Accuracy of routine data on MMR vaccination coverage and validity of parental recall of vaccination.

A study was conducted to re-validate the measles, mumps and rubella (MMR) coverage and measles susceptibility in children resident in Bolton and born between 1991 and 1995; also to examine the accuracy of parental recall of children's MMR vaccination status. Using general practitioner/practice held records, a validation survey was conducted on a random sample of 171 reportedly unvaccinated children. A questionnaire survey was used to assess parental recall. 'Records' to 'recall' agreement was examined. One- and two-dose population coverage according to routine data was 94.6% and 75.2% respectively, with measles susceptibility of 8.1%. Based on the 'records' survey, one- and two-dose population coverage estimates were 97.4% (+2.8%) and 78.9% (+3.7%), with a revised susceptibility estimate of 5.2%, respresenting a 36% reduction in the susceptibility fraction. Routine data may modestly underestimate vaccination coverage and significantly overestimate measles susceptibility. Many parents of MMR unvaccinated children believe that their children have been vaccinated.

Susceptibility of young offenders to measles and rubella: an antibody prevalence study using oral fluid samples.

Three hundred and two oral fluid samples collected in January 2001 from male young offenders aged 18 to 21 years at HMP Hindley were screened for measles and rubella specific IgG, using 'in-house' amplified ELISA assays. MMR (measles/mumps/rubella) vaccine, offered at the same time, was accepted by 68.1% of the prisoners (92.7% of those agreeing to oral fluid antibody testing). Antibody prevalences were 92.1% for rubella and 80.8% for measles. This may, however, underestimate true prevalence as the sensitivity of the oral fluid assays used may be relatively poor in adults. Susceptibility levels are theoretically high enough to result in outbreaks should either infection enter this closed institutional environment. Since a measles outbreak would carry significant morbidity and rubella would represent a risk to associated non-immune women of childbearing age, it is suggested that prison health services should offer MMR to young offenders on entry.

Prevention of pneumococcal disease in young children.

Increasing pneumococcal resistance to antimicrobial drugs, and the rapid spread of resistant strains throughout the world, underlines the importance of vaccination. Although traditional polysaccharide pneumococcal vaccines have been in use since the 1980s, and are effective among certain age groups, they do not provide protection to children under the age of two years who are at highest risk of infection from pneumococcal disease. The new conjugate vaccine is expected to contribute to a decreased need for antibiotic treatment, fewer GP visits and reduced hospitalisation rates.

Vaccination in older people.

Vaccinating older people remains a priority in the specialty of public health. However, medical practitioners, pharmacists, carers and family members and the media need to make a special effort to inform and educate older people of the need to protect themselves against certain diseases, such as influenza, pneumococcal pneumonia and tetanus. These conditions pose particular health risks to older people in terms of the high risk of developing serious complications.

Acceptability of health status questionnaire screening and vaccination among Kosovar refugees.

During 1999, following the war in Kosovo, thousands of Kosovar refugees were received by different countries. The public health reception protocols for these refugees varied widely between and within reception countries. We surveyed a group of Kosovar refugees received in Bolton to assess the acceptability of vaccination and health status questionnaire screening. We also examined the refugees' opinions about the confidentiality of their medical history and their willingness to take part in research. Fifteen out of 29 people responded. Most respondents were happy with the interventions.

Effect of a boil water notice on behaviour in the management of a water contamination incident.

In late 1998, the water supply of 878 households was affected by possible sewage contamination. A notice was issued to advise residents of the need to boil their water. This provided an opportunity to assess to what extent the boil water notice led people to avoid activities that might put them at risk of waterborne infection. A postal questionnaire sent to 350 randomly selected households in the affected area asked about timing and mode of receipt of the notice, risk behaviour (boiling water, brushing teeth, washing dishes, drinks for pets, preparation of food), and subsequent changes in drinking water consumption habits. Eighty-one per cent of the households surveyed engaged in behaviour likely to increase the risk of waterborne infection. Comments were collected from consumers on how to improve the management of future water contamination incidents.

Cross-linked growth hormone dimers have enhanced biological activity.

In this study we have investigated the effect on the bioactivity of pituitary-derived human growth hormone (hGH) and recombinant bovine (b) GH after the addition of various concentrations of the water soluble cross-linking agent 1-ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC; 6.25-100 mg/ml). The biological activity of resulting cross-linked reactions were determined by its ability to promote incorporation of 35SO4(2-) into costal cartilage of hypopituitary Snell dwarf mice in vivo. Administration of EDC-treated hGH solutions resulted in a significant enhancement of hormone activity in vivo compared with non-cross-linked samples. A similar significant enhancement of bGH activity in vivo was also observed when solutions containing recombinant bGH were cross-linked using EDC. For both hGH and bGH the degree of enhancement appears to be dose-dependent for the concentration of EDC (6.25-100 mg/ml for hGH; 6.25-50 mg/ml for bGH) present in the cross-linking reactions. SDS-PAGE analysis of EDC cross-linked solutions containing hGH and bGH spiked with 125I-hGH and 125I-bGH respectively revealed that dimeric GH was the primary cross-linked component. Increasing the concentration of EDC in cross-linking reactions resulted in increased formation of dimeric hGH and bGH. There was a significant correlation between the amount of GH dimer present and the increase in biological activity, suggesting that GH dimers were responsible for the enhanced biological activity. This was confirmed by the enhanced biological activity of a purified preparation of EDC cross-linked dimeric hGH. In conclusion, covalently cross-linked GH dimers reported here have enhanced bioactivity in vivo. However, since naturally occurring GH dimers are known to have reduced biological activity, this work suggests that the structure of EDC cross-linked GH dimers differs fundamentally from that of native dimeric hGH.

