Antagonism by NKP608 of substance P-induced plasma protein exudation in a novel preparation of perfused trachea in rats and guinea-pigs in vivo

Objective: To examine whether NKP608, a novel 1-benzoyl-2-benzyl-4-aminopiperidine NK1 receptor antagonist, inhibits substance P (SP)-induced airway plasma protein exudation in vivo. Material: Anaesthetised English shorthair guinea-pigs and Wistar rats. Treatment: Tachykinin peptides were applied topically onto the trachea and antagonists administered intravenously. Methods: Tracheal segments isolated in situ were perfused with saline and plasma-derived protein assayed in the perfusate. Results: SP (1 muM) caused plasma protein exudation, which was abolished by an NK1 antagonist (RP 67580, 1.75 mumol/kg) but unaffected by an NK2 antagonist (SR 48968, 1.75 mumol/kg) indicating the response is NK1-receptor-mediated. This was confirmed with a response to an NK1 agonist ([Sar(9), Met(O-2)(11)]-SP, 1 muM) but none to an NK2 agonist ([betaAla(8)]-neurokinin A(4-10), 1 muM). NKP608 inhibited SP responses with estimated ID50 values (mumol/kg) of 0.0044 (guinea-pigs) and 0.19 (rats). Conclusions: NKP608 is an antagonist in vivo of NK1 receptor-induced tracheal plasma protein exudation and is more potent in guinea-pigs than rats.

Polyspecific malaria antibodies present at the time of infection inhibit the development of immunity to malaria but antibodies specific for the malaria merozoite surface protein, MSP1, facilitate immunity

Serum taken from mice immune to malaria as a result of infection and drug cure, or from mice immunized with a recombinant form of the merozoite surface protein, MSP1, can provide passive protection of recipient mice against the lethal parasite, Plasmodium yoelii YM. However, recipients of MSP1-immune serum go on to develop long-term immunity, whereas recipients of serum from mice naturally immune to malaria rapidly lose their resistance to infection. We demonstrate that 'infection/cure' serum suppresses the development of both antibody and cell-mediated parasite-specific responses in recipients, whereas these develop in recipients of MSP1-specific antibodies. These data have profound implications for our understanding of the development of malaria immunity in babies who passively acquire antibodies from their mothers.

The influence of increased line-fishing mortality on the sex ratio and age of sex reversal of the venus tusk fish

The age of sex reversal of the venus tusk fish Choerodon venustus, caught by line fishing at various locations on the southern Great Barrier Reef, indicated that C. venustus is capable of modifying its life cycle in response to increased mortality. The evidence suggests Masthead Reef fish, which experience the highest mortality, underwent sex reversal at a smaller size and younger age than at the other sites. The largest female fish, sexually transitional fish and males were smaller at Masthead Reef than at the Swains Reefs or One Tree Reef at Masthead Reef. There was also considerable overlap in the size of males and females within the exploited populations indicating that sex reversal is not initiated at a particular length but may have a social cause. The sex ratio of fish was essentially the same for fish fully susceptible to line fishing in the Swains and Masthead samples. Circumstantial evidence suggested that the absence of large males in a population may initiate sex reversal, indicating the maintenance of a constant sex ratio may have a social basis. (C) 2002 The Fisheries Society of the British Isles.

The effect of sodium chloride supplementation on the milk production of grazing Holstein Friesian cows during summer and autumn in a humid sub-tropical environment

Two studies were conducted to examine the effects of including NaCl at various rates in grain-based supplements for Friesian cows grazing established, dominant (>90%), rainfed kikuyu (Pennisetum clandestinum cv. Common) pastures during summer and autumn in a humid sub-tropical environment. In study 1 (19 January-27 March 1998), 48 cows (36 multiparous, 12 primiparous; 27-96 days postpartum) were allocated to one of four groups based on genetic merit, milk production, liveweight (LW) and days postpartum. They were fed (2.7 kg dry matter (DM) per cow, twice-a-day) one of four isoenergetic and isonitrogenous barley grain-based concentrates containing NaCl at concentrations (% as-fed) of either 0 (SC1), 1.1 (SC2), 2.2 (SC3) or 3.3 (SC4). Maximum temperature humidity index (THImax) was greater than or equal to78 during 50% of the experimental period. Concentrate NaCl content had no effect (P>0.05) on daily milk yield or LW change but daily yields of 4% fat corrected milk (FCM), fat and protein were higher (P0.05) among treatments at 7.6+/-1.24 kg DM per cow. In study 2 (18 January 1999-1 March 1999), 48 cows (32 pluriparous, 16 primiparous: 32-160 days postpartum) were fed (2.7 kg DM per cow twice-a-day) one of two isoenergetic and isonitrogenous barley grain-based concentrates containing NaCl at concentrations (% as-fed) of 0 (control) or 2.2 (HSC). THImax was greater than or equal to78 during 34% of days in the experimental period. Yields of milk, FCM, fat and protein were lower (P0.05) by concentrate NaCl content. These studies indicate that NaCl supplementation can be beneficial in terms of milk production during warm, humid conditions as opposed to milder conditions. (C) 2002 Elsevier Science B.V. All rights reserved.

