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Biomedical subjects

M McIntyre

Publications and source records attributed to M McIntyre.

At least 19 recordsLinked to original sources

Growth physiology and dimorphism of Mucor circinelloides (syn. racemosus) during submerged batch cultivation.

Mucor circinelloides is being investigated as a possible host for the production of heterologous proteins. Thus, the environmental conditions defining the physiology and morphology of this dimorphic fungus have been investigated in submerged batch cultivation. The optimal conditions for growth of each form have been defined. Pure cultures of the multi-polar budding yeast form could be obtained under anaerobic conditions (with 70% N2/30% CO2 or 100% N2 as the sparge gas and without aeration). The highest maximum specific growth rate (0.30 h(-1)) was obtained in anaerobic cultivation, the yield of biomass on glucose (Y(SX)) was 0.12 (c-mole basis). A high maximum specific growth rate was obtained when the organism grew as the filamentous form under aerobic conditions (0.25 h(-1)), with a Y(SX) of 0.24 (c-mole basis). The maximum specific growth rates achieved are comparable to most industrial filamentous fungi under similar growth conditions. High levels of ethanol were observed with all growth conditions. The overriding effector of morphological development was found to be oxygen. In batch cultures it was therefore possible to induce the dimorphic shift by controlling the influent gas atmosphere. A specific growth rate of 0.19 h(-1) was maintained during the shift from the yeast to the filamentous form.

Cell Division↗

Metabolic engineering of the morphology of Aspergillus.

The morphology of filamentous organisms in submerged cultivation is a subject of considerable interest, notably due to the influence of morphology on process productivity. The relationship between process parameters and morphology is complex: the interactions between process variables, productivity, rheology, and macro- and micro-morphology create difficulties in defining and separating cause and effect. Additionally, organism physiology contributes a further level of complexity which means that the desired morphology (for optimum process performance and productivity) is likely to be process specific. However, a number of studies with increasingly powerful image analysis systems have yielded valuable information on what these desirable morphologies are likely to be. In parallel, studies on a variety of morphological mutants means that information on the genes involved in morphology is beginning to emerge. Indeed, we are now beginning to understand how morphology may be controlled at the molecular level. Coupling this knowledge with the tools of molecular biology means that it is now possible to design and engineer the morphology of organisms for specific bioprocesses. Tailor making strains with defined morphologies represents a clear advantage in optimization of submerged bioprocesses with filamentous organisms.

Aspergillus↗

Undergraduate student nurses' expectations and their self-reported preparedness for the graduate year role.

AIMS OF THE STUDY: The study identifies third-year nurses' expectations of the graduate nurse role and ascertains how prepared they feel to fulfil this role. BACKGROUND: The literature substantiates that the university-workplace transition is marked by differences between students' expectations of the graduate year and the realities of practice they encounter in the workforce setting. Nursing professionals and health service employers continue to debate the expectations required of the new nurse graduate. Yet there is little assessment of graduate nurses' expectations of the workplace. This study describes student nurses' expectations of the graduate year and the extent to which they regard themselves as well- or ill-prepared. DESIGN: Third-year student nurses (n=105) from a 3-year Bachelor of Nursing (BN) course at a large Metropolitan University in Australia were surveyed. A group of nursing academics and their senior colleagues in the clinical setting designed a questionnaire in light of common themes derived from literature on the graduate year role. Responses were examined and analysed using descriptive statistics. RESULTS: Responses revealed that student nurses tended to favour large public hospitals, and sought a good graduate programme with associated opportunities for guidance and support. Most expected to achieve good working relationships with both professional colleagues and patients. Final year students expressed some apprehension about meeting the performance expectations of the workplace, given their self-perceived lack of clinical experience. CONCLUSION: When asked about their initial expectations of the workplace, third year student nurses expressed little apprehension and reported high levels on scales of organizational commitment and professionalism. The research literature suggests that divisions exist between students' expectations of the graduate year and the actual work experience. The expectations of the graduate year described in this study offer a student-centred perspective that contributes to future planning and policy directions of undergraduate curricula, graduate year programmes and nurse retention.

