An evaluation of knowledge and perception of pharmacology in undergraduate oral hygiene students and recent graduates at a single higher institution in Pretoria.

INTRODUCTION: Pharmacology is an increasingly important area of study for oral hygienists, as it provides the scientific basis for safe and effective oral healthcare. However, a lack of fundamental understanding of the discipline among clinical graduates can present significant challenges. Oral hygienists require pharmacological training to meet the requirements of their scope of practice. Pharmacology knowledge assists with the diagnosis and treatment of oral conditions and forms the foundation for further clinical competency development. The knowledge and perceptions of pharmacology for pharmacy, nursing and medical students have been well documented; however, little information is present for Bachelor of Oral Hygiene (BOH) students. This paper sets out to evaluate BOH students' and recent graduates' knowledge and perceptions of pharmacology at a single higher institution in Pretoria to identify possible gaps and weaknesses. METHODS: A cross-sectional study design was used to collect data using an online questionnaire. The English-language questionnaire consisted of the self-reported perceptions and knowledge and actual knowledge of pharmacology of undergraduate BOH students and recent graduates. The questionnaire consisted of multiple choice questions, true or false questions and Likert scale questions. Ethics was obtained from the institution's Research Ethics Committee (REC 350/2021). RESULTS: Overall, the participants perceived the pharmacology module positively and understood its importance. Concerns were raised about insufficient time for studying and that assessments were more aligned to gaining factual knowledge than the development of problem-solving skills. Students rated their knowledge between 57.24% and 69.44%, with BOH III students and graduates having a statistically significant greater self-rated knowledge of antivirals, antifungals and common agents used to treat oral conditions in comparison with BOH I and BOH II students. Overall, BOH students and graduates' actual knowledge was between 45.24% and 66.84%. Although not statistically significant, the total self-rated knowledge of BOH III students and recent graduates tended to be higher than their actual knowledge. Knowledge deficits were evident with some pharmacological concepts across the various BOH groups, such as pharmacokinetics, pain, drugs altering dental treatment I: central nervous system drugs, drugs altering dental treatment II: respiratory and endocrine drugs, drugs altering dental treatment III: cardiovascular drugs, drug-drug interactions and common agents used to treat oral conditions. CONCLUSION: Self-rated knowledge deficiencies were noted by students and recent graduates for certain pharmacological concepts and were supported by the measurement of their actual knowledge. Further investigation into knowledge deficiencies is needed to guide curriculum review to further strengthen oral hygienists' pharmacological competencies and ensure alignment to their scope of practice.

Investigating the effect of the heavy metals cadmium, chromium and lead, alone and in combination on an endothelial cell line.

Heavy metals are natural elements characterized by their relatively large atomic mass as well as high density. It can be introduced into the ecosystem by the mining of heavy metals from deep within the earth's crust, thereby exposing the metals into air and water systems. Cigarette smoke is another source of heavy metal exposure and has been shown to have carcinogenic, toxic and genotoxic properties. Cadmium, lead, and chromium are the most abundant metals found in cigarette smoke. In response to tobacco smoke exposure, endothelial cells release inflammatory and pro-atherogenic cytokines that are linked to endothelial dysfunction. Endothelial dysfunction is directly related to the production of reactive oxygen species, leading to endothelial cell loss through necrosis and/or apoptosis. The current study aimed to investigate the effect of cadmium, lead, and chromium, alone and as part of metal mixtures, on endothelial cells. The EA.hy926 endothelial cell line was exposed to different concentrations of each of these metals and their combinations and analyzed using flow cytometric analyses with Annexin V. A clear trend was seen with the Pb + Cr as well as the triple combination group with the significant increase of early apoptotic cells. Scanning electron microscopy was used to study possible ultrastructural effects. Morphological changes observed with scanning electron microscopy included cell membrane damage and membrane blebbing at certain metal concentrations. In conclusion, the exposure of endothelial cells to cadmium, lead, and chromium, caused a disruption in cellular processes and morphology, possibly diminishing the protective ability of endothelial cells.

Fitness costs of rapid cold-hardening in Ceratitis capitata.