A review on the role of duckweed in nutrient reclamation and as a source of animal feed

The family of lemnacae colloquially known as duckweed contains the world's smallest species of flowering plants (macrophytes). Aquatic and free-floating, their most striking qualities are a capacity for explosive reproduction and an almost complete lack of fibrous material. They are widely used for reducing chemical loading in facultative sewage lagoons, but their greatest potential lies in their ability to produce large quantities of protein rich biomass, suitable for feeding to a wide range of animals, including fish, poultry and cattle. Despite these qualities there are numerous impediments to these plants being incorporated into western farming systems. Large genetically determined variations in growth in response to nutrients and climate, apparent anti-nutritional factors, concerns about sequestration of heavy metals and possible transference of pathogens raise questions about the safety and usefulness of these plants. A clear understanding of how to address and overcome these impediments needs to be developed before duckweed is widely accepted for nutrient reclamation and as a source of animal feed.

Protection of mice from group A streptococcal infection by intranasal immunisation with a peptide vaccine that contains a conserved M protein B cell epitope and lacks a T cell autoepitope

Infection with group A streptococci (GAS) can lead to rheumatic fever (RF) and rheumatic heart disease (RHD) which are a major health concern particularly in indigenous populations worldwide, and especially in Australian Aboriginals. A primary route of GAS infection is via the upper respiratory tract, and therefore, a major goal of research is the development of a mucosal-based GAS vaccine, The majority of the research to date has focused on the GAS M protein since immunity to GAS is mediated by M protein type-specific opsonic antibodies. There are two major impediments to the development of a vaccine-the variability in M proteins and the potential for the induction of an autoimmune response. To develop a safe and broad-based vaccine, we have therefore focused on the GAS M protein conserved C-region, and have identified peptides, J8 and the closely related J8 peptide (J14), which may be important in protective immunity to GAS infection. Using a mucosal animal model system, our data have shown a high degree of throat GAS colonisation in B10.BR mice 24 h following intranasal immunisation with the mucosal adjuvant, cholera toxin B subunit (CTB), and/or diptheria toxoid (dT) carrier, or PBS alone, and challenge with the M1 GAS strain. However, GAS colonisation of the throat was significantly reduced following intranasal immunisation of mice with the vaccine candidate J8 conjugated to dT or J14-dT when administered with CTB. Moreover, J8-dT/CTB and J14-dT/CTB-immunised mice had a significantly higher survival when compared to CTB and PBS-immunised control mice. These data indicate that immunity to GAS infection can be evoked by intranasal immunisation with a GAS M protein C-region peptide vaccine that contains a protective B cell epitope and lacks a T cell autoepitope. (C) 2002 Published by Elsevier Science Ltd.

The effects of level of dietary protein on the milk production and rumen physiology of dairy cows fed a diet based on a tropical grass hay

zFour rumen-fistulated, multiparous Holstein-Friesian cows in early lactation were offered mixed diets based on rhodes grass hay (Chloris gayana) cv. Callide containing 13, 14, 15 or 16% crude protein (CP) on a dry matter basis, in a 4 x 4 latin square design. The estimated undegradable protein concentration in these diets was similar with rumen degradable protein concentration varying. Cows fed a diet containing 13% CP had lower (P = 0.07) milk yields than cows in other treatments (20.4 vs 21.9, 22.0 and 22.2 L/d for 13, 14, 15 and 16% CP, respectively). A positive linear relationship was found (P = 0.06) between organic matter intake and dietary CP%. There were negative linear relationships between dietary CP% and digestibilities of dry matter (P = 0.09), organic matter (P = 0.06) and neutral detergent fibre (P = 0.02). Feeding a diet containing 13% CP resulted in significantly lower (P = 0.001) molar proportions (%) of rumen valerate in comparison with other treatments. The molar proportions of isovalerate differed (P = 0.001) between treatments (0.66, 0.78, 0.89 and 1.04%) for 13, 14, 15 and 16% CP, respectively). Dietary protein level had no effect on rates of passage, in situ digestion of rhodes grass hay or ratios of allantoin: creatinine in urine. These data showed that increasing the dietary CP concentration of lactating cows fed diets based on rhodes grass hay increased intakes and not significantly improved at dietary CP concentrations above 14% DM.

