 2 high; alpha V beta 3 absent). Regarding function, we initially examined the ability of CLL cells to interact with endothelium, because such interaction is the initial event determining the entry of CLL lymphocytes into tissues. The abnormal lymphocytes were shown to bind at low levels to unstimulated endothelium via beta 2/intercellular adhesion molecule (ICAM). However, when the endothelium was stimulated, markedly enhanced interaction with endothelium was observed in approximately half the cases; in these patients, the neoplastic population expressed alpha 4 beta 1, which conferred the ability to adhere strongly to stimulated endothelium via the alpha 4 beta 1 ligand, vascular cellular adhesion molecule-1 (VCAM-1). In relation to the migration of CLL cells within tissues, the abnormal lymphocytes showed differential binding to various adhesive proteins; they did not attach to basement membrane components, but displayed variable adhesion to fibronectin (FN). Finally, we examined the role of cell activation in these processes, and showed that activated CLL lymphocyte populations showed an increased capacity to adhere to both endothelium and matrix. Moreover, ex vivo CLL cells showed no capacity to migrate through endothelium/stroma, but were able to do so after cytokine stimulation. These studies show how the constitutive integrin expression/function, the intrinsic activation state of the cell, and the capacity of cytokines to modify integrin-mediated function all combine to determine the different patterns of clinical disease observed in CLL.     

Evidence for subtelomeric exchange of 3.3 kb tandemly repeated units between chromosomes 4q35 and 10q26: implications for genetic counselling and etiology of FSHD1

Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant myopathy, clinically characterized by asymmetric weakness of muscles in the face, shoulder girdle and upper arm. Deletion of an integral number of 3.3 kb repeated units within a highly polymorphic EcoRI fragment at chromosome 4q35, generating a relatively short EcoRI fragment (< 35 kb), has been shown to cause FSHD1. Probe p13E-11 detects these short fragments in FSHD1 patients, and has therefore been used for diagnostic DNA analysis. However, the reliability of this analysis has been hampered by cross-hybridization of p13E-11 to chromosome 10q26-linked EcoRI fragments of comparable size, which also contain a variable number of 3.3 kb repeated units. Recently, a BinI restriction site was identified within each of the repeated units derived from chromosome 10q26, which enables differentiation of the two polymorphic p13E-11 loci in most cases without haplotype analysis. Remarkably, applying the differential analysis to screen DNA of 160 Dutch cases referred to us for FSHD1 diagnosis, we obtained evidence for subtelomeric exchange of 3.3 kb repeated units between chromosomes 4q35 and 10q26 in affected and unaffected individuals. Subsequently, analysis of 50 unrelated control samples indicated such exchange between chromosomes 4q35 and 10q26 in at least 20% of the population. These subtelomeric rearrangements have generated a novel interchromosomal polymorphism, which has implications for the specificity and sensitivity of the differential restriction analysis for diagnostic purposes. Moreover, the high frequency of the interchromosomal exchanges of 3.3 kb repeated units suggests that they probably do not contain (part of) the FSHD1 gene, and supports position effect variegation as the most likely mechanism for FSHD1.      

Improved pulmonary nodule detection with scanning equalization radiography

The potential for improved pulmonary nodule detection with scanning equalization radiography (SER) was evaluated by means of observer performance testing during the interpretation of posteroanterior conventional radiographs and SER images of an anthropomorphic chest phantom with simulated nodules. A test set of 200 conventional and 200 SER radiographs of phantoms containing either one nodule or none was interpreted by four radiologists attempting to detect a nodule and indicate a confidence value. Their ability to detect nodules positioned over the lung was slightly improved with SER compared with conventional radiography (sensitivity, .56 vs .70); for nodules over the mediastinum or diaphragmatic areas, it was much improved (sensitivity, .29 vs .64). The results were also analyzed with receiver-operating characteristic methods, which revealed a significant improvement in lesion detect-ability over the thicker body parts with SER images. The capability of equalized chest radiographs to provide improved lesion detectability suggests that SER may set a new standard for film-based chest radiography and have a large clinical application.     

Fetal hyaloid artery: timing of regression with US

Large-aperture, dynamically focused ultrasonic imaging permits noninvasive, anatomic study of the eye at the millimeter level in the second and third trimesters of pregnancy. The authors report their observations of the hyaloid artery in 210 of 219 fetuses examined with this technique. This vessel is seen in fetuses of 20 weeks gestational age or less and regresses spontaneously at the start of the third trimester. The 210 subjects included 100 who were examined at gestational ages of 16-32 weeks or more and 85 healthy fetuses and 25 with pathologic findings at birth who were examined at 34 weeks gestational age to term. The presence of the hyaloid artery in the mid third trimester was uncommon in healthy subjects (less than 1%) and was not seen in any beyond 29.9 weeks gestational age. However, in nine of the 25 fetuses with abnormalities (five with trisomy syndromes), the vessel was seen at 30.8-36.8 weeks gestational age. The temporarily delayed regression of the hyaloid artery may occur with trisomy 21 syndrome and other forms of retarded brain development.