Identification of individual goat animals by means of near infrared spectroscopy and chemometrics analysis of commercial meat cuts.

Although the identification of animal species and muscles have been reported previously, no studies have been found on the use of NIR spectroscopy to identify individual animals from the analysis of commercial meat cuts. The aim of this study was to evaluate the use of a portable near infrared (NIR) instrument combined with classical chemometrics methods [principal component analysis (PCA) and partial least squares discriminant analysis PLS-DA)] to identify the origin of individual goat animals using the spectral signature of their commercial cut. Samples were collected from several carcasses (6 commercial cuts x 24 animals) sourced from a commercial abattoir in Queensland (Australia). The NIR spectra of the samples were collected using a portable NIR instrument in the wavelength range between 950 and 1600 nm. Overall, the PLS-DA models correctly classify 82% and 79% of the individual goat samples using either the goat rack or loin cut samples, respectively. The study demonstrated that NIR spectroscopy was able to identify individual goat animals based on the spectra properties of some of the commercial cut samples analysed (e.g. loin and rack). These results showed the potential of this technique to identify individual animals as an alternative to other laboratory methods and techniques commonly used in meat traceability.

The ability of a portable near infrared instrument to evaluate the shelf-life of fresh and thawed goat muscles.

The objective of this study was to evaluate the use of a portable near infrared (NIR) instrument to monitor the shelf-life of four goat muscles [longissimus thoracis et lumborum (LTL), semimembranosus (SM), semitendinosus (ST) and biceps femoris (BF)] stored for up to 8 days (4 °C). The NIR spectra of the muscle samples were collected at day 0, and after 1, 4 and 8 days of storage using a MicroNIR instrument (900-1600 nm). The coefficient of determination in cross-validation (R2) and the standard error in cross validation (SECV) obtained for the prediction of days of storage ranged between 0.76 and 0.86, where the SECV ranged from 0.32 to 0.41. The best statistics in cross-validation were obtained for the prediction of days of storage in the BF samples, followed by the ST and LTL muscles. Differences in the PLS loadings for the cross-validation models were observed due to the interactions between the different muscle samples and days of storage. Overall, these results showed the potential of NIR spectroscopy to identify the time of storage in four different goat muscles. Similar data and techniques could be used to predict the remaining shelf life of meat derived from different species under storage. This information can then be used as a tool to predict and guarantee the safety of meat samples to the consumer along the meat supply and value chains.

Erythrocyte membrane fatty acids in patients with multiple sclerosis.

BACKGROUND: Reports on fatty acids levels in multiple sclerosis remain inconclusive. OBJECTIVE: To determine the erythrocyte membrane fatty acid levels in multiple sclerosis patients and correlate with Kurtzke Expanded Disability Status Scale. METHODS: Fatty acid composition of 31 multiple sclerosis and 30 control individuals were measured by gas chromatography. RESULTS: The membrane phosphatidylcholine C20:4n - 6 concentration was lower in the multiple sclerosis patients when compared to that of the control group, P = 0.04 and it correlated inversely with the EDSS and FSS. CONCLUSION: Decrease in C20:4n - 6 in the erythrocyte membrane could be an indication of depleted plasma stores, and a reflection of disease severity.

Effect of palm oil on plasma lipoprotein concentrations and plasma low-density lipoprotein composition in non-human primates.

