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Biomedical subjects

J A Hargreaves

Publications and source records attributed to J A Hargreaves.

At least 19 recordsLinked to original sources

Accurate diagnosis of occlusal carious lesions--a stereo microscope evaluation of clinical diagnosis.

This study was undertaken to validate the caries status of 214 teeth by serial sectioning and microscopy after caries diagnosis using four methods. Two hundred and fourteen extracted human teeth with varying degrees of caries were mounted in the jaws of nine training manikins. All tooth surfaces were examined and recorded for caries by four dentists using bitewing radiographs, fibre-optic transillumination (FOTI), mirror alone and a mirror and sharp probe on two separate occasions. Thereafter the teeth were serially sectioned and assessed microscopically for depth of caries lesion on a graded score of 0-7. This report assessed the diagnostic outcome of 2,183 observations for occlusal surfaces. Sound diagnoses predominated over unsound until caries was present in the inner half of dentine. Specificity was between 90% and 95% and sensitivity 26% and 50% depending on which diagnostic method was used and where the sound/unsound threshold was set. Negative and positive predictive values were similarly influenced and varied between 53% and 80% and 73% and 90%, respectively. Probit analysis showed no significant differences (P < 0.05) between examiners and diagnostic methods. Diagnosis of occlusal caries undertaken in an in vitro simulated clinical situation is inaccurate until the caries lesion extends deep into the dentine no matter which of the four methods was used.

Chi-Square Distribution↗

Examiner performance with visual, probing and FOTI caries diagnosis in the primary dentition.

To compare clinical reproducibility of dental caries diagnosis in the primary dentition under field conditions, a convenience sample of 5-year-old children in a nursery school in Germiston, was examined for dental caries by four dentists using visual (mirror), visual plus tactile (mirror plus probe) and fibre-optic transillumination (FOTI) methods. Seventeen children were examined on day one and 11 re-examined on day two. Inter-examiner agreement was high, above 90%. Visual examination on its own is comparable with the traditional visual plus tactile method and to FOTI under field conditions. New caries data collected by visual diagnosis alone may, reasonably, be compared with historical data diagnosed with visual + tactile examination.

Chi-Square Distribution↗

Lack of evidence for antisense suppression in the fungal plant pathogen Ustilago maydis.

Modulation of the expression of the Ustilago maydis Pyr3 gene, encoding dihydroorotase (DHOase), through antisense RNA regulation has been explored. This was done by placing the gene in sense and antisense orientations under the control of an hsp70-like gene promoter in a high-copy number autonomously replicating expression vector. Cells transformed with the antisense vector contained similar levels of DHOase activity to those found in cells harboring the expression vector alone. Transformants containing the antisense vector did not exhibit uridine-dependent growth, which would be expected for a Pyr3-deficient phenocopy. This was despite detection of high levels of antisense RNA transcripts in cells transformed with the Pyr3 antisense vector.

Blotting, Northern↗

Trauma to primary teeth of South African pre-school children.

A clinical survey of 1466 children of typical South African communities showed a prevalence rate of primary tooth trauma of 15%. Trauma was least common at age 1-2 years (10.7%) and most common at 4-5 years (20.6%). The commonest trauma seen was fracture of enamel only (71.8%) followed by fracture of dentine (11.2%), tooth loss (8.2%) and discolouration without other sign of injury (5.6%).

Age Distribution↗

Isolation of an Ustilago maydis ERG11 gene and its expression in a mutant deficient in sterol 14 alpha-demethylase activity.

A gene (ERG11) encoding cytochrome P450 sterol 14 alpha-demethylase (P450(14DM)) was isolated from the maize pathogen, Ustilago maydis, by amplifying part of the coding region of the gene using PCR and by employing the amplified DNA fragment as a hybridization probe to recover the complete gene from an U. maydis lambda EMBL3 genomic library. The deduced amino acid sequence of the U. maydis gene showed homology to P450(14DM)s from other organisms and contained specific motifs which were hallmarks of P450s. Expression of the gene in an U. maydis mutant (A20) deficient in P450(14DM) led to only a partial restoration of P450(14DM) activity. Accumulation of ergosta-7,22-dienol and ergost-7-enol in A20 transformants containing the ERG11 gene implied that an additional mutation affecting sterol delta 5,6-desaturase activity accompanied the P450(14DM) lesion.

Amino Acid Sequence↗

Defence-related gene activation during an incompatible interaction between Stagonospora (Septoria) nodorum and barley (Hordeum vulgare L.) coleoptile cells.

