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Biomedical subjects

R G Knowlton

Publications and source records attributed to R G Knowlton.

At least 37 records · Page 2Linked to original sources
PubMed1993

Spondyloepiphyseal dysplasia and precocious osteoarthritis in a family with an Arg75-->Cys mutation in the procollagen type II gene (COL2A1).

Direct sequencing of polymerase chain reaction (PCR)-amplified genomic DNA from a patient with spondyloepiphyseal dysplasia and precocious osteoarthritis revealed a single-base change in exon 11 of the type II procollagen gene (COL2A1), which produces an Arg-->Cys mutation in one allele. The proband is a member of a large Chilean kindred presenting with chondrodysplasia of the hips, knees, shoulders, elbows, and spine associated with severe, early-onset osteoarthritis. All affected individuals exhibit mildly short stature; in addition, five out of seven affected family members display shortened metacarpals or metatarsals. DNA from affected and unaffected family members was PCR-amplified and analysis of restriction digests of the products determined that the mutation segregated with the disease with a lod score of 2.2 at zero recombination. The mutation, which resides in the triple-helical region of type II procollagen at amino acid position 75, is the second example of an Arg-->Cys mutation in the COL2A1 gene in heritable cartilaginous disease and is the first example of a point mutation in the amino terminal region of the alpha 1(II) chain, that results in a spondyloepiphyseal dysplastic phenotype.

Adolescent↗
PubMed1993

Re-examination of the incidence of exercise-induced hypoxaemia in highly trained subjects.

The purpose of this study was to examine the occurrence of exercise-induced hypoxaemia (EIH) during maximal exercise in highly trained athletes. Eleven trained cyclists (mean(s.d.) age 23(3.5) years; mean(s.d.) VO2max 66.9(4.8) ml kg-1min-1) performed a continuous, multistage (270 kpm min-1) cycle ergometer test to exhaustion. Measurements of arterial oxygen-haemoglobin saturation (%HbO2) were obtained simultaneously at rest, every 2 min during exercise, and at maximum exercise capacity from arterial blood sampling (%SaO2) and ear oximetry (%SpO2). Exercise induced hypoxaemia (%HbO2 < or = 91%) was present in 64% of the athletes examined when EIH was determined using pulse oximetry, whereas none of the subjects exhibited EIH when %HbO2 was determined using arterial blood. At rest the values for %HbO2 were similar with mean(s.d.) %SaO2 being 97.3(0.6)% and mean(s.d.) %SpO2 being 96.5(1.6)%. During exercise, statistically significant differences were found for %HbO2 between arterial blood and ear oximetry at the 6-min, 8-min, and maximal exercise sampling times (repeated measures analysis of variance, P < 0.05). The results indicate that ear oximetry overestimates the incidence of EIH and underestimates the oxyhaemoglobin saturation in highly trained cyclists during exercise in comparison with those measurements made from arterial blood.

Adult↗
PubMed1993

Epidermolytic hyperkeratosis (bullous congenital ichthyosiform erythroderma). Genetic linkage to chromosome 12q in the region of the type II keratin gene cluster.

Epidermolytic hyperkeratosis (EHK) is an autosomal dominant genodermatosis characterized by hyperkeratosis and blistering of the skin. Histopathology demonstrates suprabasilar blister formation with aggregation of tonofilaments. In this study, we tested the hypothesis that the EHK phenotype is linked to one of the suprabasilar keratins (KRT10 or KRT1) present in the types I and II keratin gene clusters in chromosomes 17q and 12q, respectively. For this purpose, Southern hybridizations were performed with DNA from a large kindred with EHK, consisting of 11 affected individuals in three generations. Segregation analysis with markers flanking the keratin gene clusters demonstrated linkage (Z = 3.61 at theta = 0) to a locus on 12q, while markers on 17q were excluded. These data implicate KRT1, the type II keratin expressed in suprabasilar keratinocytes, as a candidate gene in this family with EHK.

