1/7. A novel transthyretin mutation associated with familial amyloidotic polyneuropathy.We characterized the mutation associated with familial amyloidotic polyneuropathy in a Japanese patient. sequence analysis of polymerase chain reaction-amplified exons of the transthyretin gene revealed a novel point mutation resulting in a substitution of arginine for glycine at position 47. The mutation was confirmed using allele-specific olgonucleotide hybridization procedures. This most likely represents a de novo mutation since neither parent carries the mutant allele.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
2/7. A new glucocerebrosidase-gene missense mutation responsible for neuronopathic gaucher disease in Japanese patients.We have identified a new T-to-A single-base substitution at nucleotide 3548 (in the genomic sequence) in exon 6 in the glucocerebrosidase gene from a patient with gaucher disease type 3. This mutation caused a substitution of isoleucine for phenylalanine at amino acid residue 213 (of 497 residues in the mature protein). By in vitro expression study in cultured mammalian cells, this mutation resulted in deficient activity of glucocerebrosidase. By allele-specific oligonucleotide hybridization of selectively PCR-amplified dna from eight unrelated Japanese gaucher disease patients, this mutant allele was observed in other neuronopathic Japanese gaucher disease patients, in moderately frequent occurrence (three of six neuronopathic patients). This observation suggests that this allele was one of severe [corrected] alleles which were related to the development of neurological manifestations of gaucher disease.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
3/7. Two novel variants of transthyretin identified in Japanese cases with familial amyloidotic polyneuropathy: transthyretin (Glu42 to Gly) and transthyretin (Ser50 to Arg).Two mutant genes coding for two different variants of transthyretin were identified in two independent kindreds with familial amyloidotic polyneuropathy. A single base change from A to G was identified in exon 2 of transthyretin gene in two brothers from the first kindred. This base change led to replacement of glutamate by glycine at position 42 of 127-residue molecule. In a patient from the second kindred, T to G transversion in exon 3 of transthyretin gene led to replacement of Ser by Arg at position 50. The two mutants were discovered by randomly sequencing recombinant clones containing the entire length of each one of the four exons selectively amplified by polymerase chain reaction. The base change produced a new restriction site for Hae III and Cfr 13 I in the exon 2 and for Mva I in the exon 3, respectively. Restriction fragment length polymorphisms and allele-specific oligonucleotide hybridizations confirmed the base changes. The accurate detection of the new mutant genes is hereafter possible by these procedures.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
4/7. HTLV-I and chronic nervous diseases: present status and a look into the future.Three entities--multiple sclerosis, tropical spastic paraparesis, and human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM)--may represent manifestations of the same disease, with HTLV-I-like virus playing a role in their etiology. Tests for the presence of antibodies reacting with either HTLV-I-like virions or with p24 (gag) antigen, expression of HTLV-I antigen by cells of peripheral blood lymphocytes or cerebrospinal fluid, and viral sequences detected by in situ hybridization are essential to establish the role of HTLV-I-like virus in the disease. It is not yet known whether an incomplete form of the virus persists in the tissue following initial infection or whether the virus in question shares the gag protein with HTLV-I but carries the envelope of a different virus. It is recommended that investigative units comprising neurologists and laboratory workers be established as soon as possible to pursue vigorously the leads that may throw some light on the etiology of chronic neurological diseases such as multiple sclerosis.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |
5/7. Molecular cloning of human T-cell lymphotrophic virus type I-like proviral genome from the peripheral lymphocyte dna of a patient with chronic neurologic disorders.Human T-cell lymphotropic virus type 1 (HTLV-I), the etiologic agent of human T-cell leukemia, has recently been shown to be associated with neurologic disorders such as tropical spastic paraparesis, HTLV-associated myelopathy, and possibly with multiple sclerosis. In this communication, we have examined one specific case of neurologic disorder that can be classified as multiple sclerosis or tropical spastic paraparesis. The patient suffering from chronic neurologic disorder was found to contain antibodies to HTLV-I envelope and gag proteins in his serum and cerebrospinal fluid. lymphocytes from peripheral blood and cerebrospinal fluid of the patient were shown to express viral rna sequences by in situ hybridization. Southern blot analysis of the patient lymphocyte dna revealed the presence of HTLV-I-related sequences. Blot-hybridization analysis of the rna from fresh peripheral lymphocytes stimulated with interleukin 2 revealed the presence of abundant amounts of genomic viral rna with little or no subgenomic rna. We have cloned the proviral genome from the dna of the peripheral lymphocytes and determined its restriction map. This analysis shows that this proviral genome is very similar if not identical to that of the prototype HTLV-I genome.- - - - - - - - - - ranking = 2keywords = hybridization (Clic here for more details about this article) |
6/7. The polymerase chain reaction in the diagnosis of lyme neuroborreliosis.The polymerase chain reaction is sensitive and specific in the detection of defined dna sequences and holds promise for diagnosing the presence of fastidious microorganisms in human infectious diseases. We developed a methodology for nested polymerase chain reaction and hybridization analysis of the cerebrospinal fluid using primers from a genomic borrelia burgdorferi sequence and applied it to the cerebrospinal fluid (CSF) of patients suspected of having lyme neuroborreliosis and other diseases. polymerase chain reaction and hybridization demonstrated extremely high sensitivity for spirochetal dna, and was highly specific, with a false-positivity rate of less than 3%. However, the results were negative or indeterminate in 54% of CSF samples from patients with definite or probable disease, indicating an absence, or extremely low level, of spirochetes or spirochetal dna in a significant percentage of patients with lyme neuroborreliosis. polymerase chain reaction and hybridization of the CSF can thus be considered a useful adjunct in diagnosis, but its negativity does not rule out lyme neuroborreliosis.- - - - - - - - - - ranking = 3keywords = hybridization (Clic here for more details about this article) |
7/7. epidermodysplasia verruciformis with neurological manifestations.BACKGROUND: epidermodysplasia verruciformis (EV) is a rare, inherited disorder in which there is widespread and persistent infection by multiple subtypes of human papilloma virus, tinea versicolor-like lesions and plaques, and frequently malignant manifestations. MATERIALS AND methods: We report two cases of EV-a sister and brother aged 14 and 18 years respectively. Both had classical skin lesions together with neurological manifestations and deafness. In addition the man had plantar hyperkeratosis. They were treated with etretinate. CONCLUSIONS: PCR and dna hybridization of skin lesions from the man contained HPV-20 and HPV-57. He was treated with long-term oral acitretin; the warty lesions became partly or wholly flattened and the plantar hyperkeratosis showed a remarkable improvement. The woman died 10 years later as a result of metastasizing breast cancer.- - - - - - - - - - ranking = 1keywords = hybridization (Clic here for more details about this article) |