Control of hypercalcaemia of parathyroid carcinoma by immunisation

BACKGROUND: Patients with parathyroid tumours can develop extreme hypercalcaemia and osteitis fibrosa cystica. Clinical features result from the action of parathyroid hormone (PTH) on bone receptors. Because this hormone is produced in microgram quantities, inhibition of its metabolic effects with potent PTH antibodies should be possible. We tested whether an immunisation with synthetic human and bovine PTH peptides could stimulate autoantibodies against PTH. METHODS: A patient with metastatic parathyroid carcinoma in the lungs and pleura developed severe bone disease and extreme hypercalcaemia that proved resistant to conventional therapy. She was immunised with 200 microg human and bovine PTH peptides and 50 microg human PTH. Booster doses were also given at 4 weeks and 11 weeks. The patient was then seen every week. FINDINGS: Antibodies against PTH were produced within 4 weeks of initial immunisation and titres increased with repeated doses of immunogens. Total serum calcium concentrations, which had ranged from 3.5 mmol/L to 4.2 mmol/L over the previous 18 months, fell to between 2.5 mmol/L and 3.0 mmol/L over 6 months of therapy. This fall was accompanied by striking clinical improvement. INTERPRETATION: We believe this is the first use of immunotherapy to control remote, non-metastatic complications of malignant disease. B-cell tolerance to human PTH was broken by immunisation with PTH peptides in adjuvant. This therapeutic approach could be used to control excess hormone production in several types of endocrine tumour and may have applications in other diseases.     

A method and device for measuring force transfers between the deep flexors in the musician''s hand

The efficacy of passive immunisation against tick-transmitted Lyme disease spirochaetal infection was determined in relation to the duration of previous feeding of infected vector ticks. Thus, mice challenged with spirochaete-infected unfed or partially fed nymphal ticks were passively immunised with monoclonal and polyclonal antibodies against the Lyme disease spirochaete (Borrelia burgdorferi) at various intervals after tick attachment. Spirochaetal infection in challenged mice and engorged ticks was verified by xenodiagnosis and indirect immunofluorescent antibody assay, respectively. Although tick-transmitted spirochaetal infection could be aborted by anti-OspA antibodies and hyperimmune antiserum, nearly all immunised mice challenged with infected ticks that had previous 36-h attachment became infected. More than 72% of the nymphal ticks used in this challenge retained their B. burgdorferi infection after engorgement on mice immunised with anti-spirochaete antibodies, and their subsequent infectivity to mice remained effective. It is concluded that a higher efficiency of transmission by partially fed infected nymphs and a lower efficacy of passive immunisation against infection result from an effect of previous feeding of infected ticks that activates antigenic change and enables the spirochaetes to circumvent OspA-based humoral immunity.     

A peptide derived from a B cell epitope of Plasmodium falciparum rhoptry associated protein 2 specifically raises antibodies to rhoptry associated protein 1

Mice immunised with a recombinant form of malarial antigen rhoptry-associated protein 2 (RAP2) produce antisera which recognise the native protein by indirect immunofluorescence and immunoblotting. Purified IgG components of the antisera partially inhibit erythrocyte invasion in vitro. This response was obtained only if the recombinant immunogen was presented to the mice in the presence of reducing and denaturing agents. An 8-mer epitope in RAP2 was recognised by antibodies in three of the antisera: E25TEFSKLY32. Immunisation with this octapeptide raised antibodies which strongly recognised reduced RAP2 in seven out of eight mice. However, this antisera either failed to recognise (five out of eight mice), or only weakly (three out of eight mice) recognised nonreduced RAP2. Examination of disulphide bonds in native RAP2 showed that the 4 cysteines of RAP2 form two disulphide bridges: Cys24-Cys88, and Cys277-Cys376. One of these (Cys24-Cys88) is adjacent to the octapeptide in the native protein. Surprisingly, seven out of eight mice immunised with the octapeptide also raised antibodies against native rhoptry-associated protein 1 (RAP1). The raising of antibodies which recognise RAP1 was induced specifically by the RAP2 octapeptide rather than the carrier protein used for immunisation. The epitope in RAP1 recognised by the antibodies was identified and shown not to be the result of any shared contiguous homologous sequence between the two proteins, but to shared homologous amino acids in critical positions within the epitope. Purified IgG components from the antisera of mice immunised with the octapeptide gave partial inhibition of erythrocyte invasion in vitro.     

