Developmental programming of cardiovascular dysfunction by prenatal hypoxia and oxidative stress

Fetal hypoxia is a common complication of pregnancy. It has been shown to programme cardiac and endothelial dysfunction in the offspring in adult life. However, the mechanisms via which this occurs remain elusive, precluding the identification of potential therapy. Using an integrative approach at the isolated organ, cellular and molecular levels, we tested the hypothesis that oxidative stress in the fetal heart and vasculature underlies the molecular basis via which prenatal hypoxia programmes cardiovascular dysfunction in later life. In a longitudinal study, the effects of maternal treatment of hypoxic (13% O(2)) pregnancy with an antioxidant on the cardiovascular system of the offspring at the end of gestation and at adulthood were studied. On day 6 of pregnancy, rats (n = 20 per group) were exposed to normoxia or hypoxia ± vitamin C. At gestational day 20, tissues were collected from 1 male fetus per litter per group (n = 10). The remaining 10 litters per group were allowed to deliver. At 4 months, tissues from 1 male adult offspring per litter per group were either perfusion fixed, frozen, or dissected for isolated organ preparations. In the fetus, hypoxic pregnancy promoted aortic thickening with enhanced nitrotyrosine staining and an increase in cardiac HSP70 expression. By adulthood, offspring of hypoxic pregnancy had markedly impaired NO-dependent relaxation in femoral resistance arteries, and increased myocardial contractility with sympathetic dominance. Maternal vitamin C prevented these effects in fetal and adult offspring of hypoxic pregnancy. The data offer insight to mechanism and thereby possible targets for intervention against developmental origins of cardiac and peripheral vascular dysfunction in offspring of risky pregnancy.     

Lifespan ontogeny of breeding and reproductive success in Mongolian gerbils

An analysis was made of demographic data collected from a colony of Mongolian gerbils (Meriones unguiculatus). The data on births of over 3800 gerbils provide evidence on several aspects of development and fecundity. Approximately 4% more females than males survived to weaning. Mean number of pups per dam per year was estimated to be 48.3, greater reproductive success than reported previously. The shortest period of gestation resulting in viable pups was 25 days. Implantation was delayed by the presence of suckling pups. When gestation was prolonged, there was a direct relationship between the number of pups being suckled concurrently and the length of the delay. Length of gestation did not affect the size of the subsequent litter. Female gerbils gave birth as early as 72 days of age. Gerbils continued to reproduce until after 2 years of age in some cases. There were fewer pups per litter born to older females, fewer pups per litter with increasing parity, and longer intervals between litters with increasing age.     

Design and statistical methods in studies using animal models of development

Experiments involving neonates should follow the same basic principles as most other experiments. They should be unbiased, be powerful, have a good range of applicability, not be excessively complex, and be statistically analyzable to show the range of uncertainty in the conclusions. However, investigation of growth and development in neonatal multiparous animals poses special problems associated with the choice of "experimental unit" and differences between litters: the "litter effect." Two main types of experiments are described, with recommendations regarding their design and statistical analysis: First, the "between litter design" is used when females or whole litters are assigned to a treatment group. In this case the litter, rather than the individuals within a litter, is the experimental unit and should be the unit for the statistical analysis. Measurements made on individual neonatal animals need to be combined within each litter. Counting each neonate as a separate observation may lead to incorrect conclusions. The number of observations for each outcome ("n") is based on the number of treated females or whole litters. Where litter sizes vary, it may be necessary to use a weighted statistical analysis because means based on more observations are more reliable than those based on a few observations. Second, the more powerful "within-litter design" is used when neonates can be individually assigned to treatment groups so that individuals within a litter can have different treatments. In this case, the individual neonate is the experimental unit, and "n" is based on the number of individual pups, not on the number of whole litters. However, variation in litter size means that it may be difficult to perform balanced experiments with equal numbers of animals in each treatment group within each litter. This increases the complexity of the statistical analysis. A numerical example using a general linear model analysis of variance is provided in the Appendix. The use of isogenic strains should be considered in neonatal research. These strains are like immortal clones of genetically identical individuals (i.e., they are uniform, stable, and repeatable), and their use should result in more powerful experiments. Inbred females mated to males of a different inbred strain will produce F1 hybrid offspring that will be uniform, vigorous, and genetically identical. Different strains may develop at different rates and respond differently to experimental treatments.     

