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Biomedical subjects

B Houston

Publications and source records attributed to B Houston.

At least 19 recordsLinked to original sources

Chromosomal localization of the chicken and mammalian orthologues of the orphan phosphatase PHOSPHO1 gene.

PHOSPHO1 is a recently identified phosphatase expressed at high levels in the chicken growth plate and which may be involved in generating inorganic phosphate for skeletal matrix mineralization. Using a degenerate RT-PCR approach a fragment of human PHOSPHO1 was cloned. This enabled the identification of the human orthologue on HSA17q21, and the mouse orthologue on a region of MMU11 that exhibits conservation of synteny with HSA17q21. Chicken PHOSPHO1 was mapped by SSCP analysis to position 44 cM on GGA27, adjacent to the HOXB@ (44 cM) and COL1A1 (36 cM) loci. Comparison of genes on GGA27 with their orthologues on the preliminary draft of the human genome identifies regions of conserved synteny equivalent to 25 Mb on HSA17q21.2-23.3 and approximately 20 Mb on GGA27 in which the gene order appears to be conserved. Mapping of the PHOSPHO1 genes to regions of HSA17q21.3, MMU11 and GGA27 that exhibit conservation of synteny provides strong evidence that they are orthologous.

Amino Acid Sequence↗

Unusual cerebellar ataxia: "worm wobble" revisited.

We present an unusual case of cerebellar ataxia in a 2 year old girl several days after treatment with piperazine citrate for suspected worm infestation. This is the first reported case of delayed onset neurotoxicity following the therapeutic administration of piperazine in a previously well child.

Antinematodal Agents↗

Regulation of chondrocyte terminal differentiation in the postembryonic growth plate: the role of the PTHrP-Indian hedgehog axis.

Chondrocyte differentiation during embryonic bone growth is controlled by interactions between PTHrP and Indian hedgehog. We have now determined that the major components of this signaling pathway are present in the postembryonic growth plate. PTHrP was immunolocalized throughout the growth plate, and semiquantitative RT-PCR analysis of maturationally distinct chondrocyte fractions indicated that PTHrP, Indian hedgehog, and the PTH/PTHrP receptor were expressed at similar levels throughout the growth plate. However, patched, the hedgehog receptor, was more highly expressed in proliferating chondrocytes. Although all fractionated cells responded to PTHrP in culture by increasing thymidine incorporation and cAMP production and decreasing alkaline phosphatase activity, the magnitude of response was greatest in the proliferative chondrocytes. Bone morphogenetic proteins are considered likely intermediates in PTHrP signaling. Expression of bone morphogenetic protein-2 and 4--7 was detected within the growth plate, and PTHrP inhibited the expression of bone morphogenetic protein-4 and 6. Although organ culture studies indicated a possible paracrine role for epiphyseal chondrocyte-derived PTHrP in regulating growth plate chondrocyte differentiation, the presence within the postembryonic growth plate of functional components of the PTHrP-Indian hedgehog pathway suggests that local mechanisms intrinsic to the growth plate exist to control the rate of endochondral ossification.

Animals↗

Expression patterns of chondrocyte genes cloned by differential display in tibial dyschondroplasia.

Tibial dyschondroplasia (TD) appears to involve a failure of the growth plate chondrocytes within growing long bones to differentiate fully to the hypertrophic stage, resulting in a mass of prehypertrophic chondrocytes which form the avascular TD lesion. Many biochemical and molecular markers of chondrocyte hypertrophy are absent from the lesion, or show reduced expression, but the cause of the disorder remains to be identified. As differentiation to the hypertrophic state is impaired in TD, we hypothesised that chondrocyte genes that are differentially expressed in the growth plate should show altered expression in TD. Using differential display, four genes, B-cadherin, EF2, HT7 and Ex-FABP were cloned from chondrocytes stimulated to differentiate to the hypertrophic stage in vitro, and their differential expression confirmed in vivo. Using semi-quantitative RT-PCR, the expression patterns of these genes were compared in chondrocytes from normal and TD growth plates. Surprisingly, none of these genes showed the pattern of expression that might be expected in TD lesion chondrocytes, and two of them, B-cadherin and Ex-FABP, were upregulated in the lesion. This indicates that the TD phenotype does not merely reflect the absence of hypertrophic marker genes, but may be influenced by more complex developmental mechanisms/defects than previously thought.