Passive protection of neonatal calves against bovine coronavirus-induced diarrhea by administration of egg yolk or colostrum antibody powder

The protective effect of egg yolk and colostrum powders prepared from hens and cows vaccinated with inactivated bovine coronavirus (BCV) antigen was evaluated in a challenge model with a virulent BCV strain. Twenty three calves from BCV-free herds were randomly divided into control and several treatment groups. All calves were orally challenged with 1 x 10(9) TCID50 of the virulent Kakegawa strain of BCV at 24 to 36 h after birth. Calves in treatment groups received either egg yolk powder or cow colostrum containing BCV specific antibodies. Daily treatment with these antibody preparations started 6 h until 7 days post-challenge. Control calves which received no antibody had severe diarrhea and all died within 6 days after infection. In contrast, calves fed milk containing egg yolk or colostrum with neutralization titers of 1:2560 or 1:10,240 respectively all survived and had positive weight gain unlike the other treatment groups. These results indicate that the orally administered egg yolk and colostrum powders protected against BCV-induced diarrhea in neonatal calves and that the egg yolk used provided a higher degree of protection compared to colostrum powder on a titer basis. Treatment with whole egg yolk from immunized hens therefore provides a more efficacious alternative to the existing methods of specific passive protection against BCV.     

An assessment of mucosal immunisation in protection against Streptococcus equi ('Strangles') infections in horses

The ability of mucosally administered antigen to provide protection against Streptococcus equi ('Strangles') infections in horses was examined. First, an enzyme linked immunosorbent assay (ELISA) was developed to detect the immune status of horses to S. equi. This assay was used to select Strangles-naive horses for the study and also to monitor their response to immunisation. Potential vaccine candidates were: (a) orally administered paraformaldehyde killed S. equi; (b) intraperitoneally (IP) administered paraformaldehyde killed S. equi in a non-inflammatory adjuvant; (c) orally administered live avirulent S. equi; (d) orally administered microencapsulated streptococcal M protein. The latter three preparations were first assessed in a rat model, using rate of lung bacterial clearance following intratracheal inoculation of live virulent bacteria as an indication of efficacy. Candidates (a) and (b) were then assessed in an equine model. IP immunisation of horses was shown to effectively induce production of specific antibody in mucosal and systemic sites. Four weeks after initial immunisation, horses were challenged intranasally with live virulent S. equi. Both groups of immunised horses demonstrated partial protection following vaccination. Of the IP immunised horses, only two out of four developed clinical signs of Strangles following live challenge. The orally immunised horses all developed submandibular abscesses containing S. equi. However, none of the immunised horses became as ill as the control horses in terms of fever, anorexia, loss of condition and general malaise.     

Immunogenicity of purified lipopolysaccharide or protein-oligosaccharide conjugates of Pasteurella multocida type 6:B in mice

Purified lipopolysaccharide (LPS) of Pasteurella multocida type 6:B, while toxic at higher doses, was protective at lower dose levels against experimentally-induced pasteurellosis in mice. However, the observed protection was abrogated if such LPS was digested with proteinase K prior to use in immunisation. The O-antigen polysaccharide side-chain (OS) of LPS did not appear to contribute to the observed protection as judged by the fact that immunisation of mice with purified OS or OS-protein conjugates, all of which were nontoxic, failed to confer protection against challenge with homologous virulent organisms. This was despite generation of significant levels of OS-specific antibodies, predominantly either of the IgM or IgG isotypes, in immunised mice.     

