Thyroid function of steatitis-affected Mozambique tilapia Oreochromis mossambicus from a sub-tropical African reservoir.

Thyroid function and nutritional indicators were measured in obese, steatitis-affected Mozambique tilapia Oreochromis mossambicus from Loskop Reservoir (LR), South Africa. Plasma thyroid hormones (especially T3) and thyroid follicle histomorphology revealed high levels of activity in every aspect of the thyroid cascade measured in fish from LR compared to a reference population of steatitis-free fish. Concurrent measurements of nutritional state including plasma lipids, liver lipid content and hepatocyte size showed that fish from LR had significant energy stores indicative of abundant nutritional intake. There were distinct sex and seasonal differences, with the highest plasma lipids and T3 levels observed in steatitis-affected females during spring and summer. Positive correlations were observed between plasma lipids (especially cholesterol) and T3 concentrations in fish from both populations, indicating a link between lipid metabolism and thyroid function. There was no direct evidence of thyroid disruption, but this cannot be ruled out until further research determines the factors that underlie the homeostatic shift leading to elevated plasma and liver lipids and T3 levels in steatitis-affected tilapia.

Pansteatitis in polluted Olifants River impoundments: nutritional perspectives on fish in a eutrophic lake, Lake Loskop, South Africa.

This study compares the aetiology of pansteatitis in Lake Loskop, relative to two other impoundments along the Olifants River. Macroscopic and microscopic pathology, age determination and analysis of stomach content, fatty acids and stable isotopes explain the high prevalence of pansteatitis in Oreochromis mossambicus (Peters) and several other species in Lake Loskop. All the dietary indicator comparisons between pansteatitis-affected and healthy fish fail to support a systemic cause. Pansteatitis in Lake Loskop was linked to size and weight of O. mossambicus, but not to ontogenic age. Fish in Lake Loskop showed abnormally high omega-3 to omega-6 fatty acid ratios normally only found in marine fish with no significant difference in degree of assimilation of these fatty acids between pansteatitis-affected and healthy fish. This explains the vulnerability to, but not the occurrence of, pansteatitis. As a cause for the pansteatitis, these results point towards sporadic vitamin E-depleting trigger events, known sporadic fish die-off occurrences that provide surviving fish with a rich source of rancid fats on which to scavenge. The mechanism ties pansteatitis to eutrophication and trophic cascade effects, the intrinsic drivers of the disease and suggests an adaptive management strategy that might be applied by relevant conservation authorities.

Milk composition of free-ranging red hartebeest, giraffe, Southern reedbuck and warthog and a phylogenetic comparison of the milk of African Artiodactyla.

The composition of major nutrients and fatty acids of the milk of three species, red hartebeest, Southern reedbuck and warthog, and milk fatty acids of giraffe, that have not been published before, are reported, and together with the same parameters of 11 species previously published, were incorporated in a phylogenetic comparison. Unique properties of milk composition have been observed. Southern reedbuck milk seems to have a complex casein composition, similar to that of sheep. Milk composition varies between species. Although some differences may be ascribed to biological condition, such as stage of lactation, or ecological factors, such as availability of certain nutrients, the contribution by evolutionary history is not well documented and the emphasis is usually on the composition of the macro nutrients. Phylogenetic comparisons often lack representatives of multiple species of taxonomic groups and sub-groups. To date phylogenetic comparisons of milk composition have been carried out by using data from different publications. The problem with this approach is that the ecological factors cannot be completely ruled out. A statistical phylogenetic comparison by PCA between 15 species representing 7 different suborders, families or subfamilies of African Artiodactyla was carried out. The phylogenetic properties showed that the milk composition of the Bovinae, represented here by the subfamilies Bovini and Tragelaphini, differs from the other taxonomic groups, in that the Alcelaphinae had a high milk fat content of the medium chain length fatty acids C8-C12 (>17% of total fatty acids) and the Hippotraginae high amounts of oligosaccharides (>0.4%).

