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Biomedical subjects

P J Pretorius

Publications and source records attributed to P J Pretorius.

At least 19 recordsLinked to original sources
PubMed2003

Effect of acute exposure to 3660 m altitude on orthostatic responses and tolerance.

Orthostatic reflexes were examined at 375 m and after 60 min of exposure in a hypobaric chamber at 3660 m using a 20-min 70 degrees head-up tilt (HUT) test. Mean arterial blood pressure, R wave-R wave interval (RRI), and mean cerebral blood flow velocity (MFV) were examined with coarse-graining spectral analysis. Of 14 subjects, 7 at 375 m and 12 at 3660 m were presyncopal. Immediately on arrival to high altitude, breathing frequency and MFV increased, and endtidal PCO2, RRI, RRI complexity, and the parasympathetic nervous system indicator decreased. MFV was similar in HUT at both altitudes. The sympathetic nervous system indicator increased with tilt at 3660 m, whereas parasympathetic nervous system indicator decreased with tilt at both altitudes. Multiple regression analysis of supine variables from either 375 or 3660 m and the time to presyncope at 3660 m indicated that, after 1 h of exposure, increased presyncope at altitude was the result of 1). ineffective peripheral vasoconstriction, despite increased cardiac sympathetic nervous system activity with HUT, and 2). insufficient cerebral perfusion owing to cerebral vasoconstriction as the result of hypoxic hyperventilation-induced hypocapnia.

Adaptation, Physiological↗
PubMed2003

VP2 gene phylogenetic characterization of field isolates of African horsesickness virus serotype 7 circulating in South Africa during the time of the 1999 African horsesickness outbreak in the Western Cape.

We present the first VP2-gene phylogenetic analysis of African horsesickness (AHS) viruses within a serotype. Thirteen AHSV 7 isolates were obtained from cases that occurred in South Africa during 1998-1999, and three were historical AHSV 7 isolates. The goals were to start a database of isolates of known location and time of isolation and to determine if we could identify the origin of an AHS outbreak in the surveillance area in the Western Cape. We prepared full-length cDNA copies of the VP2-genes of the isolates. Nucleic acid sequence data of a 786 bp region was used to characterize the genetic relationships between the isolates. The nucleic acid identities between the isolates ranged from 95.5 to 100%. Isolates from common geographical regions grouped together. Characterization of field isolates revealed the presence of two AHSV 7 lineages in South Africa during this period. The grouping of the viruses into two clades accurately reflected the geographical groupings of the isolates. The average nucleic acid divergence between the clades was 4.3%. Within the clades the divergence was 0.5 and 0.1%, respectively. The data suggests that the AHS outbreak in the Western Cape could have been an incursion from the Kwazulu Natal Province.

African Horse Sickness↗
PubMed2003

A first full outer capsid protein sequence data-set in the Orbivirus genus (family Reoviridae): cloning, sequencing, expression and analysis of a complete set of full-length outer capsid VP2 genes of the nine African horsesickness virus serotypes.

The outer capsid protein VP2 of African horsesickness virus (AHSV) is a major protective antigen. We have cloned full-length VP2 genes from the reference strains of each of the nine AHSV serotypes. Baculovirus recombinants expressing the cloned VP2 genes of serotypes 1, 2, 4, 6, 7 and 8 were constructed, confirming that they all have full open reading frames. This work completes the cloning and expression of the first full set of AHSV VP2 genes. The clones of VP2 genes of serotypes 1, 2, 5, 7 and 8 were sequenced and their amino acid sequences were deduced. Our sequencing data, together with that of the published VP2 genes of serotypes 3, 4, 6 and 9, were used to generate the first complete sequence analysis of all the (sero)types for a species of the Orbivirus genus. Multiple alignment of the VP2 protein sequences showed that homology between all nine AHSV serotypes varied between 47.6 % and 71.4 %, indicating that VP2 is the most variable AHSV protein. Phylogenetic analysis grouped together the AHSV VP2s of serotypes that cross-react serologically. Low identity between serotypes was demonstrated for specific regions within the VP2 amino acid sequences that have been shown to be antigenic and play a role in virus neutralization. The data presented here impact on the development of new vaccines, the identification and characterization of antigenic regions, the development of more rapid molecular methods for serotype identification and the generation of comprehensive databases to support the diagnosis, epidemiology and surveillance of AHS.

