Immunity and vaccine control of Echinococcus granulosus infection in animal intermediate hosts

Much progress has been made with characterisation of the EG95 vaccine which can be used to prevent hydatid infection in animal intermediate hosts of Echinococcus granulosus. The vaccine comprises a single recombinant oncosphere antigen and the adjuvant Quil A. It induces complement-fixing antibodies that kill the invading oncosphere early in an infection. In the majority of vaccinated animals, no hydatid cysts occur following a challenge infection. However, a small number of viable cysts may occur in some vaccinated animals. The vaccine has proved effective in vaccine trials carried out in sheep in New Zealand, Australia, Argentina, Chile and China as well as in goats and cattle. Investigations of the genetic diversity of the gene encoding EG95 have identified no unequivocal variation within the G1 strain parasites; however DNA sequence diversity within the EG95 family of genes has been found in G6/G7 parasites. GMP production scale-up of the vaccine has been undertaken in New Zealand and China and it is expected that the vaccine will be become available through these sources for implementation as part of hydatid control programs worldwide.     

Induction of tumor cell apoptosis in vivo increases tumor antigen cross-presentation,cross-priming rather than cross-tolerizing host tumor-specific CD8 T cells

Cross-presentation of cell-bound Ags from established, solid tumors to CD8 cells is efficient and likely to have a role in determining host response to tumor. A number of investigators have predicted that when tumor Ags are derived from apoptotic cells either no response, due to Ag "sequestration," or CD8 cross-tolerance would ensue. Because the crucial issue of whether this happens in vivo has never been addressed, we induced apoptosis of established hemagglutinin (HA)-transfected AB1 tumors in BALB/c mice using the apoptosis-inducing reagent gemcitabine. This shrank the tumor by approximately 80%. This induction of apoptosis increased cross-presentation of HA to CD8 cells yet neither gross deletion nor functional tolerance of HA-specific CD8 cells were observed, based on tetramer analysis, proliferation of specific CD8 T cells, and in vivo CTL activity. Interestingly, apoptosis primed the host for a strong antitumor response to a second, virus-generated HA-specific signal in that administration of an HA-expressing virus after gemcitabine administration markedly decreased tumor growth compared with viral administration without gemcitabine. Thus tumor cell apoptosis in vivo neither sequesters tumor Ags nor cross-tolerizes tumor-specific CD8 cells. This observation has fundamental consequences for the development of tumor immunotherapy protocols and for understanding T cell reactivity to tumors and the in vivo immune responses to apoptotic cells.     

Estimating the latitudinal origins of migratory birds using hydrogen and sulfur stable isotopes in feathers: influence of marine prey base

Hydrogen stable isotope analysis of feathers is an important tool for estimating the natal or breeding latitudes of nearctic-neotropical migratory birds. This method is based on the latitudinal variation of hydrogen stable isotope ratios in precipitation in North America (deltaD(p)) and the inheritance of this variation in newly formed feathers (deltaD(f)). We hypothesized that the typically strong relationship between deltaD(p) and deltaD(f) would be decoupled in birds that forage in marine food webs because marine waters have relatively high deltaD values compared to deltaD values for local precipitation. Birds that forage on marine prey bases should also have feathers with high delta(34)S values, since delta(34)S values for marine sulfate are generally higher than delta(34)S values in terrestrial systems. To examine this potential marine effect on feather stable isotope ratios, we measured deltaD and delta(34)S in the feathers of nine different species of raptors from both inland and coastal locations across North America. Feathers from coastal bird-eating raptors had consistently higher deltaD and delta(34)S values than feathers from inland birds. Birds that had high delta(34)S values also deviated strongly from the typical relationship between deltaD(p) and deltaD(f). We recommend measuring both sulfur and hydrogen stable isotope ratios in feathers when some members of a migrant population could potentially forage in marine habitats. We suggest using a practical cutoff of delta(34)S >10 per thousand to remove marine-foraging birds from a migrant sample when using stable isotopes of hydrogen to estimate the latitudinal origins of migrants because high deltaD(f) values for marine-foraging birds could potentially distort estimates of origins.     

Lipid biosynthesis as a target for antibacterial agents

Fatty acid biosynthesis, the first stage in membrane lipid biogenesis, is catalyzed in most bacteria by a series of small, soluble proteins that are each encoded by a discrete gene (Fig. 1; Table 1). This arrangement is termed the type II fatty acid synthase (FAS) system and contrasts sharply with the type I FAS of eukaryotes which is a dimer of a single large, multifunctional polypeptide. Thus, the bacterial pathway offers several unique sites for selective inhibition by chemotherapeutic agents. The site of action of isoniazid, used in the treatment of tuberculosis for 50 years, and the consumer antimicrobial agent triclosan were revealed recently to be the enoyl-ACP reductase of the type II FAS. The fungal metabolites, cerulenin and thiolactomycin, target the condensing enzymes of the bacterial pathway while the dehydratase/isomerase is inhibited by a synthetic acetylenic substrate analogue. Transfer of fatty acids to the membrane has also been inhibited via interference with the first acyltransferase step, while a new class of drugs targets lipid A synthesis. This review will summarize the data generated on these inhibitors to date, and examine where additional efforts will be required to develop new chemotherapeutics to help combat microbial infections.     