Characterization of casein and alpha lactalbumin of African elephant (Loxodonta africana) milk.

The current research reports partial characterization of the caseins and α-lactalbumin (α-LA) of the African elephant with proposed unique structure-function properties. Extensive research has been carried out to understand the structure of the casein micelles. Crystallographic structure elucidation of caseins and casein micelles is not possible. Consequently, several models have been developed in an effort to describe the casein micelle, specifically of cow milk. Here we report the characterization of African elephant milk caseins. The κ-caseins and ß-caseins were investigated, and their relative ratio was found to be approximately 1:8.5, whereas α-caseins were not detected. The gene sequence of ß-casein in the NCBI database was revisited, and a different sequence in the N-terminal region is proposed. Amino acid sequence alignment and hydropathy plots showed that the κ-casein of African elephant milk is similar to that of other mammals, whereas the ß-casein is similar to the human protein, and displayed a section of unique AA composition and additional hydrophilic regions compared with bovine caseins. Elephant milk is destabilized by 62% alcohol, and it is speculated that the ß-casein characteristics may allow maintenance of the colloidal nature of the casein micelle, a role that was previously only associated with κ-casein. The oligosaccharide content of milk was reported to be low in dairy animals but high in some other species such as humans and elephants. In the milk of the African elephant, lactose and oligosaccharides both occur at high levels. These levels are typically related to the content of α-LA in the mammary gland and thus point to a specialized carbohydrate synthesis, where the whey protein α-LA plays a role. We report the characterization of African elephant α-LA. Homology modeling of the α-LA showed that it is structurally similar to crystal structures of other mammalian species, which in turn may be an indication that its functional properties, such as lactose synthesis, should not be impaired.

Effect of dietary conjugated linoleic acid supplementation on the technological quality of backfat of pigs.

Pigs were fed diets containing 0, 0.25, 0.5 and 1% conjugated linoleic acid (CLA). Compared to controls, backfat from CLA fed pigs was firmer and extracted lipid contained increasing amounts of CLA, but a ±11% overall decrease in unsaturated fatty acids and a ±5% overall increase in each of C16:0 and C18:0 saturated fatty acids were noted. This resulted in a change in the melting properties of fat. The onset setting temperature increased from ±14°C to ±18°C for lipid of backfat of pigs from the 0.25 and 0.5% CLA supplementation groups, and to ±26°C for lipid from the 1% CLA supplementation group. The final melting temperatures increased from ±37°C to ±43°C and ±45°C, respectively. The presence of ß'-crystals of C18:0-C16:0-C18:1c9 triacylglycerides in fat from CLA fed pigs and ß-crystals in fat from 1% CLA fed pigs was observed. Fatty acid and melting point results explained the improvement in the technological quality of backfat as a result of dietary CLA supplementation.

Comparison of the lipid properties of healthy and pansteatitis-affected African sharptooth catfish, Clarias gariepinus (Burchell), and the role of diet in pansteatitis outbreaks in the Olifants River in the Kruger National Park, South Africa.

Pansteatitis has been identified in wild populations of sharptooth catfish, Clarias gariepinus (Burchell), and Nile crocodiles, Crocodylus niloticus Laurenti, inhabiting the same waters in the Olifants River Gorge in the Kruger National Park, South Africa. Mesenteric and pectoral fat tissue was investigated microscopically and by fatty acid analysis in healthy and pansteatitis-affected catfish from both captive and wild populations. Variation in fatty acid composition between pectoral and mesenteric fat was noted. Composition of mesenteric fat differed between fish from various localities as a result of differences in diet. Pansteatitis in the captive population, resulting from ingestion of high amounts of dietary oxidized fat, reflected higher levels of unsaturated fatty acids within the mesenteric fat. Mesenteric fat of pansteatitis-affected wild catfish was characterized by an increase in moisture content, a decrease in fat content and a decrease in stearic and linoleic acids. The n-3 to n-6 fatty acid ratio of mesenteric fat was higher in pansteatitis-affected wild catfish than in healthy catfish from the same locality, reflecting higher polyunsaturated fat intake by pansteatitis-affected fish. The possible role of alien, invasive, phytoplankton-feeding silver carp, Hypophthalmichthys molitrix (Valenciennes), in the aetiology of pansteatitis in both catfish and crocodiles in the Olifants Gorge is discussed.

