Genetic and functional analysis of common MRC1 exon 7 polymorphisms in leprosy susceptibility

The chromosomal region 10p13 has been linked to paucibacillary leprosy in two independent studies. The MRC1 gene, encoding the human mannose receptor (MR), is located in the 10p13 region and non-synonymous SNPs in exon 7 of the gene have been suggested as leprosy susceptibility factors. We determined that G396S is the only non-synonymous exon 7-encoded polymorphism in 396 unrelated Vietnamese subjects. This SNP was genotyped in 490 simplex and 90 multiplex leprosy families comprising 704 patients (47% paucibacillary; 53% multibacillary). We observed significant under-transmission of the serine allele of the G396S polymorphism with leprosy per se (P = 0.036) and multibacillary leprosy (P = 0.034). In a sample of 384 Brazilian leprosy cases (51% paucibacillary; 49% multibacillary) and 399 healthy controls, we observed significant association of the glycine allele of the G396S polymorphism with leprosy per se (P = 0.016) and multibacillary leprosy (P = 0.023). In addition, we observed a significant association of exon 7 encoded amino acid haplotypes with leprosy per se (P = 0.012) and multibacillary leprosy (P = 0.004). Next, we tested HEK293 cells over-expressing MR constructs (293-MR) with three exon 7 haplotypes of MRC1 for their ability to bind and internalize ovalbumin and zymosan, two classical MR ligands. No difference in uptake was measured between the variants. In addition, 293-MR failed to bind and internalize viable Mycobacterium leprae and BCG. We propose that the MR–M. leprae interaction is modulated by an accessory host molecule of unknown identity.     

The dual function of barred plumage in birds: camouflage and communication

A commonly held principle in visual ecology is that communication compromises camouflage: while visual signals are often conspicuous, camouflage provides concealment. However, some traits may have evolved for communication and camouflage simultaneously, thereby overcoming this functional compromise. Visual patterns generally provide camouflage, but it was suggested that a particular type of visual pattern – avian barred plumage – could also be a signal of individual quality. Here, we test if the evolution of sexual dimorphism in barred plumage, as well as differences between juvenile and adult plumage, indicate camouflage and/or signalling functions across the class Aves. We found a higher frequency of female- rather than male-biased sexual dimorphism in barred plumage, indicating that camouflage is its most common function. But we also found that, compared to other pigmentation patterns, barred plumage is more frequently biased towards males and its expression more frequently restricted to adulthood, suggesting that barred plumage often evolves or is maintained as a sexual communication signal. This illustrates how visual traits can accommodate the apparently incompatible functions of camouflage and communication, which has implications for our understanding of avian visual ecology and sexual ornamentation.     

Simplified method for the detection of apo(a) isoforms.

Apolipoprotein a, is a high molecular weight glycoproteic component of Lp(a), a molecule associated with coronary arterial disease. Apo(a) exhibits considerable size heterogeneity due to variable repetitions of the carbohydrate-containing structural unit, termed kringle. There are five different kringle forms and 10 different kringle 4 types. Apo(a) polymorphism and molecular weight depend on the number of copies of kringle 4 type 2. In this paper we describe a modified 3.75% and 6% discontinuous polyacrylamide gel system and Western-blot technique that shortness the assay time and improves the identification of apo(a) isoforms with a theoretical error of less than 1 kringle. The assay uses a standard curve prepared with five different recombinant apo(a) molecules, detected up to 50 ng of protein in Lp(a), showed a maximal resolution of 2 kringles and, with the use of third degree polynominal regression analysis, had an error of 0.01275. The inter-assay coefficient of variation was 1.7, 2, and 1.4 for the 14 K, 18 K, and 22 K phenotypes, whereas the intra-assay coefficient of variation was 0.32%, 0.18%, and 0.17%, respectively. It is possible that this modified method will diminish the number of putative null alleles so far detected in various studies, but most of all, we are certain that it can be of use in epidemiological studies due to its ease of use, speed, low cost, and enhanced number of samples that can be tested.     

Purification of benzylpenicillin filtered broths by ultrafiltration and effect on solvent extraction

Purification of benzylpenicillin filtered broths obtained from fermented broths produced at CIPAN, SA Pilot Plant Fermentation was carried out by ultrafiltration with diafiltration. This study was done using an ultrafiltration laboratory/pilot installation from Paterson Candy International Ltd. whose module has a tubular configuration. Membranes with molecular weight cut-offs of 8000 (polysulfone), 20?000 (polysulfone) and 100?000 (PVDF) were assessed. Comparison between solvent extraction of benzylpenicillin from ultrafiltered broths and from filtered broths using flocculants was also done. Ultrafiltration with diafiltration of benzylpenicillin filtered broths was used successfully to remove proteins, colored substances and other impurities giving high benzylpenicillin recovery in the permeate, as an alternative to the use of flocculants and anti-emulsion agents to obtain good phases separation in benzylpenicillin solvent extraction.     