Blotting, Southern↗
PubMed1993

Localization of the Krabbe disease gene (GALC) on chromosome 14 by multipoint linkage analysis.

The gene responsible for Krabbe disease, an autosomal recessive disorder caused by deficiency of galactocerebrosidase (GALC), was localized by multipoint linkage analysis on chromosome 14. Eight mapped dinucleotide repeat polymorphisms were tested for linkage to GALC. Two-point linkage analysis demonstrated close linkage of GALC and D14S48, with Z = 13.69 at theta = 0. Multipoint analysis yielded strong support for this finding, with maximum likelihood for GALC located within 1 cM of D14S48. This analysis also identified markers that clearly flank the GALC locus, as the map order of D14S53-GALC-D14S45 is favored by odds greater than 10(6):1. Additional support for close linkage of GALC and D14S48 comes from the apparent linkage disequilibrium between these two loci in a consanguineous Druze community in Israel. These data localize GALC to 14q24.3-q32.1.

Chromosome Mapping↗
PubMed1993

A second mutation in the type II procollagen gene (COL2AI) causing stickler syndrome (arthro-ophthalmopathy) is also a premature termination codon.

Genetic linkage analyses suggest that mutations in type II collagen may be responsible for Stickler syndrome, or arthro-ophthalmopathy (AO), in many families. In the present study oligonucleotide primers were developed to amplify and directly sequence eight of the first nine exons of the gene for type II procollagen (COL2A1). Analysis of the eight exons in 10 unrelated probands with AO revealed that one had a single-base mutation in one allele that changed the codon of -CGA- for arginine at amino acid position alpha 1-9 in exon 7 to a premature termination signal for translation. The second mutation found to cause AO was, therefore, similar to the first in that both created premature termination signals in the COL2A1 gene. Since mutations producing premature termination signals have not previously been detected in genes for fibrillar collagens, the results raise the possibility that such mutations in the COL2A1 gene are a common cause of AO.

Adolescent↗
PubMed1992

Effect of physostigmine on plasma lactate and pyruvate in untrained/trained rats.

Physostigmine (Phy) is metabolized to eseroline, a phenolic compound that appears to alter mitochondrial functions. The effect of Phy on recovery from exercise and on time course of plasma lactate and pyruvate levels following an acute bout of exercise (AE) was examined in untrained and trained (ET) rats. Phy alone elicited significantly higher plasma lactate and pyruvate levels than sedentary control. AE + Phy had a significantly higher plasma lactate and pyruvate levels compared to AE 2 min postadministration. From 5-30 min postexercise, lactate and pyruvate levels did not differ between these two acutely exercised groups. ET + Phy exhibited significantly lower levels of plasma lactate and pyruvate from 5-60 min postexercise compared to ET. The data show that the "additive" effect of Phy on postexercise plasma lactate and pyruvate levels can be attenuated by an enhanced fitness level in these rats.

Animals↗
PubMed1992

Genetic linkage of type VII collagen (COL7A1) to dominant dystrophic epidermolysis bullosa in families with abnormal anchoring fibrils.

Epidermolysis bullosa (EB) in a group of genodermatoses characterized by the fragility of skin. Previous studies on the dystrophic (scarring) forms of EB have suggested abnormalities in anchoring fibrils, morphologically recognizable attachment structures that provide stability to the association of the cutaneous basement membrane to the underlying dermis. Since type VII collagen is the major component of the anchoring fibrils, we examined the genetic linkage of dominant dystrophic EB (EBDD) and the type VII collagen gene (COL7A1) locus, which we have recently mapped to chromosome 3p, in three large kindreds with abnormal anchoring fibrils. Strong genetic linkage of EBDD and COL7A1 loci was demonstrated with the maximum logarithm of odds (LOD) score of 8.77 at theta = 0. This linkage was further confirmed with two additional markers in this region of the short arm of chromosome 3, and these analyses allowed further refinement of the map locus of COL7A1. Since there were no recombinants between the COL7A1 and EBDD loci, our findings suggest that type VII collagen is the candidate gene that may harbor the mutations responsible for the EB phenotype in these three families.

