Masters Degrees (Physiology)

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Cardiopulmonary exercise testing for high-risk South African surgical patients.
(2007) Biccard, Bruce McClure.
Aim: To determine the prognostic value of cardiopulmonary exercise testing (CPET) for major vascular surgery in South African patients. Methods: CPET has been used in Durban since October 2004 to predict cardiac risk for high-risk patients undergoing major vascular surgery. A submaximal 'anaerobic threshold' (AT) test was conducted on all high-risk patients. Patients were classified into two groups: 'low AT' where the oxygen consumption at the AT was <1 lml.kg^.min"1 for cycling or < 9ml.kg"1.mkf1 for arm cranking and 'high AT' when the patient surpassed these targets. Analysis of all in-hospital deaths following surgery was conducted by two independent assessors blinded to the CPET test result. Deaths classified as primarily 'cardiac in origin' have been used in this retrospective cohort analysis. Results: The AT measured during CPET was not a statistically significant pre-operative prognostic marker of cardiac mortality. However, the survivors of the patients with a 'low AT' may be identified by their response to increasing metabolic demand between 5 and 7 ml.kg^.min"1. Survivors were more dependent on increasing heart rate, while non-survivors were more dependent on oxygen extraction. When this information is added to the AT, CPET was the only test statistically associated with cardiac mortality, in comparison to Lee's Revised Cardiac Risk Index and the resting left ventricular ejection fraction which were not statistically associated with cardiac death. A hundred percent of patients with a positive test died of cardiac causes, while 11% of the patients with a negative test had cardiac deaths. The risk ratio associated with cardiac death following a positive test was 8.00 [95% CI 3.8-16.9]. The sensitivity was 0.25 [95% CI 0.04-0.64], the specificity was 1.00 [95% CI 0.90-1.00], the positive predictive value was 1.00 [95% CI 0.20-0.95] and the negative predictive value was 0.88 [95% CI 0.74-0.95]. Conclusions: CPET provides valuable prognostic information in our surgical population.
Oxidative stress of tissue in hypertensive rats.
(2006) Govender, Melvin M.; Nadar, Anand.
Oxidative stress, resulting from an antioxidant/free radical imbalance, is considered to be an important etiologic factor in the patho-physiological changes associated with salt sensitive hypertension. An important unresolved issue in hypertension research is the mechanism for organ damage during the development of the syndrome. Reactive oxygen species (ROS) such as the superoxide radical (02) , hydrogen peroxide (H202), and the hydroxyl radical (OH), may playa critical role in the pathogenesis of hypertension by targeting the very tissue that is responsible for regulating blood pressure, during the hypertensive state. Thus, this study was undertaken to evaluate the antioxidant and free radical status in the DSS rat strain, which has been shown to be an excellent model of salt sensitive hypertension. The antioxidant status was evaluated on the basis of the vascular superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, and the free radical status was evaluated on the basis of the plasma H20 2 concentration. The levels of malonyldialdehyde (MDA), which is a bio-marker for lipid peroxidation was used to determine the level of oxidative stress in the kidney, liver and brain. The kidney and liver were also subjected to an induced free radical mediated lipid peroxidation, by exposing the tissue to increasing known concentrations of H202 (2.5mM - 15mM). The level of lipid peroxidation was used to assess the tissues antioxidant buffering capacity to an induced free radical "attack". The results have shown that the DSS strain may have a compensatory increase in vascular SOD levels, to counter an increase in 02-. SOD levels were significantly lower during salt loading. The GPx levels were significantly lower in the DSS strain, and showed a slight increase during salt loading. The results demonstrate that the DSS strain has a compromised antioxidant status compared to the DSR strain. The plasma H202concentration displayed non-significant changes in the DSS strain, however salt loading did result in a non-significant increase in the plasma H202 concentration in the DSS strain. The GPx : HZ02 ratio, demonstrated an inadequate increase in GPx levels during salt loading to neutralise this non-significant increase in HzOz concentration. The kidney showed an increased level of in vivo lipid peroxidation, which could implicate increased tissue damage, and thus confirm the kidney as being a target organ during the hypertensive state. The liver and brain showed non-significant differences in the level of in vivo lipid peroxidation and are therefore thought not to be target tissue in the hypertensive state. The kidney displayed a decreased antioxidant buffering capacity to the induced free radical "attack", thereby demonstrating the tissue's decreased ability to neutralise an increased free radical level. Although the liver displayed a "normal" level of in vivo lipid peroxidation, it also displayed a decreased antioxidant buffering capacity to an induced free radical "attack", showing that the liver is able to cope with in vivo free radical levels, but at higher free radical levels, its loses its ability to quench a free radical "attack" and thereby minimise lipid peroxidation. The in vivo lipid peroxidation levels of the kidney, liver and brain have shown that tissues have varying abilities to cope with tissue oxidative stress, and behave differently, in their free radical quenching abilities. These results have shown that a compromised free radical and antioxidant status results in oxidative damage to the tissue responsible for regulating blood pressure.
The kind of society required for human flourishing : a critical comparison of the formation of ethical character in Aristotelian and African ethics.
(2005) Oguamanam, Eugene Ezenwa.; Roberts, D.
One thing that ethics attempts to determine is the right way to live in order to attain human flourishing. Both Aristotelian and African ethics give us communitarian accounts of how flourishing is attained by individuals who are brought up to have the right sorts of character. I argue that there are significant similarities between the accounts of the formation of ethical character in Aristotelian and African ethics. I aim to show that through a critical comparison of these two accounts, an account of the kind of society required from human flourishing can be developed. This can then be used to critique a dominant view of human flourishing: that of contemporary individualism. First I set out the Aristotelian account showing how it depends on a certain conception of the nature of persons. Second, I explore the African account of ethics and ethical character and show how this account is based on a similar communitarian conception of the nature of persons. In both Aristotelian and African ethics, society and upbringing play a crucial role in the attainment of human flourishing. Thus, third, I examine in detail the kind of society required for the formation of ethical character according to Aristotelian and African ethics respectively. I argue that there are many fruitful structural similarities between the two accounts. Lastly, I use the work done in the third chapter, as well as the work of certain prominent communitarian theorists, to critique a contemporary individualist view of human flourishing.