African Horse Sickness↗
PubMed2000

Interaction of Ap1, Ap2, and Sp1 with the regulatory regions of the human pro-alpha1(I) collagen gene.

In the pro-alpha1(I) collagen gene a number of cis-regulatory elements, which interact with a variety of trans-acting factors, are present in the promoter and first intron. We have undertaken a comprehensive study of Sp1, Ap1, and Ap2 binding in the region spanning -442 to +1697 nt. DNase I footprinting analysis revealed these factors bind with varying affinities to some of the potential sites: Sp1 binds to 16 of 34 potential sites, Ap2 binds to 22 of 40 potential binding sites, and Ap1 binds to its only potential site. The Sp1 sites were mostly clustered in the intron region, while the Ap2 sites were clustered in the promoter region. Transmission electron microscopic analysis of DNA-protein complexes not only confirmed these results, but also clearly showed that heterologous and/or homologous protein-protein interactions between Sp1 and/or Ap2 bring the promoter and intron in contact with each other, with the resulting looping out of the intervening DNA. This strongly suggests that the DNA-looping model is an explanation for the orientation preference of the enhancing element in the first intron as these interactions possibly create an optimum environment for the binding of the rest of the transcriptional machinery.

Base Sequence↗
PubMed2000

Characterization of a novel transcription factor binding to the regulatory regions of the human pro-alpha1(I) collagen gene.

We have identified a novel transcription factor TGP (TG Binding Protein) that binds to the consensus sequence 5'-TGTGGGGTGG-3' in the promoter and intron of the human pro-alpha1(I) collagen gene. This recognition sequence, or sequences closely resembling these sequences, was also identified in the pro-alpha1(I) and pro-alpha2(I) collagen genes of other species. Competition experiments revealed that TGP is related to but distinguishable from the Ap4/5 family of transcription factors and that it can be separated from Ap4/5 according to size.

Animals↗
PubMed2000

The utilization of alanine, glutamic acid, and serine as amino acid substrates for glycine N-acyltransferase.

The conjugation of benzoyl-CoA with the aliphatic and acidic amino acids by glycine N-acyltransferase, as well as the amides of the latter group, was investigated. Bovine and human liver benzoyl-amino acid conjugation were investigated using electrospray ionization tandem mass spectrometry (ESI-MS-MS). Bovine glycine N-acyltransferase catalyzed conjugation of benzoyl-CoA with Gly (Km(Gly) = 6.2 mM), Asn (Km(Asn) = 129 mM), Gln (Km(Gln) = 353 mM), Ala (Km(Ala) = 1573 mM), Glu (Km(Glu) = 1148 mM) as well as Ser in a sequential mechanism. In the case of the human form, conjugation with Gly (Km(Gly) = 6.4 mM), Ala (Km(Ala) = 997 mM), and Glu was detected. The presence of these alternative conjugates did not inhibit bovine glycine N-acyltransferase activity significantly. Considering the relatively low levels at which these conjugates are formed, it is unlikely that they will have a significant contribution to acyl-amino acid conjugation under normal conditions in vivo. However, their cumulative contribution to acyl-amino acid conjugation under metabolic disease states may prove to have a useful contribution to detoxification of elevated acyl-CoAs.

Acyltransferases↗
PubMed2000

Antibiotic-resistant gram-negative bacteria in a virtually closed water reticulation system.

