Ab initio modeling of small, medium, and large loops in proteins.

This study presents different procedures for ab initio modeling of peptide loops of different sizes in proteins. Small loops (up to 8--12 residues) were generated by a straightforward procedure with subsequent "averaging" over all the low-energy conformers obtained. The averaged conformer fairly represents the entire set of low-energy conformers, root mean square deviation (RMSD) values being from 1.01 A for a 4-residue loop to 1.94 A for an 8-residue loop. Three-dimensional (3D) structures for several medium loops (20--30 residues) and for two large loops (54 and 61 residues) were predicted using residue-residue contact matrices divided into variable parts corresponding to the loops, and into a constant part corresponding to the known core of the protein. For each medium loop, a very limited number of sterically reasonable C(alpha) traces (from 1 to 3) was found; RMSD values ranged from 2.4 to 5.9 A. Single C(alpha) traces predicted for each of the large loops possessed RMSD values of 4.5 A. Generally, ab initio loop modeling presented in this work combines elements of computational procedures developed both for protein folding and for peptide conformational analysis.     

Effects of early malnutrition on Swiss white mice: avoidance learning after rearing in large litters

Mice were undernourished by rearing in litters of 20 pups per mother, weaned onto stock diet and their shuttle-box avoidance performance measured at 2 months of age. The mice from these large litters learned the avoidance response more slowly than controls (mice reared in litters of 5). In a second experiment, avoidance performance was measured after nutritional rehabilitation until 2, 4 or 6 months old. No significant improvement was observed in the previously malnourished mice. In the controls, performance was high at 2 and 4 months but by 6 months had fallen almost to the same level as the test group.     

Assessment of endothelial function of large, medium, and small vessels: a unified myograph

Endothelial dysfunction precedes the development of morphological atherosclerotic changes and can also contribute to lesion development in cardiovascular diseases. Currently, there is a lack of a single method to determine endothelial function of the entire range of vessel dimensions from aorta to arterioles. Here we assessed endothelial function of a large range of size arteries using a unified isovolumic myograph method. The method maintains a constant volume of fluid in the lumen of the vessel during contraction and relaxation, which are characterized by an increase and a decrease of pressure, respectively. Segments of six aortas, six common femoral arteries, and six mesenteric arteries from rats; six carotid arteries from mice; and six coronary and carotid arteries from pigs were used. The endothelium-dependent dose-response vasorelaxation was determined with endothelium-dependent vasodilators while arterial preconstriction was induced with vasoconstrictors at a submaximal dose. The circumferential midtension during vascular reactivity varied from 43.1 ± 7.9 to 2.59 ± 0.46 mN/mm (from large to small arteries), whereas the circumferential midstress showed a much smaller variation from 217 ± 23.5 to 123 ± 15.3 kPa (in the same range of vessels). We also found that overinflation and axial overelongation compromised endothelium-dependent vasorelaxation to underscore the significance of vessel preload. In conclusion, an isovolumic myograph was used to unify arterial vasoreactivity from large to small arteries and shows the uniformity of wall stress and %tension throughout the range of vessel sizes.     

Expression of the hepatitis delta virus large and small antigens in transgenic mice.

Simultaneous infection with hepatitis delta virus (HDV) and hepatitis B virus (HBV) in humans is often associated with severe viral liver disease including fulminant hepatitis. Since HBV is thought to be noncytopathic to the hepatocyte, the enhanced disease severity observed during dual infection has been attributed to either simultaneous immune responses against the two viruses or direct cytotoxic effects of HDV products on the hepatocyte or both. To examine these alternate possibilities, we produced transgenic mice that express the small and large delta antigens (HDAg) in hepatocyte nuclei at levels equal to those observed during natural HDV infection. No biological or histopathological evidence of liver disease was detectable during 18 months of observation, suggesting that neither the large nor small form of HDAg is directly cytopathic to the hepatocyte in vivo.     