Two previously unidentified cDNA clones (bsi1 and bpr1-1) were isolated by differential hybridization from a cDNA library of Stagonospora (Septoria) nodorum (Berk) Castellani & E.G. Germano (teleomorph Phaeosphaeria (Leptosphaeria) nodorum (E. Muller) Hedjaroude-challenged barley (Hordeum vulgare L.) coleoptiles. bsi1 encoded a cysteine-rich protein containing 89 amino acids (aa) with a relative molecular mass (M(r)) of 9405. Protein sequence homologies showed that Bsi1 was very similar to an aluminium-induced protein from wheat and indicated that it was related to the Bowman-Birk-type proteinase inhibitors (BB-PIs). The predicted aa sequence of Bsi1 contained an N-terminal secretory signal sequence which implied that the protein was exported. The other clone, bprl-1, which was truncated at the 5' end, encoded a type-1 pathogenesis-related (PR-1) protein. The complete sequence of bpr1-1 was obtained after cloning a barley genomic DNA fragment and was shown to encode a basic protein containing 174 aa with a M(r) of 18 859. The deduced aa sequence of bpr1-1 contained both an N-terminal secretory signal sequence and a charged C-terminal extension. This latter sequence may represent a vacuolar targeting signal. bsil and bpr1-1 and four other defence-related genes (encoding 1,3-beta-glucanase, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, a homologue of a putative wheat peroxidase, and barley leaf-specific thionin), showed increased transcription levels in S. nodorum-challenged coleoptiles, although their pattern of accumulation varied after inoculation (a.i.). The potential role of these induced genes in defence against fungal attack is discussed.

Amino Acid Sequence↗

A gene encoding gamma-adaptin is required for apical extension growth in Ustilago maydis.

The nucleotide (nt) sequence of an Ustilago maydis (Um) gene that complemented a partially osmotic-remedial temperature-sensitive (or-ts) mutant defective in apical extension growth has been determined. It contained a continuous open reading frame (OFR) predicted to encode a protein of 853 amino acids (aa). The deduced aa sequence was homologous to gamma-adaptin, a component of clathrin-coated vesicles derived from the Golgi.

Adaptor Protein Complex gamma Subunits↗

Discussion: diet and nutrition in dental health and disease.

The recognition in 1993 by the International Association of Dental Research (IADR) for the initiation of a Nutrition Research Group within the Association prompted this nutrition symposium related to diet and nutrition in health and disease. The IADR Nutrition Research Group is encouraged to collaborate with other nutritional research groups in important areas of medicine, food science technology, and agriculture to target health issues influenced by dietary choices, nutritional adequacy, and dental health status.

Dental Caries↗

Energy intake, dental caries and periodontal disease in 11-year-old black children in two regions of Southern Africa: KwaZulu and Namibia.

The study examined energy intake in relation to dental caries and periodontal disease in 11-yr-old rural and urban black children in low fluoride areas of KwaZulu and Namibia (0.15 ppm F) and one higher fluoride area of Namibia (1.56 ppm F). Twenty-four hour dietary recalls were conducted by trained interviewers and daily energy intake estimated using the MRC dietary analysis programme. DMFS was recorded according to WHO criteria: periodontal disease was measured using CPITN and SAS was used for statistical analysis. The prevalence of dental caries and periodontal disease (using CPITN), were compared within three energy groupings; < or = 850 kcal/day; > 850, < 1400 kcal/day and > or = 1400 kcal/day. The rural low fluoride Namibian children had the lowest mean energy intake (616 kcal/day), which also was the grouping with highest healthy periodontal prevalence (65%). The urban groups had higher energy intakes than the rural communities. Statistically significant effects on caries prevalence were seen for country and fluoride grouping; for periodontal disease, significant effects were noted for country, fluoride group and environment. Energy intake had no statistically significant effect, so this is not a risk marker for the disease.

Black or African American↗

Isolation of the ERG2 gene, encoding sterol delta 8-->delta 7 isomerase, from the rice blast fungus Magnaporthe grisea and its expression in the maize smut pathogen Ustilago maydis.

The Magnaporthe grisea ERG2 gene, encoding delta 8-->delta 7 sterol isomerase, was isolated from a genomic library by heterologous hybridization to a fragment of the Ustilago maydis ERG2 gene. The isolated gene contained a reading frame of 745 bp which encoded a protein of 221 amino acids. The coding region was interrupted by a single putative 79-bp-long intron. The deduced amino-acid sequence exhibited similarity to the ERG2 gene products of U. maydis and of Saccharomyces cerevisiae, particularly in the central region of the proteins. The NH2-terminal of all three proteins contained a long stretch of amino acids that were strongly hydrophobic, suggesting that they may function by anchoring the protein to a membrane surface. The M. grisea ERG2 gene complemented a U. maydis deletion mutant in which the ERG2 gene had been removed using a one-step gene replacement procedure. The delta 8-->delta 7 sterol isomerase produced by the M. grisea ERG2 gene exhibited a level of sensitivity to the sterol biosynthesis inhibitor, tridemorph, similar to that of the enzyme derived from the U. maydis ERG2 gene.