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.     

Warming and grazing enhance litter decomposition and nutrient release independent of litter quality in an alpine meadow

增温和放牧对高寒草甸凋落物分解及其养分释放的影响不依赖于凋落物品质在放牧生态系统中,增温、放牧和凋落物品质共同决定着凋落物分解和养分释放。然而,在以往的研究中这些因子的效应通常被单独地研究。在本研究中,我们在青藏高原高寒草甸开展了一个昼夜非对称增温和中度放牧两因子的凋落物分解试验。从每个处理中收集了凋落物样品,这些凋落物一部分放在它们的来源处理小区,另一部分放在其他处理小区以此来探究增温、放牧以及凋落物品质对凋落物分解和养分释放的影响。研究结果表明,增温而不是放牧显著增加了凋落物质量的损失、单位面积全碳、全氮以及全磷含量的损失,这主要是因为增温增加了凋落物生物量和分解速率。然而,尽管同时增温放牧处理也加快了凋落物分解速率,但由于降低了凋落物生物量,所以增温放牧处理并没有显著影响单位面积的凋落物碳和养分释放量。相比木质素含量和碳氮比而言,季节性土壤平均温度能够更好地预测凋落物分解速率。增温和放牧对凋落物分解存在交互作用,但它们和凋落物品质对凋落物的影响均不存在交互作用。单位面积的总氮释放的温度敏感性要高于总磷。因此,我们的结果表明,增温对凋落物分解以及养分释放的影响要显著大于凋落物品质变化对其分解的影响。在高寒草甸,氮释放的增加可能会间接导致土壤磷有效性的缺乏。     

Modelling sub-binomial variation in the frequency of sex combinations in litters of pigs

In some samples of pig litters, the number of males per litter exhibits significant sub-binomial dispersion: there are fewer unisexual and more "sex-balanced" litters than expected under a binomial model. It is argued here that the explanation is that the sex of zygotes may be subject to hormonal control with the consequence that, within a litter, the zygotes do not all have the same probability of being male. This variation in probability is approximated by a quadratic function of the time during oestrus that a zygote is formed so as to parallel the variation in maternal hormone levels. Other explanations are examined briefly, but each seems to have serious weaknesses.     

A generalized estimating equations approach to quantitative trait locus detection of non-normal traits

To date, most statistical developments in QTL detection methodology have been directed at continuous traits with an underlying normal distribution. This paper presents a method for QTL analysis of non-normal traits using a generalized linear mixed model approach. Development of this method has been motivated by a backcross experiment involving two inbred lines of mice that was conducted in order to locate a QTL for litter size. A Poisson regression form is used to model litter size, with allowances made for under- as well as over-dispersion, as suggested by the experimental data. In addition to fixed parity effects, random animal effects have also been included in the model. However, the method is not fully parametric as the model is specified only in terms of means, variances and covariances, and not as a full probability model. Consequently, a generalized estimating equations (GEE) approach is used to fit the model. For statistical inferences, permutation tests and bootstrap procedures are used. This method is illustrated with simulated as well as experimental mouse data. Overall, the method is found to be quite reliable, and with modification, can be used for QTL detection for a range of other non-normally distributed traits.     