Palm oil (PO) contains approximately 43% of palmitic acid. It is the most abundant saturated fatty acid in the diet and it is generally considered the primary cholesterol (C)-raising fatty acid. However, the effect of palmitic acid on plasma cholesterol appears to depend on the cholesterol content of the diet. The aim of this study was to determine the effect of PO with either a high-fat, high-C or moderate-fat, moderate-C diet on lipoprotein C and low-density lipoprotein (LDL) composition. Fifty adult, male vervet monkeys were randomly assigned to the high-fat diet group (HFD: 35%E fat, approximately 0.106 mg C/kJ; n = 30) and the moderate-fat diet group (MFD: 30%E fat, approximately 0.027 mg C/kJ; n = 30). Baseline LDL-C, high-density lipoprotein (HDL)-C and body weight were used to stratify the vervets into comparable experimental groups within each dietary group. The HFD group was divided into two groups of 10 each: one group continued with the HFD in which 8.1%E was derived from lard (AF); in the other group, AF was substituted isocalorically with PO. The MFD group was divided into three groups of 10 each: one group continued with the MFD in which 11.8%E was derived from AF; in the other two groups, the AF was substituted isocalorically with either sunflower oil (SO) or PO. This article presents preliminary results on plasma lipoproteins and LDL composition after 6 months of dietary intervention. Plasma total and LDL-C was higher in all the groups, but the mean changes elicited by PO with either the HFD or MFD were no different from that observed with AF and SO. There was no difference in the mean change of LDL molecular weight within the HFD and MFD. It is concluded that PO is no different from AF (HFD and MFD) or SO (MFD) in its cholesterolaemic effect.

The effects of B-sitosterol (BSS) and B-sitosterol glucoside (BSSG) mixture on selected immune parameters of marathon runners: inhibition of post marathon immune suppression and inflammation.

A pilot study was undertaken to investigate the effects of the intake of capsules containing the plant sterols and sterolins (BSS:BSSG mixture) on selected immune parameters of volunteers participating in an ultra-marathon in Cape Town, South Africa. Those runners having received active capsules (n=9) showed less neutrophilia, lymphopenia and leukocytosis when compared to their counterparts having received placebo capsules (n=8): the placebo treated individuals showed significant increases in their total white blood cell numbers as well as in their neutrophils (p=0.03 and 0.03 respectively). Furthermore, statistically significant increases within lymphocyte subsets were observed in the runners having received the active capsules: CD3+ cells increased (p=0.02) as did CD4+ cells (p=0.03). In parallel, the BSS:BSSG capsules decreased the plasma level of IL6 in the runners using the active capsules (p=0.08) and significantly decreased the cortisol: DHEAs ratio (p=0.03), suggesting that these volunteers had less of an inflammatory response and were less immune suppressed during the post-marathon recovery period. These findings justify further investigations into the use of the phytosterols to prevent the subtle immunosuppression associated with excessive physical stress.

Effect of catfish supplementation on the fatty acid status and growth of undernourished rural preschool children under 6 years of age: An intervention trial in Lebowa, South Africa.

A group of 102 preschool children aged 13-69 months from a rural area of Lebowa were selected from a cross-sectional study of 659 children for an intervention trial (12 months) to study the effect of catfish (Clarias gariepinus) supplementation on their plasma phospholipid fatty acid status and growth. They were classified into undernourished and control groups according to their weight-for-age. The undernourished children (n = 52) received 43 g fish and 7.5 g sunflower cooking oil per day, whereas a matched (age and sex) well-nourished control group (n = 50) was not supplemented. At baseline, after 6 months and after 12 months of the study, anthropometry, haematology, blood biochemistry and plasma phospholipid fatty acid analyses were done. In the undernourished group, high baseline oleic acid (18:1 9) levels in plasma phosphatidylcholine (PC) were replaced by docosahexaenoic acid (22:6 3) with supplementation. In plasma PC, this reduction in 18:1 9 and increase in 22:6 3 was associated with significant increases in weight-for-age Z-scores, P = 0.0378 and P = 0.0415, respectively. The fish supplement and cooking oil that supplied additional 7% energy (7% E) and nutrients promoted growth of undernourished children, although this was inadequate for sustained growth during the second 6 months of intervention.

Plasma and red blood cell total phospholipid fatty acid status of nonpregnant female Vervet monkeys (Cercopithecus aethiops) on a high carbohydrate maintenance diet.