Development of snake antivenom antibodies in chickens and their purification from yolk

Adult white leghorn hens hyperimmunised with Brazilian snake venoms of the genus Bothrops and/or Crotalus produced antibodies capable of recognising, combining with and neutralising the toxic and lethal components of the venoms. The antibodies were first detected by an enzyme-linked immunosorbent assay two weeks after starting the immunisation schedule, reached the highest titres by the third week and remained high for at least 24 weeks. These antibodies are transferred to the egg yolk from which they were isolated as enriched IgY preparations by a combination of methods using positive and negative precipitation with sodium sulphate and/or caprylic acid. The yolk-derived IgY preparations contained antibodies which blocked the phospholipase A2-dependent haemolytic activity of both venoms and the haemorrhagic activity of Bothrops venom, and neutralised the toxic lethal activities of the venoms with good efficacy. The median effective dose (ED50) of the IgY anti-Bothrops venom was 592.5 microliters/2LD50 and, 1.0 ml neutralised 0.0675 mg of venom. The ED50 of the IgY anti-Crotalus venom was 457.5 microliters/3LD50 and 1.0 ml neutralised 0.075 mg of venom.     

Amino acid supplementation of turkey breeder rations

Beltsville Small White turkey hens (60) were randomized into four groups of 15 hens each, weighed and placed in individual laying cages. Immediately after lighting to initiate production, they were placed on a practical type turkey breeder diet calculated to contain 18.26% protein, 2893 Cal. M.E./kg., 3.13% Ca, 0.80% available P, 1.04% arginine, 0.86% lysine, 0.50% methionine & cystine and 0.24% tryptophane. Amino acids were added to this diet as follows: (1) none; (2) 0.1% lysine; (3) 0.05% methionine and (4) 0.1% lysine & 0.05% methionine. Lysine (0.1%) had no effect on reproductive performance when added to the basal ration or to the ration containing additional methionine. Methionine addition (0.05%) improved production significantly (P less than or equal to .005), improved feed efficiency, and increased egg size.     

Effect of hen production age on egg composition and embryo development in commercial Pekin ducks

At 26, 31, 36, and 42 wk of age, eggs were collected from the same duck breeder flock to study the effects of hen production age on egg composition and embryo development. At each hen age, yolk and albumen measurements were made on a random subsample of unincubated eggs. Embryo and yolk sac measurements were made at 20, 22, 24, 26, 27, and 28 d of incubation. Egg weight was significantly affected by hen production age, but most of the observed age effects occurred between 26 and 31 wk with minimal age differences thereafter due to a quantitative feed restriction for egg weight. Yolk weight increased significantly with hen production age, with the largest increase, 6.6 g, also occurring between 26 and 31 wk of age. Yolk-free duckling weight increased with hen production age at 27 d of incubation. The yolk sacs of embryos from the 31-, 36-, and 42-wk-old hens were heavier at 20 and 22 d, before the differentiation in embryo weight. Differences in yolk sac weight were not consistently affected by hen production age between 26 and 28 d of incubation. The eggs from the 42-wk-old hens weighed 3.3 g less than those from the 31-wk-old hens, but yolk-free duckling weight at 27 d was 2.9 g heavier from the 42-wk-old hens.     

Poly(lactide-co-glycolide) microencapsulation of vaccine antigens

Fimbriae from Bordetella pertussis have been encapsulated in poly(lactide-co-glycolide) (PLG) microspheres of a size appropriate for oral administration. The binding of antibodies which react with conformational or linear fimbrial epitopes, to fimbriae released from microspheres, suggested that the process of was not detrimental to the native integrity of the protein. Mice were immunised by oral gavage with a single dose of microencapsulated fimbriae, or with fimbriae adsorbed onto alhydrogel and administered by intraperitoneal injection. The resulting immune responses in serum were comparable but only oral administration of microencapsulated fimbriae elicited specific immune responses in external secretions. Six weeks after immunisation, both groups of immunised animals were protected against challenge with live B. pertussis.     