Developmental toxicity of orally administered 2',3'-dideoxycytidine in mice

2',3'-Dideoxycytidine (DDC), a potent inhibitor of human immunodeficiency virus (HIV), is presently undergoing clinical trials as a promising anti-AIDS drug. Since there are very limited published animal toxicity data available, and nucleoside analogues are being considered for treatment of HIV-infected pregnant women, a study was conducted in mice to investigate the potential adverse developmental effects of this drug. DDC, suspended in 0.5% methyl cellulose, was administered via gavage twice per day during gestation days (gd) 6 through 15 to C57Bl/6N mice in a total dose of 0, 200, 400, 1,000, or 2,000 mg/kg/day. Maternal weight gain during the gestation and treatment period, as well as gravid uterine weight, decreased significantly in the 2,000 mg group, but weight gain, corrected for gravid uterine weight, was not affected by DDC. The percent resorptions per litter increased significantly in the highest dose group, and there were fewer live litters because of complete litter resorption in six dams. Among litters with live fetuses, the mean litter size was significantly reduced in the 2,000 mg group. Average fetal body weight per litter decreased significantly in the 1,000 and 2,000 mg groups. The number of fetuses with any malformation, the number of litters with one or more malformed fetuses and the percent of malformed fetuses per litter increased significantly in the 1,000 and 2,000 mg groups. There was an increase in malformations at 400 mg/kg/day; however, it was not statistically significant. In conclusion, DDC produced developmental toxicity (malformations, reduced fetal body weight, and resorptions) in the absence of overt maternal toxicity except for body weight changes due to resorptions and reduced fetal weights.     

Placental insulin‐like growth factor II (IGF‐II) and its relation to litter size in the common marmoset monkey (Callithrix jacchus)

The primate placenta produces a wide variety of hormones throughout gestation that regulate placental function and fetal growth. One such hormone is insulin‐like growth factor‐II (IGF‐II), a peptide implicated in cell division, differentiation, and amino acid transport. IGF‐II concentrations were measured in 23 common marmoset (Callithrix jacchus) term placentas from twin and triplet litters in order to determine whether previously described differences in fetoplacental phenotype such as placental and litter mass and placental surface area were related to differences in endocrine function. IGF‐II was extracted from frozen tissue samples and measured using an enzyme‐linked immunosorbent assay kit designed for human tissue, which was validated for marmoset placenta. IGF‐II concentrations were not related to placental or litter mass, and twin and triplet placentas did not differ in total concentration. However, per individual fetus, triplets were associated with a significant 42% reduction in IGF‐II concentration (P=0.03), and IGF‐II concentration per gram of fetal mass was a third lower in triplet litters. The triplet placenta exhibits a global expansion of the surface area which was contrasted by a per unit area reduction in IGF‐II concentration (r=?0.75, P=0.01), a pattern that explains why twin and triplet placentas overall did not differ in concentration. Per fetus, triplet pregnancies are associated with relatively less maternal mass, placental mass and microscopic surface area suggesting that the intrauterine growth of triplets is supported by systems that increase the efficiency of nutrient transfer. The finding that individual triplet fetuses are also associated with significantly lower IGF‐II concentrations is consistent with the view that the marmoset fetoplacental unit exhibits a flexible pattern of placental allocation and metabolism. Plasticity in placental endocrine and metabolic function is likely to play an important role in the ability of the fetus to sense and accommodate the intrauterine environment and, by extension, the external ecology. Am. J. Primatol. 71:969–975, 2009. © 2009 Wiley‐Liss, Inc.     

Analysis of litter size and average litter weight in pigs using a recursive model

An analysis of litter size and average piglet weight at birth in Landrace and Yorkshire using a standard two-trait mixed model (SMM) and a recursive mixed model (RMM) is presented. The RMM establishes a one-way link from litter size to average piglet weight. It is shown that there is a one-to-one correspondence between the parameters of SMM and RMM and that they generate equivalent likelihoods. As parameterized in this work, the RMM tests for the presence of a recursive relationship between additive genetic values, permanent environmental effects, and specific environmental effects of litter size, on average piglet weight. The equivalent standard mixed model tests whether or not the covariance matrices of the random effects have a diagonal structure. In Landrace, posterior predictive model checking supports a model without any form of recursion or, alternatively, a SMM with diagonal covariance matrices of the three random effects. In Yorkshire, the same criterion favors a model with recursion at the level of specific environmental effects only, or, in terms of the SMM, the association between traits is shown to be exclusively due to an environmental (negative) correlation. It is argued that the choice between a SMM or a RMM should be guided by the availability of software, by ease of interpretation, or by the need to test a particular theory or hypothesis that may best be formulated under one parameterization and not the other.     