Nonhuman primates are of interest as models of human physiology to study the effect of multiple pregnancies on birth weight. Reference plasma and red blood cell (RBC) total phospholipids fatty acids were established in nonpregnant breeding female Vervet monkeys. Twenty-three clinically healthy nonpregnant Vervet monkeys (Cercopithecus aethiops), contained in a controlled closed environment and consuming a high carbohydrate diet (68 E%) that contained 20 E% fat and 12 E% protein were sampled for blood during a cross-sectional study. A low intake of omega3 fatty acids was reflected by a high omega6/omega3 ratio (66:1) of the diet. Inverse relations were seen between plasma and RBC total phospholipid fatty acids, 18:2omega6, 20:3omega6, and 20:4omega6, which suggested selective incorporation in membranes. Low levels of 20:5omega3 and 22:6omega3 of plasma and RBC total phospholipids render Vervet monkeys as ideal subjects to study the effect of omega3 fatty acid supplementation on pregnancy outcomes.

Trimodality of isoniazid elimination: phenotype and genotype in patients with tuberculosis.

The study was undertaken to show that polymorphic isoniazid elimination in humans is trimodal; that the acetylator genotype and eliminator phenotype of the individual patient are concordant; and that the differences in the pharmacokinetic parameters of fast, intermediate, and slow eliminator subgroups are statistically significant. Sixty adult patients of both sexes and of mixed race with tuberculosis participated in the trial. The apparent elimination rate constant (k, h(-1)) and the area under the isoniazid concentration-time curve (AUC, mg/L/h), over the interval 2 to 6 h after oral isoniazid were determined in all patients; NAT2 allele composition was determined in 47 patients. Serum INH concentrations were determined by HPLC and genotypes by PCR/restriction enzyme analysis. Three eliminator phenotypes could be distinguished, and concordance between the phenotype and the genotype of the individual could be demonstrated. The isoniazid concentration-time profiles of the three eliminator subgroups were significantly different (p < 0.05). The NAT2*12A allele, which codes for fast acetylation, has a high frequency in the population studied, the intermediate acetylator genotype is constituted of codominant fast and slow alleles, and the distribution of phenotypes/genotypes in the population is consistent with Hardy-Weinberg predictions. The therapeutic implications of polymorphic isoniazid metabolism are discussed.

Are phosphorylated tyrosine residues of certain receptors involved in inducing transitory covalent protein cross-linking?

Following ligand binding a number of cell-surface receptors become phosphorylated at tyrosine residues of their cytosolic domains. These phosphorylations are associated with initiation of a signalling programme involving a sequence of tyrosine-phosphorylated protein-protein interactions. In the recognition process between phosphorylated proteins, electrostatic interactions between negatively charged phosphorylated tyrosines, serine and threonine residues and positively charged lysines play an important role as well as hydrophobic and H-bonding reactions. We suggest in this paper that the fairly high-energy phosphate bond of certain protein phosphorylated tyrosines are possibly involved in inducing transitory protein cross-linking reactions. Through a process involving transfer of an activated phosphate of phosphorylated tyrosine to a side-chain carboxyl group of the receptor or next protein of the signalling sequence, an acyl phosphate is formed. This then acylates a hydroxyl group on a serine, threonine or tyrosine residue of the protein not carrying the carboxyl phosphate to give an ester linkage, thus cross-linking the two proteins of the signalling pathway. The covalent ester linkage is labile to hydrolysis and depending on the protein-protein molecular environment it might have a finite half-life. On hydrolysis, the transitory covalent linkage is broken with separation of the proteins. It is suggested therefore that formation of a protein-protein ester linkage introduces a type of timing device into the system. Breakdown of the original protein-phosphorylated tyrosine in this case therefore does not involve a phosphatase enzyme.

Vervet monkeys and whole-food diets for studying the effects of dietary lipids on plasma lipoprotein metabolism and atherosclerosis.

It is well established that some species of nonhuman primates are models of choice for polygenic hyperlipoproteinaemia and atherosclerosis induced and promoted by diets as occur in man. The Vervet monkey (Cercopithecus aethiops) has proved to be one such model. Our group has used this model extensively to determine the effects of a variety of dietary lipid components on plasma lipoprotein metabolism and atherosclerosis against a background of a Western atherogenic or prudent diet. The diets fed in all these studies were formulated entirely from cooked foods that are normal components of Westernised diet with no extra synthetic cholesterol added. This model has been used successfully to evaluate the effect of fish oil, amount and degree of dietary fat unsaturation and w-6/w-3 fatty acid ratio and lipid-lowering agents on plasma lipoprotein metabolism and atherosclerosis. Dietary manipulation in this model is simple, relatively inexpensive and offers almost unlimited options for future dietary intervention studies.