Rapid cold-hardening (RCH) is a unique form of phenotypic plasticity which confers survival advantages at low temperature. The fitness costs of RCH are generally poorly elucidated and are important to understanding the evolution of plastic physiology. This study examined whether RCH responses, induced by ecologically relevant diel temperature fluctuations, carry metabolic, survival, or fecundity costs. We predicted that potential costs in RCH would be manifested as differences in metabolic rate, fecundity, or survival in flies which have hardened versus those which have not, or flies that have experienced more RCH events would show greater costs than those which have experienced fewer events. One group of flies cooled to 10°C for 2 h for 11 consecutive days experienced daily RCH (Hardened), whereas the other group exposed to 15°C for the same 2-h period each day formed a Control group. Hardened flies had higher survival at -5°C for 2 h than control flies (69 ± 9% vs. 44 ± 19%, P = 0.04). Hardened flies showed no metabolic or fecundity costs, but had reduced average survival (P = 0.0403). Thus, a major cost to repeated low temperature exposures in Ceratitis capitata is through direct mortality caused by chilling injury, although this appears not to be a direct cost of RCH.

Respiratory pattern transitions in three species of Glossina (Diptera, Glossinidae).

Glossina exhibit cyclic ((CYC)GE) or continuous gas exchange ((CON)GE) patterns at rest. However, the factors influencing the transition from one pattern to another are not well understood for these or other insect species. This study examines which factors could aid in predicting the presence or absence of (CYC)GE in adults of three Glossina species: G. palpalis, G. brevipalpis and G. austeni. We report the results of temperature effects on VCO(2), pattern type and the proportion of a population showing (CYC)GE, and the prediction of (CYC)GE versus (CON)GE in Glossina. First, we investigated the influence of temperature on VCO(2) and found significant elevation in resting metabolic rate (RMR) with higher temperature in all three species (P<0.001). Temperature-induced increases in VCO(2) were modulated by increased burst volume and by cycle frequency, except in G. brevipalpis which only appeared to modulate burst volume. These results are largely in keeping with VCO(2) modulation reported for other Glossina species previously. Second, elevating temperature resulted in significantly reduced numbers of individuals showing (CYC)GE (P<0.001 for all three species) contrary to previous reports for other Glossing species. Finally, we examined a range of variables as potential predictors of presence or absence of (CYC)GE in these three species. Using an information theoretic approach (Akaike weights) to select the best explanatory combination of variables which predicts likelihood of (CYC)GE, we found that results varied among species. When species were pooled, the simplest, best-fit model (ΔAIC<2 from the best model, 44.4% probability of being the best model) for predicting pattern type variation was RMR. Overall these results suggest that RMR is a key variable driving pattern type and that elevated temperature reduces the number of individuals showing cyclic patterns through elevation of RMR in these species. This study supports the idea that an interaction between cellular metabolic demand, morphological features of the gas exchange system (e.g. tracheal and spiracular conductances), and CO(2) buffer capacity likely determine gas exchange pattern variation over short time-scales.

Upregulation of pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) activity in strawberry.

Pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) is a cytosolic enzyme catalyzing the first committed step in glycolysis by reversibly phosphorylating fructose-6-phosphate to fructose-1,6-bisphosphate. The position of PFP in glycolytic and gluconeogenic metabolism, as well as activity patterns in ripening strawberry, suggest that the enzyme may influence carbohydrate allocation to sugars and organic acids. Fructose-2,6-bisphosphate activates and tightly regulates PFP activity in plants and has hampered attempts to increase PFP activity through overexpression. Heterologous expression of a homodimeric isoform from Giardia lamblia, not regulated by fructose-2,6-bisphosphate, was therefore employed to ensure in vivo increases in PFP activity. The coding sequence was placed into a constitutive expression cassette under control of the cauliflower mosaic virus 35S promoter and introduced into strawberry by Agrobacterium tumefaciens-mediated transformation. Heterologous expression of PFP resulted in an up to eightfold increase in total activity in ripe berries collected over two consecutive growing seasons. Total sugar and organic acid content of transgenic berries harvested during the first season were not affected when compared to the wild type, however, fructose content increased at the expense of sucrose. In the second season, total sugar content and composition remained unchanged while the citrate content increased slightly. Considering that PFP catalyses a reversible reaction, PFP activity appears to shift between gluconeogenic and glycolytic metabolism, depending on the metabolic status of the cell.

Metabolic responses of Glossina pallidipes (Diptera: Glossinidae) puparia exposed to oxygen and temperature variation: implications for population dynamics and subterranean life.

