Biomedical subjects

P Carango

Publications and source records attributed to P Carango.

8 recordsLinked to original sources

PubMed1995

Effect of myotonic dystrophy trinucleotide repeat expansion on DMPK transcription and processing.

The myotonic dystrophy (DM) mutation has been identified as an unstable, expanded (CTG)n repeat in the 3' untranslated region of a gene designated DM protein kinase (DMPK). Both decreased and increased levels of mutant DMPK mRNA as well as decreased levels of protein have been variously reported and invoked to explain disparate molecular bases of this dominantly inherited disease. Most recently, increased nucleosome binding to such expanded repeats has been interpreted as support for transcriptional repression. A quantitative allele-specific RT-PCR procedure was developed and applied to a spectrum of patient tissue samples and cell lines. Equal levels of unprocessed pre-mRNA were produced by the wildtype (+) and disease (DM) alleles in skeletal muscle and cell lines of heterozygous DM patients. Thus, any increased nucleosome binding had no effect at the level of transcriptional initiation and transcription of the mutant DMPK locus. In contrast, processed mRNA levels from the DM allele were reduced relative to the+allele as the size of the expansion increased. The unstable repeat, therefore, impairs post-transcriptional processing of DM allele transcripts. This phenomenon has profound effects on overall DMPK locus steady-state transcript levels in cells missing a wildtype allele and does not appear to be mediated by imprinting, decreased mRNA stability, generation of aberrant splice forms, or absence of polyadenylation of the mutant allele.

Adolescent↗

PubMed1995

Overexpression of DM20 messenger RNA in two brothers with Pelizaeus-Merzbacher disease.

Pelizaeus-Merzbacher disease is a rare, sex-linked recessive, dysmyelinating disease of the central nervous system that has been associated with mutations in the myelin proteolipid protein (PLP) gene. Only 25% of patients studied with Pelizaeus-Merzbacher disease have exonic mutations in this gene, the underlying cause of the disease in the remaining patients is unknown. The PLP gene encodes two major alternatively spliced transcripts called PLP and DM20. PLP messenger RNA is specifically expressed in central nervous system tissue, whereas DM20 messenger RNA is found in central nervous system, cardiac, and other tissues. We studied cultured skin fibroblasts from 2 brothers with Pelizaeus-Merzbacher disease who exhibited no detectable exonic mutation of the PLP gene. Examination of RNA from these cells showed that the level of DM20 messenger RNA is elevated sixfold relative to male control skin fibroblasts. An unrelated female carrier, also with no detectable exonic mutation, showed a threefold increase in DM20 messenger RNA in cultured skin fibroblasts. Our findings suggest that in some patients, Pelizaeus-Merzbacher disease is caused by overexpression of PLP gene transcripts, and that in these families a 50% increase of DM20 messenger RNA in females, relative to the increase in affected males, can identify a female carrier.

Adolescent↗

PubMed1993

Absence of myotonic dystrophy protein kinase (DMPK) mRNA as a result of a triplet repeat expansion in myotonic dystrophy.

Myotonic dystrophy is an autosomally dominant inherited disease in which system-wide abnormalities are caused by a triplet repeat expansion within the 3' untranslated region of the myotonic dystrophy protein kinase (DMPK) gene. To determine the effect an expanded repeat region has on DMPK expression, we have separated the chromosome 19 homologues from a 36-year-old woman with myotonic dystrophy into different cell lines by way of somatic cell hybridization. Hybrid DM9101 contains the normal DMPK allele (13 repeats), whereas hybrid DM1115 harbors the mutant allele (approximately 133 repeats). Reverse transcription/polymerase chain reaction (RT/PCR) amplification of coding sequences from the DMPK gene has shown both reduced levels of primary DMPK transcripts and impaired processing of these transcripts in hybrid cell line DM1115. These findings suggest that the presence of a large number of repeats in the 3' untranslated region of the DMPK gene reduces both the synthesis and the processing of DMPK mRNA, resulting in undetectable levels of processed DMPK mRNA from the mutant allele.

Adult↗

PubMed1992

Creatine kinase activity is required for mineral deposition and matrix synthesis in endochondral growth cartilage.

In earlier studies, we have drawn attention to the unique changes in energy metabolism that accompany the maturation of epiphyseal growth plate chondrocytes. The objective of this investigation was to examine the importance of the ATP generating enzyme creatine kinase (CK), in the development and mineralization of the growth plate. We inhibited CK function by administering beta-guanidinopropionic acid (beta-GPA) to rats in vivo and to cultured chick chondrocytes in vitro. We found that this agent inhibited normal development of cartilage. Disorganization of chondrocytes in the proliferative and hypertrophic zones, poor vascular invasion, and retention of calcified cartilage occurred in the long bones of beta-GPA-fed rats. beta-GPA caused a change in the electrophoretic mobility of type II and type X collagens. Inhibition of apatite formation in the bones of shell-less chick embryos was accompanied by a CK isoenzyme shift from a bone-specific phenotype to a CK isozyme profile similar to that of cartilage. The results of these studies indicate that CK activity is required for normal development of the growth plate and that interference with creatine phosphate metabolism results in profound changes in the synthesis of cartilage and the maturational activities of chondrocytes.