The Use of an enzyme-linked immunosorbent assay (Elisa) for the determination and characterization of antiendotoxin antibodies.
(1994) Badsha, Nasima.; Gaffin, Stephen L.
Recent clinical studies have highlighted the effectiveness of immunotherapy for Gram-negative bacteraemia in humans. Studies in America, undertaken on patients with Gram-negative bacteraemia, have shown that mortality was reduced by virtually 50% in patients who received specific antiendotoxin antiserum. In India, mortality from pseudomonas septicaemia was significantly reduced by the administration of small quantities of a anti-pseudomonas immunoglobulin. The antibodies in those studies were raised by vaccination of healthy volunteers with heat-killed Gram-negative bacteria or vaccines containing endotoxin. Adverse side effects in volunteers as well as logistic and legal problems make it difficult to produce antiserum on a large scale, in this manner. In Israel, S.L. Gaffin and coworkers found that approximately 7% of plasma units in a blood bank had antiendotoxin antibody concentrations of 40 ).1g/m1 or greater. This high titre human plasma significantly protected cats from lethal endotoxic shock secondary to haemorrhage. The immunoprecipitin technique used by them to measure antiendotoxin antibody concentrations was unsuitable for screening large numbers of blood samples. To overcome this problem we have devised an enzyme-linked imounosorbent assay (ELISA) for determining the level of antiendotoxin immunoglobulin G in human plasma. The assay, which is suitable for large scale use, was found to be specific for antiendotoxin antibodies. It was calibrated using a serum sample of specific antibody concentration as determined by an ilununoprecipitin assay. Serum samples found to be high in antiendotoxin titres (> 40_ug/m1) were tested for their specificity towards endotoxins from 12 bacterial iv strains and species. While each sample was found to have its own characteristic specificities, most were found to react strongly with Sh. flexneri, S. typhimurium and S. enteritidis. The Natal Blood Transfusion Service has found that in Natal, blood units containing high concentrations of specific antibodies occur with a frequency of 3,6% among all White donors and 10,35% among all African donors. They found that African females, in turn, had almost twice the frequency of high titre serum as African males. In this study, Indian female hospital patients did not have a statistically higher frequency of high-titre serum than Indian male patients. Blood units donated to the Natal Blood Transfusion Service are now routinely screened by ELISA for antiendotoxin antibodies and those units with high concentrations (> 40 ug/ml) of antibody were pooled and fractionated to obtain a gamma globulin, Lot LG-l. The binding capacity of the LG-1 antibodies towards 12 endotoxins was examined. Binding was found to be highest with endotoxin from Sh. flexneri, S. abortus equi and S. typhimurium and intermediate with S. enteritidis and E.coli 026:B6. Binding with the other endotoxins tested was relatively low. Differential absorption experiments showed that LG-1 was made up of a mixture of cross-reacting as well as specific antibodies For example, the antibodies binding Sh. flexneri endotoxin were mainly specific. Those binding E. coli 026:B6 endotoxin were specific and cross-reacting in almost equal proportions. Antibodies to the endotoxins from the salmonella strains tested were mainly cross-reacting. The specificities of the LG-1 antibodies towards endotoxins from the various Gram-negative bacteria did not in most cases reflect the incidence of these organisms in blood cultures taken from hospital patients. V The activity of LG-1 antibodies was compared to that of normal human immunoglobulin preparations obtained from the National Blood Fractionation Centre, Pinetown and to an anti-pseudomonas immunoglobulin prepared by Wellcome Laboratories, England. The binding capacity of the antibodies in the standard globulin preparations towards most of the endotoxins tested was less than 15% of that of the LG-1 antibodies. The anti-pseudomonas immunoglobulin was shown to bind poorly to most of the endotoxins tested in comparison with binding by LG-1 antibodies.
Mycotoxins in food with particular reference to fumonisin B1 : their health impact on a Kranskop rural community, KwaZulu Natal.
(1998) Chelule, Paul Kiprono.; Dutton, Michael Francis.; Gqaleni, Nceba.
The use of the multi-mycotoxin screen based on dialysis to analyze foods and feeds for mycotoxins, is well documented. This study investigated the possibility of incorporating FB I into the screen. Maize meal (25g) was spiked with AFB I , CPA, FB1, ST and ZEA and extraction was done using acetonitrile/4% potassium chloride (90:10 v/v). The recoveriesof the mycotoxins were 77.4, 61.5, 97.4, 79.8 and 98% respectively on analysis by HPLC. Fumonisin B1 could not be completely incorporated into the screen due to its reaction with sodium hydrogen carbonate, which is a component in the method. Thus, FB I was determined in a separate portion of the extract. The high cost of FBI standards which are often of inferior purity necessitated that FB I standards be locally produced in the laboratory using Fusarium moniliforme MRC 826, a good producer of FB 1 . In this study, production of FB I was carried out using a stirred jar fermenter and patty cultures. The yields were 160mg/1 and 6mg/g of FB I for the two methods respectively. Methyl esterification of tricaballylic acid moieties of FB I was done for effective clean-up. This was achieved by derivatizing FBI, with diazomethane. It was found that other functional groups besides the tricaballylic acid moieties of FB I were undesirably methylated as well, which made cleanup by this method difficult as shown by electrospray mass spectrometric analysis. Attempts to de-methylate FBI methyl esters with esterase was not successful. Analysis of human faecal samples was carried out with the view of developing a short term marker for assessing human exposure to FB I . Faeces from rural (20) and urban (23) volunteers were analyzed by high performance liquid chromatography. The results showed that 35% of the rural samples and 9% of the urban volunteers had detectable amounts of FB I ranging from 0.600 to 19.56 mg/kg. There was a significant difference (p = 0.04)between the two population groups. A study was carried out to assess the occurrence of FBI in a rural area of Tugela valley in Kranskop magisterial district of KwaZulu Natal. A questionnaire was administered to gather information on the family health and nutrition. Raw (stored) and processed foods and faeces, were collected for analysis of FB1. A similar control study was carried out in the urban area of Durban Metro. Homes were mapped out using the GIS for easy follow up. Oesphageal cancer (OC) incidence from the local hospital and weather data for the study area were collected from South African Weather Bureau, Johannesburg. The questionnaire results showed that the common diseases were mainly of respiratory origin (24% and 26%) from both rural and urban groups respectively. Food analysis (by HPLC) showed that the number of maize samples with FB I were higher in the rural area (31.9%) in comparison to the urban samples (6.1%). The level ranged from 0.092-22.225 mg/kg in food and 0.513-39 mg/kg in faeces. The mean concentration of FB i in the faeces and maize samples showed a similar significant difference of 0.014 between the two groups. However, these concentrations were much lower than those of high OC area in Transkei (117 mg/kg). There was no detection of FBI in fermented food products.