Animals↗

Identification of a family of noncanonical ubiquitin-conjugating enzymes structurally related to yeast UBC6.

Ubiquitin-conjugating enzymes (UBCs) selectively target proteins for proteasomal degradation by the covalent attachment of ubiquitin moieties. Yeast UBC6 is unusual in having an active site distinct from all other UBCs and in possessing a transmembrane domain that anchors it to the cytoplasmic surface of the endoplasmic reticulum. During a differential display analysis on chick growth plate chondrocytes we isolated a cDNA encoding a noncanonical ubiquitin-conjugating enzyme (NCUBE1) structurally similar to yeast UBC6. Chick NCUBE1 transcripts were detected in all tissues examined and decreased threefold during chondrocyte terminal differentiation. Database searches identified other related proteins; the human and mouse orthologues of NCUBE1, a second human homologue of yeast UBC6 (NCUBE2), and related proteins from S. pombe, C. elegans, and P. mariana. Together with yeast UBC6 these proteins constitute a distinct family of UBCs sharing a conserved noncanonical active site sequence and a C-terminal transmembrane domain. By analogy with yeast UBC6 they are likely to be localised to the endoplasmic reticulum where they may be involved in targeting retrotranslocated, ER-associated proteins for proteasomal degradation.

Amino Acid Sequence↗

Identification and cloning of a novel phosphatase expressed at high levels in differentiating growth plate chondrocytes.

Growth plate chondrocytes progress through a proliferative phase before acquiring a terminally-differentiated phenotype. In this study we used Percoll density gradients to separate chick growth plate chondrocytes into populations of different maturational phenotype. By applying agarose gel differential display to these populations we cloned a cDNA encoding a novel 268 amino acid protein (3X11A). 3X11A contains two peptide motifs that are conserved in a recently identified superfamily of phosphotransferases. It is likely that 3X11A is a phosphatase, but its substrate specificity remains uncertain. 3X11A expression is upregulated 5-fold during chondrocyte terminal differentiation and its expression is approximately 100-fold higher in hypertrophic chondrocytes than in non-chondrogenic tissues. This suggests that 3X11A participates in a biochemical pathway that is particularly active in differentiating chondrocytes.

Amino Acid Sequence↗

Firearm storage practices and children in the home, United States, 1994.

OBJECTIVES: To estimate the national prevalence of firearm ownership and storage practices in the home, to compare storage practices in homes with and without children, and to analyze demographic characteristics related to firearm storage practices in homes with children. DESIGN: A 1994 random-digit dialing telephone survey. We weighted the data to provide national estimates. PARTICIPANTS: English- and Spanish-speaking adults in households in 50 states and Washington, DC. MAIN OUTCOME MEASURES: Ownership of working powder firearm(s) in home and/or vehicle and firearm storage practices in the home. RESULTS: Of 5238 households surveyed, one third kept at least 1 firearm in the home and/or vehicle. Of 1598 households with firearm(s) in the home and known firearm storage practices, 21.5% kept at least 1 gun loaded and unlocked in the home, 30.0% stored all firearms unloaded and locked, and 48.5% stored firearms in a manners classified between these 2 practices. Households with children were more likely than households without children to store all firearms unloaded and locked (41.5% vs 20.9%); households without children were more likely than households with children to store at least 1 firearm loaded and unlocked (29.8% vs 11.1%). Among households with children and firearms, there were regional differences with respect to storage practices. CONCLUSIONS: These prevalence data show that children are potentially exposed to firearms in many households. This health threat illustrates the need for education about the issue of pediatric firearm injuries and for interventions to minimize associated risks. Health care providers should take advantage of opportunities to counsel patients regarding firearm safety in the home.

