Determinants of potency on alpha-conotoxin MII, a peptide antagonist of neuronal nicotinic receptors

Alpha-conotoxin MII, a peptide toxin isolated from Conus magus, antagonizes a subset of neuronal nicotinic receptors. Rat alpha3beta2 receptors, expressed in Xenopus oocytes, are blocked with an IC(50) of 3.7 +/- 0.3 nM. To identify structural features that determine toxin potency, a series of alanine-substituted toxins were synthesized and tested for the ability to block the function of alpha3beta2 receptors. Circular dichroism and protein modeling were used to assess the structural integrity of the mutant toxins. Three residues were identified as major determinants of toxin potency. Replacement of asparagine 5, proline 6, or histidine 12 with alanine resulted in >2700-fold, 700-fold, and approximately 2700-fold losses in toxin potency, respectively. A decrease in pH improved toxin potency, while an increase in pH eliminated toxin blockade, suggesting that, in the active form of the toxin, histidine 12 is charged. The imidazole ring of histidine 12 protrudes from one side, while asparagine 5 and proline 6 are located at the opposite end of the toxin structure. The side chains of these three residues are exposed on the surface of the toxin, suggesting that they directly interact with the alpha3beta2 receptor.     

Infrapopulation dynamics of a wild strain of Taenia crassiceps (WFU) (Cestoda: Taeniidae) in BALB/cJ mice

Taenia crassiceps cysticerci form large infrapopulations that persist in the tissues of their rodent hosts. Early infrapopulation growth appears inhibited and is followed by rapid increases that appear not to be controlled by the host immune response. This investigation was undertaken to examine the infrapopulation growth dynamics of a normally developing strain (WFU) of T. crassiceps during a 60-day primary intraperitoneal (i.p.) infection. Three, 6, 9, 14, 28, and 60 days after i.p. inoculation of 5 cysticerci, mice were killed, and the numbers of larvae, developmental stage, and buds per larva were recorded. Larval infrapopulation abundance increased exponentially beginning on day 6 postinoculation (PI), indicating an initial lag in reproduction. A stage-structured exponential growth model, assuming no mortality, fits the larval infrapopulation dynamics in terms of the numbers of larvae in reproductive and nonreproductive stages, indicating that cysticerci evade or suppress (or both) host immune mechanisms that are parasite restrictive after the first week of infection.     

Posters

Introduction  Fine needle aspiration (FNA) cytology of the thyroid is a well-established test in the clinical work-up of patients with solitary nodules of the thyroid. Thyroid FNA does however have limitations and audit of diagnostic performance is important.
Methods  The histopathology archives of the Royal Victoria Hospital were searched for all thyroid resections and the histopathological diagnosis was correlated with the pre-operative cytological diagnosis, where available. Special emphasis was placed on the accuracy of tumour diagnosis.
Results  A total of 173 cases were identified during the 2-year period, of these 93 had available pre-operative FNA. A total of 57 tumours were identified. A small number (six of 57) of significant discrepancies were identified. These included a malignant lymphoma diagnosed as Hashimoto's thyroiditis, a metastasis which the FNA had suggested was a medullary carcinoma and an insular carcinoma diagnosed as medullary carcinoma on FNA. False positives included a colloid cyst diagnosed as suspicious of malignancy and a cytological diagnosis of papillary carcinoma not confirmed on histology.
Discussion  At present, the majority of thyroid FNAs in our clinics are performed by surgeons and material is not routinely available for immunocytochemistry. In spite of these limitations, there were few major discrepancies. These might be reduced if pathologist aspirators were able to perform FNAs and collect material for further studies, where necessary. This would allow identification of medullary carcinomas and malignant lymphomas.
Conclusion  FNA of thyroid lesions is a useful investigation in our clinical setting, however, some areas of potential for improvement have been identified.     

Dependence of electroosmotic flow in capillary electrophoresis on Group I and II metal ions

The effect of Group I and II metal ions on electroosmotic flow in capillary electrophoresis in fused-silica capillaries is characterized. The electroosmotic mobility of aqueous mobile phases of lithium, sodium, potassium, calcium and barium acetates in fused-silica capillaries is measured as a function of pH at constant voltage. Cross contamination is avoided by using separate columns for each study and pH control is maintained with the aid of He sparging. The shape of a plot of pH vs. electroosmotic mobility depends on the particular cation used which in turn depends on the surface sorption properties of the ions. Column history is demonstrated to have an effect on electroosmotic flow and therefore retention times. The resolution of a test mixture is optimal in the lithium-based buffer.     