Mammalian reproductive strategies: A generalized relation of litter size to body size

Summary A generalized relationship of litter size to mammalian body size was predicted by a graph model. The model was used to generate hypotheses explaining specific features of variation in gestation time, relative litter weight, birth weight, and reproductive capacity. The predictions were tested by means of data from the literature.Mammals were assumed to maximize neonatal survival of offspring to the limits allowed by litter weight per female body weight. Gestation time correlated negatively with the foetal growth rate of relative litter weight. Gestation time did not correlate with the foetal growth rate of individual offspring.Relative litter weight correlated negatively with adult body weight. This relationship was explained by the higher assimilation rate per unit weight relative to metabolic rate in small mammals.Birth weight correlated positively with body weight. However, small mammals produce larger offspring than predicted by the linear relationship of birth weight to body weight in large mammals. There is obviously a minimum birth weight which cannot be decreased without special arrangements for parental care.The prediction of the relationship of litter size to body size was derived from the relations of relative litter weight and birth weight to body weight. In small mammals (less than 1 kg) litter the correlation was negative. When litter size was compared with body length, the correlation was positive in small mammals (less than 30 cm) and negative in large mammals. In both sets of data there was a negative overall correlation between litter size and body size.Reproductive capacity, defined as the number of offspring per season, correlated negatively with life-span.     

Reciprocal effects of litter from exotic and congeneric native plant species via soil nutrients

Invasive exotic plant species are often expected to benefit exclusively from legacy effects of their litter inputs on soil processes and nutrient availability. However, there are relatively few experimental tests determining how litter of exotic plants affects their own growth conditions compared to congeneric native plant species. Here, we test how the legacy of litter from three exotic plant species affects their own performance in comparison to their congeneric natives that co-occur in the invaded habitat. We also analyzed litter effects on soil processes. In all three comparisons, soil with litter from exotic plant species had the highest respiration rates. In two out of the three exotic-native species comparisons, soil with litter from exotic plant species had higher inorganic nitrogen concentrations than their native congener, which was likely due to higher initial litter quality of the exotics. When litter from an exotic plant species had a positive effect on itself, it also had a positive effect on its native congener. We conclude that exotic plant species develop a legacy effect in soil from the invaded range through their litter inputs. This litter legacy effect results in altered soil processes that can promote both the exotic plant species and their native congener.     

Effects of alcohol (ethanol) on the fetus

Alcohol (ethanol) use during pregnancy can produce a wide spectrum of effects in the developing embryo/fetus that are dependent on the maternal drinking pattern. The effects of chronic ethanol exposure on the developing conceptus are reviewed with primary focus on ethanol teratogenesis, manifesting in the human as the fetal alcohol syndrome or fetal alcohol effects. The effects of acute ethanol exposure on the near-term fetus are described, including suppressed fetal breathing movements, electrocorticographic (ECoG) activity and electrooculographic (EOG) activity. The ethanol-induced suppression of fetal breathing movements is a very sensitive index of acute exposure of the near-term fetus to ethanol, and appears to involve a direct mechanism of action rather than an indirect mechanism involving suppression of electrocortical activity. The disposition of ethanol and its pharmacologically active proximate metabolite, acetaldehyde, and the activity of alcohol dehydrogenase and aldehyde dehydrogenase in the near-term maternal-fetal unit are described, and a pharmacokinetic model is proposed. The effects of short-term ethanol exposure on the near-term fetus include the development of tolerance to the ethanol-induced suppression of fetal breathing movements, low-voltage ECoG activity and EOG activity. The development of tolerance occurs more rapidly to the latter two fetal biophysical activities. The mechanism of tolerance development appears to be pharmacodynamic (functional) in nature, as there is no increase in the rate of ethanol elimination from the maternal-fetal unit. The role of prostaglandins (PGs) in the mechanism of the ethanol-induced suppression of fetal breathing movements is described. In the near-term fetus, there is a direct relationship between fetal blood ethanol concentration and fetal plasma PGE2 concentration, and an inverse relationship between the incidence of fetal breathing movements and each of fetal plasma and fetal cerebrospinal fluid (CSF) PGE2 concentrations. Indomethacin, a PG synthetase inhibitor, selectively blocks and reverses the ethanol-induced suppression of fetal breathing movements. These data support the postulates that the ethanol-induced suppression of fetal breathing movements is mediated by increased PGE2 concentration in the near-term fetus and that the ability of indomethacin to antagonize the ethanol-induced suppression of fetal breathing movements is due to its biochemical action to decrease fetal PGE2 concentration.(ABSTRACT TRUNCATED AT 400 WORDS)     