Effect of vaccination of hens with an avirulent strain of Salmonella typhimurium on immunity of progeny challenged with wild-Type Salmonella strains

The avirulent Salmonella typhimurium chi3985 was used to vaccinate white leghorn chickens at 16 and 18 weeks of age, and the effect of maternal antibody on Salmonella colonization of progeny of vaccinated hens was assessed with S. typhimurium F98 or chi3985. Progeny of hens that had been vaccinated at 1 and 3 or 2 and 4 weeks of age with chi3985 were used to determine the effect of maternal immunity on vaccine efficacy. Vaccination of hens induced long-lasting Salmonella-specific antibodies which were transferred into eggs and were detected as immunoglobulin G (IgG) in the egg yolk. Maternal antibody was detected in the progeny of vaccinated birds as IgG and IgA in serum and intestinal fluid, respectively. The titer of maternally transmitted IgG or IgA was highest in the first week of life of the progeny and declined with age. Maternal antibodies prevented colonization of the chicks by S. typhimurium chi3985 and reduced colonization by S. typhimurium F98. Overall, chicks from vaccinated hens had significantly higher antibody responses than did the progeny of nonvaccinated hens after oral infection with Salmonella strains. Maternal antibody reduced the efficacy of vaccination of progeny with chi3985 at 1 and 3 weeks of age. But vaccination at 2 and 4 weeks of age induced excellent protection against challenge with S. typhimurium F98 or S. enteritidis 27A PT 8 in birds from vaccinated hens and in specific-pathogen-free chickens. Vaccination of chickens at 2 and 4 weeks of age has been shown to protect the birds against challenge with homologous and heterologous Salmonella serotypes. A combination of vaccination of adult animals and use of the progeny of vaccinated birds will enhance effective control of Salmonella infections in the poultry industry. This will complement the present control of Salmonella-associated food poisoning caused by Salmonella enteritidis in eggs because the avirulent S. typhimurium vaccine strain chi3985 induced excellent protection against S. enteritidis in chickens.     

Identification of mammosomatotrophs in the turkey hen pituitary: increased abundance during hyperprolactinemia

We have previously reported that the hyperprolactinemia in incubating turkey hens is associated with recruitment of lactotrophs in the pituitary gland. In this study we have used double immunofluorescence and in situ hybridization histochemistry to 1) identify mammosomatotrophs in the anterior pituitary gland of egg-laying turkey hens and incubating hens, and 2) verify PRL gene expression within mammosomatotrophs by colocalizing PRL messenger RNA in GH-immunoreactive (ir) cells. The pituitaries of laying and incubating turkey hens were collected, and the midsagittal sections were dual labeled for either PRL and GH or PRL messenger RNA and GH. The plasma PRL concentrations were higher in incubating hens (231 +/- 10.6 ng/ml) than in laying hens (43 +/- 7.4 ng/ml; P < 0.01). In the midsagittal pituitary sections, mammosomatotrophs were predominantly found scattered in the caudal lobe of the anterior pituitary gland, in the ventral half of the cephalic lobe, and at the junction of cephalic and caudal lobes. In incubating hens, the proportion of mammosomatotrophs was 7.4 +/- 1.52% (mean +/- SEM) of the total number of GH-ir and/or PRL-ir cells counted, which was significantly higher (P < 0.05) than that found in laying hens (0.6 +/- 0.23%). Furthermore, PRL gene expression was observed in many GH-ir cells in the incubating hen pituitary gland. These data suggest that 1) mammosomatotrophs are present in the turkey pituitary gland, and 2) there is an increased abundance of mammosomatotrophs in the incubating turkey hen that may contribute to hyperprolactinemia.     