Different antiamebic antibody isotype patterns in the large and small intestine after local and systemic immunization of mice with glutaraldehyde fixed Entamoeba histolytica trophozoites

We have determined the major immunoglobulin isotypes (IgG, IgA, IgM) of antiamebic antibodies induced in the serum and in the large and small intestine after local (oral and rectal) or systemic (intraperitoneal and intramuscular) immunization of mice with glutaraldehyde-fixed Entamoeba histolytica trophozoites (GFT). IgA predominated in the small intestine after immunization through all routes, whereas in the large intestine similar antibody levels of the major isotypes were induced by rectal, intraperitoneal and intramuscular immunization. The intramuscular route elicited intestinal responses lower than those induced by the rectal and intraperitoneal routes, but higher than the slight IgA antibody increase observed after oral immunization. The differences in antiamebic antibody response patterns at the large and small intestine suggest that there are different mucosal effector compartments. They also indicate that isotype analysis of mucosal antibodies from the sites where an infectious agent resides is needed to evaluate whether a vaccine candidate induces responses of higher protective value in the appropriate site, and that the study of antibody responses must not be limited to sampling the serum or mucosal sites distant to the relevant one.     

Kin selection, local competition, and reproductive skew

In a spatially structured population, limited dispersal gives rise to local relatedness, potentially favoring indiscriminate helping behavior. However, it also leads to local competition, which reduces the benefits of helping local kin. This tension has become the focus for a growing body of theoretical work. Existing models, however, have focused chiefly on the net impact of limited dispersal on cooperative or competitive effort in a homogeneous population. Here, I extend existing models of kin selection in a group-structured population to allow for asymmetries in expected fecundity and reproductive success among group members. I explore the consequent impact of limited dispersal on the evolution of helping and harming behavior, and on the degree of reproductive inequality or skew. I show that when individuals in a group differ in their expected fecundity, limited dispersal gives rise to kin selection for harming behavior on the part of more fecund individuals, and for helping behavior on the part of less fecund individuals. As a result, philopatry tends to exaggerate differences in reproductive success, and so promotes greater reproductive skew.     

Physicochemical properties and structure of large, medium and small granule starches in fractions of normal barley endosperm

Normal barley grain was milled to flour with a machine used to polish brewers' rice from the surface layer to the center. Large (18.4 microm, median size), medium (12.3 microm) and small (2.2 microm) granule starches were isolated from classified flours. Their physicochemical properties and fine structure were investigated. The percentage (w%) of large granules decreased from the surface layer to the center, while the amounts of medium and small granules increased. Although all the starch granules were an A-type crystal, the relative crystallinity varied from 22.0 to 27.4%. The DPn of the amyloses was around 1600 and similar for all the samples. But the amylose content of the starches varied from 21.9 to 26.4%. Also, the amylopectins showed differences in DPn (around 5700-7900) and chain-length distribution between granule size or fractions. The transition temperature ranges and the enthalpy values of the starch granules differed with granule size. The gelatinization properties showed no correlation with any of the parameters, except the enthalpy value and relative crystallinity (gamma = +0.73). The findings suggested that the structural characteristics of the starches in classified flours of normal barley differed essentially from those of waxy barley.     

Anomalous litters in hybrid mice and the retention of spermatozoa in the female tract.

Suspected superfetation was investigated in a Glasgow hybrid stock of mice. The male was removed either (i) a few days before parturition, or (ii) immediately after mating and on 23 and 25 occasions, respectively, a second litter was born. Members of the anomalous litters were inbred for 10 generations, but the incidence of supernumerary litters did not increase beyond 2-5%. The anterior part of over 500 reproductive tracts, at various stages of pregnancy and after parturition, were serially sectioned but a second set of embryos was not found. The second gestation was of normal length and superfetation was not therefore considered to be the cause of the anomalous litters. In two females, one non-pregnant and one pregnant, spermatozoa were found in the uterus and oviducts 8 days after mating and in distended uterine glands 15 days after mating respectively. It is concluded that the anomalous litters were derived from the fertilization of eggs ovulated at the post-partum oestrus by spermatozoa which had been retained in the female tract for at least 23 days.     