Understanding the factors affecting insect gas exchange in subterranean environments is critical to understanding energy budgets and predicting mortality under field conditions. Here, we examine the metabolic rate (MR) responses of tsetse puparia, which remain underground for ca. 1 month in this life-stage, to varying oxygen and temperature. First, the effects of temperature and oxygen on puparial MR were investigated by ramping temperature from 15 to 35°C under 10, 21 or 40% O(2). Overall, temperature was the dominant effect on puparial MR although O(2) had small but significant impacts. Second, critical O(2) concentration (P(CRIT)) for MR of puparia was examined across a range of oxygen concentrations (0-40%). P(CRIT) was 6% O(2) which is similar to P(CRIT) in other basal arthropods but relatively high for inactive or subterranean insects. Third, we asked if puparia exposed to anoxia might experience oxygen debt, potentially indicative of anaerobic metabolism or cellular repair. Metabolic responses to anoxia were limited or insignificant, but MR was marginally elevated (∼ 15%) in anoxia-exposed (4h) puparia by 12h post-anoxia. Finally, we examined the ability of puparia to withstand water submersion, thus simulating flooding conditions frequently experienced in tropical soil habitats. Puparia were unable to survive submersion for >24h suggesting limited flooding tolerance. These novel results suggest that soil conditions experienced by puparia should not be limiting for MR, except possibly under high temperature-low O(2) conditions. Due to a large safety margin between P(CRIT) and soil oxygen levels and limited effects of oxygen on metabolism during temperature ramping experiments, we suggest that Glossina pallidipes puparia are not particularly susceptible to oxygen availability in their natural environment. However, soil flooding associated with tropical rainfall likely imposes strong selection on tsetse populations and may have had important effects for tsetse energy budgets and evolution.

Probability of assertive behaviour, interpersonal anxiety and self-efficacy of South African registered dietitians.

OBJECTIVES: There is little information on the probability of assertive behaviour, interpersonal anxiety and self-efficacy in the literature regarding dietitians. The objective of this study was to establish baseline information of these attributes and the factors affecting them. METHOD: Questionnaires collecting biographical information and self-assessment psychometric scales measuring levels of probability of assertiveness, interpersonal anxiety and self-efficacy were mailed to 350 subjects, who comprised a random sample of dietitians registered with the Health Professions Council of South Africa. RESULTS: Forty-one per cent (n=145) of the sample responded. Self-assessment inventory results were compared to test levels of probability of assertive behaviour, interpersonal anxiety and self-efficacy. The inventory results were compared with the biographical findings to establish statistical relationships between the variables. The hypotheses were formulated before data collection. CONCLUSION: Dietitians had acceptable levels of probability of assertive behaviour and interpersonal anxiety. The probability of assertive behaviour was significantly lower than the level noted in the literature and was negatively related to interpersonal anxiety and positively related to self-efficacy.

Statin therapy and stroke prevention.

Hypercholesterolemia has not traditionally been considered an important risk factor in the pathogenesis of stroke. However, recent studies show that statin therapy significantly reduces ischemic stroke for patients with established coronary artery disease. Statin therapy may reduce stroke through amelioration of precerebral atherosclerosis in the carotid artery and the aorta. Anti-atherosclerotic, anti-inflammatory, and antithrombotic actions of statins occur within the blood and in plaque. Statins may also protect against cerebral ischemia through beneficial modulation of the brain endothelial nitric oxide system. Ongoing studies are exploring the role of statin therapy in the primary prevention of stroke and in the prevention of cognitive decline and multi-infarct cerebrovascular disease.

Do statins afford neuroprotection in patients with cerebral ischaemia and stroke?

An emerging body of evidence indicates that beta-hydroxy-beta-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or 'statins', provide neuroprotection in addition to reducing ischaemic stroke. Statins reduce the incidence of ischaemic stroke by stabilising atherosclerotic plaques in the precerebral vasculature and through antithrombotic actions, and the neuroprotective effects of statins may confer significant clinical benefit. Some of these neuroprotective effects are likely to be cholesterol independent and mediated by the interruption of isoprenoid biosynthesis. Therapy with statins may modulate endothelial function and preserve blood flow to regions exposed to an ischaemic insult. In particular, statin-mediated preservation of endothelial nitric oxide synthase activity in cerebral vasculature, especially in the ischaemic penumbra, may limit neurological deficit. Moreover, putative anti-inflammatory and antioxidant properties of statins may confer additional neuroprotection. Further large clinical trials are necessary to address the role of statin therapy in the primary prevention of stroke, small vessel cerebrovascular disease and vascular dementia.