The effect of the effluent from a chicken meat-processing plant on the antibiotic-resistant bacterial profile was investigated in an almost closed water reticulation system. Of the 273 faecal coliform isolates 256 (93%) were resistant to one or more of the eight antibiotics tested. The most prevalent isolates were for the beta-lactam antibiotics ampicillin and cephalothin followed by the sulphonamides sulphatriad and cotrimoxazole. Eleven different resistance patterns were identified with a single pattern, comprising of ampicillin-, cephalothin-, streptomycin-, sulphatriad-, cotrimoxazole- and tetracyclin-resistant isolates, dominating the meat-processing effluent. An apparent correlation was observed between the specific use of certain antibiotics and the prevalence of the corresponding resistant bacterial isolates. The drugs used to treat the occasional infections, belonging to the beta-lactam and sulphonamide group of antibiotics, seemed to have a more pronounced effect on the antibiotic-resistant bacterial profile in the primary water source than those drugs used as feed additives, oxytetracyclin and the aminoglycoside flavomycin.

Ampicillin↗
PubMed1999

Haemodynamic changes in the cardiovascular system during the early phases of orthostasis.

In this study, we monitored the changes in arterial blood pressure continuously in two groups of Caucasian men during normal passive orthostasis as well as reversed passive orthostasis. Group A consisted of a group of 23 younger men (16 +/- 0.5 years) and group B consisted of 21 older men (62.9 +/- 2.7 years). The normal passive orthostatic test and the reversed passive orthostatic test were used to induce blood pressure changes. We found that the temporary and initial changes in blood pressure during the normal and reversed orthostatic tests were significantly lower in the older group. Heart rate increases were also lower in the older group. These findings could be explained in terms of a reduced compliance of the thin walled venous blood vessels in the elderly.

Adolescent↗
PubMed1998

Carnitine palmitoyltransferase I activity monitoring in fibroblasts and leukocytes using electrospray ionization mass spectrometry.

Carnitine palmitoyltransferase I (CPT I) is one of the enzymes associated with normal mitochondrial membrane transport of certain metabolites. The importance of the enzyme in normal energy production is well illustrated during fasting conditions when a large flux of long-chain fatty acids must be transported over the mitochondrial membrane to undergo beta-oxidation. Up to now CPT I activity has been assayed in various tissues, including liver, leukocytes, platelets, and fibroblasts by the use of an isotope exchange forward assay which measures the rate of palmitoyl-l-[methyl-3H]carnitine formation from palmitoyl-CoA and l-[methyl-3H]carnitine. We have developed an electrospray ionization mass spectrometric method for detecting palmitoylcarnitine formation from palmitoyl-CoA and carnitine, thus avoiding the use of radiolabeled isotopes. In this assay, time-dependent conversion of free carnitine by CPT I to palmitoylcarnitine is measured quantitatively, relative to isotopically labelled palmitoylcarnitine, by parent ion monitoring of fragment ion m/z 85. The specific activity of CPT I in fibroblasts and leukocytes compared well with the activity determined with the isotope exchange method, however, the combination of high sensitivity and selectivity of tandem mass spectrometry along with the environment-friendly nature of the electrospray method makes it an ideal technique to measure CPT I activity.

Carnitine O-Palmitoyltransferase↗
PubMed1997

Circadian blood pressure and systemic haemodynamics during 42 days of 6 degrees head-down tilt.