Amino Acid Sequence↗

Seasonal variation of vitamin A (retinol) status in older men and women.

OBJECTIVE: The present study was undertaken to determine vitamin A status in 59 free-living (26 males, 33 females) healthy older persons (65-74 years) in winter and summer. DESIGN: Three-day dietary intake data for vitamin A along with carbohydrate, lipid and protein were collected during the summer (June-September) and again during the winter (November-March). In addition, retinol and its carrier proteins, retinol-binding protein (RBP) and transthyretin (TTR), were measured in the plasma in each season. RESULTS: The mean vitamin A intake met the Canadian Recommended Intake (RNI) for both gender and season. However, probability analysis of dietary data revealed that 7 and 11% of males, and 8 and 14% of females, in summer and winter, respectively, were at risk of deficiency. None of the subjects in the present study exhibited biochemical evidence of vitamin A deficiency as determined by plasma levels of retinol and its transport proteins. Overall, the mean intake of vitamin A was significantly higher in males than in females; no seasonal effect was observed. On the other hand, the plasma levels of retinol and its carrier proteins were significantly lower in winter season than in summer, without any gender variation effect. CONCLUSION: Although mean values for dietary intake and plasma concentration of vitamin A may indicate nutritional adequacy, a small proportion of an older population may be at nutritional risk. The prevalence of risk appears to be generally higher in the winter than in the summer season and in females than in males.

Aged↗

Isolation of an Ustilago maydis gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase and expression of a C-terminal-truncated form in Escherichia coli.

A gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase was isolated from the maize fungal pathogen Ustilago maydis. This was accomplished by identifying cDNA and genomic clones that hybridized to an internal fragment of the gene, amplified from U. maydis genomic DNA by PCR. The nature of the gene was determined by nucleotide sequence analysis, and by comparing the derived amino acid sequence of the gene with HMG-CoA reductases from yeast, and from other organisms. The hydrophobic nature of the N-terminal region of the deduced protein sequence also supported the view that this gene encoded HMG-CoA reductase. A C-terminal-truncated fragment of the U. maydis HMG-CoA reductase gene was shown to be expressed in Escherichia coli in a catalytically active form. The expressed protein was also shown to be sensitive to an inhibitor of mammalian HMG-CoA reductase activity.

Amino Acid Sequence↗

Dental caries and social factors in 12-year-old South African children.

Dental caries prevalence (percentage caries-free) and experience (DMFS) were recorded, in 414 12-yr-old Indian and 401 white children living in adjacent urban communities with the same fluoride concentration in the drinking water (0.21-0.33 ppm) using WHO (11) criteria. Details of social factors: education level, family income, home space and occupants and parental occupation were obtained by questionnaire. Dental caries was significantly worse in the Indian children with regard to numbers caries-free (30%--white and 40%--Indian) and DMFS mean (sd) (3.65 (3.98) and 2.66 (3.49) working group, respectively). Social class, white or blue collar, family income and room to person ratio were significantly associated with dental caries in the white children but there were no significant associations in the Indian children. Multiple regression analysis showed race and sex to be significant factors.

Chi-Square Distribution↗

Associations of dental caries with salivary mutans streptococci and acid producing bacteria in 5-year-old children from KwaZulu and Namibia.

Two hundred and eighty-five, 5-year-old children from rural and urban KwaZulu and Namibia were examined for dental caries, salivary mutans streptococci and salivary acid producing bacteria. The findings showed statistically significant correlations between salivary mutans streptococci counts and ds and dmfs scores in all groups of children, the highest values being in Namibian children. Salivary acid forming bacteria had low correlations with few statistically significant groups. While salivary mutans streptococci counts may be a useful caries screening method in children of this age, salivary acid forming bacterial counts appear unsuitable.

Acids↗

Dental caries and fluorosis in 7-9 and 11-14 year old children who received fluoride supplements from birth.

One hundred and sixty children who had lived from birth in a region with low fluoride levels in the drinking water, and who had been offered sodium fluoride supplementation in the form of drops for daily use, were examined to evaluate dental caries and dental fluorosis. Two age ranges were selected: 7-9 years and 11-14 years. In addition to the dental examinations, questionnaires were mailed to the parents, followed-up by telephone interviews, to gather information on compliance with the fluoride supplementation program. The results showed no statistically significant differences in dental caries activity between the regular and irregular users of fluoride supplementation. Considerable dental fluorosis was found in both regular and irregular user groups of the fluoride supplement (38 to 63 per cent of the children seen), however, with no statistical difference between the user groups. Most of the fluorosis detected was of a mild degree. Fluoride supplementation under the daily control of a parent or child is not recommended because of the difficulty in maintaining regular compliance and the risk of fluorosis.

Adolescent↗