Prenatal alcohol exposure shortens life span in rats

In a study of the effects of in utero alcohol exposure on life span in rats, pregnant rats were intubated twice daily with 3.5 gm/kg alcohol on gestational days 11-21 or with an isocaloric sucrose solution. These latter animals were pair-fed and pair-watered to alcohol-treated animals. A third group served as nontreated ad lib-fed controls. At birth, all offspring were removed from their biological mothers, culled to eight per litter, and placed with nontreated surrogate dams. Alcohol-exposed animals died at a significantly younger age than pair-fed and ad lib controls and never attained the same maximum body weights as control animals. For females prenatally exposed to alcohol, life span was shortened by about 20 weeks; in male cohorts, life span was shortened by about 2.5-7 weeks.     

Scintillation autoradiography at the light microscopic level: A review

Summary This article reviews the reported attempts to expose autoradiographs, intended for examination by light microscopy, in the presence of scintillation fluid in order to increase efficiency and thus shorten exposure times. The scintillation process is reviewed, together with the use of comparable techniques in the autoradiography of chromatograms and electrophoresis gels. The conflicting reports on the usefulness of scintillation autoradiography in light microscopy are then discussed, and explanations sought for the wide diversity of claims made for the technique. Many of the more optimistic claims can be explained on the basis that the normal autoradiographs used for comparison had unduly low efficiencies caused by inadequate drying of the emulsion and consequent latent image fading. The techniques used are unlikely to have produced any increase in efficiency through the scintillation process. Optimal conditions for obtaining such increases are derived from theoretical considerations.     

Social stress shortens lifespan in mice

Stress and low socioeconomic status in humans confer increased vulnerability to morbidity and mortality. However, this association is not mechanistically understood nor has its causation been explored in animal models thus far. Recently, cellular senescence has been suggested as a potential mechanism linking lifelong stress to age‐related diseases and shorter life expectancy in humans. Here, we established a causal role for lifelong social stress on shortening lifespan and increasing the risk of cardiovascular disease in mice. Specifically, we developed a lifelong chronic psychosocial stress model in which male mouse aggressive behavior is used to study the impact of negative social confrontations on healthspan and lifespan. C57BL/6J mice identified through unbiased cluster analysis for receiving high while exhibiting low aggression, or identified as subordinate based on an ethologic criterion, had lower median and maximal lifespan, and developed earlier onset of several organ pathologies in the presence of a cellular senescence signature. Critically, subordinate mice developed spontaneous early‐stage atherosclerotic lesions of the aortic sinuses characterized by significant immune cells infiltration and sporadic rupture and calcification, none of which was found in dominant subjects. In conclusion, we present here the first rodent model to study and mechanistically dissect the impact of chronic stress on lifespan and disease of aging. These data highlight a conserved role for social stress and low social status on shortening lifespan and increasing the risk of cardiovascular disease in mammals and identify a potential mechanistic link for this complex phenomenon.     

Aneuploidy shortens replicative lifespan in Saccharomyces cerevisiae

Aneuploidy and aging are correlated; however, a causal link between these two phenomena has remained elusive. Here, we show that yeast disomic for a single native yeast chromosome generally have a decreased replicative lifespan. In addition, the extent of this lifespan deficit correlates with the size of the extra chromosome. We identified a mutation in BUL1 that rescues both the lifespan deficit and a protein trafficking defect in yeast disomic for chromosome 5. Bul1 is an E4 ubiquitin ligase adaptor involved in a protein quality control pathway that targets membrane proteins for endocytosis and destruction in the lysosomal vacuole, thereby maintaining protein homeostasis. Concurrent suppression of the aging and trafficking phenotypes suggests that disrupted membrane protein homeostasis in aneuploid yeast may contribute to their accelerated aging. The data reported here demonstrate that aneuploidy can impair protein homeostasis, shorten lifespan, and may contribute to age‐associated phenotypes.     

Nitrate deficiency shortens the circadian period in gonyaulax

The circadian rhythms in bioluminescence and photosynthesis in Gonyaulax polyedra suspended in unsupplemented sea water have been compared to the same rhythms in f/2, an enriched seawater medium. Cells suspended in sea water for 2 days in continuous light (450 microwatts per square centimeter) showed significantly shorter circadian periods and lower amplitudes than did cells in f/2 medium (a period of 22.2 hours as compared to 23.5 hours). Both period and amplitude changes could be completely reversed by the addition of nitrate at one-fourth or more of the concentration in f/2 medium (0.88 millimolar). The addition to autoclaved seawater of phosphate, vitamins, minerals, or soil extract in concentrations present in f/2 medium had no effect. Thus, the shortening of the circadian period is the consequence of reduced nitrogen supply. Since both the rhythms in bioluminescence and photosynthesis showed similarly shortened circadian periods and lower amplitudes, it is probable that the depletion of nitrate directly affects the circadian clock.     