Doppler velocity measurements from large and small arteries of mice

With the growth of genetic engineering, mice have become increasingly common as models of human diseases, and this has stimulated the development of techniques to assess the murine cardiovascular system. Our group has developed nonimaging and dedicated Doppler techniques for measuring blood velocity in the large and small peripheral arteries of anesthetized mice. We translated technology originally designed for human vessels for use in smaller mouse vessels at higher heart rates by using higher ultrasonic frequencies, smaller transducers, and higher-speed signal processing. With these methods one can measure cardiac filling and ejection velocities, velocity pulse arrival times for determining pulse wave velocity, peripheral blood velocity and vessel wall motion waveforms, jet velocities for the calculation of the pressure drop across stenoses, and left main coronary velocity for the estimation of coronary flow reserve. These noninvasive methods are convenient and easy to apply, but care must be taken in interpreting measurements due to Doppler sample volume size and angle of incidence. Doppler methods have been used to characterize and evaluate numerous cardiovascular phenotypes in mice and have been particularly useful in evaluating the cardiac and vascular remodeling that occur following transverse aortic constriction. Although duplex ultrasonic echo-Doppler instruments are being applied to mice, dedicated Doppler systems are more suitable for some applications. The magnitudes and waveforms of blood velocities from both cardiac and peripheral sites are similar in mice and humans, such that much of what is learned using Doppler technology in mice may be translated back to humans.     

Rate,composition and efficiency of growth in mice selected for large and small body weight

Summary Mice selected for high (H₆) and low (L₆) 6-week body weight and a randombred control population (C₁) were characterized for rate, composition and efficiency of growth. Individual body weights were obtained from birth to 8 weeks of age on 682 mice representative of the three lines. Individual whole carcass determinations of water, fat, ash and protein (residual) were obtained for 180 mice sampled weekly from 3–8 weeks of age. Efficiency of feed utilization was estimated from individual body weight and feed consumption data obtained on 189 mice from 3–8 weeks of age. Growth curves for body weight and gain in body weight, constructed by line and sex, showed a temporary retardation of maximum growth rate in the L₆ line, which was attributed in part to an extended depression in growth following weaning. The composition of growth yielded no evidence that the more rapid growth rate in the H₆ line resulted from an increase in fat deposition relative to the other carcass components. A decrease in fat percent at 7 weeks of age in the H₆ and C₁ lines was not evident in the L₆ line until 8 weeks of age. Females had a higher percentage carcass fat than did males during the 4–7 weeks growth period, but this difference was essentially reduced to zero by 8 weeks of age. Percentage water was highly correlated negatively with percentage fat. Percentages protein and ash were essentially constant across lines and ages. A positive relation between rate and efficiency of growth was observed between lines. Consistent sex differences, males more efficient than females, were observed prior to 6 weeks of age, but were not evident in the later (6–8 week) data.

---

Zusammenfassung Bei Mäusen, die auf hohes (H₆) und geringes (L₆) 6 Wochen-Körpergewicht selektiert waren, und einer unselektierten Kontrollpopulation (C₁) wurde die Zuwachsrate, die Veränderung des Wasser-, Fett-, Protein- und Aschegehaltes während des Wachstums sowie die Wuchsleistung untersucht. Das Körpergewicht von der Geburt bis zum Alter von 6 Wochen wurde an 682 einzelnen Mäusen, die repräsentativ für die drei Linien sind, festgestellt. Die Ermittlung des Wasser-, Fett-, Asche- und (Rest-)Proteingehaltes der Tiere erfolgte wöchentlich für 180 Mäuse im Alter von 3–8 Wochen. Die Futterverwertung (Wuchsleistung) wurde für 189 drei bis acht Wochen alte Mäuse auf Grund von Einzelgewichten und Futterverbrauch geschätzt.Die Wachstumskurven für Körpergewicht und Gewichtszunahme, nach Linien und Geschlecht zusammengestellt, zeigten bei der L₆-Linie eine zeitweilige Verzögerung der maximalen Zuwachsrate, was z. T. einer längeren Wachstumsdepression nach der Entwöhnung zugeschrieben wird. Es ergab sich kein Beweis dafür, daß die schnellere Wachstumsrate bei der H₆-Linie auf eine Erhöhung der Fetteinlagerung im Verhältnis zu den anderen Komponenten zurückzuführen ist. Ein Rückgang des Fettgehalts, der bei den H₆- und C₁-Linien im Alter von 7 Wochen festzustellen war, wurde bei der Linie L₆ erst nach 8 Wochen sichtbar. Weibliche Tiere hatten im Alter von 4–7 Wochen einen höheren Körperfettgehalt als männliche, im Alter von 8 Wochen war dieser Unterschied aber nicht mehr vorhanden. Der Wassergehalt war in hohem Maße negativ mit dem Fettgehalt korreliert. Die Protein- und Ascheanteile waren im wesentlichen für die Linien und untersuchten Wuchsperioden konstant. Zwischen den Linien wurde eine positive Relation hinsichtlich Wuchsrate und Wuchsleistung festgestellt. Deutliche Geschlechtsunterschiede wurden bis zum Alter von 6 Wochen beobachtet, und zwar wuchsen die Männchen schneller als die Weibchen, später (6.–8. Woche) waren keine Unterschiede mehr feststellbar.