Adult↗

Morphological characterization of Aspergillus nidulans: growth, septation and fragmentation.

The influence of the sepA gene on the growth of Aspergillus nidulans has been investigated by characterizing and comparing the parental strain A28 (pabaA6 biA1) with the sepA null mutant (sepA4DeltaBM:). The sepA gene is known to affect the septation process in A. nidulans, therefore the sepA4DeltaBM: strain does not produce any septa during the first hours of growth. During batch cultivations sepA4DeltaBM: shows an abrupt decrease in specific growth rate and more pronounced fragmentation (in response to elevated stirrer speed) than the parental strain. Higher specific fragmentation rates (q(frag)) were obtained for the sepA4DeltaBM: strain. The physiological reasons for the differences have been investigated by employing fluorescent stains. Computerized image analysis revealed that the more pronounced fragmentation in the mutant was due to the lower number and irregular spacing of septa (visualized by calcofluor white staining), which resulted in a weaker hyphal structure that is more vulnerable to shear stress and fragmentation than the parental strain. This led to a loss of active biomass (determined by Mag fura staining) from the hyphae of the mutant, which had failed to compartmentalize by formation of septa, in turn resulting in decreased specific growth rates for the culture.

Aspergillus nidulans↗

Quantification of autolysis in Penicillium chrysogenum by semiautomated image analysis.

An image analysis method is described for the characterization of empty (autolyzed and inactive) regions within the mycelia of filamentous fungi. It extends a previous method that characterized only regions filled with cytoplasm or vacuoles (i.e., the active biomass). The method is semiautomatic, requiring some manual editing before automated measurements. When the method was used for samples from a batch fermentation of an industrial strain of Penicillium chrysogenum, the empty regions were observed to constitute up to 15% (by projected area) of the biomass during the growth phase. After nutrient exhaustion, however, the proportion of empty regions rose rapidly, eventually representing more than 50% of the biomass by the end of fermentation. The increase in the percentage of empty regions coincided with a decrease in biomass (as measured by dry cell weight) and a fall in penicillin titre. Further morphological analysis revealed that fragmentation of mycelia, particularly clumps, coincided with increases in the levels of empty regions. This new image analysis method gave additional information on hyphal differentiation and a measure of autolysis. It was also a useful indicator of the processes leading to autolysis.

Biomass↗

Superoxide excess in hypertension and aging: a common cause of endothelial dysfunction.

There is evidence in humans that hypertension and aging similarly impair endothelial function, although the mechanism remains unclear. Superoxide anion (O(2)(-)) is a major determinant of nitric oxide (NO) bioavailability and thus endothelial function. We sought to determine the relationship between endothelial function, O(2)(-), and age in normotensive Wistar-Kyoto (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP). Aortic rings were removed from female WKY and SHRSP at 3 to 4 months (young) and 9 to 12 months (old). O(2)(-) generation by aortic rings was measured before and after removal of the endothelium or incubation with N(G) nitro-L-arginine methyl ester, diphenyleneiodonium, or apocynin. Levels of p22phox were studied with immunohistochemistry and used as a marker of NAD(P)H oxidase expression. NO bioavailability was significantly lower in old WKY compared with young WKY (P=0.0009) and in old SHRSP compared with young SHRSP (P=0.005). O(2)(-) generation was significantly greater in old WKY compared with young WKY (P=0.0001). Removal of the endothelium and N(G) nitro-L-arginine methyl ester treatment resulted in a significant reduction in O(2)(-) generation in old SHRSP (P=0.009 and 0.001, respectively). Diphenyleneiodonium significantly reduced O(2)(-) generation in 12-month WKY (P=0.008) and 12-month SHRSP (P=0.009). Apocynin attenuated O(2)(-) generation by older WKY (P=0.038) and SHRSP (P=0.028). p22phox was increased in older animals compared with young. We conclude that NO bioavailability decreases with age in female WKY and SHRSP. O(2)(-) generation increases with age in WKY and is higher in SHRSP and may contribute to the reduced NO by scavenging. NAD(P)H oxidase may contribute to the age-related increase in O(2)(-).