Adjuvancy and reactogenicity of N-acetylglucosaminyl-N-acetylmuramyl-dipeptide (GMDP) orally administered just prior to trivalent influenza subunit vaccine. A double-blind placebo-controlled study in nursing home residents.

One hundred and fifty-three nursing home residents received 0, 5, 25 or 50 mg N-acetylglucosaminyl-N-acetylmuramyl-dipeptide (GMDP) orally, and trivalent influenza subunit vaccine intramuscularly. One day after intervention, there was a strong increase of total leucocytes, monocytes and neutrophils in the groups receiving 25 or 50 mg GMDP. A GMDP dose dependent increase in systemic, but not in local, vaccine side-effects was observed. No significant differences in post-vaccination haemagglutination inhibiting serum antibody titres were observed between the four groups, indicating that oral administration of GMDP together with influenza vaccination, does not lead to a higher vaccine efficacy.

Assessing the completeness of tuberculosis notification in a health district.

We have estimated the completeness of tuberculosis notification in Bolton health district by combining notification data with information from prescriptions issued by general practitioners and hospital admission data. From 1 January 1991 to 31 December 1993, 307 cases of tuberculosis were notified in Bolton. Thirty-four cases were identified using general practitioners' prescriptions: 27 of these had been notified between January 1991 and December 1993, and seven had not been notified during this period. Although prescriptions may be used as a marker for tuberculosis, they would not be an adequate alternative to notification of cases. One hundred and forty-eight cases of tuberculosis were identified between 1 October 1992 and 30 September 1993 by combining data from the three sources: 138 cases had been notified (93.2%), 28 identified from prescriptions (23.0%), and 54 identified from hospital data (36.5%).

Tetracyanoquinodimethane mediated glucose sensor based on a self-assembling alkanethiol/phospholipid bilayer.

An amperometric tetracyanoquinodimethane (TCNQ) mediated biosensor for glucose is described, based on a self-assembling alkanethiol/phospholipid bilayer laid down onto a gold surface. Gold was sputter deposited onto chromium coated silicon wafers to a thickness of 200 nm. A monolayer of alkanethiol was allowed to self-assemble from an ethanolic solution of dodecanethiol onto a freshly cleaned gold electrode in an overnight incubation. The monolayer was characterized by ellipsometric, impedance and cyclic voltammetry measurements (capacitance = 1.60 +/- 0.06 microF/cm2, and thickness 1.34 +/- 0.15 nm). A mixture of phospholipid liposomes containing free amino groups was placed on the monolayer and allowed to incubate overnight. The self-assembly of a phospholipid monolayer and allowed to incubate overnight. The onto the alkanethiol monolayer, resulted in the formation of a bilayer. The formation of bilayer was again characterized by impedance and cyclic voltammetry measurements (capacitance = 0.98 +/- 0.09 microF/cm2, and thickness = 1.85 +/- 0.22 nm). TCNQ has been incorporated into the liposomes before the formation of the bilayer. Glucose oxidase was cross-linked with the amino-groups of the phospholipids using bis [2-(sulphosuccinimiidooxicarbonyloxy) ethyl] sulphone. TCNQ which was incorporated in the bilayer acted as an efficient mediator to regenerate glucose oxidase. Cyclic voltammetry of the modified electrode and a response curve for the glucose sensor are reported.

The modulation of tumour necrosis factor-alpha, interleukin-1 alpha and glucose levels with GMDP and other analogues of muramyl dipeptide.

Immunomodulatory agent muramyl dipeptide (MDP) and seven of its analogues were tested for ability to counteract the toxic actions of lipopolysaccharide (LPS) in experimental mouse models. Female BALB/c mice were presensitized with Corynebacterium parvum (P. acnes) and given MDP or equimolar doses of one of its analogues after 2 weeks, followed by intravenous challenge with LPS 18 h later. This treatment produced a sharp increase in blood cytokine (TNF-alpha, IL-1 alpha) levels 4 h after LPS administration followed by a decline to control values after 6 h. Four analogues, GMDP, threonylMDP, GMDPBenz and GMDPOBut, were able to reduce the level of cytokines induced with LPS. For most of the analogues, the higher doses reduced the levels of TNF-alpha but slightly increased the concomitant IL-1 alpha levels. GMDP was the most effective compound tested in terms of reduction of TNF-alpha and IL-1 alpha levels, as well as for reduction of the hypoglycaemia caused by the administration of LPS.