A randomised placebo-controlled trial of the efficacy of beta-sitosterol and its glucoside as adjuvants in the treatment of pulmonary tuberculosis.

OBJECTIVE: To evaluate the adjuvant effect of beta-sitosterol and its glucoside in the treatment of culture proven pulmonary tuberculosis (PTB). DESIGN: A blinded randomised placebo-controlled trial in culture proven drug sensitive PTB. Patients were hospitalised for the duration of treatment and evaluated at monthly intervals with regard to sputum culture positivity, chest radiography, weight gain, Mantoux test response, routine haematology and liver functions. STATISTICAL EVALUATION: General linear models for repeated measures (SAS GLM package) compared the interaction effects, group effects and time effects of findings in 19 patients receiving sitosterols with those in 18 patients receiving a placebo (talcum powder). Absolute values and change from baseline values were evaluated, although only the latter are reported. RESULTS: Weight gain was significantly greater in the sitosterol group (mean weight gain 8.9 kg) than the placebo group (mean gain 6.1 kg) (P = 0.0023 group effects; P = 0.0001 for time effects). Speed of achieving culture negativity, radiological improvement and induration on Mantoux testing was similar in the two groups. Change in lymphocyte counts from baseline was significantly higher in the sitosterol group (P = 0.0001 and P = 0.0001 for group and time effects) as was the increase in eosinophil counts (P = 0.0001 and P = 0.0137 for group and time effects). CONCLUSION: The study has shown significantly improved weight gain and higher lymphocyte and eosinophil counts in PTB patients receiving sitosterols in addition to an efficacious antituberculosis regimen. Sitosterols and their possible mode of action should now be evaluated in larger numbers of tuberculosis patients and in diseases with a similar immunopathogenesis.

beta-Sitosterol and beta-sitosterol glucoside stimulate human peripheral blood lymphocyte proliferation: implications for their use as an immunomodulatory vitamin combination.

The phytosterols, beta-sitosterol (BSS), and its glucoside (BSSG) enhance the in vitro proliferative response of T-cells stimulated by sub-optimal concentrations of phytohaemagglutinin (PHA) several fold at extremely low concentrations (femtogram level). A 100:1 (mass:mass) ratio of BSS:BSSG (termed essential sterolin formulation, ESF) showed higher stimulation than the individual sterols at the same concentration. In vivo activity of ESF was also demonstrated when volunteers ingested ESF for 4 weeks. Proliferation of their T-cells, stimulated maximally with PHA, was significantly enhanced (20-920%) when compared to baseline values. In vitro, ESF (1 microgram.ml) was able to significantly enhance the expression of CD25 and HLA-Dr activation antigens on T-cells and increased the secretion, into the medium, of IL-2 and gamma interferon. NK-cell activity was also increased by BSS and BSSG alone, but with EST a higher activity was always found at different effector:target ratios (100:1 12:1).

Pharmacokinetic behaviour and cardiovascular effects of intravenously administered hypoxoside and rooperol.

This study concerns the pharmacokinetic behaviour and cardiovascular effects of rapid infusions of hypoxoside (CAS 83643-94-1) and rooperol (CAS 83644-00-2) in anaesthetised Chacma baboons. Institutional approval was obtained and animal care conformed to international guidelines. Hypoxoside (500 mg) and rooperol (240 mg) dissolved in isotonic saline were infused during 15 min. Concentration-time data from high performance liquid chromatography of arterial blood samples were subjected to non-linear curve-fitting to obtain two-compartment mammillary pharmacokinetic models. Mean values were: [Table: see text] Hypoxoside was eliminated without significant metabolite formation and it revealed no cardiovascular effects. Rooperol was metabolized rapidly with formation of nine metabolites of which the major three were the diglucuronide, disulphate and mixed glucuronide sulphate. Rooperol caused moderate, transient increased cardiac output, stroke volume and vascular pressures without increased heart rate or filling pressures, suggestive of increased myocardial contractility probably allied to its catechol structure.