Adolescent↗

Microtubules are potential regulators of growth-plate chondrocyte differentiation and hypertrophy.

Terminal differentiation of growth-plate chondrocytes is accompanied by the acquisition of a spherical morphology and a large increase in cell volume. These changes are likely to be associated with rearrangement of the cytoskeleton, but little information on this aspect of chondrocyte hypertrophy is available. We report a role for microtubules in the control of chondrocyte maturation and hypertrophy. Chick growth-plate chondrocytes were fractionated into five maturationally distinct populations by Percoll density gradient centrifugation, and agarose gel differential display analysis was performed. We identified a 1200 bp cDNA fragment derived from a transcript that was most highly expressed in the hypertrophic chondrocytes. After cloning and sequencing, FASTA and BLAST analysis revealed 100% identity to chick beta7-tubulin. Differential expression was confirmed in a reverse transcription-polymerase chain reaction (RT-PCR) assay using specific primers for a 343 bp fragment from the 3' untranslated region of beta7-tubulin. Beta7-tubulin was upregulated three-fold in fully hypertrophic chondrocytes compared with the other four fractions, which all had similar levels of expression. Immunocytochemical localization of beta-tubulin in chick growth-plate sections demonstrated little staining in the chondrocytes of the proliferating zone, but intense cytoplasmic staining was present in the large hypertrophic chondrocytes. In cell culture studies, the addition of colchicine (10(-6) mol/L) resulted in a higher rate of [3H]-thymidine uptake (36.0%; p < 0.001), but lower amounts of alkaline phosphatase activity (69.1%; p < 0.001), collagen (49.1%; p < 0.01), and glycosaminoglycan (43.3%; p < 0.01) accumulation within the cell-matrix layer. Further evidence for the involvement of microtubules in chondrocyte differentiation and hypertrophy was obtained by morphological assessment of colchicine-treated growth-plate explant cultures. A partial failure of chondrocyte hypertrophy was observed, although collagen type X immunoreactivity was noted within the interstitial matrix. Further studies are required to identify the exact role of microtubules in chondrocyte hypertrophy, but the results presented here suggest that upregulation of beta-tubulin may be required for increased microtubule synthesis during changes in cell size during the hypertrophic process. In addition, as cell-matrix interactions are required for chondrocyte maturation, microtubules may promote the differentiated phenotype as a result of their role in Golgi-mediated secretion of matrix proteins.

Animals↗

A novel integral membrane protein is differentially expressed in the chick growth plate and maps to chromosome 1.

The growth plate is a specialised region of cartilage located at the growing ends of long bones in higher vertebrates. It is responsible for longitudinal bone growth and is under the control of many local and systemic factors. The growth plate consists of an orderly arrangement of small proliferative and larger mature hypertrophic chondrocytes. This paper describes the isolation by differential display of a 988-bp cDNA fragment derived from a transcript that is more highly expressed in proliferating rather than hypertrophic chondrocytes of the chick growth plate. Using 3' RACE, a further 939 bp of cDNA sequence was obtained. The 1.9 kb sequence contains a 924-bp open reading frame encoding an unknown 308 amino acid protein. This protein has a putative transmembrane domain near its N-terminus and three dileucine motifs at its carboxy tail. This gene was expressed in all other tissues examined. A polymorphism was identified by SSCP analysis and the gene was mapped to the centromeric region of the short arm of chicken chromosome 1, close to the locus for autosomal dwarfism.

5' Untranslated Regions↗

A spontaneous corneal change in juvenile Wistar rats.