Chromosome Mapping↗
PubMed1992

Genetic linkage of recessive dystrophic epidermolysis bullosa to the type VII collagen gene.

Generalized mutilating recessive dystrophic epidermolysis bullosa (RDEB) is characterized by extreme skin fragility owing to loss of dermal-epidermal adherence. Immunohistochemical studies have implicated type VII collagen, the major component of anchoring fibrils, in the etiology of RDEB. In this study, we demonstrate genetic linkage of the type VII collagen gene and the generalized mutilating RDEB phenotype. We first identified a Pvull polymorphic site by digestion of an amplified product of the type VII collagen gene, which was shown to reside within the coding region. Genetic linkage analysis between this marker and the RDEB phenotype in 19 affected families which were informative for this polymorphism showed no recombination events, and gave a maximum lod score of 3.97 at a recombination fraction (theta) of 0, demonstrating that this DNA region is involved in this form of RDEB. These data provide strong evidence that the type VII collagen gene, which has also been linked with the dominant form of the disease, harbors the mutation(s) causing the generalized mutilating form of RDEB in these families, thus underscoring the major functional importance of type VII collagen in basement membrane zone stability.

Base Sequence↗
PubMed1992

Accuracy of RPE from graded exercise to establish exercise training intensity.

The purpose of this investigation was to examine the accuracy of an exercise intensity prescription based upon perceptual responses obtained during a graded exercise test. Fifteen physically active men completed a graded exercise test (GXT) on a motor driven treadmill. Heart rate (HR), oxygen uptake (VO2), and RPE were measured each minute. An RPE intensity prescription was calculated as 75% of heart rate reserve from the GXT heart rate and RPE data. A minimum of 48 h later the subjects completed 10 min of exercise (EXT) on a level treadmill at the prescribed RPE. The subjects set the treadmill speed to elicit an exercise intensity equal to the prescribed perception of effort. There were significant mean differences (P < 0.05) in heart rate between the GXT (161.8 +/- 1.3) and EXT (154.9 +/- 4.5). However, by minute 6 the subjects were within four beats.min-1 of the target heart rate. There were no significant differences (P > 0.05) between GXT and EXT for VO2 (36.1 +/- 5.2, 33.1 +/- 6.4) and VE (64.1 +/- 10.8, 58.4 +/- 13.5) respectively. The present investigation demonstrates that a subject's perceptual response to a GXT can be used to accurately prescribe exercise intensity during level treadmill running. The intensity selected was within a typical range used for exercise prescription. The advantage of RPE as a method of exercise prescription is that an individual does not need to stop during exercise and measure a heart rate, but can make pace adjustments while exercising based solely upon the perception of effort.

Adult↗
PubMed1992

Influence of age on caloric expenditure during exercise.

Young (147 +/- 2 g) and adult (322 +/- 7 g) male Sprague Dawley rats were subjected to an incremental treadmill protocol and assessed via the Oxyscan Metabolic System in order to determine if differences exist relative to oxygen consumption (VO2, expressed in ml/kg/min), respiratory exchange ratio (RER) and caloric expenditure of two different age groups. The young rats attained a higher VO2 (81.55 +/- 1.22) compared to adult rats (68.97 +/- 2.05) at a maximal level of exercise. The adult rats became dependent upon carbohydrates as the primary source of energy until reaching 52% VO2 max, whereas the young rats did not resort to carbohydrate utilization as a primary fuel source until reaching 87% VO2max. At peak exercise, the adult animals burned only 15% less kcal/kg/h than the young rats, but the adult rats burned 37% more carbohydrate than the young animals. These data suggest that as exercise intensity increases, younger rats can more readily maintain a higher level of oxygen consumption accompanied by a more efficient use of fat as an energy source compared to adult rats. The clinical implication of this finding suggests that during the growth and development years of life, fat plays a significant role in energy expenditure needs and may therefore be more important than currently believed in young children's nutritional intake.