Acute exposure to low-dose radiation disrupts reproduction and shortens survival of Wasmannia auropunctata (Hymenoptera: Formicidae) queens

Irradiation is a postharvest quarantine treatment option to control ants and other hitchhiker pests on fresh horticultural products exported from Hawaii. The radiotolerance of the invasive little fire ant, Wasmannia auropunctata (Roger) (Hymenoptera: Formicidae: Myrmicinae), was studied to determine a dose sufficient for its control. Queens from each of five laboratory microcolonies started from five geographic locations in Argentina were irradiated at 20, 50, 70, or 100 Gy or left untreated as controls and then followed for 13 wk to observe colony growth. In general, queen survivorship, and the number of eggs, larvae, and pupae observed in the microcolonies decreased with increasing irradiation dose. In the 50-, 70-, and 100-Gy treatments, the number of eggs observed was reduced by 68, 66, and 76%, respectively, compared with untreated control microcolonies. The number of larvae in the 50-, 70-, and, 100-Gy treatments was reduced by 99.6%, and only one pupa was observed in the 50-Gy treatment and none in the 70- and 100-Gy treatments during the 13-wk experiment. Queens in the 100-Gy treatment had significantly reduced longevity compared with queens in the other treatments. Radiation doses > or = 70 Gy stopped reproduction in W. auropunctata queens and should be sufficient as a phytosanitary treatment. Information from additional invasive ants in Myrmicinae and other subfamilies is needed before recommending a generic irradiation treatment for ants.     

Vitrification of mouse oocytes using short cryoprotectant exposure: effects of varying exposure times on survival.

The effects on oocyte viability of varying the duration of exposure to cryoprotectants before rapid cooling to -196 degrees C were examined, using the vitrification protocol of Nakagata. A very short exposure (15 sec) was found to be optimal, resulting in an overall rate of development from vitrified oocytes to hatching blastocysts of 31.8%. Very high rates of survival (77-89%) of oocytes exposed to the cryoprotectant media, but without the vitrification, together with extreme variability in results between straws in the vitrified groups, suggest that losses in viability during vitrification may result from ice damage during devitrification of the medium.     

Automated image analysis in autoradiography

Light microscope autoradiographs can be analysed with the aid of the Quantimet, a commercially available image scanning instrument. Standardization of the performance of the instrument was obtained by performing on-line data reduction and display with a small digital computer. The number of silver grains over individual cells can be rapidly and routinely determined, and excellent agreement can be achieved between visually determined and machine generated grain counts.     

Unsteady near wall residence times and shear exposure in model distal arterial bypass grafts

Building on previous studies of unsteady flow within model distal bypass grafts we analyse the near wall residence times and shear exposure in a 45 degrees anastomosis under symmetrical and symmetry breaking geometric configurations. We define residence time as the minimum time for a particle to exit a spherical region and shear exposure as a temporal integral of the Huber-Henky-von-Mises criterion along a particle path over a fixed time interval. Decomposing the pulsatile cycle into four equal intervals we find that the interval of peak residence time in the host vessel is from mid-deceleration to peak diastole and peak diastole to mid-acceleration. The asymmetric model is shown to have a significantly lower residence time during these intervals. Considering the shear exposure prior to the residence time evaluation we determine that a higher average shear exposure exists in the asymmetric model associated with the upstream geometry modification. Analysis of the regions of high residence time and shear exposure suggests that the "toe" region and the interface between the "heel" and bulk flow are more significant than the bed and heel region. Although the asymmetric model considered in this study reduces residence times in the host artery, the product of the measure of shear exposure and residence time is not found to be preferable. If shear exposure were to be considered as an important factor in particle activation, the findings imply that for junction optimisation, greater consideration needs to be given both to the local junction asymmetry and upstream influence on the shear history.     

Application of MOSFET detectors for dosimetry in small animal radiography using short exposure times

Digital subtraction angiography (DSA) X-ray imaging for small animals can be used for functional phenotyping given its ability to capture rapid physiological changes at high spatial and temporal resolution. The higher temporal and spatial requirements for small-animal imaging drive the need for short, high-flux X-ray pulses. However, high doses of ionizing radiation can affect the physiology. The purpose of this study was to verify and apply metal oxide semiconductor field effect transistor (MOSFET) technology to dosimetry for small-animal diagnostic imaging. A tungsten anode X-ray source was used to expose a tissue-equivalent mouse phantom. Dose measurements were made on the phantom surface and interior. The MOSFETs were verified with thermoluminescence dosimeters (TLDs). Bland-Altman analysis showed that the MOSFET results agreed with the TLD results (bias, 0.0625). Using typical small animal DSA scan parameters, the dose ranged from 0.7 to 2.2 cGy. Application of the MOSFETs in the small animal environment provided two main benefits: (1) the availability of results in near real-time instead of the hours needed for TLD processes and (2) the ability to support multiple exposures with different X-ray techniques (various of kVp, mA and ms) using the same MOSFET. This MOSFET technology has proven to be a fast, reliable small animal dosimetry method for DSA imaging and is a good system for dose monitoring for serial and gene expression studies.     