Essential fatty acids and modern lifestyle: a reappraisal.

Controversy surrounds the effects of dietary fish oil supplementation on atherosclerosis. Three studies were undertaken, where Vervet monkeys were fed either a Western atherogenic diet (WAD) or a high carbohydrate diet (HCD). The first study indicated that enhanced atherosclerosis may be the result of an imbalance of fatty acids in plasma and tissue lipids as eicosapentaenoic acid (EPA; C20:5 ω3) was increased with fish oil (FO) supplementation at the expense of arachidonic acid (AA; C20:4 ω6). The second study investigated the effect of diet on the metabolism of EPA. Disappearance of EPA, after EPA loading, was delayed in Vervets on the WAD in comparison with those on the HCD. Results of this study indicate that diet is able to modulate EPA metabolism, and that the beneficial effects of a HCD on plasma lipoprotein concentrations can be augmented by EPA supplementation. The third study investigated the combined effect of a supplement that contained different ratios and dosages of gamma-linolenic acid (GLA; C18:3 ω6) and EPA during ingestion of the WAD. Based on a favourable response to plasma lipoprotein cholesterol and a phosphatidylcholine fatty acid metabolism with increases in both EPA and dihomogamma-linolenic acid (DGLA; C20:3 ω6), we conclude that a 4:1 ω6/ω3 fatty acid supplement at 200 mg/day would be the optimum supplement in our animal model. The long-term effects of this supplement on lipoprotein metabolism and atherosclerosis in the non-human primate model, is currently under investigation.

High-performance liquid chromatographic determination of isoniazid, acetylisoniazid and hydrazine in biological fluids.

The basic principle of derivatization of a hydrazide moiety with an aldehyde as applied in the method developed by Lacroix et al. [J. Chromatogr., 307 (1984) 137-144] for the quantitation of isoniazid and acetylisoniazid was improved by modification, standardization and extension to allow quantitation of hydrazine in patient samples. It could be shown that 40 microliters of 1% methanolic cinnamaldehyde per 200 microliters of deproteinized analysate gave maximal chromophoric isoniazid-cinnamaldehyde conjugate, read at 340 nm. The hydrolytic loss of isoniazid, crucial to the quantitation of acetylisoniazid, could be compensated for by introduction of an appropriate set of calibration curves. Although the method described here allows quantitation of monoacetylhydrazine and diacetylhydrazine, in addition to hydrazine, in mono-spiked samples, the method cannot be used for the quantitation of the acetylated metabolites of hydrazine in patient samples because of a lack of specificity. Linear calibration curves in the range 1-25 micrograms/ml for isoniazid and acetylisoniazid, 10-400 ng/ml for hydrazine and 50-1000 ng/ml for monoacetylhydrazine and diacetylhydrazine, could be constructed; analyte recoveries approaching 100% could be achieved in all instances.

The pharmacokinetic behaviour of hypoxoside taken orally by patients with lung cancer in a phase I trial.

OBJECTIVE: To study the pharmacokinetic behaviour of hypoxoside taken orally by 24 patients with lung cancer. DESIGN: Randomised open study with three single doses of 1,600, 2,400 and 3,200 mg standardised Hypoxis plant extract (200 mg capsules) and a multiple-dose study on the first 6 patients taking 4 capsules 3 times daily for 11 days. PARTICIPANTS AND SETTING: Patients with histologically proven squamous, large-cell or adenocarcinoma were hospitalised at the Radiation Oncology Ward, Karl Bremer Hospital, Bellville, W. Cape. METHODS: Blood was drawn at regular intervals up to 75 hours after single doses and the concentrations of metabolites of the aglucone of hypoxoside, rooperol, were measured with a high-performance liquid chromatography method. For the multiple-dose study blood was drawn before the first dose each day. Concentration-time relationships were analysed according to a conventional single open-compartment model and also by using the NONMEM digital computer programme. RESULTS: Neither hypoxoside nor rooperol appear in circulation. This is due to complete phase II biotransformation to diglucuronide, disulphate and mixed glucuronide-sulphate metabolites, of which the latter is the major component. Considerable interpatient variation in concentration-time relationships was found in the single-dose studies. It was due to an active enterohepatic recirculation in some patients and a distinct lag phase in others together with zero-order rate of formation of rooperol in the colon. Computer modelling indicated a single open-compartment model in which the mass of the patient did not influence volume of distribution and clearance because formation of the metabolites is dependent on the metabolising capacity of the patient. However, the elimination of the metabolites follows first-order kinetics with half-lives ranging from 50 hours for the major metabolite to 20 hours for the two minor metabolites. Multiple-dose studies also showed large interpatient variation. CONCLUSION: In order to reach metabolite levels near 100 micrograms/ml, which have been shown to be tumouricidal after enzymatic deconjugation to rooperol, maintenance doses need to be individualised for each patient. For most patients, however, a daily dose of 2,400 mg was sufficient.