Characterization of a spray-dried soymilk powder and changes observed during storage.

Physical characterization of a soymilk powder was carried out by electron microscopy. Chemical characterization was analyzed by proximate analysis, mineral composition by atomic absorption spectrometry, fatty acid composition by gas chromatography and protein composition by electrophoresis. The powder consists of large granules of 60-80 µm, which may be hollow, with smaller granules of 10-20 µm attached to them. Powder particles are covered by a layer of fat. During storage at 25 °C fat is spreading over the surface, while at -12 °C the fat is contracting. This change affected chemical stability, resulting in high level of fat oxidation when stored at 4 °C or 25 °C as well as a decrease in unsaturated fatty acids. Storage also affected the chemical properties of the re-constituted soymilk; the pH of a 12% soy powder suspension increased from 6.68 ± 0.05 to 7.06 ±0.08 after 12 months of storage. Storage temperature did not affect the pH of the suspension and this change could also not be ascribed to protein aggregation.

Comparison of the milk composition of free-ranging blesbok, black wildebeest and blue wildebeest of the subfamily Alcelaphinae (family: Bovidae).

The proximate composition of milk from free ranging blesbok, black wildebeest and blue wildebeest is reported, with detailed analysis of fatty acid composition and protein identification by electrophoresis. The milk of the blesbok, black- and blue wildebeest contain respectively 5.6 +/- 0.3, 4.3 +/- 0.7 and 4.1 +/- 0.9 g/100 g protein, 8.6 +/- 1.1, 5.5 +/- 1.5 and 7.5 +/- 2.1 g/100 g fat, and 4.9 +/- 0.4, 4.1 +/- 0.6 and 5.3 +/- 1.6 g/100 g lactose, and are comparable with ovine milk. The milk contains high levels of saturated fatty acids, respectively, 82.5 +/- 1.7 g/100 g, 75.2 +/- 1.5 g/100 g and 88.2 +/- 1.8 g/100 g milk fat. The medium carbon chain length saturated fatty acids 8:0, 10:0 and 12:0 occur in the highest amounts recorded so far in ruminant milk, and the 14:0 content is the highest recorded in any milk. This suggests a genetic characteristic of the Alcelaphinae that reflects on the specificity and activity of the thioesterase involved in the fatty acid synthesis. By electrophoresis the kappa- and gamma-caseins are separated into multiple bands and the alpha-caseins of the wildebeest into two bands.

Milk composition of captive vervet monkey (Chlorocebus pygerythrus) and rhesus macaque (Macaca mulatta) with observations on gorilla (Gorilla gorilla gorilla) and white handed gibbon (Hylobates lar).

The nutrient content and fatty acid composition of vervet monkey milk has been determined and is compared with rhesus macaque, and two hominoid apes, the white handed gibbon and gorilla. With 15.7+/-4.1 g protein, 33.1+/-9.4 g fat, and 85.1+/-7.5 g lactose per kg milk, vervet monkey milk does not differ from that of rhesus macaque, and is within the range of other primates. Small amounts (>1 g kg(-1)) of oligosaccharides, glucose, galactose and fucose were noted. In comparison, gorilla milk has a low fat content of 13.8 g kg(-1), but contains high levels of oligosaccharides at 7.0 g kg(-1) milk. The hominoid partner, the white handed gibbon, contains no oligosaccharides and a milk fat content similar to other hominoid species. Differences between vervet monkey and rhesus macaque milks were observed in the electrophoretic pattern of the milk proteins, mainly amongst the kappa- and gamma-caseins, which also differ from that of the hominids. The fatty acid contents of these milks differ from studies where a natural diet of leafy material was available in that a low content of alpha-linolenic acid (18:3n-3) was noted. A phylogenetic effect is observed for the content of 8:0, 10:0 fatty acids between the Cercopithecidae and Hominoidea, and a further phylogenetic effect suggested between the Hylobatidae and Hominidae.