Aging↗
PubMed1991

Stop codon in the procollagen II gene (COL2A1) in a family with the Stickler syndrome (arthro-ophthalmopathy).

Linkage analysis with restriction fragment length polymorphisms for the gene for type II procollagen (COL2A1) was carried out in a family with the Stickler syndrome, or arthro-ophthalmopathy, an autosomal dominant disorder that affects the eyes, ears, joints, and skeleton. The analysis demonstrated linkage of the disease and COL2A1 with a logarithm-of-odds score of 1.51 at zero recombination. A newly developed procedure for preparing cosmid clones was employed to isolate the allele for type II procollagen that was linked to the disease. Analysis of over 7000 nucleotides of the gene revealed a single base mutation that altered a CG dinucleotide and converted the codon CGA for arginine at amino acid position alpha 1-732 to TGA, a stop codon. From previous work on procollagen biosynthesis, it is apparent that the truncated polypeptide synthesized from an allele with a stop codon at alpha 1-732 cannot participate in the assembly of type II procollagen, and therefore that the mutation would decrease synthesis of type II procollagen. It was not apparent, however, why the mutation produced marked changes in the eye, which contains only small amounts of type II collagen, but relatively mild effects on the many cartilaginous structures of the body that are rich in the same protein.

Base Sequence↗
PubMed1991

Exclusion of COL1A1, COL1A2, and COL3A1 genes as candidate genes for Ehlers-Danlos syndrome type I in one large family.

Ehlers-Danlos syndrome (EDS) type I is a generalized connective tissue disorder, the major manifestations of which are soft, velvety hyperextensible skin and moderately severe joint hypermobility. The gene defect or defects causing EDS type I have not yet been defined, but previous observations suggested that the syndrome may be caused by mutations in the genes for type-I collagen (COL1A1 and COL1A2) or type-III collagen (COL3A1). Here, we performed linkage studies for these three genes in large Azerbaijanian family with EDS type I. Three polymorphisms in the COL3A1 gene, two in the COL1A1 gene, and one in the COL1A2 gene were tested using the polymerase chain reaction. The data obtained excluded linkage of any of the three genes to EDS type I in the family.

Base Sequence↗
PubMed1991

Bullous pemphigoid antigens (BPAGs): identification of RFLPs in human BPAG1 and BPAG2, and exclusion as candidate genes in a large kindred with dominant epidermolysis bullosa simplex.

Bullous pemphigoid antigens (BPAGs) are defined as autoantigens in a blistering skin disease, bullous pemphigoid. Two of the BPAGs, a 230-kD (BPAG1) and a 180-kD (BPAG2) protein, have been localized to hemidesmosomes, attachment structures at the basal keratinocyte-basement membrane interphase. We have recently cloned cDNAs corresponding to human BPAG1 and BPAG2, and mapped the corresponding genes to human chromosomes 6p and 10q, respectively. These cDNAs have now been used in a search for RFLPs in the corresponding genes. Southern hybridizations of genomic DNA from normal unrelated individuals with a BPAG1 cDNA detected an informative MspI RFLP, and similar hybridizations with a BPAG2 cDNA revealed an informative TaqI RFLP. These RFLPs were applied to a large kindred with epidermolysis bullosa simplex (EBS), generalized (Koebner) type, consisting of 14 affected and 12 unaffected individuals in three generations. Linkage analysis excluded the EBS locus in this pedigree approximately 9 cM and approximately 5 cM on either side of the BPAG1 and BPAG2 loci, respectively, when a lod score of -2.0 was taken as the limit of exclusion. This study demonstrates that mutations in the BPAG1 or BPAG2 genes are not the primary genetic defect in this family with EBS.