Food choices and eating difficulty among elderly edentate patients in Greece

Aims: To evaluate food choice and eating difficulty experienced by older Greek edentate patients. Subjects: Three samples of patients seeking provision of replacement dentures were studied. A primary study of urban mainland U1 (n=54) and island rural R1 samples (n=84) was followed by a mainland urban U2 sample (n=119) in the Secondary study. Setting: Greek dental clinics. Intervention: Semi‐structured interviews (SSI) were employed, using both open and closed questions. Design: The primary study used SSI to identify eating difficulty experienced by two culturally different Greek groups. The secondary study established patterns of difficulty for comparisons between an urban Greek population and northern European urban studies. Main Outcome Measures: The prevalence of eating difficulty, the degree of difficulty eating specified foods and the exclusion of foods because difficult. Results: Most patients expressed difficulty eating at least one type of food, a high percentage of these patients were willing to eat foods found difficult, while others use particular methods of food preparation that make food easier to eat. Chicken illustrated the importance of specifying the method of cooking when questioning eating difficulties. Roast meats provided insight into the more difficult end of the food range. Raw vegetables were rated difficult. Apples and oranges were also food of particular interest. Conclusions: The semi‐structured interview method provides a succesful method to identify eating difficulty and food choices by older Greek complete denture wearers. Differences, probably largely cultural, were identified between Greek island rural and mainland urban communities. Greek food choices differed favourably from an English sample, strikingly in that Greek patients report continuing to eat difficult foods despite difficulty eating them. This may be relevant to health data on Greek populations that show better mortality statistics despite adverse factors such as high prevalence of smoking.     

Validation of indices for functional assessment of dentures

Aims: To validate an assessment of qualities of complete dentures that may relate to functional success. Subjects: A consecutive sample of 131 complete denture wearers took part in the study. Setting: a Greek department of prosthetic dentistry. Intervention : All participants and their dentures underwent clinical examination. Design: The Functional Assessment of Dentures (FAD) 10‐item criteria¹ describing clinical factors of denture quality was used. The rationale for selecting these particular criteria was to include a range of clinical factors to produce an overall assessment of complete dentures. The 10‐item scale was examined for Internal Consistency and Construct Validity providing a summated rating scale whose total score could be used as an index for the quality of dentures. Results: Both intra‐ and inter‐examiner agreement for all 10 dichotomous scale criteria were all good or very good; the intra‐examiner Kappa values were 0.72 to 0.91 and the inter‐ examiner values were 0.72 to 0.99. Statistical analysis revealed that a 7‐item summated rating scale could be constructed which has marginally better internal consistency (reliability) and Construct Validity with fewer questions. Conclusions: The FAD criteria¹ can be used to give good repeatability. The adjustments made to the original summation scale, if confirmed by new studies, would further increase the practical utility of a FAD index for general dentists. In due course with data from future studies, discrimination on weighting of items will be appropriate to relate the impact of denture quality on quality of life.     

The development of a method for functional assessment of dentures

Aims: To design and validate a method of assessing complete dentures from a functional standpoint. Subjects: A random sample of 40 complete denture wearers took part in the study. Setting: A university clinical department of prosthetic dentistry. Intervention: We undertook a pilot study to refine the protocol and criteria. All participants and their dentures were examined by two authors independently, with no prior knowledge of the patients'complaints. Design: We defined nine clinical factors of functional quality and applied criteria with binary scoring. We analysed the scores for these factors for inter‐rater reliability. Results: The method proved simple to apply and took less than 5 minutes to complete. The inter‐examiner agreement for all factors was 86% to 100% giving Kappa scores of 0.64 to 1.00 (all Good or Very Good). Conclusions: This study successfully demonstrates that the technique, which we call the Functional Assessment of Dentures (FAD), can give good inter‐examiner reliability. It can therefore be used separately as a routine diagnostic tool and to investigate the relationship between denture qualities and functional ‘outcome’ such as difficulty eating or dietary selection.     

Development of a bioartificial nerve graft. II. Nerve regeneration in vitro

A promising alternative for the repair of peripheral nerve injuries is the bioartificial nerve graft, or BNG, comprised of a tubular conduit preseeded with Schwann cells, which are an effective substrate for enhancing nerve regeneration. The physical properties of the conduit, porosity and wall thickness, as well as the Schwann cell seeding density, were tested for their effect on axon growth using rat dorsal root ganglia. These parameters can influence the amount of nutrients and growth factors made available to the neural tissue. Results show that a greater wall thickness and lower porosities have a detrimental effect on the growth of the axons. Over a four week period, axons extended 3.2 mm for the optimum case (DeltaR = 0.82 mm, epsilon = 0.75) compared to 1.8 and 1.6 mm for a lower porosity (0.55) and a greater wall thickness (1.4 mm), respectively. A maximum in the growth rate occurs at a porosity of 75% for Schwann cell seeded conduits but not for unseeded ones. When compared to mass transfer predictions, the results suggest that, at higher porosities, more growth factors diffuse out of the conduit, while at low porosities there is competition for nutrients. Increasing the Schwann cell seeding density enhances growth but also leads to an increase in the number of axons along the length of the conduit. This is indicative of branching of the axons, which requires additional resources to maintain and can lead to painful neuroma formation. Wall thickness and porosity were found not to have any significant effect on the axon number sprouting from the dorsal root ganglia and the mean diameter (p > 0.05). Considerations need to be made, not just on the polymer used, but also on its porosity, wall thickness, and Schwann cell seeding density. These parameters can be adjusted to create a bioartificial nerve graft that provides the optimal environment for nerve growth.