Biosynthetic relationship between the major matrix proteins of adrenal chromaffin granules

The matrix of the chromaffin granule contains a family of acidic proteins, collectively known as the chromogranins. It has been suggested that this family results from protease action on the major component, chromogranin A. Evidence for this has now been obtained from in vitro translation of adrenal medullary messenger RNA and immunoprecipitation of translation products using an antiserum directed against chromogranin A, but which also recognises other chromogranins.     

Scent‐Mark Identification and Scent‐Marking Behaviour in African Wild Dogs (Lycaon pictus)

Scent‐marking is common in mammals, but where signals are carried by urine and faeces, distinguishing between scent‐marking and mere elimination is problematic. To do so, we documented behaviours and context variables associated with urination and defecation in free‐ranging endangered African wild dogs (Lycaon pictus) and tested whether these were related to the responses of other dogs to deposits. We found that distinct postures were almost uniquely associated with deposits by dominant wild dogs, were more common during urination than defecation, and increased the likelihood that these deposits would be investigated by other wild dogs. The likelihood of investigation depended on the sex and dominance status of the depositor, the type of deposit and the substrate. Urine from dominant females was more likely to be investigated by other wild dogs than any other deposits, and deposits placed on vegetation were more likely to be investigated than those on bare ground. The likelihood that a deposit would be overmarked was affected by the deposit type and the sex and dominance status of the last depositor. Collectively, these results suggest that dominant wild dog urine is of greatest interest to other dogs. Our results show that some deposits by African wild dogs are not scent‐marks and that detailed observations of behaviours and context variables during elimination events can be used to distinguish deposits that are likely to be of communication value.     

The discovery of the benzazepine class of histamine H3 receptor antagonists

This Letter describes the discovery of a novel series of H₃ receptor antagonists. The initial medicinal chemistry strategy focused on deconstructing and simplifying an early screening hit which rapidly led to the discovery of a novel series of H₃ receptor antagonists based on the benzazepine core. Employing an H₃ driven pharmacodynamic model, the series was then further optimised through to a lead compound that showed robust in vivo functional activity and possessed overall excellent developability properties.     

A light microscope-based double retrograde tracer strategy to chart central neuronal connections

This protocol describes a double retrograde tracing method to chart divergent projections in the CNS using light microscope techniques. It is based on immunohistochemical visualization of retrograde transport of cholera toxin b-subunit (CTb) and silver enhancement of a gold-lectin conjugate. Production of the gold-lectin is explained in detail, and a technique is offered to record through the injection pipettes, to help guide accurate placement of injections. Visualization of the two tracers results in light brown staining of CTb-labeled neurons and labeling by black particles of gold-lectin-containing neurons. Both types of label are easily recognized in the same neuron. The labeling is permanent and is well suited for studies in which large areas of the brain need to be surveyed. The whole procedure (excluding survival time) takes approximately 5-7 d to complete.     

Correlates of Protective Cellular Immunity Revealed by Analysis of Population-Level Immune Escape Pathways in HIV-1

HLA class I-associated polymorphisms identified at the population level mark viral sites under immune pressure by individual HLA alleles. As such, analysis of their distribution, frequency, location, statistical strength, sequence conservation, and other properties offers a unique perspective from which to identify correlates of protective cellular immunity. We analyzed HLA-associated HIV-1 subtype B polymorphisms in 1,888 treatment-naïve, chronically infected individuals using phylogenetically informed methods and identified characteristics of HLA-associated immune pressures that differentiate protective and nonprotective alleles. Over 2,100 HLA-associated HIV-1 polymorphisms were identified, approximately one-third of which occurred inside or within 3 residues of an optimally defined cytotoxic T-lymphocyte (CTL) epitope. Differential CTL escape patterns between closely related HLA alleles were common and increased with greater evolutionary distance between allele group members. Among 9-mer epitopes, mutations at HLA-specific anchor residues represented the most frequently detected escape type: these occurred nearly 2-fold more frequently than expected by chance and were computationally predicted to reduce peptide-HLA binding nearly 10-fold on average. Characteristics associated with protective HLA alleles (defined using hazard ratios for progression to AIDS from natural history cohorts) included the potential to mount broad immune selection pressures across all HIV-1 proteins except Nef, the tendency to drive multisite and/or anchor residue escape mutations within known CTL epitopes, and the ability to strongly select mutations in conserved regions within HIV''s structural and functional proteins. Thus, the factors defining protective cellular immune responses may be more complex than simply targeting conserved viral regions. The results provide new information to guide vaccine design and immunogenicity studies.     