The effects of nerve stimulation on pacemaking activities of biological tissues.
(1973) Bhagat, Chotoo Ichharam.; Reid, John Victor Oswald.
The effects on the cardiac cycle length of stimulating the vagus nerves with single supramaximal electrical shocks depended upon when they were stimulated during the cycle. A maximum prolongation of the cardiac cycle was obtained when the vagi were stimulated 167 msec (SD±64) after the peak of an electrocardiogram P wave. The interval between a P wave and the subsequent vagal stimulation was called Pl-St interval. Pl-St(max) was the Pl-St interval at which maximum prolongation of the cardiac cycle occurred. Pl-St(max) increased significantly (p (0.001) with longer cardiac cycles. When the Pl-St intervals were shorter or longer than 167 msec (SD±64) the effects of vagal stimulation were less. The latent period for the effects of vagal stimulation was 195 msec (SD±32) The latent period also increased significantly (p(O.Ol) with longer cardiac cycles. The rise time of the vagal effect, obtained by subtracting (Pl-St(max)+ latent period) from the control cardiac cycle length, was 124 msec (SD+31) and occurred between Pl-St intervals of 167 msec (SD±64) and 291 msec (SD±70). The rise time did not vary with cardiac cycle length (p) 0.1), but the magnitude of the maximum response to vagal stimulation was inversely proportional to rise time (p <. 0.02). The peak response to vagal stimulation must have occurred when the vagal effects pegan somewhere in the middle of diastolic depolarization of the pacemaker cells in the S-A node. The reasons for this were discussed. The half-decay time for the effects of vagal stimulation was 210 msec (SD±102). The slope of the curve relating the prolongation of the cardiac cycle length to Pl-St is positive at Pl-St intervals less than 167 msec (SD±64) and negative at Pl-St intervals between 167 msec (SD±64) and 291 msec (SD±90). The positive slope ranged from 0.13 to 0.48 with a mean of 0.23. The paradoxical responses of the S-A node to vagal inhibitory input obtained by Reid (1969), Levy et al (1969)and Dong and Reitz (1970) would be explained by the dependence of the cardiac cycle length upon the time of arrival of vagal stimulus in relation to the previous P wave and upon the slope of the curve relating the prolongation of the cardiac cycle length to Pl-St interval being positive and between zero and two at Pl-St intervals less than 167 msec (SD±64. The effects of single shock stimulation of the vagus nerves persisted for 3.890 sec (SD+l.255)7 the number of cardiac cycles involved varied between 5 and 11. The duration of the effects of vagal stimulation did not depend upon when during the cardiac cycle the vagi were stimulated. A "dip" in the response to vagal stimulation was present in all the experiments. The possibility of the "dip" phenomenon being due to simultaneous stimulation of the sympathetic fibres in the vago-sympathetic trunk was ruled out. It is suggested that the "dip" phenomenon may be due to transient accumulation of K+ in the interstitial fluid surrounding the pacemaker cells in the S-A node.There was no paradoxical response of the smooth muscle in the distal colon of the adult rabbit when the frequency of sympathetic inhibitory input was continuously increased. A paradoxical response in the frequency but not in the size of the contraction of the smooth muscle was obtained when the sympathetic nerves were stimulated with bursts of stimuli, each burst consisting of 5-40 impulses, 10 msec apart. One may conclude from this that the delay of the next spontaneous contraction but not the inhibition of the size of smooth muscle contraction is dependent upon the arrival time of a burst of stimuli during a contraction cycle. This was confirmed in an experiment when the sympathetic nerves were stimulated with single bursts of stimuli applied at different times during the contraction cycle. It is unlikely that such a paradoxical response would occur under physiological conditions as this would require the natural sympathetic efferent discharges to the smooth muscle to occur in regular bursts, each burst consisting of impulses at a high frequency. Stimulation of the sympathetic nerves at 3, 5, 10 and 25 PPS caused an inhibition of the size and frequency of smooth muscle contraction in the distal colon of the newborn rabbit. Assuming that the cholinergic fibres are excitatory there is therefore no evidence for the sympathetic fibres to the distal colon being cholinergic in the newborn rabbit. This is contrary to Burn's (1968) report of the sympathetic fibres being motor and cholinergic to the small intestinal smooth muscle in the newborn rabbit.The heart rate increased rapidly at the onset of exercise and then more gradually over the rest of the exercise period. The initial increase in the heart rate during exercise was not affected by adrenergic blockade but the subsequent increase in heart rate was significantly reduced by adrenergic blockade. Hence the increase in heart rate at the onset of exercise is due primarily to a decrease in the cardiac vagal efferent discharge, whereas the subsequent increase in heart rate is due to both a further decrease ln vagal discharge and an increase in sympathetic discharge to the S-A node. In almost all the sub jects there was initially a rapid decline in the heart rate in the post-exercise period, but subsequently the heart rate returned to resting levels in a variety of ways. These were classified into 5 types. Of particular interest to the present study was the Type V pattern of heart rate change. This was characterised by an increase in heart rate of 6 beats or more per minute during the post-exercise period, with or without superimposed arrhythmia. The Type V pattern may be the equivalent of the paradoxical responses to inhibitory input demonstrated in animal experiments i.e. an increase in the heart rate with increasing vagal stimulation frequency. Type V pattern occurred more frequently at mild exercise levels (4 out of 14) than at moderate exercise level (lout of 14) and also more frequently in adrenergic blocked individuals (11 out of 28) than in control subjects (5 out of 28) It is suggested that the sympathetic effects on the P-R interval and arterial baroreceptor modulation of vagal efferent discharge protect again st the occurrence of paradoxical responses to vagal inhibitory input. They may do so by confining the vagal discharge to the rise time of vagal effect during the cardiac cycle. On the other hand the Type V pattern in p-adrenergic blocked individuals may be due to a decrease in the vagal discharge, in which case Type V pattern would not be a paradoxical response. The changes in minute ventilation in the post-exercise period were also variable. Besides a gradual decline in minute ventilation there were also gradual increases and sudden increases and decreases in minute ventilation. These may represent a form of paradoxical response to increasing inhibitory input and decreasing excitatory input to the respiratory neurones in man. However, all the changes in minute ventilation could also be explained by fluctuating excitatory and inhibitory neural input to the respiratory neurones.