Structural characterization of oligosaccharides in the milk of an African elephant (Loxodonta africana africana).

The oligosaccharides present in the milk of an African elephant (Loxodonta africana africana), collected 4 days post partum, were separated by size exclusion-, anion exchange- and high-performance liquid chromatography (HPLC) before characterisation by (1)H NMR spectroscopy. Neutral and acidic oligosaccharides were identified. Neutral oligosaccharides characterised were isoglobotriose, Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc, Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc, Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc and a novel oligosaccharide that has not been reported in the milk or colostrum of any other mammal: Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc. Acidic oligosaccharides that are also found in the milk of Asian elephant were Neu5Ac(alpha2-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-6)Gal(beta1-4)Glc, Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]Glc, Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc and Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3){Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)}Gal(beta1-4)Glc, while Neu5Gc(alpha2-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)[Fuc(alpha1-3)]Glc, Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)[Gal(beta1-4)GlcNAc(beta1-6)]Gal(beta1-4)Glc and Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3){Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)}Gal(beta1-4)Glc have not been found in Asian elephant milk. The oligosaccharides characterised contained both alpha(2-3)- and alpha(2-6)-linked Neu5Ac residues. They also contain only the type II chain, as found in most non-human, eutherian mammals.

Milk composition of three free-ranging African elephant (Loxodonta africana africana) cows during mid lactation.

Data are presented that indicate the dynamic changes of nutrients in milk from three free ranging African elephant (Loxodonta africana africana) cows during lactation. At the respective collection times of 12, 14 and 18 months of lactation the nutrient content was 47.3, 52.0 and 68.6 g protein; 60.7, 87.4 and 170.8 g fat; 1.6, 2.1 0.5 g lactose and 20.9, 21.5 and 8.6 g oligosaccharides per kg milk. The protein fraction respectively consisted of 18.0, 31.7 and 45.9 g caseins/kg milk and of 29.3, 20.3 and 22.7 g whey proteins/kg milk. Electrophoresis and identification of protein bands showed that polymorphs of one whey protein may be present in elephant's milk similar to polymorphs of alpha-lactalbumin found in cow's milk. From the middle of the lactation time lactose was replaced by oligosaccharides as major carbohydrate, and the major compound of these was identified as isoglobotriose by 1H NMR spectroscopy. The lipid fraction contains a high content, of capric and lauric acids, approximately 70% of the total fatty acids, and low content of myristic, palmitic and oleic acids. During these lactation times the content of short chain fatty acids, capric and caprylic acids increased, while fatty acids lauric acid and longer decreased.

The composition of serval (Felis serval) milk during mid-lactation.

Milk was obtained from three captive servals. The average nutrient content was 158.3+/-44.4 g protein; 152.6+/-62.3 g fat; and 68.7+/-31.4 g lactose per kg milk. The protein fraction respectively consisted of 117.7+/-44.8 g caseins per kg milk and of 40.6+/-6.7 g whey proteins per kg milk. Electrophoresis and identification of protein bands showed a similar migrating sequence of proteins as seen in cheetah and cat milk, with small differences in the beta-caseins. The lipid fraction contains 313.3+/-18.8 g saturated and 338.6+/-11.9 g mono unsaturated fatty acids per kg milk fat respectively. The high content of 292.4+/-24.9 g kg(-1) milk fat of polyunsaturated fatty acids is due to a high content in linolenic acid. No short chain fatty acids, but substantial levels of uneven carbon chain fatty acids were observed. In general, serval milk has a higher protein and fat content than that of the domestic cat and cheetah, and a lower content of unsaturated fatty acids than that of the domestic cat.

Milk composition of free-ranging springbok (Antidorcas marsupialis).