Corneal changes were seen in a shipment of male and female Charles River Wistar (Crl:(WI)BR) albino rats aged 33 days. Seventeen weeks later, varying degrees of corneal roughness were observed ophthalmoscopically in 40 of 43 males and 26 of 29 females. Corneal lesions, described clinically as multiple punctate or particulate opacities, were most often bilateral and located paracentrally. Histologically, lesions were observed in only 21 of 40 males and in 15 of 26 females which had shown clinical changes. The lesions consisted of basophilic particulate material, which occasionally formed linear plaques, lining the basement membrane of the corneal epithelium. Ultrastructurally, irregularly distributed electron-dense deposits, occasionally assuming a concentric laminar appearance, were observed. X-ray microanalysis revealed that the granules consisted predominantly of calcium and phosphorus, with trace amounts of iron, zinc, sodium and aluminium. These mineral deposits in the corneas of Wistar rats were similar to those described in other species. Awareness of this spontaneously occurring disease in Wistar rats is of importance in relation to toxicological investigations in which the cornea may be a target organ.

Aging↗

Cloning differentially regulated genes from chondrocytes using agarose gel differential display.

The technique of RNA differential display has been used extensively to clone differentially expressed genes from a wide variety of cells and tissues. Recently, a simplified method of cloning differential display products, separated on agarose gels, was described. Here we report an adaption of this method, using total RNA, to clone differentially expressed genes. The approach is simple and rapid, and requires only small quantities of total RNA. Utilising this approach, we have cloned three differentially regulated genes from chondrocytes stimulated to hypertrophy in vitro, and confirmed their pattern of expression by Northern blotting. These gene fragments were sequenced and found to correspond to known genes, although only one has previously been isolated from chondrocytes.

Animals↗

Dog bites: how big a problem?

OBJECTIVE: To estimate the magnitude of the dog bite problem in the US. METHODS: Data on dog bites were gathered as part of a 1994 national telephone survey of 5,238 randomly dialed households. Data were weighted to provide national estimates. RESULTS: The weighted total number of dog bites was 4,494,083 (estimated incidence = 18/1,000 population); of these, 756,701 persons sustained bites necessitating medical attention (incidence rate = 3/1,000). Children had 3.2 times higher medically attended bite rates than adults (6.4/1,000 children v 2/1,000 adults). CONCLUSIONS: More attention and research needs to be devoted to the prevention of dog bites. Potential prevention strategies include: educational programs on canine behavior, especially directed at children; laws for regulating dangerous or vicious dogs; enhanced animal control programs; and educational programs regarding responsible dog ownership and training. Unfortunately, the relative or absolute effectiveness of any of these strategies has not been assessed. Continuing surveillance for dog bites will be needed if we are to better understand how to reduce the incidence of dog bites and evaluate prevention efforts.

Accident Prevention↗

Bicycle helmet use among American children, 1994.

OBJECTIVE: To estimate ownership and use of bicycle helmets among children in the US in 1994. METHODS: As part of a 1994 national telephone survey of 5,238 randomly dialed households, adult respondents reported data on bicycle helmet ownership and helmet use among 1,645 child bicyclists. Data were weighted to provide national estimates. RESULTS: It is estimated that 72.7% of children 5-14 year olds ride bicycles, that is, 27.7 million child bicyclists. Of the bicyclists, 50.2% have a helmet and 25.0% reportedly always wore their helmet when cycling. Reported helmet ownership and use increased with income and educational level and decreased with age. Among regions of the US, those with the highest proportion of states with helmet use laws in 1994 also had the highest proportion of helmet use among children. Among child bicyclists who had been seen by a health care provider in the preceding 12 months, 43.9% of those counseled to wear a bicycle helmet were reported to comply compared with 19.1% of those seen by a provider but not so counseled (p < 0.001). CONCLUSIONS: To meet the year 2000 objective of 50% of bicyclists wearing helmets, use among American children will have to double. Concerted and increased efforts to promote the wearing of bicycle helmets are necessary.

Adolescent↗

Chondroid syringoma of the head and neck: clinical management and literature review.

