Evolution of morphological integration in the skull of Carnivora (Mammalia): Changes in Canidae lead to increased evolutionary potential of facial traits

Morphological integration refers to the fact that different phenotypic traits of organisms are not fully independent from each other, and tend to covary to different degrees. The covariation among traits is thought to reflect properties of the species' genetic architecture and thus can have an impact on evolutionary responses. Furthermore, if morphological integration changes along the history of a group, inferences of past selection regimes might be problematic. Here, we evaluated the stability and evolution of the morphological integration of skull traits in Carnivora by using evolutionary simulations and phylogenetic comparative methods. Our results show that carnivoran species are able to respond to natural selection in a very similar way. Our comparative analyses show that the phylogenetic signal for pattern of integration is lower than that observed for morphology (trait averages), and that integration was stable throughout the evolution of the group. That notwithstanding, Canidae differed from other families by having higher integration, evolvability, flexibility, and allometric coefficients on the facial region. These changes might have allowed canids to rapidly adapt to different food sources, helping to explain not only the phenotypic diversification of the family, but also why humans were able to generate such a great diversity of dog breeds through artificial selection.     

Histoincompatibility reactions in the hydrocoral Millepora dichotoma

The xenogeneic- and allogeneic immunological specificity of the hydrocoral Millepora dichotoma has been investigated. Xenogeneic histoicompatibility reactions have been observed between this hydrocoral and a series of species belonging to the Demospongiae and to the Anthozoa (both Hexacorallia and Octocorallia). The xenogeneic histoincompatibility reactions proceed in the following sequence: (a) Species-unspecific sensitization; (b) necrosis formation, which is very likely due to an autolytic process; (c) callus formation, due to an hyperplastic growth of stolons; and (d) formation of a contact barrier in form of a barrier layer or a restored stolonial layer. Allogeneic histoincompatibility reactions are restricted to those regions of the coral which are interspersed with polyps; allogenic fusion is observed between branches, with a high density of polyps and the “foot”-region, which is characterized by a low polyp density.     

Influence of molecular weight of DNA on the determination of anti-DNA antibodies in systemic lupus erythematosus (SLE) sera by radioimmunoassay.

Using a radioimmunoassay (RIA) based on the Farr technique with radioactively labeled 3-H-DNA for quantitative measurements of anti-DNA antibodies in sera of patients with systemic lupus erythematosus (SLE), the influence of molecular weight of DNA (ranging from 0.1 times 10-6 to 22.0 times 10-6 daltons) on binding and precipitation in this system has been investigated. Comparing our results with mathematical models it follows that one antibody molecule is fixed on the average to a statistical DNA segment of 2 times 10-6 to 4 times 10-6 daltons. Furthermore binding capacity of the DNA was found to be independent of the molecular weight, as demonstrated in a double label experiment using 14-C and 3-H-labeled DNA of different size. However, the amount of radioactivity precipitated was found to depend on the molecular weight of the labeled DNA following a non-linear function. It was calculated that a minimal ratio of fixed antibody molecules per a certain size of DNA was necessary for precipitation. The mathematical treatment of the observed non-linear precipitation dependence will be discussed using various statistical models. Our results indicate that the quantitative measurements of anti-DNA antibodies with the Farr technique e.g. for diagnosis and control of SLE in clinical immunology is highly dependent on the molecular weight of the labeled DNA used in the assay system and reliable results are only obtained with DNA of a sufficiently high molecular weight.     

The biological activity of bacteriophage DNA, prepared by the cationic detergent dilution technique

The preparation of phage lambda DNA infecting E. coli K 12 with cationic detergent is described. This DNA infects E. coli spheroblasts with the same efficiency as DNA prepared by phenol methods.     

Human serum deoxyribonuclease assay in (3H)DNA-polyacrylamide gels without staining artifacts

A method is described for detecting and evaluating serum deoxyribonuclease (DNase) activities after their separation by disc electrophoresis in polyacrylamide gels containing radioactively labeled DNA.After the electrophoretic run the gels are sliced, incubated in an appropriate buffer, and the amount of diffusible radioactive DNA fragments formed by the action of DNases in the incubation buffer is determined.The method has a high sensitivity as well as a quantitative reproducibility within ±5% even at low enzyme activities down to 10 pg Worthington DNase I equivalents.This method has been found superior to procedures that use staining of the gels for unhydrolyzed DNA, where irrelevant stained bands may invalidate the results. Thus, we get meaningful results even with human serum.     

Different deoxyribonucleases in human lymphocytes

The distribution pattern of deoxyribonuclease activities in human lymphocytes has been examined by micro-disc-electrophoresis. Four groups of deoxyribonuclease activities, differing in their electrophoretic mobility, in the nature of their optimal substrate and in their optimal incubation conditions, are characterized. There are two alkaline DNase-activities. One corresponds to DNase I (EC 3.1.4.5), the other having pH optimum of about pH 9.0, prefers denatured DNA as substrate and is not dependent on divalent cations. The fractions with an acid pH optimum can be subdivided into two groups, which differ in their activity towards native DNA, towards denatured DNA, in their activity when succinate is present and in their pH optimum.     

Size determination of poly(A) after in vitro methylation with radioactive dimethyl sulfate

A simple method for the size determination of poly(A) using in vitro labeling by methylation with [³H]dimethyl sulfate is described. After methylation, modified poly (A) has the same mobility, using polyacrylamide gel electrophoresis, as has the unmodified polymer, thus showing that the methylation does not cause degradation. Therefore the method is a sensitive assay to size the poly(A) segments from in vivo unlabeled tissue. The method was applied to determine the size of poly(A) sequences on mRNA from mouse L5178Y cells and from rat liver.     

Lectin, a possible basis for symbiosis between bacteria and sponges.

From the marine sponge Halichondria panicea a lectin was isolated and characterized. The homogeneous lectin (composed of protein to 80.7% and of neutral carbohydrates to 14.1%) had a molecular weight of 78,000 (determined by gel filtration) and consisted of four subunits with a molecular weight of 21,000 each (determined by gel electrophoresis in the presence of sodium dodecyl sulfate). The hemagglutinating activity was only slightly dependent upon ionic strength and incubation temperature and did not require divalent cations, but it was inhibited by reagents for thiol groups. The Halichondria lectin was completely inhibited in hemagglutination competition experiments in the presence of fetuin, D-galacturonic acid, D-glucuronic acid, polygalacturonic acid, or L-fucose. The purified Halichondria lectin did not cause reaggregation of dissociated H. panicea cells. From the same sponge species bacteria were isolated and identified as Pseudomonas insolita. These bacteria were cultivated in marine broth 2216. Under these culture conditions the bacteria grew only in the presence of the homologous lectin; the lectin-caused effect was not abolished by D-glucuronic acid or D-galacturonic acid. However, after addition of a polysaccharide-containing fraction isolated from P. insolita, the lectin-caused, growth-promoting effect was abolished. Other lectins were found to exhibit no growth-promoting effect. On the basis of colony counts, P. insolita was the predominant bacterial species in the sponge extract; 1.9 X 10(6) Pseudomonas colonies were measured in extracts isolated from 1 g of sponge. The assumption of an interrelationship between the sponge and the bacterium is supported by the results indicating that the Halichondria lectin has no effect on the growth of such bacteria isolated from six other marine sponge species. Evidence is presented which indicates that the Halichondria lectin is not utilized during growth of the Pseudomonas species. Lectin activity was detected on the surface of mucoid cells from H. panicea. From the data obtained the possibility is discussed that the Halichondria lectin is a basis for a symbiotic relationship between the sponge and the bacterium.