Angiotensinogen genotype, plasma protein and mRNA concentration in isolated systolic hypertension

Isolated systolic hypertension (ISH) occurs predominantly in the elderly, with a considerable morbidity and mortality. Its etiology is unknown but is likely to involve a significant genetic component. The aim of this study was to examine the angiotensinogen gene in ISH. The M235T and G(- 6)A polymorphisms were genotyped by polymerase chain reaction (PCR) in 86 ISH patients and 120 normotensive controls. Plasma angiotensinogen concentration was determined in 198 subjects by an indirect radioimmunoassay technique. Angiotensinogen mRNA concentration was determined by quantitative competitive reverse transcription (RT)-PCR in subcutaneous adipose tissue from a subset of these patients (n = 8) and controls (n = 6). Both the M235T (p = 0.0015) and G(- 6)A (p = 0.029) polymorphisms were associated with ISH. Plasma angiotensinogen concentration was higher in patients than controls (p < 0.0001), but was not associated with genotype. Angiotensinogen mRNA concentration in adipose tissue from ISH subjects was significantly lower than in adipose tissue from normotensive subjects (p = 0.033). The association of angiotensinogen gene variants with ISH and the elevation of plasma angiotensinogen concentration in these patients suggests a role of the angiotensinogen gene in this form of hypertension. Angiotensinogen gene expression may be altered in ISH, but this requires further examination.

Effects of dietary level of protein, lysine and methionine and strain of bird on production and egg yolk cholesterol

The effects of dietary level of protein (151, 181 g/kg), lysine (nil, 10g L-lysine hydrochloride/kg) and methionine (nil, 5g DL-methionine/kg) on the production performance and egg yolk cholesterol of two strains of birds were studied for 12 weeks. Birds fed on the high protein diet had higher body weight gain, feed conversion ratio (FCR), rate of lay, egg weight and mass and yolk weight and mass. A high lysine diet decreased feed intake and improved FCR. High dietary level of methionine increased egg yolk cholesterol. There were differences between strains of laying bird in feed intake, rate of lay, egg and yolk weights and egg cholesterol content. It is concluded that strain of bird and dietary level of protein and lysine influenced the production performance of birds. Whilst, egg yolk cholesterol was not reduced by any of the factors studied.

Practical and economic advantages of choice feeding systems for laying poultry

Poultry can be managed under different feeding systems, depending on the husbandry skills and the feed available. These systems include the following: (1) a complete dry feed offered as a mash ad libitum; (2) the same feed offered as pellets or crumbles ad libitum; (3) a complete feed with added whole grain; (4) a complete wet feed given once or twice a day; (5) a complete feed offered on a restricted basis; (6) choice feeding. Of all these, an interesting alternative to offering complete diets is choice feeding which can be applied on both a small or large commercial scale. Under choice feeding or free-choice feeding birds are usually offered a choice between three types of feedstuffs: (a) an energy source (e.g. maize, rice bran, sorghum or wheat); (b) a protein source (e.g. soyabean meal, meat meal, fish meal or coconut meal) plus vitamins and minerals and (c), in the case of laying hens, calcium in granular form (i.e. oyster-shell grit). This system differs from the modern commercial practice of offering a complete diet comprising energy and protein sources, ground and mixed together. Under the complete diet system, birds are mainly only able to exercise their appetite for energy. When the environmental temperature varies, the birds either over- or under-consume protein and calcium. The basic principle behind practising choice feeding with laying hens is that individual hens are able to select from the various feed ingredients on offer and compose their own diet, according to their actual needs and production capacity. A choice-feeding system is of particular importance to small poultry producers in developing countries, such as Indonesia, because it can substantially reduce the cost of feed. The system is flexible and can be constructed in such a way that the various needs of a flock of different breeds, including village chickens, under different climates can be met. The system also offers a more effective way to use home-produced grain, such as maize, and by-products, such as rice bran, in developing countries. Because oyster-shell grit is readily available in developing countries at lower cost than limestone, the use of cheaper oyster-shell grit can further benefit small-holders in these countries. These benefits apart, simpler equipment suffices when designing and building a feed mixer on the farm, and transport costs are lower. If whole (unground) grain is used, the intake of which is accompanied by increased efficiency of feed utilisation, the costs of grinding, mixing and many of the handling procedures associated with mash and pellet preparation are eliminated. The choice feedstuffs can all be offered in the current feed distribution systems, either by mixing the ingredients first or by using a bulk bin divided into three compartments.