This report describes the case of a chondroid syringoma occurring in the nasofacial groove of a 60-year-old woman. This benign, mixed epithelial tumor is infrequently seen by the otolaryngologist-head and neck surgeon, and therefore may not be included in the differential diagnosis of a nodular lesion on the skin of the head and neck. However, the most frequent site of occurrence for these relatively rare cutaneous lesions is the head and neck region. Histologically, these tumors are quite similar to pleomorphic adenomas of salivary gland origin, and optimal surgical management similarly requires removal of a cuff of normal tissue, rather than a simple shelling out of the lesion. The clinical presentation, pathology and recommended management of this rare tumor are discussed.

Adenoma, Pleomorphic↗

Expression of genes encoding bone morphogenetic proteins and sonic hedgehog in talpid (ta3) limb buds: their relationships in the signalling cascade involved in limb patterning.

The chicken mutant talpid3 (ta3) has polydactylous limbs with up to 7-8 morphologically similar digits. This lack of antero-posterior polarity in digit pattern is correlated with symmetrical expression of genes of the HoxD complex. We determined the distribution of polarizing activity in limb buds of the chick mutant ta3 by assessing the ability of mesenchyme from various positions along the antero-posterior axis to induce digit duplications when grafted anteriorly into a normal limb. Cells with highest polarizing activity were found at the posterior margin of the wing as in the polarizing region of normal limb buds. However, in contrast to normal limb buds, ta3 anterior mesenchyme also had low polarizing activity. Application of retinoic acid or a polarizing region graft to the anterior of ta3 limb buds changed digit morphology but did not induce digit duplications or digits with any characteristic a-p pattern. To determine which genes are associated with polarizing activity and which are associated with patterning of the digits, we examined expression of the genes Sonic hedgehog (shh), Bmp-2, and Bmp-7, whose expression is normally confined to the posterior margin of the early wing bud and is associated with the polarizing region. In addition, we determined the distribution of Fgf-4 transcripts which in normal limb buds are restricted to the posterior part of the apical ectodermal ridge. In ta3 limb buds, shh expression is restricted to the posterior limb mesenchyme, which has high polarizing activity, but is not expressed in regions which have low polarizing activity. In contrast, Bmp-2 and Bmp-7 are expressed uniformly along the a-p axis. Fgf-4 transcripts are present throughout the apical ectodermal ridge in ta3 limb buds. In the ta3 mutant, there is both an abnormal distribution of signalling activity and response to polarizing signals. In addition, the dissociation between the expression of shh and Bmps suggests distinct roles for the encoded molecules in signalling and response in a-p patterning of limb buds.

Animals↗

Molecular cloning and expression of bone morphogenetic protein-7 in the chick epiphyseal growth plate.

Longitudinal bone growth occurs in the epiphyseal growth plate and is regulated by a network of paracrine and autocrine interactions. Bone morphogenetic proteins (BMPs) are a family of growth factors whose potent osteogenic properties suggest that they may play an important role within this network, but direct evidence for this is lacking. To address this question, a cDNA encoding chick BMP-7 was cloned from a chick embryo cDNA library. Sequence homology and evolutionary arguments strongly suggested that we had cloned the chicken BMP-7 homologue. Using a reverse transcription-PCR assay, BMP-7 expression was readily detected in bone, growth plate cartilage, brain and heart, and was just detectable in liver, skeletal muscle and adipose tissue. In contrast to the pattern of BMP-7 expression in the rat and mouse, no BMP-7 expression was detected in the chick kidney. In situ hybridization was used to locate the site of BMP-7 expression more precisely within the growth plate. BMP-7 expression was confined to hypertrophic chondrocytes adjacent to and at the tips of the metaphyseal vessels. No expression was detected in the reserve zone or in proliferating chondrocytes. These results point to a specific role for BMP-7 in the growth plate, possibly in osteoblast activation or as a chemotactic agent for the metaphyseal vessels.

Amino Acid Sequence↗