Tumors and the heart: molecular genetic advances.

Recent molecular genetic investigations of primary cardiac tumors (myxomas, lipomas, rhabdomyomas, and fibromas) have provided insight into fundamental mechanisms of cardiac cell growth. Myxomas are the most common adult cardiac tumor, and familial cardiac myxomas are now appreciated to be caused by mutations in the PRKAR1alpha gene that encodes a regulatory subunit of protein kinase A. Cytogenetic studies have targeted candidate chromosomal loci that may be perturbed during cardiac lipoma pathogenesis. Rhabdomyomas, the most common pediatric cardiac neoplasm, are frequently associated with tuberous sclerosis, caused by mutations in the TSC-1 and TSC-2 genes. The study of Gorlin syndrome has shed light on the etiology of cardiac fibromas. This disorder is caused by mutation of the PTC gene, which regulates cell growth, commitment and differentiation. In the future, manipulation of PRKAR1alpha-, TSC-, and PTC-dependent pathways may foster new strategies to regenerate myocardium in the ischemic or myopathic heart.

Identification of a chromosome 11q23.2-q24 locus for familial aortic aneurysm disease, a genetically heterogeneous disorder.

BACKGROUND: Aortic aneurysms cause significant mortality, and >20% relate to hereditary disorders. Familial aortic aneurysm (FAA) has been described in such conditions as the Marfan and Ehlers-Danlos type IV syndromes, due to defects in the fibrillin-1 and type III procollagen genes, respectively. Other gene defects that cause isolated aneurysms, however, have not thus far been described. METHODS AND RESULTS: We studied 3 families affected by FAA. No family met the diagnostic criteria for either Marfan or Ehlers-Danlos syndrome. Echocardiography defined involvement of both the thoracic and abdominal aorta. In family ANA, candidate gene analysis excluded linkage to loci associated with aneurysm formation, including fibrillin-1, fibrillin-2, and type III procollagen, and chromosome 3p24.2-p25. Genome-wide linkage analysis identified a 2.3-cM FAA locus (FAA1) on chromosome 11q23.3-q24 with a maximum multipoint logarithm of the odds score of 4.4. In family ANB, FAA was linked to fibrillin-1. In family ANF, however, FAA was not linked to any locus previously associated with aneurysm formation, including fibrillin-1 and FAA1. CONCLUSIONS: FAA disease is genetically heterogeneous. We have identified a novel FAA locus at chromosome 11q23.3-q24, a critical step toward elucidating 1 gene defect responsible for aortic dilatation. Future characterization of the FAA1 gene will enhance our ability to achieve presymptomatic diagnosis of aortic aneurysms and will define molecular mechanisms to target therapeutics.

Familial thoracic aortic aneurysms and dissections: genetic heterogeneity with a major locus mapping to 5q13-14.

BACKGROUND: Aneurysms and dissections affecting the ascending aorta are associated primarily with degeneration of the aortic media, called medial necrosis. Families identified with dominant inheritance of thoracic aortic aneurysms and dissections (TAA/dissections) indicate that single gene mutations can cause medial necrosis in the absence of an associated syndrome. METHODS AND RESULTS: Fifteen families were identified with multiple members with TAAs/dissections. DNA from affected members from 2 of the families was used for a genome-wide search for the location of the defective gene by use of random polymorphic markers. The data were analyzed by the affected-pedigree-member method of linkage analysis. This analysis revealed 3 chromosomal loci with multiple markers demonstrating evidence of linkage to the phenotype. Linkage analysis using further markers in these regions and DNA from 15 families confirmed linkage of some of the families to 5q13-14. Genetic heterogeneity for the condition was confirmed by a heterogeneity test. Data from 9 families with the highest conditional probability of being linked to 5q were used to calculate the pairwise and multipoint logarithm of the odds (LOD) scores, with a maximum LOD of 4.74, with no recombination being obtained for the marker D5S2029. In 6 families, the phenotype was not linked to the 5q locus. CONCLUSIONS: A major locus for familial TAAs and dissections maps to 5q13-14, with the majority (9 of 15) of the families identified demonstrating evidence of linkage to this locus. The condition is genetically heterogeneous, with 6 families not demonstrating evidence of linkage to any loci previously associated with aneurysm formation.

TBX5 transcription factor regulates cell proliferation during cardiogenesis.