A phase I trial of hypoxoside as an oral prodrug for cancer therapy--absence of toxicity.

OBJECTIVE: To assess the toxicity of hypoxoside taken orally by 24 patients with lung cancer. DESIGN: Open study with patients taking 1,200-3,200 mg standardised Hypoxis plant extract (200 mg capsules) per day divided in 3 doses in order to maintain metabolite blood levels near 100 micrograms/ml. PARTICIPANTS AND SETTING: Patients with histologically proven squamous, large-cell or adenocarcinoma were hospitalised initially at the radiation oncology ward, Karl Bremer Hospital, Bellville, W. Cape. Thereafter they returned every 2 weeks for full clinical examinations. METHODS: Routine biochemical and haematological measurements were done. Patients underwent regular full clinical examinations including radiographs and computed tomography scanning according to the discretion of the principal investigator. RESULTS: Nineteen patients on hypoxoside therapy survived for an average of 4 months with progression of their primary tumours and metastases, while 5 survived for more than a year. One of them survived for 5 years and histological examination of the primary lesion showed absence of cancer. No toxic effects, in clinical examinations or biochemical or haematological measurements, were found that could be ascribed to the ingestion of hypoxoside. Only one occasion of possible drug intolerance, with anxiety, nausea, vomiting and diarrhoea, was noted. CONCLUSION: The absence of toxicity warrants further investigation of hypoxoside as an oral prodrug, especially in patients with slow-growing necrotising tumours that are inoperable and have high concentrations of beta-glucuronidase and sulphatase as well as a high sensitivity for rooperol.

The effect of iron fortification on the fatty acid composition of plasma and erythrocyte membranes in primary school children with and without iron deficiency.

An intervention study was designed to evaluate the fatty acid (FA) status of children aged 6-11 years before and after iron fortification. Iron-deficient (ID) and matched controls without ID (n = 30) were selected. All children received soup (160 ml) fortified with 20 mg iron and 100 mg vitamin C for 15 weeks on school days. Measurements before and after intervention included dietary intake, haematological and iron status and FA composition of plasma and erythrocyte membranes (EMBs). The prevalence of low plasma ferritin concentration and transferrin saturation decreased in the ID children by 40% and 56%, respectively, with intervention. Plasma FAs reflected dietary FA intake. In comparison with controls, the ID group presented with increased percentage total saturated FAs (SFAs; p = 0.0002) in their EMB phosphatidylcholine (PC) and reduced percentage total polyunsaturated FAs (PUFAs; p = 0.0037) before intervention. Lower total n-3 FAs (p = 0.0070), including eicosapentenoic acid (EPA; p = 0.0034), docosapentenoic acid (DPA; p = 0.0048) and docosahexenoic acid (DHA; p = 0.0058), were observed in the ID group. The EMB phosphatidylethanol-amine (PEA) of the ID children presented with lower percentages of alpha-linolenic acid (ALA; p = 0.0001), EPA (p = 0.0051) and DHA (p = 0.0084) compared to controls before intervention. Iron intervention was associated with an increase (p < 0.05) in the percentage of n-3 FAs in the EMB-PC and EMB-PEA of the ID group to percentages comparable to that in the control group. It appears that iron status can influence FA metabolism of specific n-3 FAs in the EMBs of young children.