Effects of in ovo exposure to 2,3,7,8-TCDD on F1 generation adult chickens (Gallus gallus)

White Leghorn chickens (Gallus gallus) were used as surrogate species for the resident wild turkeys found on the Times Beach, Missouri, Superfund site. Parental chickens were injected with concentrations of 2,3,7,8-TCDD which modeled soil concentrations before (200 ppb) and after remediation (1ppb)[1]. Offspring were followed through development to assess alterations in reproductive maturity through the use of a four-way breeding study. F1 adult females exposed to a maternal dose of 8.6 ng/day began egg production approximately two weeks later than did F1 control adult females. By week eight, however, egg production between groups was equivalent. No differences were observed in eggshell gland estrogen or progesterone receptor levels.     

Future vaccines and a global perspective

Advances in medical biotechnology mean that vaccines to prevent more than 75 infectious diseases are being or have been developed. Vaccination is unfortunately not reliant purely on biotechnology but also on politics and resources. Countries with the greatest demand for vaccines have the least ability to pay for or produce them. Health-care Infrastructure and diagnostic facilities also hamper immunisation projects in developing countries. Charitable organisations are relied on heavily to support such projects but the challenge to ensure all infants are immunised against the most common infections of childhood is still enormous. Difficulties that present themselves now should not prevent us looking into future possibilities such as immunisation during pregnancy and targeting of children for immunisation against sexually transmitted diseases. Other avenues for research are in administration of vaccines. A move to mucosal immunisation rather than use of the syringe and needle would be positive both economically and from the point of view of risk of needle contamination. Plant science may also provide a new vehicle for vaccines by engineering plants such as the banana tree to be naturally bioencapsulated vaccines. Prospects for control and eradication of infectious disease in the next century are certainly good.     

Comparison of serological tests for detection of Mycoplasma gallisepticum antibodies in eggs and chicks hatched from experimentally infected hens

Specific pathogen free hens and males were experimentally infected with Mycoplasma gallisepticum. Eggs were then collected, and a part was incubated and set for hatching. Mycoplasma cultures were performed on infected adults and antibodies to MG were analysed by use of slide agglutination (SA) test and commercial ELISA tests on adults and chicks sera and on yolks from non incubated eggs. Both ELISA tests could detect antibodies in yolks from non incubated eggs laid three weeks after infection. SA and the three ELISA tests revealed positive sera in chicks hatched from eggs laid as soon as one week after infection.     

Destruction of allogeneic tumour cells by peritoneal macrophages

The population of peritoneal macrophages from mice immunised with allogeneic tumor cells contains two types of cytotoxic cell. One lyses the specific target cell, the other initiates activation of macrophages and hence leads to inhibition of growth of the specific target cell. The yield of lytic effector macrophages was found to depend on the route and the nature of the cell used for immunisation and the condition of the mice. The yield correlated with the yield of complement-dependent cytotoxic antibodies. In contrast, production of specific "activator" macrophages did not depend critically on these factors. The results underline the difference between the two types of cell and suggest that they are produced independently of one another.     

Duration of fertility in ad libitum and feed-restricted caged broiler breeders

It has been shown in previous studies that fertility can be reduced in overweight broiler breeder (BB) flocks. In an effort to determine the effect of ad libitum feeding on the duration of fertility in BB hens, 60 52-wk-old Shaver Starbro hens were randomly assigned to one of two treatments, ad libitum feeding (F) or restricted feeding (R) to maintain breeder target weights. All hens were reared to 52 wk under conditions of feed restriction. All birds were weighed individually on a weekly basis. At the beginning of each of two 4-wk study periods (56 to 60 wk and 60 wk to 64 wk), all birds were inseminated on 2 consecutive d with 0.05 mL pooled BB semen. All eggs were weighed and placed in a forced air incubator the same day that they were laid. After 7 to 10 d of incubation, the eggs were broken out and scored macroscopically as fertile with live embryo, fertile with dead embryo (early embryonic death), or clear (assumed infertile). The duration of fertility was defined as the number of days from the day after the second insemination to the last fertile egg before two consecutive interfile eggs. Hen BW were significantly different between treatments within each of the two 4-wk studies. The mean BW of the F hens was 4,261 g in Study 1 and 4,448 g in Study 2. The BW of the R hens were 3,459 g in Study 1 and 3,565 g in Study 2. Egg production levels and average egg weight was not different between treatments in either study. In Study 1, the duration of fertility for the F hens (12.7 d) and the R hens (12.7 d) were not different. In Study 2, the durations of fertility were significantly higher (P < 0.05) in the R hens (12.7 d) than in the F hens (10.0 d). These results support the theory that overweight BB have a reduced duration of fertility that may contribute to a reduced fertility in artificially inseminated and naturally mated flocks.     