The effect of birthweight on tooth-size variability in twins.

Studies indicate that low birthweight (LBW) children display reduced deciduous tooth size but there is little information about permanent tooth size. It has also been shown that dental fluctuating asymmetry (FA) increases in response to various environmental influences, but the relationship between birthweight and FA remains unclear. The aim of this study was to compare tooth size and asymmetry, according to birthweight, in the deciduous and permanent dentitions of a sample of Australian twins. The study sample comprised 436 twins, classified into 2 groups: normal birthweight (NBW > 2500 g) and low birthweight (LBW < or = 2500 g). For each individual it was generally possible to measure maximum mesiodistal crown diameters of both deciduous and permanent central incisors from serial dental models. Correlations were calculated between tooth-size variables and birthweight; subsequently comparisons of tooth size and FA were made between the LBW and NBW samples using Student's t tests. Small positive correlations (around .1) were noted between birthweight and tooth-size variables. There was no evidence of tooth-size reduction in the LBW male sample, but the LBW females displayed tooth-size reduction of approximately 2-3% for both deciduous and permanent incisors, compared to the NBW females. There was no evidence of increased FA in the LBW individuals of either sex. These findings indicate that developing teeth are generally well-protected from developmental disturbances during prenatal and perinatal periods. Further research is needed to clarify the biological basis of an apparently true but weak association between tooth size and birthweight.     

Unusual redox behaviour of cytochrome b-561 from bovine chromaffin granule membranes

(1) Redox titrations of cytochrome b-561 have been performed with the purified cytochrome and with intact and detergent-solubilized chromaffin-granule membranes. (2) The midpoint redox potential of the cytochrome is 100–130 mV; this depends upon the composition of the buffer, but is independent of pH in the range 5.5–7.5; partial proteolysis of the cytochrome raises the midpoint potential to 160 mV. (3) The Nernst plots of titration data have slopes of 75–115 mV, and are in some cases sigmoid in shape. This may be explained by negative cooperativity during redox transitions in oligomeric cytochrome b-561. (4) Measurements of the haem and cytochrome content of chromaffin granule membrane suggest a haem content of 1 mol/mol protein. (5) Chemical crosslinking of cytochrome b-561 suggests that it may exist as an oligomer of 4–6 polypeptide chains within the chromaffin granule membrane. Aggregation of purified cytochrome b-561 was shown by gel filtration studies and by immunological methods in SDS-polyacrylamide gels. Studies of the molecular weight of the aggregates suggest that the monomer has a molecular weight close to 22 000, but migrates anomalously slowly during electrophoresis.     

Anatomical and physiological foundations of cerebellar information processing

A coordinated movement is easy to recognize, but we know little about how it is achieved. In search of the neural basis of coordination, we present a model of spinocerebellar interactions in which the structure-functional organizing principle is a division of the cerebellum into discrete microcomplexes. Each microcomplex is the recipient of a specific motor error signal - that is, a signal that conveys information about an inappropriate movement. These signals are encoded by spinal reflex circuits and conveyed to the cerebellar cortex through climbing fibre afferents. This organization reveals salient features of cerebellar information processing, but also highlights the importance of systems level analysis for a fuller understanding of the neural mechanisms that underlie behaviour.     

Some examples of the use of computer-produced contour plots in the fitting of enzyme rate equations to reaction-velocity measurements

The use of computer-based isotach plots, relating reaction velocity to simultaneous variation of two substrates or effectors of an enzyme, in producing estimates of the parameters of enzyme rate equations was investigated. The computer program (;SYMAP') incorporates an interpolation algorithm, and the superiority of this over visual estimation in producing interpolated velocity values for the estimation of parameter values by conventional double-reciprocal plots is described. The usefulness of the SYMAP program in monitoring the process of fitting data obtained by simultaneous changes in two experimental variables is also described. It is shown that if the residual errors are weighted by a procedure described elsewhere (Ottaway, 1971b, 1973), the percentage error of the computed velocity is distributed evenly over a plot which contains a 100-fold variation in the concentration of one substrate and a 500-fold variation in the concentration of Mg(2+), and in which the velocity of the reaction (that catalysed by NAD kinase) varies over a 60-fold range. The two-dimensional percentage error plot was used to assess the limits within which an incomplete inhibition equation is valid, and to detect a discrepancy in an expected good fit, caused by an impurity in one of the substrates.