Milk was obtained from three free-ranging springbok ewes of the Karoo, South Africa. The nutrient content was 74.4+/-13.8 g protein; 145.2+/-4.5 g fat; and 42.3+/-16.4 g lactose/kg milk. Small amounts of glucose, galactose and fucose were noted, and 0.3+/-0.4 g oligosaccharides. The protein fraction respectively consisted of 60.0+/-13.7 g caseins/kg milk and of 14.1+/-4.5 g whey proteins/kg milk. The lactation stage of the springbok ewes was not known, but variation in milk composition among individuals indicates that they were at different stages. Electrophoresis and identification of protein bands showed a similar migrating sequence of proteins as seen in caprine milk. The lipid fraction contains 604.0+/-26.5 g saturated fatty acids/kg milk fat, and 278.2+/-20.5 and 45.2+/-3.6 g/kg mono and poly-unsaturated fatty acids respectively. Compared to domesticated dairy species, a low content of short chain length fatty acids was observed, while stearic acid was at higher, and arachidonic acid at lower levels. Substantial levels of uneven carbon chain fatty acids were also observed. Springbok milk is much more concentrated than the milks of most ruminants, with higher fat and oligosaccharide contents.

The composition of cheetah (Acinonyx jubatus) milk.

Milk was obtained from two captive bred cheetahs. The nutrient content was 99.6 g protein; 64.8 g fat; and 40.21 g lactose per kg milk. Small amounts of oligosaccharides, glucose, galactose and fucose were noted. The protein fraction respectively consisted of 34.2 g caseins per kg milk and of 65.3 g whey proteins per kg milk. Very little variation in milk composition among the individual cheetahs was noted. Electrophoresis and identification of protein bands showed a similar migrating sequence of proteins as seen in lion's and cat's milk, with small differences in the beta-caseins. The lipid fraction contains 290.4 g saturated and 337.3 g mono-unsaturated fatty acids per kg milk fat respectively. The high content of 279.5 g kg(-1) milk fat of polyunsaturated fatty acids is due to a high content in alpha-linolenic acid. No short chain fatty acids, but substantial levels of uneven carbon chain fatty acids were observed.

Milk composition of a free-ranging African elephant (Loxodonta africana) cow during early lactation.

Only one study previously reported comprehensively on the composition of African elephant's (Loxodonta africana) milk that was collected from 30 dead animals. In the current study milk was obtained from a tame but free-ranging African elephant cow without immobilization during the period when she was 4-47 days postpartum. At the respective collection times the nutrient content was 21.8 and 25.0 g protein; 56.0 and 76.0 g fat; 71.1 and 26.0 g sugars per kilogram of milk. The protein fraction, respectively, consisted of 10.0 and 14.0 g caseins/kg milk and of 11.8 and 11 g whey proteins/kg milk. During lactation the lactose content dropped from 52.5 to 11.8 g/kg milk, while the oligosaccharide content increased from 11.8 to 15.2 g/kg milk. The oligosaccharide was characterized as a galactosyllactose, which is digestible by cellulase. Electrophoresis and identification of protein bands showed a similar migrating sequence of proteins as seen in cow's milk, but some of the corresponding proteins were less negatively charged. The lipid fraction contains a high content of capric and lauric acids, approximately 60% of the total fatty acids, and low content of myristic, palmitic and oleic acids.

Purification and characterization of a metalloprotease from Chryseobacterium indologenes Ix9a and determination of the amino acid specificity with electrospray mass spectrometry.