南滑蜥繁殖行为和繁殖策略的研究

南滑蜥Scincella reevesii 系温暖气候区卵胎生蜥蜴,每年繁殖1次,繁殖季节为5月下旬到6月上旬.平均每只繁殖雌蜥产仔6(3～9)条.雌蜥分娩持续时间为30～ 60 min,仔蜥在产出时包被卵膜.南滑蜥雌体采取高投入的繁殖策略,平均繁殖投入为0.359,平均窝仔投人为0.247.相关性回归分析表明,窝仔数、窝仔重与雌蜥体长呈正相关关系,幼仔重与雌蜥体长不相关;窝仔数与雌蜥的繁殖投入及窝仔投入均成正相关,与分娩时的能耗投入不相关,与幼仔投入成显著负相关.     

Ground layer heterogeneity and hardwood regeneration in mixed oak forest

Question: What are the effects of small‐scale litter disturbances (simulating large vertebrate scratching and foraging), on Quercus alba, Q. velutina, Fagus grandifolia, and Acer rubrum seedling recruitment? Location: Southeastern Ohio, USA. Methods: Two mixed oak forests containing four experimental management units (burned, thinned, thinned & burned, and un‐manipulated control) were utilized in this study. Silvicutural treatments were applied in the spring of 2001. A small scale disturbance experiment was initiated in the spring of 2002 (Trial 1) and was replicated again in the spring of 2003 (Trial 2). Experimental hardware cloth exclosures, each containing a control (ambient litter) and experimentally scratched (litter removed to bare soil) compartment, were erected in each management unit (N= 8 exclosures per unit). Results: Acer rubrum, a species that occupies a wide germination niche, produced the most numerous seedlings. Nut‐producing species established more readily in compartments with ambient leaf litter, while A. rubrum was unresponsive to scratching treatment. Units burned (surface leaf litter removed at a broad scale) had the lowest rate of seedling recruitment in Trial 1. Conclusions: These data suggest that an adequate cover of leaf litter is needed to promote optimal recruitment. Control units had the greatest rate of seedling recruitment in Trial 2. The development of a dense understory layer (promoting excessive shading) between Trial 1 and Trial 2 may have affected the recruitment rate in the thinned and thinned & burned units.     

NIH/Ola: a highly productive inbred strain of laboratory mouse

According to historical records the NIH/Ola strain was developed from outbred 'Swiss' mice imported into the USA by Dr C. Lynch in 1926. A comparison of biochemical markers in strain NIH/Ola and other strains such as SJL and SWR derived from the same foundation stock supports the historical records. Data on litter size, length of gestation, bodyweight to 70 days of age, and reproductive performance when housed in polygamous groups of up to 10 females per male were compared with similar data on inbred CBA/CaOla and C57BL/10ScSnOla mice. NIH/Ola inbred mice had an exceptional reproductive performance, producing about 1.8 young weaned/female/week when housed as monogamous pairs over a period of 20 weeks, compared with less than 1.0 young/female/week with the other 2 strains. NIH/Ola mice were also extremely tolerant to mating in polygamous groups of up to about 8 females per male. Mating ratios of over 2-3 females per male resulted in a marked decline in the total number of litters produced per female in C57BL/10ScSnOla and CBA/CaOla, but this reduction was not nearly so marked in NIH/Ola. It is concluded that the NIH/Ola inbred strain may be particularly useful in studies such as teratology where a high reproductive performance needs to be combined with the advantages of a fully inbred strain.     