Head-down tilted bedrest is a ground-based microgravity simulation model. Since in this position the influence of chief external determinants of circadian blood pressure variation, i.e. activity and posture, are reduced, it may reveal endogenous oscillatory factors. The effects of 42 days of 6 degrees head-down tilt on the circadian profiles of continuous finger blood pressure, heart rate, stroke volume, cardiac output and total peripheral resistance were analysed. In seven healthy volunteers (25-31 years) twelve 22 h Portapres registrations were performed: two in an ambulatory baseline period, eight during 42 days of head-down tilt, and two during recovery. Stroke volume was estimated by a pulse contour method ('Modelflow') from the finger arterial blood pressure tracing. Head-down tilt rapidly reduced circadian BP variation, especially for diastolic blood pressure. No effect of long-term head-down tilt on blood pressure level was observed. The day-night difference in heart rate was essentially unaffected. Cardiac output was maintained through an increase of heart rate and simultaneous decline of stroke volume. Our observations confirm the overriding importance of physical activity and orthostatic load on the diurnal variation of BP. The time-frame of the changes in stroke volume and heart rate during head-down tilt might point to a contribution of other factors besides a reduction of circulating blood volume affecting cardiovascular performance under these conditions.

Adult↗
PubMed1996

Contribution of regions 3' and 5' to the hIL-5 gene on the expression of rhIL-5 in CHO-cells.

The effect of the presence of the regions 5' and 3' of the hIL-5 gene on the expression of recombinant hIL-5 in CHO-cells was investigated. The 3.2 kb hIL-5 gene-fragment was cloned and four different dexamethasone-inducible rhIL-5 mammalian expression-vectors were constructed, each containing a different part of the gene-fragment. Results indicated that deletion of the region 5' to the gene increases production three-fold whereas a four-fold decrease in production was observed with deletion of the region 3' to the gene. Deletion of both regions increased rhIL-5 production by only one and a half-fold. These results indicate that there are elements in a 928 bp region 3' to the gene, including the 3'-NTR, that have an enhancing or stabilizing effect on expression of hIL-5 in CHO-cells.

Animals↗
PubMed1993

A cysteine for glycine substitution at position 175 in an alpha 1 (I) chain of type I collagen produces a clinically heterogeneous form of osteogenesis imperfecta.

The molecular basis for Osteogenesis Imperfecta in a large kindred with a highly variable phenotype was identified by sequencing the mutant pro alpha 1 (I) protein, cDNA and genomic DNA from the proband. Fibroblasts from different affected individuals all synthesize both normal Type I procollagen molecules and abnormal Type I procollagen molecules in which one or both pro alpha 1 (I) chain(s) contain a cysteine residue within the triple helical domain. Protein studies of the proband localized the mutant cysteine residue to the alpha 1 (I) CB 8 peptide. We now report that cysteine has replaced glycine at triple helical residue 175 disrupting the invariant Gly-X-Y structural motif required for perfect triple helix formation. The consequences include post-translational overmodification, decreased thermal stability, and delayed secretion of mutant molecules. The highly variable phenotype in the present kindred cannot be explained solely on the basis of the cysteine for glycine substitution but will require further exploration.

Amino Acid Sequence↗
PubMed1990

A foundation for physiology.

From a study of the literature on the Philosophy of Physiology it became clear that a crisis exists. This crisis was studied by means of a classification of philosophical views. Two main categories, namely Positivistic and Transcendentalistic, frameworks of thought were used. Under the Positivistic category such models as radical materialistic, mechanistic and cybernetic were placed. Under the Transcendentalistic category were classified holistic, humanistic, mystic and eastern ideas. Paradoxic ideas were discussed. None of the frameworks of thought supply a good foundation for Physiology. A liberating foundation for Physiology is proposed. A model of man as a personal unity in a created environment is described. This model does not contain either the contradictions nor polarizations of the positivistic versus transcendentalistic frameworks of thought.