Acetophenones↗

Acid-fast bodies in faecal smears stained by the modified Ziehl-Neelsen technique.

The modified Ziehl-Neelsen (ZN) stain has proved useful in the laboratory diagnosis of cryptosporidiosis and, more recently, for the laboratory diagnosis of cyclospora. Apart from cryptosporidia and cyclospora, many other organisms and artefacts are present in faeces, and may be seen in faecal smears stained by the modified ZN method. Described here is the presence of such organisms and artefacts in faecal samples submitted to the routine microbiology laboratory of a district general hospital. Over 6000 faecal smears were examined using the modified ZN method, with an incidence of cyclospora and cryptosporidium of approximately 0.1% and 1%, respectively. Other organisms and artefacts were observed, with an incidence ranging from 0.1% to 1%. It is emphasised that the identification of known gastrointestinal tract pathogens should not rely solely on the results of their staining reactions. It is essential that criteria such as morphology and size be taken into account to differentiate organisms with similar staining reactions.

Animals↗

Trans fatty acids and cardiovascular risk.

Trans fatty acids are found in partially hydrogenated vegetable oil, in meats, and in dairy products. Their effect on blood cholesterol concentrations was examined decades ago, but recently there has been renewed interest in understanding how trans fatty acids affect blood lipids and lipoprotein cholesterol concentrations. Current advice to reduce cardiovascular disease (CVD) risk includes decreasing the consumption of saturated and total fat to help manage blood cholesterol concentrations. Saturated fat contributes significantly to total fat intake and markedly raises blood cholesterol concentrations. Trans fatty acids, which are consumed in much smaller quantities, have been shown to be modestly hypercholesterolemic in studies that have substituted hydrogenated vegetable oils for unhydrogenated oils. In contrast, when partially hydrogenated vegetable oils containing trans fatty acids are substituted for cholesterol-raising saturated fats, blood cholesterol levels are reduced. Partially hydrogenated vegetable oils are used in place of saturated fat in many food products. These foods can help consumers lower their saturated fat intake to achieve dietary recommendations. The following review critically examines the role of hydrogenated fats in the food supply, the metabolism of trans fatty acids, and the scientific literature surrounding the effects of partially hydrogenated vegetable oils and trans fatty acids on blood cholesterol concentrations and cardiovascular disease risk.

Cardiovascular Diseases↗

The intersection of relational and cultural narratives: women's abortion experiences.

Although a great deal has been written on women's personal deliberations in deciding to have and seeking an abortion, little has been written on the experience of living with this decision or on the meanings an abortion generates in the context of a particular life. In this hermeneutic phenomenological study, 14 women aged 19-44 expressed a need for professionals and others to accept the reality of unplanned pregnancy and to acknowledge that the choice of abortion cannot be fully understood in isolation from women's other reproductive choices. Using a feminist analysis of the tensions and contradictions in the women's accounts, the authors explore the intersections between their experiences and the cultural narratives in which they are situated, in a search for new possibilities for women and new understandings of women's experience of abortion.

Abortion, Legal↗

Role of proteases in autolysis of Penicillium chrysogenum chemostat cultures in response to nutrient depletion.

An industrial strain of Penicillium chrysogenum was subjected to carbon or nitrogen limitation in a chemostat and the response monitored in terms of the "classical" indicators of autolysis (biomass decline and ammonia release), culture degradation (as measured by image analysis) and by obtaining profiles for three classes of proteases implicated in autolysis. Under both sets of conditions (carbon or nitrogen limitation), once started, autolysis involved a succession of different protease activities. The first stages of the process of autolysis in starved chemostat cultures was associated with peaks in the activities of both serine and aspartyl proteases, coinciding with the mobilisation of endogenous energy reserves. Conversely, a peak in the activity of metalloproteases was associated with the later stages of autolysis, perhaps occurring in response to depletion of endogenous energy reserves; the activity of these enzymes led to gross culture degradation, disintegration of ordered mycelial structures and signalled the end of metabolic activity (respiration) within the culture. These findings indicate that strategies intended to control/regulate autolysis in large-scale industrial fungal cultures might profitably be focused on regulation of the activity of key classes of proteases involved in the series of events leading to culture degradation.