The trade-off between fleeing and tonic immobility behaviors in an ectothermic animal

acta ethologica - When a prey is perceived and attacked, it may adopt direct anti-predator strategies, such as fleeing and tonic immobility (TI). During this latter, individuals remain in a...     

Inherited hemoglobin disorders in an Afro-Amazonian community: Saracura

The most common hemoglobinopathies, viz, hemoglobins S and C, and α- and β-thalassemias, were investigated through the molecular screening of 116 subjects from the community of Saracura, comprising fugitive African slaves from farms of the municipality of Santarém, in the west of Pará State, Brazilian Amazon. The observed frequency of the HBB*S gene (0.9%) was significantly lower than that encountered in other Afro-derived communities in the region. Concomitantly, the absence of the HBB*C allele has been reported for most of the Afro-Amazonian communities thus far studied. As remnant populations of quilombos are generally small, the heterogeneous distribution of HBB*S and HBB*C alleles among them is probably due to genetic drift and/or founder effect. The observed frequency of 3.7 kb deletion in Saracura (8.5%) was consistent with the African origin of the population, with a certain degree of local differentiation and admixture with individuals of Caucasian ancestry, placed in evidence by the occurrence of - -(MED) deletion (1.2%), a common mutation in Mediterranean regions. As regards β-thalassemia, among the seven different mutations found in Saracura, three β^o and two β⁺ mutations were of Mediterranean origin, and two β⁺ of African. Thus, only 28% of the local β-thalassemia mutations found in Saracura were of African origin.     

Immobilization of amyloglucosidase on alkylamine derivatives of metal-link-activated inorganic supports

A new process to couple amyloglucosidase (AG) to inorganic supports is described. The technique consists in activating the support with a transition metal salt according to the metal-link method and subsequent amination and linkage of the alkylamine derivative using glutaraldehyde. The various parameters susceptible of influencing the properties of the immobilized enzyme (IME) preparation are investigated. The best result are obtained when 100 mg of 1000-Å controlled porous glass (CPG) are treated with 45 mg of TiCl₄ and the activated carrier aminated using a 10-g/L solution of hexamethylenediamine (HMDA) in carbon tetrachloride (10 Ml/100 mg). Preparation obtained according to the process here described show operational stabilities much superior to those of AG immobilized on the same support by the traditional metal-link method or its variations. The mechanism involved in the preparation of the amino derivative of CPG is proposed.     

Sexual Signals as Advertisers of Resistance to Mistakes

To be evolutionarily stable, sexual signals need to convey information regarding the quality or motivation of individuals. This can be achieved through direct or indirect signal costs that guarantee honest communication or through signal designs that facilitate the assessment of quality. Here, I present the case for an alternative hypothesis: that signalling exposes informative mistakes. Signalling mistakes range from occasional to frequent and from slight distortions of typical signals to grossly atypical signals. Their occurrence may be enhanced by disease or stress, thus revealing individual quality or motivation, and receivers typically respond negatively to them. By this mechanism, honest communication is due to costs of developing resistance to mistakes. Therefore, the hypothesis can function independently of signal design costs, although it can also be enhanced by signal design costs when those increase the occurrence of mistakes. This hypothesis widens the scope of signals expected to be sexually selected and creates new approaches to research in sexual selection and animal communication.     

Aprotinin binding to amyloid fibrils.

Different low molecular mass ligands have been used to identify amyloid deposits. Among these markers, the dyes Thioflavin T and Congo Red interact specifically with the beta-sheet structure arranged in a cross-beta conformation, which is characteristic of amyloid. However, the molecular details of this interaction remain unknown. When labelled with technetium-99m, the proteinase inhibitor aprotinin has been shown to represent a very important radiopharmaceutical agent for in vivo imaging of extra-abdominal deposition of amyloid in amyloidosis of the immunoglobulin type. However, no information is available as to whether aprotinin binds other types of amyloid fibrils and on the nature and characteristics of the interaction. The present work shows aprotinin binding to insulin, transthyretin, beta-amyloid peptide and immunoglobulin synthetic amyloid fibrils by a specific dot-blot ligand-binding assay. Aprotinin did not bind amorphous precipitates and/or the soluble fibril precursors. A Ka of 2.9 microM-1 for the binding of aprotinin to insulin amyloid fibrils was determined by Scatchard analysis. In competition experiments, analogues such as an aprotinin variant, a spermadhesin and the soybean trypsin inhibitor were tested and results suggest that both aprotinin and the spermadhesin interact with amyloid fibrils through pairing of beta-sheets of the ligands with exposed structures of the same type at the surface of amyloid deposits. An electrostatic component may also be involved in the binding of aprotinin to amyloid fibrils because important differences in binding constants are observed when substitutions V15L17E52 are introduced in aprotinin; on the other hand beta-sheet containing acidic proteins, such as the soybean trypsin inhibitor, are unable to bind amyloid fibrils.