Optimisation of an analytical method for the analysis of folic acid derivatives in biological materials.
(2007) Khanyi, Purity Duduzile.
Folic acid is a water-soluble, B-complex vitamin influencing a number of biological processes in humans and particularly important in the prevention of neural tube defects (associated with spinal bifida) in unborn children. Reliable analytical methods are therefore needed for quantisation of the amount of total folic acid (FA) in biological materials of quality assurance and regulatory purposes. What is particularly needed are rapid and reliable methods for ensuring that the correct amount of FA is consumed and the degradation rates of these compound is kept at minimum during the extraction process. Analytical methods for determination of folic acid in biological materials have been around for decades and the most common procedures include microbiological assay; biomolecular interaction analysis (BIA); immunoassay; conventional chromatographic procedure such as thin-layer column chromatography (TLC) and high performance liquid chromatography (HPLC). These procedures were replaced by HPLC, which is more rapid and in many instances yields a better resolution. Current HPLC methods uses C-18 column and reverse phase conditions in combination with ion-pair or ion suppression techniques; fluorescence or electrochemical detector, unfortunately, excitation and emission of folic acid is found not sufficiently to allow physiological levels of the form of the vitamin to be detected. In addition, ion-pair reagent nullifies the mobile phase and interferes with the absorption! fluoresce spectrum resulting in poor separation. Therefore this study was carried out to address and improve the problems that are in the existing HPLC methods. Currently scarce information is available on the determination of folic acid in biological materials by HPLC with UV detection. Serum samples were spiked with folic acid standard to check the efficiency of the method. Other wavelengths from 200 nm to 300 nm were attempted for detection of folic acid, in which the wavelength 250 nm was found to have better absorbance compared to other wavelengths. Folic acid was detected at 250 nm wavelength under isocratic elution using a mobile phase consisting of citrate phosphate buffer: acetic acid: methanol. Folic acid in maize meal was detected at 290 nm using mobile phase containing potassium phosphate containing ascorbic acid/sodium ascorbate mixture and 2-mercaptoethanol under gradient elution. The mobile phase used for gradient and isocratic elution was suitable for separation of folic acid from other compounds with flow rate of 3 ml/min modified to Iml/mim to avoid overloading of the column under isocratic elution. For good separation of folic acid under gradient elution the flow rate was set at 0.8ml/min with pH of mobile phase modified from pH 2.2 to pH 2.5. The recovery of folic acid added to human serum was 91% -100% and recovery of folic acid added in unfermented maize meal and fermented maize meal ranged from 55% - 73%. Folic acid level from unfermented maize meal and fermented maize meal ranged between 1.29 - 1.3 [!g/g and 1 - 2.1 [!g/g respectively. In conclusion the optimised method in this study gives better analytical results when compared with earlier HPLC method in terms of efficiency, reproducibility and sensitivity for folic acid in human serum and maize meal. However, there is a need to minimise the loss of folic acid during the sample treatment. The outcome of this work indicated that more work has to be done to improve extraction procedure for specific foods with minimum time preparation to sample analysis.
A comparative assessment of local, commercial and homemade amahewu with respect to nutritional value, hygiene, and other health benefits to the community.
(2003) Mbongwa, Hlengiwe Prosperity.; Gqaleni, Nceba.
Fermentation is a process by which primary food products are modified biochemically by the action of microorganisms and/or their enzymes. Several societies have, over the years, intentionally carried it out to enhance the taste, aroma, shelf-life, texture, nutritional value and other properties of food. It is used in many parts (lithe world. However, there are regional differences in use and these depend on the availability of raw materials, consumption habits. and other socio-cultural factors. This study was aimed at (comparatively) assessing, local commercial and homemade amahewu with respect to nutritional value, hygiene and other health benefits to the commirn ity. Methods employed were Thin Layer Chromatography (TLC) (mycotoxins), High Perliffmance Liquid Chromatography (HPLC) (mycotoxins, sugars and amino acids), Dumas (proteins), SOxhlet (lipids) and intubation technique (metabolisable energy) to analyse maize meal and amahewu samples from various regions. The regions sampled included mal3heleni (South Coast) and kwaNgcolosi (North Coast) villages. Commercial amahewu was analysed with kind permission from Clover SA. Species from the following genera were isolated and identified from amahewu samples: Lactobacillus, Saccharonivccs, Lcuconostoc, Lactococcus, Panioca, Entcrobacter and kleb•iella. Saccharotnyces was detected in commercial samples only. Gram-negative strains were identified in most of manheleni village samples. No traceable amounts of aflatoxin BI (AFB1), fumonisin B 1 (FBI) and zearalenone (ZEA) were found in Clover SA samples. AFB I was detected in 40% of both maize meal and amahewu samples from maBheleni (range 0.55 — 0.84ng/g and 8.3x10 5 — 9.1x10-5ng/g respectively). From the same village, 100% of the maize meal and 80% of the amahewu samples were contaminated with FBI (range 4.1 47.2ng/g and 1.4 ---- 6.9ng/g respectively). ZEA was detected in all maize meal samples (range 0.9 — 4.3ng/g). None of the amahewu samples contained detectable levels of ZEA. All maize meal and amahewu samples from kwaNgcolosi were contaminated with AF13 1 (range 8.3 — 30.I ng/g and 0.04 - 0.102ng/g respectively). FB I was detected in 75% of both maize meal and amahewu samples from the same village (range 0.5 — 4.1ng/g and 0.04 0.56ng/g respectively). ZEA was also found in all maize meal