Weekly bull exchange shortens postpartum anestrus in suckled beef cows

The duration of anestrus in cattle was usually shortened when cows were exposed to bulls. The objective of the present experiment was to determine if weekly bull exchange accelerated the resumption of cyclicity in postpartum suckled beef cows. We tested the hypothesis that exposure of postpartum, anestrous, suckled beef cows (extensively managed) to weekly exchange of bulls, accelerates the resumption of cyclicity, compared to continuous exposure to the same bulls. Ninety-one multiparous suckled Hereford and Hereford x Angus cows, <60d postpartum, were assigned to two homogeneous groups. Beginning on December 1st (late spring), the control group (C, n=45) remained with one pair of bulls throughout the breeding period (7 weeks), whereas in the "exchanged" (E, n=46) group two pairs of bulls were exchanged weekly. Based on weekly ultrasonographic examinations of all cows, none had a CL at the start of the experiment and for 2 weeks after the start of bull exposure. However, the accumulated frequency of cows with a CL was greater in group E than in group C cows on week 4 (P=0.024), as well as on weeks 5-7 (P<0.001) after the start of bull exposure. Furthermore, in group E versus group C, there was a higher pregnancy rate 30d after the end of bull exposure (26 of 46, 56.2% vs. 16 of 45, 35.6%; P=0.045). In conclusion, weekly exchange of two pairs of bulls shortened postpartum anestrus in suckled multiparous cows, compared to continuous exposure to a single pair of bulls.     

Phenidone-ascorbic acid development in electronmicroscopic autoradiography

Summary Phenidone-ascorbic acid development was calibrated for electronmicroscopic autoradiography, using Ilford L4 as photographic emulsion and microdol-x as reference developer. Grain yield and efficiency were studied on pale gold sections of uniformly labeled tritium methacrylate. For determination of the resolution, a radioactive line source was prepared by crosssectioning of an epon-embedded film of tritium labeled albumin. The spatial relationship between silver grains and silver bromide crystals was investigated by shadowing the emulsion with platinumcarbon before development. In shadowed autoradiographs both, silver grains and silver bromide crystal were visible.Phenidone was about twice as sensitive as microdol-x and had a half distance value (Salpeter et al., 1969) of 175 mm. Most of the silver grains of both developers were located within the perimeters of their parent silver bromide crystals. In the case of phenidone more than 80% of the excited crystals gave rise to just one silver deposit. These parameters, together with grain size and shape, and counting feasibility make phenidone a useful developer for quantitative EM-autoradiography.     

Telomerase enzymatic component hTERT shortens long telomeres in human cells

Telomere lengths are tightly regulated within a narrow range in normal human cells. Previous studies have extensively focused on how short telomeres are extended and have demonstrated that telomerase plays a central role in elongating short telomeres. However, much about the molecular mechanisms of regulating excessively long telomeres is unknown. In this report, we demonstrated that the telomerase enzymatic component, hTERT, plays a dual role in the regulation of telomere length. It shortens excessively long telomeres and elongates short telomeres simultaneously in one cell, maintaining the optimal telomere length at each chromosomal end for efficient protection. This novel hTERT-mediated telomere-shortening mechanism not only exists in cancer cells, but also in primary human cells. The hTERT-mediated telomere shortening requires hTERT’s enzymatic activity, but the telomerase RNA component, hTR, is not involved in that process. We found that expression of hTERT increases telomeric circular DNA formation, suggesting that telomere homologous recombination is involved in the telomere-shortening process. We further demonstrated that shelterin protein TPP1 interacts with hTERT and recruits hTERT onto the telomeres, suggesting that TPP1 might be involved in regulation of telomere shortening. This study reveals a novel function of hTERT in telomere length regulation and adds a new element to the current molecular model of telomere length maintenance.     

临近闪烁分析技术在生命科学研究中的新进展

起源于放射免疫分析的临近闪烁分析法(scintillation proximity assay,SPA)是一种均相、灵敏、快速和简便的基于闪烁载体的分析平台.放射性标记分子和亲和标签分子的多样化和商业化,以及液闪计数器和液相操作等技术的发展,使得SPA操作具有无需分离、被测目标分子易于固定和检测的优势.SPA已成为高通量检测的重要方法,广范应用于筛选药物靶点及先导化合物筛选、药物作用机理研究,以及病毒、肿瘤标志物等抗原的高通量检测.本文在简要介绍SPA技术的原理及系统分析SPA操作中环境、条件对测试结果影响的基础上,比较全面地介绍了SPA技术在生物学研究中的应用,尤其是在膜蛋白和细胞水平上的应用进展,介绍了应用实例,归纳了国内研究人员开展相关技术应用与技术创新的制约因素,给出了未来的发展趋势.基于SPA的生物学应用技术创新必然提升我们对细胞体系生物学的全面理解.