Animals↗

PubMed1991

Spinal decompensation in Cotrel-Dubousset instrumentation.

Forty-one patients with idiopathic scoliosis having a primary right thoracic and a compensatory left lumbar curve underwent posterior spinal fusion of the primary curve only. Twenty-four patients had instrumentation with a Harrington rod or variant, and 17 patients underwent Cotrel-Dubousset instrumentation. Decompensation occurred postoperatively when the apex of the thoracic curve was located on or to the left of the center sacral line. There was no statistically significant change in the lumbosacral portion of the lumbar curve from the apex to the lumbosacral junction in both groups. Curve correction occurred cephalad to the apex of the lumbar curve, and not along the center sacral line. The decompensation rate was 4% for Harrington rod instrumentation and variants and 41% for Cotrel-Dubousset instrumentation. Cotrel-Dubousset instrumentation translated the apex of the thoracic curve 1.5 cm farther to the left than Harrington rod instrumentation and variants. When the apex of the lumbar curve is 2 cm or greater to the left of the center sacral line, the patient's spine will decompensate to the left, centered over the apex of the lumbar curve.

Arthrodesis↗

PubMed1990

Occipito-atlantal translation in Down's syndrome.

Several cases of occipito-atlantal instability have been reported in patients with Down's syndrome. However, standards for radiographic measurement and normal values for translational motion at the occiput-C1 level have not been established for the Down's patient. A retrospective analysis of 102 flexion and extension lateral cervical spine radiographs of 73 patients with Down's syndrome was performed. Patients with congenital cervical spine anomalies, C1-C2 instability, or previous neck surgery were excluded. Occiput-C1 translation was measured by the technique described by Wiesel and Rothman. In normal adult patients, occiput-C1 translation should be no more than 1 mm by this method. Analysis of the data demonstrated a range of 0 to 10 mm, with a mean of 2.62 mm and a standard deviation of +/- 1.94 mm. Only 30 sets of films (29%) representing 27 patients (37%) showed anteroposterior translation of 1 mm or less. This laxity was not dependent on patient age in the current study group. A review of these patients' medical records did not disclose any evidence of impaired neurologic function related to cervical spine instability. The authors' data suggest that the prevalence and magnitude of occipito-atlantal instability in Down's syndrome is greater than previously appreciated.

Adolescent↗

PubMed1988

Induction of a 58,000 dalton protein during goldenrod gall formation.

Despite the widespread occurrence of plant-gallmaker interactions, little is known about the actual mechanisms of gall formation. To further characterize this type of parasite-host interaction, the mechanism of gall formation in Solidago altissima, tall goldenrod, by the larva of the tephritid fly Eurosta solidaginis was studied. Proteins produced by galled and ungalled tissues were examined, and the hyperinduction of a 58 kilodalton protein was observed in galled tissues for the second and third week of gall growth. The presence of this protein suggests that a substance secreted by the larva may function as a trans-acting gene regulator.

Electrophoresis, Polyacrylamide Gel↗

PubMedDate not supplied

Effect of triplet repeat expansion on chromatin structure and expression of DMPK and neighboring genes, SIX5 and DMWD, in myotonic dystrophy.

Myotonic dystrophy (DM), an autosomal dominant neuromuscular disease, is associated with expansion of a polymorphic (CTG)n repeat in the 3'-untranslated region of the DM protein kinase (DMPK) gene. The repeat expansion results in decreased levels of DMPK mRNA and protein, but the mechanism for this decreased expression is unknown. Loss of a nuclease-hypersensitive site in the region of the repeat expansion has been observed in muscle and skin fibroblasts from DM patients, indicating a change in local chromatin structure. This change in chromatin structure has been proposed as a mechanism whereby the expression of DMPK and neighboring genes, sine oculis homeobox (Drosophila) homolog 5 (SIX5) and dystrophia myotonica-containing WD repeat motif (DMWD), might be affected. We have developed a polymerase chain reaction (PCR)-based method to assay the chromatin sensitivity of the region adjacent to the repeat expansion in somatic cell hybrids carrying either normal or affected DMPK alleles and show that hybrids carrying expanded alleles exhibit decreased sensitivity to PvuII digestion in this region. Semiquantitative multiplex reverse transcriptase PCR (RT/PCR) assays of gene expression from the chromosomes carrying the expanded alleles showed marked reduction of DMPK mRNA, partial inhibition of SIX5 expression from a congenital DM chromosome, and no reduction of DMWD mRNA. Nested RT/PCR analysis of DMPK mRNA from somatic cell hybrids carrying the repeat expansions revealed that most of the DMPK transcripts expressed from the expanded alleles lacked exons 13 and 14, whereas full-length transcripts were expressed predominantly from the normal alleles. These results suggest that the CTG repeat expansion leads to a decrease in DMPK mRNA levels by affecting splicing at the 3' end of the DMPK pre-mRNA transcript.

Cells, Cultured↗