Differential effects of small tumour necrosis factor-alpha peptides on tumour cell cytotoxicity, neutrophil activation and endothelial cell procoagulant activity.

Tumour necrosis factor-alpha (TNF-alpha) is a pluripotent cytokine with its receptors distributed throughout many different cell types. Because of the diverse effects of the cytokine, it is difficult to clearly define its role in infection and immunity, and appreciate its clinical therapeutic value. We have identified peptides derived from the primary amino acid sequence of human TNF-alpha that have neutrophil-stimulating activity, as measured by enhanced chemiluminescence and superoxide production, and peptides which are both directly cytotoxic for tumour cells (WEHI-164) in vitro and also prevent TNF binding to tumour cells. However, only one of these neutrophil-stimulating peptides was toxic for tumour cells in vitro. Our results indicate that the region of amino acids 54-94 of human TNF-alpha has previously undescribed human neutrophil-stimulatory activity, while peptides encompassing the regions 43-68 and 132-150, which are in close proximity, as indicated in the recently determined three-dimensional structure of human TNF-alpha, have in vitro anti-tumour activity. These peptides also slowed tumour growth or induced tumour regression in WEHI-164 tumour-bearing mice. The peptide 73-94, which activated neutrophils but which was not cytotoxic for tumour cells in vitro, also caused in vivo tumour regression, presumably by activating neutrophils with the consequent release of free radicals at the tumour site. Peptide 63-83, which was able to activate neutrophils in vitro, did not possess tumour regression activity in vivo. The TNF peptides described in this report did not elicit procoagulant activity in cultured bovine aortic endothelial cells and as such are devoid of at least one of the potentially lethal side-effects of elevated TNF levels in vivo.

The enhancement of human tumor necrosis factor-alpha antiviral activity in vivo by monoclonal and specific polyclonal antibodies.

This report describes the potentiation of the antiviral effects of human tumour necrosis factor-alpha (TNF-alpha) in vivo by specific antibodies. Complexes of TNF with either an anti-TNF monoclonal or polyclonal antibody at optimal doses induced a significantly greater antiviral (vaccinia virus) state in CBA/H mice than TNF alone. Furthermore, an antiviral synergy between murine gamma-interferon and TNF was found in vivo when the latter was in complex with enhancing antibody. Two other inbred mouse strains, C57/B6 and BALB/c, were also examined under these conditions. These antibodies greatly increase the binding of TNF to the surface of cells, which may account for the observed enhancement of TNF activity. Such antibodies may be of value clinically in viral diseases and cancer therapy where an increase in TNF activity, in the absence of side effects, would lead to more effective use of this cytokine in human therapy.

JAK2, a third member of the JAK family of protein tyrosine kinases.

We have isolated cDNA clones encoding a third, widely expressed, member of the JAK family of protein tyrosine kinases (PTKs). The anticipated amino acid sequence of JAK2 predicts the presence of two kinase-related domains, a feature characteristic of this family of PTKs. The structural similarity of JAK2 to the other members of this family extends towards their N-termini, beyond the two kinase-related domains, and reveals five further domains of substantial amino acid similarity. The C-terminal portion of one of these domains, the JH4 domain, bears an intriguing, albeit tenuous, similarity to the core element of the SH2 domain, whereas the remaining JAK homology domains do not appear to be a feature of other known proteins.

Selective enhancement of the tumour necrotic activity of TNF alpha with monoclonal antibody.

The binding and biological activity of human TNF alpha on endothelial and tumour cells has been studied in the presence of monoclonal antibodies (MAbs). In particular, one monoclonal antibody to TNF alpha (MAb 32) has been identified which failed to inhibit binding and cytotoxicity of TNF alpha on WEHI-164 tumour cells but which was a potent inhibitor of TNF alpha-induced endothelial cell procoagulant activity on bovine aortic endothelial cells. The ability of MAb 32 to inhibit selectively the actions of TNF alpha on endothelial cells but not on tumour cells suggests a mechanism for enhancement of the anti-tumour action of TNF alpha in vivo when in complex with this antibody. Treatment of tumour bearing mice (WEHI-164 and Meth A fibrosarcoma) with TNF alpha-MAb 32 complex resulted in a 5- to 10-fold enhancement in the potency of the cytokine in comparison to free TNF alpha. Complexes between this cytokine and other MAbs generally resulted in either no effect or inhibition of TNF alpha activity in vivo and in vitro. Neither intact MAb 32 nor FAb' fragments of MAb 32 showed any tumour regressive activity in the absence of TNF alpha. The FAb' fragments were equipotent to the bivalent form of the antibody in enhancing TNF alpha activity. These data provide evidence that it is possible to segregate the individual biological activities of TNF alpha with concomitant enhancement of the tumour regressive activity of the cytokine in vivo.