Growth and Body Mass Index of Brazilians Ages 9 Through 17 Years

Obesity during adolescence is considered a strong predictor of adult obesity, and obesity and overweight have been increasing among Brazilian adults. To gauge the relative frequency of overweight among adolescents in Brazil, we compared the distributions of body mass index (kg/m²) and stature in national population based samples of the U.S. and Brazil. U.S. adolescents were on average about 10 cm taller than Brazilians, although growth spurts occurred at the same age for both populations. Brazilian adolescents were leaner than their U.S. counterparts. This difference was reduced among girls in the postpubertal period. At age 17 years, U.S. boys were about 10 kg heavier than Brazilian boys, but the difference among girls was only 2 kg. In families above the poverty level in the more developed South region, body mass index distribution for boys was closer to that of the U.S., and older girls tended to have higher body mass index than U.S. girls. Within Brazil, body mass index varied by ethnicity with Mulattos, but not Blacks, of both sexes having lower body mass index than Whites of the same age. Urban adolescents had higher body mass index than those living in rural areas. In general, the patterns seen among Brazilian adults were found among children. Among girls, in particular, overweight has become an identifiable problem during adolescence.     

Tracer diffusion coefficients of oxyhemoglobin a and oxyhemoglobin s in blood cells as determined by pulsed field gradient NMR

It is demonstrated that tracer diffusion coefficients can be determined for oxyhemoglobin A (HbA-O₂) and oxyhemoglobin S (HbS-O₂) in intact blood cells by means of pulsed field gradient NMR (PFG-NMR). This is possible because the method discriminates between both rapidly moving water molecules and molecules having small proton transverse relaxation times (T₂). The results indicate that only hemoglobin molecules contribute to the echo signals when large field gradients are used. The dependence of the measured diffusion coefficients on osmolarity and pH are attributed to changes in hemoglobin concentration resulting from changes in cell volume.     

Implications of climate change for Daphnia in alpine lakes: predictions from long-term dynamics, spatial distribution, and a short-term experiment

Alpine lakes may be particularly useful as sentinels of climate change because they are highly sensitive to environmental conditions. To explore the potential biotic consequences of climate change in these systems, we conducted paleo- and neoecological observational studies, as well as a short-term experiment to examine Daphnia responses to changing environmental conditions in Rocky Mountain alpine lakes. Our analysis of a sediment core from Emerald Lake representing two periods from the Holocene revealed a significant positive relationship between the abundance of Daphnia remains and fossil Aulacoseira lirata, a diatom associated with deeper mixing depths. In addition, we detected a significant increase in mean Daphnia density in the long-term record (1991–2005) from Pipit Lake, a trend that correlated well with increases in mean surface temperature. In our survey of Daphnia in 10 lakes in the Canadian Rocky Mountains, Daphnia abundance was positively correlated with both dissolved organic carbon concentration and temperature. Finally, our short-term incubation experiment demonstrated significant effects of physical conditions (i.e., temperature and/or UV radiation) and water chemistry on the juvenile growth rate of Daphnia. Overall, our findings highlight the sensitivity of Daphnia to changes in mixing depth, water temperature, and dissolved organic matter, three limnological variables that are highly sensitive to changes not only in air temperature, but also to precipitation and location of the treeline in alpine catchments. Thus, we conclude that Daphnia abundance could serve as a powerful sentinel response to climate change in alpine lakes of the Rocky Mountains.     

Proffered papers 9.30–10.00 Monday 15 September 2003

Both the original Bethesda system and the current UK classifications of cervical cytology have proved robust but each has a major weakness in the area of abnormalities of uncertain significance. Cytologists recognize that sometimes it is simply impossible to differentiate between reactive and dyskaryotic material. For this reason, the Australian version of the Bethesda system introduced a new category of 'high grade inconclusive' with a recommendation for referral to colposcopy. Approximately 60% of such cases are found to have high grade lesions at colposcopy (Schoolland M, Sterrett G, Knowles S et al .). The present UK system even with the proposed changes requires of the pathologist, a decision as to whether such cases are probably high grade (=a report of moderate dyskaryosis) or not (= a report of borderline). This continues to ignore the fact that sometimes you just cannot tell, even on review. We have taken a consecutive series of 50 referral smears, reported as moderate dyskaryosis, where the histological outcome (by loop cone) is known. These cases were rescreened and then reviewed blind by a pathologist with extensive experience of the Australian NH & MRC modified Bethesda system. On review, the material was reclassified along NH & MRC lines. The results were compared with the biopsy findings in order to determine whether the category of 'inconclusive' might be of value in the context of the NHSCSP.