---

---

Paper number 2640 of the Journal Series of the North Carolina State University Agricultural Experiment Station, Raleigh, North Carolina. Supported by Public Health Service Grant GM 11546-05. Computation was supported by NIH Grant No. FR-00011. The senior author was supported by NIH Training Grant No. 2-Tl-GM-296.     

Sex allocation in a polygynous mammal with large litters: the wild boar

Predictions from Trivers & Willard's (1973, Science, 179, 90-92) hypothesis of sex-biased maternal investment in polygynous species do not apply well to species where mothers produce more than one offspring per reproductive attempt. First, as litter size increases, the benefits to the mother of adjusting sex ratio decrease because (1) she could benefit more by adjusting litter size and (2) sex differences in reproductive potential are negatively related to litter size. Second, testing sex-biased investment in these species requires predictions about the simultaneous adjustment of sex ratio and litter size. The wild boar, Sus scrofa, although polygynous, produces large litters. Here we present data for 58 litters from a free-ranging wild boar population in central Spain. Maternal expenditure per individual offspring, as measured by piglet weight, was higher for male than female fetuses. In more than 81% of cases the heaviest fetus in the litter was a male regardless of the quality of the mother; this might have influenced his ranking within the 'teat order' and consequently his development and survival. Mother quality (size and weight) appeared to be related to litter size but not to the sex ratio of the litter. However, it was highly related to a variable that combined the effects of litter size and sex ratio within the litter, thus supporting Williams' (1979, Proceedings of the Royal Society of London, Series B, 205, 567-580) hypothesis that mothers should adjust both litter size and offspring sex. Copyright 1999 The Association for the Study of Animal Behaviour.     

Activity-dependent local protection and lateral inhibition control synaptic competition in developing mitral cells in mice

1. Download : Download high-res image (182KB)
2. Download : Download full-size image

     

Development of 1-cell embryos from different strains of mice in CZB medium

One-cell embryos from several different strains of mice have been cultured to the blastocyst stage in CZB medium. CZB medium can be used to culture CF1 x B6SJLF1/J 1-cell embryos to the blastocyst stage provided glucose is introduced into the medium on Day 3 of culture. The amount of glucose required for embryo development was titrated using a concentration range of 5.5 to 49.5 mM. With the exception of the highest concentration, all glucose levels tested supported 65-85% development to the morula and blastocyst stages. Variations of CZB medium were tested for their ability to support the development of 1-cell embryos from 4 strains of mice. For embryos from CF1 and DBA/2J (both x B6SJLF1/J) mice, which exhibit a "2-cell block" to development in vitro, CZB medium containing glutamine with the addition of glucose on Day 3 supported optimum development from the 1-cell stage to morula and blastocysts (79% and 87%). For embryos from B6D2F1/J and CD1 female mice (both x B6SJLF1/J males), which do not exhibit a "2-cell block" to in vitro development, optimum development to morula and blastocyst stages (95% and 50%) was in CZB medium containing both glutamine and glucose from the start of culture.