Transfer of maternal plasma free fatty acids into the rat fetus.

The amount of maternal plasma free fatty acids passing to the fetus has been determined to be 0.09 mumoles fatty acids per min per each litter. Taking account of the increase of the total fetal fatty acid pool due to the fetal growth (0.2 mumoles fatty acids per min for each litter) we conclude that the maternal circulation is the source of about half of fetal fatty acids on day 21 of pregnancy.     

Diet and breeding performance in cats

A conventional cat breeding colony with 70 queens (female cats) was studied during a 4 year period 1979-1982. During that time the fat content in the diet was increased from 15% to 27% of dry matter. An increase in the number of kittens per litter (from 4.5 to 5.5) and in the annual number of litters per queen (from 1.4 to 2.3) was found. In addition, the mortality decreased from over 20% to 9%. Bodyweight gain under the new diet was such that the males reached 2500 g in 4 months while the females showed this same weight at 5 months of age. Litter size and sex distribution as a function of queen age, litter interval and time of year are presented. It is concluded that husbandry and diet are factors which are of great importance in a cat breeding unit. It is shown that under our conditions it is possible to breed conventional cats with good results.     

Tumorigenicity Assays,Including Use of the Jackknife

The complexities of long-term tumorigenicity assays are discussed and statistical approaches suggested. Account must be taken of how tumors are discovered, and a reasonable distinction might be made between self-evidencing tumors (palpable tumors and rapidly lethal tumors) and those identified only on necropsy. Tumor autolysis can pose difficulties. The nature of the experimental design must be taken into account-the study can be completely randomized, or may involve litter-matching, or circumstances may require that all members of a litter be treated alike. The use of a logrank procedure or a closely equivalent Mantel-Haenszel procedure can provide a unifying approach, but modification needs to be made so as to take into account-data on non-self-evidencing tumors. Censoring plays a role, whether because of eventual study termination or because losses to competing risks are treated as censored observations. Where there are several non-zero dosage levels, the analysis can be modified according to the dosage metric used. Multiplicity of statistical tests can arise whether because a number of questions are asked or because a multiplicity of tumor sites have to be considered. Some of the kinds of questions raised have been addressed in earlier papers by various of the present authors, to which the reader is referred. Specifically new material is given on use of the jackknife so as to take into account that the unit of observation is the litter or even the group of litters caged together. Various problems were found to attend the application of the jackknife in such situations.     

母猪繁殖力性状影响因素分析及遗传参数估计

母猪繁殖力是影响种猪场经济效益的重要因素。文章对纯种大白猪、长白猪、杜洛克猪的8491窝产仔记录进行统计分析,建立固定效应模型对总产仔数、健仔数、初生窝重、弱仔数、死胎数、木乃伊胎数和畸形胎数共7个繁殖性状进行最小二乘分析,分析胎次、配种季节、品种对母猪繁殖性能的影响,同时比较了纯繁和杂交的效果。利用动物模型REML(约束最大似然)方法估计繁殖性状的遗传力和遗传相关。结果表明,胎次、配种季节和品种对总产仔数、健仔数、初生窝重影响极显著(P<0.001),胎次和品种对弱仔数影响极显著(P<0.001),但配种季节对弱仔数影响不显著。胎次对死胎数影响显著(P<0.05),而配种季节和品种对死胎数影响不显著,胎次、配种季节和品种对木乃伊胎、畸形胎数影响不显著。长白♂×大白♀交配组合具有最高的总产仔数、健仔数和初生窝重。繁殖性状遗传力估计结果显示,长白猪初生窝重的遗传力最高,为0.227。其余性状遗传力均在0.2以下,为低遗传力性状。3个品种母猪的健仔数与初生窝重、总产仔数与健仔数之间的遗传相关达0.96以上。研究结果为降低非传染性因素造成的种母猪产仔数低的问题以及种猪场对母猪繁殖力的选育提高提供了参考数据和理论依据。