Parenteral and mucosal delivery of a novel multi-epitope M protein-based group A streptococcal vaccine construct: investigation of immunogenicity in mice

Primary vaccine strategies against group A streptococci (GAS) have focused on the M protein-the target of opsonic antibodies important for protective immunity. We have previously reported protection of mice against GAS infection following parenteral delivery of a multi-epitope vaccine construct, referred to as a heteropolymer. This current report has assessed mucosal (intranasal (i.n.) and oral) delivery of the heteropolymer in mice with regard to the induction and specificity of mucosal and systemic antibody responses, and compared this to parenteral delivery. GAS-specific IgA responses were detected in saliva and gut upon i.n. and oral delivery of the heteropolymer co-administered with cholera toxin B subunit, respectively. High titre serum IgG responses were elicited to the heteropolymer following all routes of delivery when administered with adjuvant. Moreover, as with parenteral delivery, serum IgG antibodies were detected to the individual heteropolymer peptides following i.n. but not oral delivery. These data support the potential of the i.n. route in the mucosal delivery of a GAS vaccine. (C) 2002 Elsevier Science Ltd. All rights reserved.

Single-chain antibodies produced by phage display against the C-terminal 19 kDa region of merozoite surface protein-1of Plasinodium yoelii reduce parasite growth following challenge

Antibodies have the potential to be therapeutic reagents for malaria. Here we describe the production of a novel phage antibody display library against the C-terminal 19 kDa region of the Plasmodium yoelii YM merozoite surface protein-1 (MSP1(19)). In vivo studies against homologous lethal malaria challenge show an anti-parasite effect in a dose dependent manner, and analysis by plasmon resonance indicates binding to the antigen is comparable to the binding of a protective monoclonal antibody. The data support the lack of a need for any antibody Fc-related function and hold great significance for the development of a therapeutic reagent for malaria. (C) 2002 Elsevier Science Ltd. All rights reserved.

Enhancing the immunogenicity and modulating the fine epitope recognition of antisera to a helical group A streptococcal peptide vaccine candidate from the M protein using lipid-core peptide technology

A conserved helical peptide vaccine candidate from the M protein of group A streptococci, p145, has been described. Minimal epitopes within p145 have been defined and an epitope recognized by protective antibodies, but not by autoreactive T cells, has been identified. When administered to mice, p145 has low immunogenicity. Many boosts of peptide are required to achieve a high antibody titre (> 12 800). To attempt to overcome this low immunogenicity, lipid-core peptide technology was employed. Lipid-core peptides (LCP) consist of an oligomeric polylysine core, with multiple copies of the peptide of choice, conjugated to a series of lipoamino acids, which acts as an anchor for the antigen. Seven different LCP constructs based on the p145 peptide sequence were synthesized (LCP1-->LCP7) and the immunogenicity of the compounds examined. The most immunogenic constructs contained the longest alkyl side-chains. The number of lipoamino acids in the constructs affected the immunogenicity and spacing between the alkyl side-chains increased immunogenicity. An increase in immunogenicity (enzyme-linked immunosorbent assay (ELISA) titres) of up to 100-fold was demonstrated using this technology and some constructs without adjuvant were more immunogenic than p145 administered with complete Freund's adjuvant (CFA). The fine specificity of the induced antibody response differed for the different constructs but one construct, LCP4, induced antibodies of identical fine specificity to those found in endemic human serum. Opsonic activity of LCP4 antisera was more than double that of p145 antisera. These data show the potential for LCP technology to both enhance immunogenicity of complex peptides and to focus the immune response towards or away from critical epitopes.