Humans↗
PubMed1988

Complete nucleotide sequence of the region encompassing the first twenty-five exons of the human pro alpha 1(I) collagen gene (COL1A1)

Dysfunctions of the genes coding for the two chains of the human type-I procollagen result in genetic disorders that affect the integrity of bone, ligaments, tendons, and other connective tissues. While the primary amino acid (aa) sequence of one of the two type-I subunits, pro alpha 2(I), has been derived in its entirety from the analysis of overlapping cDNAs, the sequence of the first 247 aa residues of the helical domain of the other polypeptide, pro alpha 1(I), had yet to be determined. To this end, we have sequenced nearly 4 kb of the human pro alpha 1(I) collagen gene and identified twelve open reading frames whose conceptual amino acid translation exhibits 95% homology to the first 247 aa of rat alpha 1(I) chain. Furthermore, with these and other data, some of which previously unpublished, we have derived the complete sequence of the first 7618 bp of the gene. This region comprises the 25 exons encoding the N-terminal pre-propeptide and five of the eight cyanogen-bromide-derived peptides. This information therefore represents a most useful reference for the characterization of molecular defects in individuals affected by various connective tissue disorders.

Amino Acid Sequence↗
PubMed1988

Experimental gallop rhythm in sheep with Gousiekte: correlation of changes in amplitude with haemodynamic parameters.

To investigate the correlation of haemodynamic parameters with the intensity of the gallop sound (S3), use was made of right heart catheterization with a Swan-Ganz catheter to measure the pulmonary and right atrial pressures. The cardiac output was determined with the thermodilution method. A radiocardiogram was obtained after a bolus injection of technetium pertechnetate. The cardiopulmonary flow-index was obtained from the simultaneous recordings of the radiocardiogram and an electrocardiogram. With the haemodynamic parameters, heart sounds were recorded simultaneously and externally with a microphone. Eight Merino sheep were dosed with dried Pachystigma pygmaeum (Schltr) Robyns plant material through rumen fistulas until the clinical symptoms of heart failure such as gallop sounds, systolic murmurs and haemodynamic symptoms as well as increases in end diastolic pressure and decreases in stroke volume, appeared. After the appearance of the symptoms the sheep were treated symptomatically to delay the development of the cardiomyopathy. The changes in haemodynamic parameters before and after treatment were used and correlated with the intensity of the gallop sounds on a 6 point criterium scale. The results in brief show that, for gallop sound intensities between 0 and 6 on the criterium scale, most of the haemodynamic parameters correlate with the intensity of the gallop sounds, except the cardiac output, cardiopulmonary blood volume and the systemic and pulmonary vascular resistances. The haemodynamic parameters correlate better with the intensities of the gallop rhythm between 0 and 3. It is evident from this study that the model of heart failure in sheep is useful to study heart sounds and may also be valuable in the study of the genesis of the gallop sound.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Isolation and characterization of a cDNA coding for a novel human 17.3K myelin basic protein (MBP) variant.

Human fetal spinal cord poly A (+) mRNA was found to direct the synthesis of three major myelin basic protein (MBP) variants with molecular weights of 17K, 18.5K, and 21.5K when translated in reticulocyte lysates. In order to investigate the structural relationships between these MBP variants and their corresponding mouse variants, human fetal spinal cord and mouse brain cDNA libraries were constructed and screened for MBP cDNAs. A number of MBP cDNA clones were isolated and characterized. One of these, PP535 contained the entire coding region of the mouse 14K MBP; and another mouse cDNA clone, PP1.85, was almost full-length and coded for either the 21.5K MBP or the 18.5K MBP. A human clone (KK36), 1,173 nucleotides in length, contained the entire coding region of an MBP variant with a molecular weight of 17,342. The structure of this clone within its coding region is significantly different from the corresponding mouse 17K MBP cDNA. It is missing two sequences found in the mouse 17K MBP cDNA (exons 2 and 5); and it contains a sequence (exon 6) that is missing from the mouse 17K MBP cDNA. Thus, this human 17.3K cDNA codes for a "17K" human MBP variant that is quite different from the corresponding mouse variant and is identical to the human 18.5K MBP except for a deletion of a peptide consisting of 11 amino acids that includes the single tryptophan residue of the 18.5K MBP. An analysis of the structure of this 17.3K human MBP cDNA suggests that the major pathway for splicing the primary human MBP gene product may be different from that in the mouse.

Animals↗