Mutations in human TBX5, a member of the T-box transcription factor gene family, cause congenital cardiac septation defects and isomerism in autosomal dominant Holt-Oram syndrome. To determine the cellular function of TBX5 in cardiogenesis, we overexpressed wild-type and mutant human TBX5 isoforms in vitro and in vivo. TBX5 inhibited cell proliferation of D17 canine osteosarcoma cells and MEQC quail cardiomyocyte-like cells in vitro. Mutagenesis of the 5' end of the T-box but not the 3' end of the T-box abolished this effect. Overexpression of TBX5 in embryonic chick hearts showed that TBX5 inhibits myocardial growth and trabeculation. TBX5 effects in vivo were abolished by Gly80Arg missense mutation of the 5' end of the T-box. PCNA analysis in transgenic chick hearts revealed that TBX5 overexpression does suppress embryonic cardiomyocyte proliferation in vivo. Inhibitory effects of TBX5 on cardiomyocyte proliferation include a noncell autonomous process in vitro and in vivo. TBX5 inhibited proliferation of both nontransgenic cells cocultured with transgenic cells in vitro and nontransgenic cardiomyocytes in transgenic chick hearts with mosaic expression of TBX5 in vivo. Immunohistochemical studies of human embryonic tissues, including hearts, also demonstrated that TBX5 expression is inversely related to cellular proliferation. We propose that TBX5 can act as a cellular arrest signal during vertebrate cardiogenesis and thereby participate in modulation of cardiac growth and development.

Deriving probes from large-insert clones by PCR methods.

This unit describes several polymerase chain reaction (PCR)-based methods to obtain DNA fragments from clones with large inserts without prior knowledge of the insert DNA sequence. The protocols can be categorized into three groups: (1) methods to generate DNA fragments at random representing the entire length of the cloned insert, (2) methods to generate DNA fragments representing the extremities of an insert, and (3) methods to generate complex probes suitable for fluorescence in situ hybridization. Support protocols describe direct cloning of these PCR products and the isolation of total yeast DNA from yeast artificial chromosome (YAC) clones.

Identification and localization of TBX5 transcription factor during human cardiac morphogenesis.

Mutations in the TBX5 transcription factor gene cause human cardiac malformation in Holt-Oram syndrome. To identify and localize TBX5 during cardiac morphogenesis, we performed immunohistochemical studies of TBX5 protein cardiac expression during human embryogenesis. Specific antibody to human TBX5 was generated in rabbits with a TBX5 synthetic peptide and affinity purification of antiserum. Anti-TBX5 was used in immunohistochemical analyses of human cardiac tissue. In embryonic and adult heart, TBX5 is expressed throughout the epicardium and in cardiomyocyte nuclei in myocardium of all four cardiac chambers. Endocardial expression of TBX5 is only present in left ventricle. Asymmetric left-sided transmyocardial gradients of TBX5 protein expression were observed in embryonic but not adult hearts. Human cardiac expression of TBX5 protein correlates with the cardiac manifestations of Holt-Oram syndrome. TBX5 transmyocardial protein gradients may contribute to normal patterning of the human heart during embryogenesis.

Mutations in the protein kinase A R1alpha regulatory subunit cause familial cardiac myxomas and Carney complex.

Cardiac myxomas are benign mesenchymal tumors that can present as components of the human autosomal dominant disorder Carney complex. Syndromic cardiac myxomas are associated with spotty pigmentation of the skin and endocrinopathy. Our linkage analysis mapped a Carney complex gene defect to chromosome 17q24. We now demonstrate that the PRKAR1alpha gene encoding the R1alpha regulatory subunit of cAMP-dependent protein kinase A (PKA) maps to this chromosome 17q24 locus. Furthermore, we show that PRKAR1alpha frameshift mutations in three unrelated families result in haploinsufficiency of R1alpha and cause Carney complex. We did not detect any truncated R1alpha protein encoded by mutant PRKAR1alpha. Although cardiac tumorigenesis may require a second somatic mutation, DNA and protein analyses of an atrial myxoma resected from a Carney complex patient with a PRKAR1alpha deletion revealed that the myxoma cells retain both the wild-type and the mutant PRKAR1alpha alleles and that wild-type R1alpha protein is stably expressed. However, in this atrial myxoma, we did observe a reversal of the ratio of R1alpha to R2beta regulatory subunit protein, which may contribute to tumorigenesis. Further investigation will elucidate the cell-specific effects of PRKAR1alpha haploinsufficiency on PKA activity and the role of PKA in cardiac growth and differentiation.