samples and 75% of amahewu samples (range 3.7 — 16.4ng/g and 0.03 -- 0.06ng/g respectively). MaBheleni, Clover SA and kwaNgcolosi maize meal and amahewu samples contained vitamins B1, 13 2 and B6 with a range of 0.31+0.21 - 4.48±0.81 B 1 ; 0.15±0.14 - 1.67±0.33 B2 and 0.05±0.07 - 0.77±1.45 lig/g B6. Fat levels ranged from 0.28±0.40 to 4.54±0.05 percentage by weight. The levels of proteins varied from 4.02±0.02 to 8.40±0.04 percentage by weight. Starch concentrations ranged from 31.51.5.28 to 75.911.92g/100g. Maize meal samples contained glucose and maltose, while glucose, fructose, sucrose, maltose, M-triose, DP 4 and 5 and DP >15 were detected in amahewu. Apparent and true metabolisable energy for homemade and commercial Freeze-dried amahewu was 13.194 and 13.696MJ/kg (AME N ); and 13.605 and 14.106M.Ekv ( 1 MEN ), respectively. This study has shown that lactic acid maize fermentation reduce' the levels of AF13 1 , FB I and ZEA toxins in maize meal, inhibits the growth of most Gram-negative bacteria, and in some instances, fermentation did improve the nutritional value. Metabolisable energy analysis represents an important tool to assess whether or not compounds ingested are converted to sources of energy in the body and utilised. Amahewu fermentation yielded beneficial products (probiotics: reduced mycotoxins levels and reduced starch). In conclusion, natural lactic acid maize fermentation to produce amahewu will do more good than harm to the consumer, therefore, people need to be advised on how to safely store their maize and also to be encouraged to consume their stored maize in fermented form.
The fate of mycotoxins in non-alcoholic lactic acid maize meal fermentation.
(2003) Mokoena, Mduduzi Paulos.; Gqaleni, Nceba.
This study was aimed at investigating the potential of lactic acid fermentation in reducing myco toxin concentration in maize meal products. Maize meal was spiked separately with aflatoxin Bi, fumonism Bi, and zearalenone, and fermented for four days. During this period the concentration of each toxin and the pH of the fermented maize meal were monitored. There was a significant (p= 0.000) decrease in the concentration of all the mycotoxins, with a percentage reduction of 55-69 by the third day and 68-75 by the fourth day, respectively. Commercial amahewu samples were also screened for the presence of these three mycotoxins, and the results indicated that the samples were not contaminated with detectable levels of these toxins. An attempt was made to characterise the metabolic derivatives (by-products) of each mycotoxin following lactic acid maize meal fermentation. To achieve this maize meal samples were separately spiked with each of mycotoxin, fermented for four days and screened for specific mycotoxin derivatives (by-products) using GC/MS, HPLC and relevant standards (i.e. partially hydrolysed fumonisin Bi, aflatoxin B2a, a- and Pzearalenol). None of the targeted derivatives could be detected in the fermented maize meal samples. The potential cytotoxicity of the mycotoxin-spiked fermented samples was investigated using an SNO cell line. The fermented toxin-spiked maize meal samples with a starter culture were comparatively less toxic (29 - 36%) to SNO oesophageal cells than samples spiked with toxin without a starter culture (24 - 30%). However, this observed difference was not statistically significant (p = 0.295 - 0.681). Furthermore, cells that were only inoculated with the cell culture medium had significantly (p = 0.000) high percentage cell viability. This study indicates that it is possible to significantly reduce the concentration of mycotoxins using lactic acid maize fermentation to trace levels. However, such a reduction will not significantly alter the possible chronic toxic effects of such toxins in the diet, particularly a maize based diet containing poor quality protein. The trace amounts of these toxins in fermented and unfermented maize meal should continue to be a cause for concern.
An immunohistochemical evaluation of the effect of salt (NaCI) on adrenal adrenomedullin content in Dahl rats.
(2003) Hariram, Arvind.; Harripersad, Rohan.; Somova, Liliana I.
Adrenomedullin (ADM) is a 52 amino acid vasodilator peptide isolated, in 1993, from human pheochromocytoma. It has been demonstrated in the adrenal medulla of several mammalian species, including humans and rats. There have been conflicting results of the tissue distribution in the adrenal cortex. Hypertension is a complex trait with multiple genetic and environmental influences. Furthermore, salt-sensitive hypertension is characterized by a cluster of renal, hormonal, and metabolic derangements that might favour the development of cardiovascular and renal complications. Therefore the objective of this study was to investigate the adrenal distribution of ADM as well as to semi-quantitatively assess the adrenomedullin secretory capacity of the adrenal gland in the rat model of salt sensitive hypertension. Fourty-four male weanling rats were divided into 4 experimental groups and placed on a dietary regimen for 6 weeks viz. Dahl salt sensitive (DSS) rats on a high sodium diet (8% NaCl), DSS on a normal sodium diet (1% NaCl) matched with normotensive Dahl salt resistant (DSR) rats on the same dietary treatments. Blood pressure was monitored by tail-cuff readings and by the end of the six weeks, the DSS rats developed hypertension with tachycardia irrespective of the diet they were fed. The normal sodium diet was found to delay the development of hypertension, whilst the high sodium diet exacerbated the development of hypertension. Kidney weights and heart weights were greater in DSS rats than DSR rats probably due to their renal pathology or cardiac hypertrophy. Adrenomedullin immunopositivity was found predominantly in the adrenal medulla, and to varying degrees in the zona glomerulosa and zona reticularis of the adrenal cortex. The semi-quantitative analysis indicate that there was a 6.3 fold increase in ADM content of DSS rats compared to the DSR rats, where both consumed the 1% NaCI supplemented diet (DSR : 5.98 ± 0.3 vs. DSS : 37.85 ± 0.5, P
The occurrence and detection of aflatoxin-macromolecular conjugates in humans.