Autoantigens↗
PubMed1991

Human type VII collagen: genetic linkage of the gene (COL7A1) on chromosome 3 to dominant dystrophic epidermolysis bullosa.

Epidermolysis bullosa (EB) is a heterogeneous group of heritable blistering disorders affecting the skin and the mucous membranes. Previous ultrastructural studies on the dystrophic (scarring) forms of EB have demonstrated abnormalities in the anchoring fibrils, morphologically distinct structures below the basal lamina at the dermal/epidermal basement membrane zone. Type VII collagen is the major collagenous component of the anchoring fibrils, and it is therefore a candidate gene for mutations in some families with dystrophic forms of EB. In this study, we performed genetic linkage analyses in a large kindred with dominant dystrophic EB. A 1.9-kb type VII collagen cDNA clone was used to identify a PvuII RFLP to follow the inheritance of the gene. This RFLP cosegregated with the EB phenotype in this family, strongly supporting genetic linkage (Z = 5.37; theta = .0). In addition, we assigned the type VII collagen gene (COL7A1) to chromosome 3 by hybridization to a panel of human x rodent somatic cell hybrids. These data demonstrate very close genetic linkage between the clinical phenotype in this family and the polymorphism in the type VII collagen gene mapped to chromosome 3. The absence of recombination between EB and the type VII collagen gene locus, as well as the observed abnormalities in the anchoring fibrils, strongly suggest that this collagen gene is the mutant locus in this kindred.

Blotting, Southern↗
PubMed1991

Mapping of epidermolysis bullosa simplex mutation to chromosome 12.

Epidermolysis bullosa simplex (EBS) is a dominantly inherited genodermatosis characterized by intraepidermal blister formation. Recent reports have suggested that EBS mutations may relate to keratin abnormalities. In this study, we conducted RFLP analyses to test the hypothesis that EBS is linked to one of the keratin gene clusters on chromosome 12 or chromosome 17. Although these keratin gene loci are not defined by RFLPs, several mapped RFLPs in the same chromosomal regions could be tested for linkage. A large EBS family with 14 affected and 12 unaffected individuals in three generations was analyzed for RFLP inheritance. Within this family there was no evidence for linkage of the EBS mutation to markers on chromosome 17q. However, there was evidence for close linkage to D12S17 located on chromosome 12q, with a maximum LOD score of 5.55 at theta = 0. Mapping of this mutation to chromosome 12 defines an EBS locus distinct from both EBS1 (Ogna) and EBS2 (Koebner), which are on chromosomes 8 and 1, respectively. Further mapping will determine whether this EBS locus on chromosome 12 resides within the keratin gene cluster at 12q11-q13.

Chromosome Mapping↗
PubMed1991

Effect of caffeine ingestion on alveolar ventilation during moderate exercise.

The purpose of this study was to examine the effect of caffeine ingestion on alveolar ventilation and physiological dead space ventilation during exercise in high and low caffeine users. Eleven males (mean age 26.4 +/- 5.4 years), classified as either high caffeine users (greater than 350 mg/d, n = 6) or low caffeine users (less than 50 mg/d, n = 5) performed two treadmill exercise conditions at a constant work rate (50% VO2max) 45 min after ingestion of 3.3 mg of caffeine/kg body weight or placebo using a double-blind protocol. Open circuit spirometry was used to determine ventilatory and gas exchange variables every 10 min during the 50 min of walking exercise. Analysis of variance showed that caffeine produced significant differences in alveolar ventilation (VA) and the physiological dead space ventilation/tidal volume ratio (VD/VT) with VA increased from 1.36 to 1.54 L/breath and VD/VT decreased from 22.3 to 20.5% between the placebo and caffeine treatments, respectively. Additionally, caffeine ingestion produced a significant increase in tidal volume (VT) and a significant decrease in frequency of breathing (fb). We conclude that caffeine consumed prior to exercise enhances ventilatory dynamics during exercise without regard to prior habitual caffeine consumption.

Adult↗