Loss of second and sixth conserved cysteine residues from trypsin inhibitor-like cysteine-rich domain-type protease inhibitors in Bombyx mori may induce activity against microbial proteases

Previous studies have indicated that most trypsin inhibitor-like cysteine-rich domain (TIL)-type protease inhibitors, which contain a single TIL domain with ten conserved cysteines, inhibit cathepsin, trypsin, chymotrypsin, or elastase. Our recent findings suggest that Cys^2nd and Cys^6th were lost from the TIL domain of the fungal-resistance factors in Bombyx mori, BmSPI38 and BmSPI39, which inhibit microbial proteases and the germination of Beauveria bassiana conidia. To reveal the significance of these two missing cysteines in relation to the structure and function of TIL-type protease inhibitors in B. mori, cysteines were introduced at these two positions (D36 and L56 in BmSPI38, D38 and L58 in BmSPI39) by site-directed mutagenesis. The homology structure model of TIL domain of the wild-type and mutated form of BmSPI39 showed that two cysteine mutations may cause incorrect disulfide bond formation of B. mori TIL-type protease inhibitors. The results of Far-UV circular dichroism (CD) spectra indicated that both the wild-type and mutated form of BmSPI39 harbored predominantly random coil structures, and had slightly different secondary structure compositions. SDS-PAGE and Western blotting analysis showed that cysteine mutations affected the multimerization states and electrophoretic mobility of BmSPI38 and BmSPI39. Activity staining and protease inhibition assays showed that the introduction of cysteine mutations dramaticly reduced the activity of inhibitors against microbial proteases, such as subtilisin A from Bacillus licheniformis, protease K from Engyodontium album, protease from Aspergillus melleus. We also systematically analyzed the key residue sites, which may greatly influence the specificity and potency of TIL-type protease inhibitors. We found that the two missing cysteines in B. mori TIL-type protease inhibitors might be crucial for their inhibitory activities against microbial proteases. The genetic engineering of TIL-type protease inhibitors may be applied in both health care and agricultural industries, and could lead to new methods for breeding fungus-resistant transgenic crops and antifungal transgenic silkworm strains.     

The Treatment of Missing Values in Logistic Regression

The efficiencies of the estimators in the linear logistic regression model are examined using simulations under six missing value treatments. These treatments use either the maximum likelihood or the discriminant function approach in the estimation of the regression coefficients. Missing values are assumed to occur at random. The cases of multivariate normal and dichotomous independent variables are both considered. We found that in general, there is no uniformly best method. However, mean substitution and discriminant function estimation using existing pairs of values for correlations turn out to be favourable for the cases considered.     

Sexual dimorphism in the emergence of deciduous teeth: its relationship with growth components in height

Sexual dimorphism in the emergence of the deciduous dentition of French-Canadian children may be explained by differences in recumbent length. Relative to the chronological age scale, boys are longer and their teeth emerge earlier than girls. Recumbent lengths attained at the exact age of emergence, as estimated by fifth-order polynomials fitted to each subject's serial data, are comparable between the sexes. Multi- and univariate analyses of variance show no significant sex differences in the lengths attained at the age of emergence of the deciduous teeth. These findings suggest that clinical standards for emergence of deciduous teeth scaled relative to length rather than chronological age are more accurate and efficient.     

Breeding vervet monkeys in a closed environment

A breeding group of vervet monkeys produced 95 offspring over five years. Fetal wastage for 114 pregnancies was 16.7%. Uterine sizes were recorded for 103 pregnancies in various stages of gestation. This enabled diagnosis of pregnancy, estimation of stage of gestation, and prediction of parturition to within two weeks. Detailed observations were made on dental eruption, pelage development, and growth of 57 infants.     

The mechanical significance of the occlusal geometry of great ape molars in food breakdown

Analyses of dental function are an essential component of the study of human evolution. However, with few exceptions, they have utilized the traditional analogizing method of comparative anatomy, and have assumed rather than demonstrated that proposed adaptive characters confer a performance benefit. Since food reduction is a mechanical process, it is appropriate to measure performance using mechanical parameters, specifically the ability of a given morphology to induce failure in food particle by either of the two major regimes: crush and shear, corresponding to simple stresses (tensile and compressive) and shear stress, respectively. We apply finite elements stress analysis to model the relationship between the angulation of the intercuspal occlusal surfaces in a “puncture crushing” mode of mastication. On the basis of morphological data acquired from sectioned great ape molars, we have predicted the nature, magnitude and distribution of stress in a standard food particle by models representing each morphotype. Results indicate that the blunt-cusped molars ofHomo, the gradually-sloping supporting (buccal) cusps but high-angled guiding (lingual) cusps of the lower molars ofPan, and the high angled occlusal surfaces ofGorillaare all more likely to fracture small food particles by shear, while the gradually sloping occlusal surfaces ofPongomolars are more likely to break them down by “crush”. Mechanisms of food failure induced by molars ofPanandHomowill vary according to the orientation of the tooth–food contacting surfaces, which in turn will vary according to the size of the food particle. These genera may be able to break food down either by shear or by “crush”.