(1998) Myeni, Sibongiseni Selby.; Chuturgoon, Anil Amichund.; Dutton, Michael Francis.
Aflatoxin Bi (AFBi), a highly toxic fungal metabolite (mycotoxin) of certain strains of Aspergillus, has long been known to be carcinogenic in animal species. Accumulation of epidemiological evidence led to its classification, in 1993, by the International Agency for Research on Cancer as a Group I human carcinogen. Aflatoxin Bi contaminates the food supply in most tropical and sub-tropical countries, where it is associated with increased incidence of hepatocellular carcinoma (HCC). In these countries, AFBi is also linked to kwashiorkor, jaundice, and Rey's syndrome. The biological action of AFBi is through its oxidation to AFBi-8,9 epoxide (AFBiO). This epoxide binds to macromolecules like DNA, RNA and proteins as well as amino acids to form AFBi-macromolecular adducts. Quantitation of these adducts is thought to be the most promising approach in the development of methods to measure levels of exposure to aflatoxins. Aflatoxin Bi was produced, isolated and purified using preparative thin layer chromatography (TLC). The toxin was oxidised to AFBiO using dimethyldioxirane and the UV spectra of both the AFBi and AFBiO were determined. Reaction of selected Na-acetyl amino acids (AA) with AFBiO was studied and UV spectrophotometry, TLC, high performance liquid chromatography (FfPLC) and high performance capillary electrophoresis (CE) were used to characterise the reaction products. The epoxide was also reacted with albumin and DNA. Aflatoxin Bi-albumin reaction mixture was hydrolysed and characterised by TLC. Spectrum measurement of the oxidative product of AFBi gave peaks at 266 and 367nm. Qualitative TLC and the epoxide spray reagents confirmed that epoxidation was successful. The in vivo reaction of selected Na-acetyl AA with the epoxide gave peaks between 300 and 400 nm. Naacetyl-arginine, Na-acetyl-lysine and Na-acetyl-histidine showed reaction with AFBiO with maximum wavelengths at 392, 397 and 391 nm respectively. These results strongly suggest that AFBiO is able to covalently bind to lysine, histidine and arginine in albumin. A total of twenty nine blood samples were analysed by HPLC for the presence of AFBilysyl adduct. Of the twenty nine samples, ten were from HCC patients, ten from control patients and nine from kwashiorkor patients. The results show that AFBi-lysine does occur in patients at King Edward VIII Hospital (KEH) and the highest level was detected in HCC patients followed by kwashiorkor patients.
The effect of dietary egg on human plasma cholesterol and triglyceride levels.
(1990) Raidoo, Kogie.; Burger, F. J.
No abstract available.
The interrelationship of dietary cholesterol, copper and zinc on plasma lipids and tissue copper and zinc levels in the rat.
(1992) Nadar, Anand.; Burger, F. J.
No abstract available.
Effects of plants-derived oleanolic acid in an in-vitro model hyperglycaemia-induced oxidative stress.
(2010) Dlamini, Immaculate Nonkululeko.; Zungu, M.; Essop, M. Faadiel.
Diabetes mellitus (DM) has become a global threat in developing and developed countries, where diabetic patients are more prone to cardiovascular complications, a condition called diabetic cardiomyopathy. Studies have shown a direct link between hyperglycaemia and an increase in the production of reactive oxygen species in cardiac cells leading to diabetic cardiomyopathy. This study tests oleanolic acid, a bioactive compound from the plant Syzigium aromaticum as an antioxidant which could have a potential role in management of DM. Aims i) To extract Oleanolic acid (OA) from Syzigium aromaticim, ii) Investigate the antioxidant effects of plant derived OA in an in-vitro model of hyperglycaemia induced oxidative stress. Methods The flower buds of the Syzigium aromaticim [(Linnaeus) Merrill & Perry] (Myrtaceae) plant (commonly called cloves) were used to isolate OA. The ethyl acetate solubles from the cloves were subjected to chromatographic fractionation to yield OA powder. Spectroscopic analysis was done using 1D and 2D 1H and 13C NMR techniques for the identification of the structure of the compound. This compound was then used in vitro to test for its antioxidative properties. H9C2 cardiac myoblasts were employed which were treated with normoglycaemic (5.5 mM) and hyperglycaemic (33 mM) glucose conditions. The cells were then treated with oleanolic acid to test for its antioxidant properties. We looked at a dose-dependent (0, 20, 50 μM) and time-dependent effects of OA treatment (6 and 24 hrs) following 48 hours glucose exposure. ROS levels were measured using H2DCF-DA fluorescence staining using microscopy and flow cytometry techniques for analysis. xviii Results Recrystallisation of the powder with ethanol and inspection of the 1 and 2- dimensional 1H- and 13C-NMR spectra of the compound with comparison to literature data confirmed OA molecular structure and IUPAC numbering similar to that of literature characterized and confirmed the structure of oleanolic acid. In cell specific data high glucose treatments on H9C2 cells showed increased ROS production (22 ± 6 % and 20 ± 7 % n= 3 p< 0.01) for 6 and 24 hrs treatments, respectively, compared to their normoglycaemic control groups. The 6 h OA treated group showed a decrease in ROS production with 26.6 ± 17.4 % for the 20 μM while for 50 μM there was a 37.7 ± 14.3% decrease. A ROS reduction trend was observed in the normoglycaemic group, but this was significant at 24 hrs with 46.8 ± 45.3% and 57.3 ± 9 % for both 20 and 50 μM treatments, respectively. The 24 hrs OA treated group showed a dose-dependent decrease in ROS with 50 μM more pronounced (80.7% ± 4.5 %). The 20 μM OA treatments also showed a 15.7 ± 19 % decrease in ROS. Discussion In the present study, we have evaluated the antioxidant effects of OA in vitro following extraction of the compound from Syzigium aromaticim. The oxidative stress induced by hyperglycaemia was attenuated by oleanolic acid and this also translated into decreased ROS suggesting its use as an antioxidant in alleviating cardiovascular complications associated with diabetes mellitus.