The effect of accidental pre-lighting of turkey hens on subsequent reproduction

The length of the light day was accidentally increased from nine hours to continuous light on one group of thirty-week old hens for an unknown period of not more than eleven days. Egg production, after attaining a level of twenty-five percent, ceased five weeks later as the result of restricting the length of the light day to four hours and two series of feed and water deprivations. One month later hens from the pre-lighted and control groups were brought into egg production. Pre-lighting depressed later egg production more in medium-bodied hens than in large-bodied hens. The major effect of pre-lighting was to reduce the length of a laying period included 9 weeks later with no significant effects on intensity of lay, number of broody periods or average length of the broody period. It was concluded that the egg production would not be greatly reduced by pre-lighting under the above conditions if the length of the laying period were shortened from 180 days to a normal 140-day period.     

Developmental profile of glucocorticoid binding to synaptic plasma membrane from rat brain

Plasma prolactin levels were measured in free-living breeding adult and immature (pre-breeding) macaroni (Eudyptes chrysolophus) and gentoo (Pygoscelis papua) penguins at Bird Island, South Georgia (54 degrees S, 38 degrees W). Macaroni and gentoo penguins first breed at 5-6 and 2 years of age, respectively. In adult birds, of both species, prolactin was low (< 1.0 microgram. liter-1) during the courtship period and then increased during early (gentoo) to mid (macaroni) incubation (to 3.9-4.7 and 2.4-3.5 micrograms.liter-1, respectively), remaining elevated until the creche period, by which time continuous nest attendance by the adults had ceased. This pattern is similar to that seen in other altricial species and is consistent with delayed onset of brood patch development and full incubation efficiency, which has been previously reported in penguins. Adult female macaroni penguins showed a marked, but transient, increase in prolactin concentrations within 24 hr of the first egg being laid (from 1.7 to 7.0 micrograms.liter-1), plasma levels decreasing following clutch completion (to prelaying levels) before increasing again during incubation. Elevated plasma prolactin levels occurred in all age classes of immature (nonbreeding) birds in both macaroni (1- to 5 year-olds) and gentoo (1-year-olds) penguins. However, compared to that in adult birds, the increase in prolactin was more transient in immatures, a smaller proportion of immatures had detectable prolactin levels at each stage of the breeding cycle, and, at least in 1- and 2-year-olds, absolute levels of prolactin were lower.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of irradiated fruits by gas-chromatographic methods

The purpose of this study was to investigate the effects of prolactin, both in vitro and in vivo, upon heterophil phagocytic activity of the ring dove, Streptopelia risoria. In vitro incubation of heterophils with either 0.1 or 100 micrograms/mL of ovine prolactin for 2 h-significantly increased both latex bead phagocytosis and Nitroblue tetrazolium (NBT) reduction, an index of phagocytic metabolic activity. Ring doves given antigen demonstrated an increase in latex bead phagocytosis and NBT reduction. The greatest increase in both of these parameters was observed in those birds which possessed elevated plasma prolactin concentrations, either through exogenous administration or naturally through being in the later stages of incubation. These results suggest that during the incubation period of the avian breeding cycle there is an increase in phagocytic activity which is a direct consequence of the elevation shown by these birds in the concentration of plasma prolactin.