Ammonia↗

Differential expression of the cyclin-dependent kinase inhibitor P27 in primary hepatocytes in early-mid G1 and G1/S transitions.

P27, an inhibitor of cyclin-dependent kinases, plays an important role in the control of cell adhesion and contact inhibition-dependent cell cycle regulation. Hepatocytes, maintained in primary culture, offer a model of synchronized primary epithelial cells which retain a differentiated profile while stimulated to proliferate. We therefore investigated the pattern of endogenous p27 expression in cyclin rat hepatocytes isolated by collagenase perfusion followed by mitogenic stimulation. P27 was expressed in whole normal liver and freshly isolated hepatocytes. We then observed a sharp decrease in p27 levels, concomitant with the progression in early-mid G1, followed by reaccumulation in late G1 and the G1/S transition. Immunochemistry and BrdU labelling demonstrated nuclear localization of p27 and its expression in cells engaged in both G1 and S phase. P27 was detected in late G1 in complexes containing cyclins D1, E and A. Cyclin E- and A-associated kinase activities, however, were detected at the G1/S transition and depletion experiments confirmed that most active complexes were free of p27. Phosphorylated forms of p27 were detected in unstimulated and stimulated hepatocytes in both early-mid G1 and G1/S. Finally, two-dimensional gel electrophoresis showed evidence for several forms of p27 with a distinct profile of distribution in quiescent and stimulated hepatocytes. Collectively, our data offer a model in which p27 shows a biphasic profile of accumulation, with the early decrease possibly involved in the progression through early and mid G1. In contrast with most cell types tested so far, the late G1 accumulation did not impair formation of active cyclin E- and A associated kinases, and thus G1/S transition.

Animals↗

Angiotropic large B-cell lymphoma with clinical features resembling subacute combined degeneration of the cord.

Angiotropic large cell lymphoma is a rare neoplastic disorder associated with a high mortality. The hallmark of the disease is lymphoid proliferation confined to the intravascular compartment without local tissue or vessel wall infiltration [1]. This feature is so striking that the disease was originally thought to arise from endothelial tissue and early cases were described as malignant angioendotheliomatosis. However, application of immunohistochemical methods for detection of lymphoid markers such as the CD45 and CD20 cell surface markers has confirmed its lymphoid origin, usually of B-cell lineage [2]. Clinical manifestations of the disease are protean and are due to multifocal medium and small vessel occlusion by tumour cells [3]. Characteristic sites of involvement are skin and central nervous system and although an ante-mortem diagnosis can be made from a biopsy specimen, it is often unsuspected [4]. We present a case of angiotropic large B-cell lymphoma in a 74-year-old man who presented with urinary symptoms and had a neurological picture resembling subacute combined degeneration of the cord.

Aged↗

An intelligent framework for the classification of the 12-lead ECG.

An intelligent framework has been proposed to classify an unknown 12-Lead electrocardiogram into one of a possible number of mutually exclusive and combined diagnostic classes. The framework segregates the classification problem into a number of bi-dimensional classification problems, requiring individual bi-group classifiers for each individual diagnostic class. The bi-group classifiers were generated employing Neural Networks (NN), combined with a combination framework containing an Evidential Reasoning framework to accommodate for any conflicting situations between the bi-group classifiers. A number of different feature selection techniques were investigated with the aim of generating the most appropriate input vector for the bi-group classifiers. It was found that by reducing the original input feature vector, the generalisation ability of the classifiers, when exposed to unseen data, was enhanced and subsequently this reduced the computational requirements of the network itself. The entire framework was compared with a conventional approach to NN classification and a rule based classification approach. The framework attained a significantly higher level of classification in comparison with the other methods; 80.0% compared with 66.7% for the rule based technique and 68.00% for the conventional neural approach.

Computer Simulation↗