The effects of plant-derived oleanolic acid on kidney function in male Sprague-Dawley rats and, in cell lines of the kidney and liver.
(2012) Madlala, Hlengiwe Pretty.; Musabayane, Cephas Tagumirwa.; Masola, Bubuya.
Adverse effects and increasing cost of therapeutic drugs have renewed an interest in the use of medicinal plant products for the treatment of a variety of chronic disorders. One such bioactive plant-derived compound is a pentacyclic triterpenoid, oleanolic acid (3ß-hydroxy-olea-12-en-28- oic acid, OA) present in herbs. OA possesses a variety of pharmaceutical activities and of interest in this study are the anti-diabetic properties. Diabetes is associated with disorders grouped as microvascular (retinopathy and nephropathy) and macrovascular (atherosclerotic) complications. Accordingly, this study further investigated the potential of OA in diabetes management by studying the effects of this triterpene on kidney function as well as proximal tubular Na+ handling in an effort to identify the site of action of OA. Furthermore, the study evaluated the effects of OA in kidney and liver cell lines to establish whether this triterpene exhibits any toxicity in these organs. OA was extracted using a previously validated protocol in our laboratory. Briefly, dried flower buds of Syzygium aromaticum were soaked in dichloromethane overnight, thereafter in ethyl acetate to obtain ethyl acetate solubles which contained a mixture of OA/ursolic and maslinic acid (MA). OA/MA mixture was subjected to column chromatograph and pure OA was obtained through recrystallization in methanol. The absolute stereostructure of OA was elucidated using 1H and 13C NMR spectroscopy and was comparable to previously reported data. In kidney function studies, various doses of OA (30, 60, 120 mg/kg, p.o.) were administered to male Sprague-Dawley rats twice (8h apart) every third day for five weeks. Rats administered deionised water served as controls. Measurements of body weight, food and water intake, blood pressure, Na+, K+, Cl-, urea and creatinine were taken 24 h from dosing. Renal clearance studies investigated the influence of OA on Na+ handling in the proximal tubule of anaesthetized rats using lithium clearance. Animals were given water with lithium (12mmol/l) for 48 hours following which they were anaesthetized and cannulated using a previously validated standard protocol that has been reported from our laboratories. After a 3½ h equilibration, animals were challenged with hypotonic saline for 4 h of 1 h control, 1½ h treatment and 1½ h recovery periods. OA was added to the infusate during the treatment period. In vitro effects of various OA concentrations (5, 10, 20, 40, 80 μmol/l) were investigated in HEK293, MDBK and HepG2cell lines. Cells were exposed to OA for 24, 48 and 72 h, thereafter, 3-4,5 dimethylthiazol-2-yl- 2,5diphenyltetrozolium bromide (MTT) and single cell gel electrophoresis (comet) assays were conducted. All data are presented as means ±SEM. OA significantly (p<0.05) increased urinary Na+ output from week 2 until the end of the experimental period in a dose independent manner. However, this OA-evoked natriuresis was not reflected in plasma collected at the end of the experiment as there was no change in plasma Na+ concentrations compared with control animals at the corresponding time. OA administration had no significant influence on K+ and Cl- excretion rates throughout the experiment. However, OA significantly (p<0.05) reduced plasma creatinine concentration with a concomitant increase in glomerular filtration rate (GFR). Furthermore, OA administration significantly (p<0.05) decreased mean arterial pressure from week 2 until the end of the experimental period. Intravenous infusion of OA at 90 ug/h for 1 ½ h induced a marked increase in urinary excretion rates of Na+. This increase was accompanied by concomitant increase in FENa proximal and FENa distal and FELi which persisted until the end of the experiment without any apparent changes in GFR. The cell viabilities of HepG2, HEK293 and MDBK cell lines were significantly increased after 24 h exposure, however, the viabilities of all the three cell lines dropped after 72 h exposure to values that did not achieve statistical significance in comparison to the respective controls. In addition, all OA-treated cells in the comet assay had intact DNA after exposure for 24, 48 and 72 h. Hence, the decrease in viability that was observed in the MTT assay after 72 h exposure could probably be attributed to the depletion of nutrients in the culture medium. The results of the present study, apart from confirming our previous observations of the natriuretic effects of OA in rats, indicate that this effect is in part mediated via the inhibition of proximal tubular Na+ reabsorption and increased Na+ secretion. We speculate that this increased Na+ secretion could have been due to increased tubular function and not to the toxicity of OA as indicated by MTT and comet assays. These findings suggest that OA does not exhibit toxicity in the kidney and the liver.
Determination of exposure of humans to selected mycotoxins with particular reference to aflatoxins.
(1995) Early, Deborah Angeline.; Dutton, Michael Francis.; Chuturgoon, Anil Amichund.
Mycotoxins are poisonous secondary metabolites commonly produced by fungi and are involved in human disease conditions known as mycotoxicoses. There is evidence to show that food eaten by the rural Black population of Southern Africa is contaminated with mycotoxins. A tenuous relationship exists between the occurrence of mycotoxins in foods and certain disease conditions in humans. In order to verify this relationship, efforts have, in the past, been made to detect mycotoxins and their metabolites in physiological fluids and tissues. The difficulty with this approach is that mycotoxins in the body have short half lives, being rapidly excreted or metabolised to other forms. More recently it has been shown that aflatoxin B1, as its activated epoxide, can conjugate with macromolecules such as nucleic acids and proteins. These survive for much longer than the free toxins and by suitable methods can be isolated and measured. This allows for a much better estimate of exposure of the individual to aflatoxin. This study reviews and evaluates screening methods for the detection and analysis of mycotoxin contamination in rural foodstuffs such as maize and groundnuts. Methods for the production of aflatoxin-lysine and protein adducts are motivated and developed then used in the identification of naturally occurring adducts in humans. Isolation and quantitative analysis techniques are proposed to routinely screen patients for evidence of aflatoxin exposure.
The effect of ultradistance running on premenopausal women of different ethnic groups.
(2005) McGregor, Avril.; Mars, Maurice.; Cassim, Bilkish.
No abstract available.