The heat-stable protease from Chryseobacterium indologenes Ix9a was purified to homogeneity using immobilized metal affinity chromatography. The enzyme was characterized as a metalloprotease with an approximate relative molecular mass of 24,000, a pH optimum of 6.5, and a high temperature optimum (50 degrees C). The metal chelator EDTA and the Zn2+-specific chelator 1,10-phenanthroline were identified as inhibitors and atomic absorption analysis showed that the enzyme contained Ca2+ and Zn2+. The activity of the apoenzyme could be restored with Ca2+, Zn2+, Mg2+, and Co2+. Phosphoramidon and Gly-d-Phe did not inhibit Chryseobacterium indologenes Ix9a protease. Heat inactivation did not follow first order kinetics, but showed biphasic inactivation curves. The protease has a Km of 0.813 microg. ml-1 for casein as substrate. Amino acid analysis showed that the protease contains a high amount of small amino acids like glycine, alanine, and serine, but a low concentration of methionine and no cysteine at all. Electrospray mass spectrometry of proteolysis fragments formed when insulin B chain was hydrolyzed showed cleavage at the amino terminal of leucine, tyrosine, and phenylalanine. A hydrophobic amino acid at the carboxyl donating side seems to increase the rate of reaction.

Action of lysosomal phospholipase A1 on bis(monoacylglycerol)phosphate.

Bis(monoacylglycerol)phosphate (BMP) in macrophages is known to rapidly turn over its acyl moiety(s) located at primary positions of the glycerols, yet the glycerols and phosphate remain stable within the BMP molecule. Here we examine whether the phospholipase A1 isolated from rat-liver lysosomes is capable of deacylating BMP. By comparison with the precursor of BMP, phosphatidylglycerol, BMP is a very poor substrate for the phospholipase A1. We conclude, therefore, that a direct deacylation of the acyl groups at the primary alcohol level of the glycerol probably does not occur, but postulate that transacylations may occur to account for the removal of the acyl moiety.

Metabolism of phosphatidylglycerol and bis(monoacylglycero)-phosphate in macrophage subcellular fractions.

Bis(monoacylglycero)phosphate (BMP) is synthesized from exogenous phosphatidylglycerol (PG) by macrophages (Cochran, F. R., Roddick, V. L., Connor, J. R., Thornburg, J. T., and Waite, M. (1987) J. Immunol. 138, 1877-1883). Previous work from our laboratory showed that arachidonic acid in BMP was released by the macrophages upon challenge of the cells with PMA (Cochran, F. R., Connor, J. R., Roddick, V. L., and Waite, M. (1985) Biochem. Biophys. Res. Commun. 130, 800-806). Here we extend those studies using a model cultured cell line of macrophages, RAW 264.7. When PG labeled with 32P- and [3H]glycerol in both moieties was added to the culture medium, 32P/[3H]BMP was synthesized in a time-dependent manner. Fractionation of cell homogenates on a discontinuous sucrose gradient in which the light membranes were floated from dense sucrose showed an enrichment of [3H]BMP in light membrane fractions. The precursor [3H]PG was also found in the light fractions but, relative to the [3H]BMP, was more abundant in the denser membrane fractions. The appearance of [3H]PG and [3H]BMP in the light membrane fraction was time-dependent which suggested that the initial uptake and metabolism of [3H]PG was into the denser membranes. Incubation of the light membranes under conditions that are optimal for the lysosomal phospholipase A1 led to significant metabolism of [3H]PG. Both degradation of [3H]PG to water-soluble compounds and its conversion to acylphosphatidylglycerol occurred while no lyso-PG was detected. On the other hand, little BMP was found to be degraded. From these studies we postulate that in lysosomes acylphosphatidylglycerol is a precursor of BMP and that the previously reported turnover of arachidonic acid by BMP may occur via transacylation rather than hydrolysis.

Far-UV CD spectroscopy, IR spectroscopy, and immunological properties of synthetic peptides, including an amphipathic alpha-helical peptide from bovine phospholipase A2.

The immunological properties of bovine phospholipase A2 were investigated in order to locate possible sequential epitopes. Polyvalent antiserum raised against the enzyme was tested for its interaction with synthetic peptides derived from regions adopting hydrogen-bonded secondary structures within the tertiary structure of phospholipase A2. The conformations of each peptide in various solvents were determined by CD and ir spectroscopy in order to relate immunological to structural properties. It was found that sequential epitopes are absent in this enzyme, but that region 90-109 could be a possible T-cell epitope through the formation of an amphipathic alpha-helix.