The cytotoxic effects of T-2 toxin on normal human lymphocytes.
(1998) Moodley, Therishnee.; Chuturgoon, Anil Amichund.; Dutton, Michael Francis.
T -2 toxin is an immunosuppressive mycotoxin that has been conjoined with several symptoms and diseases as early as the turn of the century, but whose mechanisms of action are still being investigated. Accordingly, this study was an attempt to determine the cytotoxic effects of T -2 toxin on normal human lymphocytes in vitro, with particular emphasis on mitochondrial viability, cellular and nuclear morphology as well as the localisation of the subcellular sites of toxin interaction. The cytotoxicity of T -2 toxin was assessed with the use of a methylthiazol tetrazolium (MTT) assay. This assay targeted the succinate dehydrogenase activity of the lymphocytic mitochondria, over a range of concentrations of T-2 toxin at various incubation times. The morphology of treated lymphocytes was analysed with the use of transmission electron microscopy and the localisation of the toxin was accomplished via immunocytochemistry. DNA fragmentation studies formed an integral part of the analyses. The cytotoxicity assay indicated that not only was cell viability inversely proportional to both the dose and exposure time, but that the eftects of the different doses were only evident at prolonged incubation times (12-24 hours). The electron microscopy studies showed that T-2 toxin (1,56 ug/ml) induced apoptosis (cell suicide) in normal human lymphocytes. This was determined by the observation of chromatin condensation and nuclear disintegration within the toxin treated lymphocytes. Apoptosis seemed to occur independently of mitochondrial damage at 6 hours of exposure to T-2 toxin. The presence of polyribosomes within the treated lymphocytes indicated that protein synthesis was not inhibited. Anti-T-2 toxin conjugated gold label was present in all areas of damage, particularly within the nuclei of the T-2 toxin treated lymphocytes. The DNA fragmentation results showed that T-2 toxin induced fragmentation in lymphocytes, the extent of which was directly proportional to the exposure time. It appears that the early signs of T-2 toxin induced apoptosis in normal human lymphocytes can be determined by damage to the nucleus.
A cytotoxic evaluation of aflatoxin B1, zearalenone and their epoxide derivatives using human cell lines.
(1996) Pillay, Dharmarai.; Chuturgoon, Anil Amichund.; Dutton, Michael Francis.
Since the discovery of mycotoxins in food, the thrust of biochemical and toxicological research has been carried out on animals which has proven to be uncoordinated and not easily extrapolated to humans. Over the last decade, there have been increasing pressures to review and reduce the use of animals in experimental toxicological studies. Consequently in this study aflatoxin B1 (AFB1), zearalenone (Zea) and their epoxide derivatives have been evaluated using in vitro assays. The HepG2, A549 and Hela cell lines were used for assessing the cytotoxicity, effects on cellular metabolism and sites of action of AFB1, Zea and their derivatives. The cytotoxicity of these mycotoxins was evaluated using the methylthiazol tetrazolium (MTT) reduction assay. Cells, treated with mycotoxins were prepared for transmission electron mlcroscopy (TEM), immunocytochemistry (ICC), scanning electron microscopy (SEM), confocal and light microscopy. From the cytotoxicity assay it was found that the epoxide derivatives were more toxic than the parent toxin when exposed to HepG2 cells with no significant differences in toxicity levels in A549 and Hela treated cells. Both epoxide derivatives displayed a regression of hepatoma cell proliferation at high doses (25ug/ml) while lower concentrations (<12.5ug/ml) enhanced cell growth. Microscopy analyses showed distinct cellular alterations. When exposed to AFB1 (12.5ug/ml) hepatoma cells showed prominent ultrastructural alterations such as areas of cytoplasmic lysis and increased numbers of secondary lysosomes while cells exposed to Zea (l2.5ug/ml) displayed numerous ovoid mitochondria and proliferation of rough endoplasmic reticulum which is indicative of enhanced protein synthesis. The presence of label in toxin treated cells is suggestive of the effects of these mycotoxins. Such cellular changes may lead to altered metabolism and cell function.
Studies on the isolation of the polymerase genes from the H1N1 influenza A virus.
(1992) Naidoo, Richard.; Manning, D.
Vaccines directed against the influenza virus become ineffective due to continuous mutation. An alternative approach might be to control replication at the genomic level by enzymatic methylation of the polymerase genes. Hence in this study, a method to locate and successfully isolate the H1N1 influenza A polymerase genes was investigated. The virus was cultured in chick embryos via the allantoic route using aseptic techniques. Following incubation, the allantoic fluid was isolated and washed to remove any contaminating blood cells. The allantoic fluid was checked for fungal and bacterial contamination using the blood agar test and the presence of the virus was established by the haemagglutination titration test. Viral particles were pelleted by ultracentrifugation. Electron microscopy verified the morphology and size of these viruses while immunofluorescence studies, using a monoclonal antibody, confirmed the influenza A strain. The ribose test verified the presence of RNA in the samples. Purified viral pellets were pooled and homogenised in buffer containing guanidine thiocyanate, mercaptoethanol and sarkosyl. The samples were incubated on ice before mechanical disruption of the virus. Viral RNA was isolated from the upper aqueous layer after a standard phenol/chloroform extraction procedure. RNA was quantified spectrophotometrically and purity assessed initially by the absorbance ratio readings at 260/280 nm. Electrophoresis of the RNA samples was performed together with RNA molecular weight markers on a 1.5% formamide agarose gel. Five bands were identified and the band containing the polymerase genes was size selected, located and excised. Purification of the polymerase genes from the agarose was achieved by using the BIO 101 RNAid kit. The three isolated polymerase RNAs were reverse transcribed using the Boehringer Mannheim cDNA synthesis kit. The results indicate that the H1N1 influenza virus was successfully grown and isolated from chick embryos. Absence of contamination and verification of viral presence at different stages of the study were indications that asepsis was successfully achieved. The RNA obtained was sufficient and suitable for cDNA synthesis. This cDNA may now be used for further molecular analysis and subsequent DNA methylation studies. Further, transfection studies may then be performed to determine, if any, the the expression of methylated and unmethylated cDNA.

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