Axial resolution enhancement of light‐sheet microscopy by double scanning of Bessel beam and its complementary beam

The side lobes of Bessel beam will create significant out‐of‐focus background when scanned in light‐sheet fluorescence microscopy (LSFM), limiting the axial resolution of the imaging system. Here, we propose to overcome this issue by scanning the sample twice with zeroth‐order Bessel beam and another type of propagation‐invariant beam, complementary to the zeroth‐order Bessel beam, which greatly reduces the out‐of‐focus background created in the first scan. The axial resolution can be improved from 1.68 μm of the Bessel light‐sheet to 1.07 μm by subtraction of the two scanned images across a whole field‐of‐view of up to 300 μm × 200 μm × 200 μm. The optimization procedure to create the complementary beam is described in detail and it is experimentally generated with a spatial light modulator. The imaging performance is validated experimentally with fluorescent beads as well as eGFP‐labeled mouse brain neurons.      

Cryopreservation of mouse embryoid bodies

Cryopreservation of embryonic stem (ES) cells is essential to establish them as a resource for regenerative therapy. We evaluated survival adhesion rate, cell structure, gene expression, and multipotency of frozen and thawed embryoid bodies (EBs) derived from mouse ES cells. EBs were cryopreserved using the BICELL and the Program Freezer. After one week the EBs were thawed and cultured. EBs prepared in the Program Freezer had the highest survival adhesion (Program Freezer; 55-69%, BICELL; 30-38%). Though many cells in the thawed EBs were damaged, some were not, especially those prepared in the Program Freezer. RT-PCR analysis showed that genes characteristic of the three embryonic germ layers were expressed in thawed EBs cultured for one week. EBs transplanted into mice formed teratomas consisting of cells derived from the three germ layers. In conclusion, EBs frozen in the Program Freezer had higher survival adhesion rates compared to the BICELL and formed differentiated cells characteristic of the three embryonic germ layers.     

Short-term effects of a "health-at-every-size" approach on eating behaviors and appetite ratings

Objective: To assess the effects of a “Health‐At‐Every‐Size” (HAES) intervention on eating behaviors and appetite ratings in 144 premenopausal overweight women. Research Methods and Procedures: Women were randomly assigned to one of the 3 groups: HAES group, social support (SS) group, and control group (N = 48 in each group). Interventions were conducted over a 4‐month period, and measurements were taken before and after this period. Eating behaviors (cognitive dietary restraint, disinhibition, and susceptibility to hunger) were evaluated by the Three‐Factor Eating Questionnaire. Appetite ratings (desire to eat, hunger, fullness, and prospective food consumption) were assessed by visual analogue scales before and after a standardized breakfast. Results: More important decreases in susceptibility to hunger and external hunger were observed in the HAES group when compared with the SS group (p = 0.05, for susceptibility to hunger) and the control group (p = 0.02 and p = 0.005, for susceptibility to hunger and external hunger, respectively). In addition, women from the HAES group had more important decreases in postprandial area under the curve for desire to eat (p = 0.02) and hunger (p = 0.04) when compared with the control group. The change in the desire to eat noted in the HAES group was also different from the one observed in SS group (p = 0.02). Women from the HAES group experienced significant weight loss at 4 months (?1.6 ± 2.5 kg, p < 0.0001), which did not differ significantly from the SS and control groups (p = 0.09). An increase in flexible restraint was significantly related to a greater weight loss in both HAES and SS groups (r = ?0.39, p < 0.01; and r = ?0.37, p < 0.05, respectively). A decrease in habitual susceptibility to disinhibition was also associated with a greater weight loss in HAES and control groups (r = 0.31, p < 0.05; and r = 0.44, p < 0.05, respectively). Discussion: These results suggest that a HAES intervention could have significant effects on eating behaviors and appetite ratings in premenopausal overweight women, when compared with an SS intervention or a control group.     

Ultrasound evaluation of visceral fat and metabolic risk factors during early pregnancy

Objective: The objective was to study the relationships between ultrasound estimated visceral fat and metabolic risk factors during early pregnancy. Research Methods and Procedures: Thirty consecutive healthy pregnant women at 11 to 14 weeks of gestation were studied. Maximum subcutaneous fat thickness (SFT) and visceral fat thickness (VFT) were successfully measured by ultrasound. Fasting plasma glucose, insulin, triglycerides, total cholesterol, high‐density lipoprotein cholesterol (HDL‐C), and blood pressure were measured. Insulin resistance was calculated by using the homeostasis model assessment (HOMA). Results: VFT significantly correlated with diastolic blood pressure (r = 0.37, p = 0.04), glycemia (r = 0.37, p = 0.04), insulinemia (r = 0.59, p = 0.001) insulin sensitivity (HOMA; r = 0.59, p = 0.001), triglycerides (r = 0.58, p = 0.03), HDL‐C (r = ?0.39, p = 0.03), and total cholesterol/HDL‐C ratio (p = 0.002), whereas SFT was significantly correlated with only diastolic blood pressure (p = 0.03). VFT better significantly correlated with the metabolic risk factors than pre‐gestational BMI [r = 0.39, p = 0.03 for insulinemia, r = 0.42, p = 0.02 for insulin sensitivity (HOMA), and r = 0.49, p = 0.01 for triglycerides and not significant for the rest]. Discussion: Visceral fat thickness can be easily measured by ultrasound at early pregnancy and correlates better than BMI with metabolic risk factors.     

Monthly change of community structure of zoobenthos in Xiangxi Bay after impoundment of the Three Gorges Teservoir, China

Xiangxi Bay, a typical bay of the Three Gorges Reservoir (TGR) in China, was formed in June 2003 after the impoundment of the TGR. We investigated the community structure of zoobenthos in Xiangxi Bay from August 2003 to August 2004. Benthic abundance and biomass were calculated per unit area of sediment. A total of 26 taxa were identified. They included two families of Lamellibranchia (7.7% of the total taxa number), two families of Oligochaeta (50%), Chironomidae (38.5%), and one other animal (3.8%). The average density and biomass were 276 ind./m² and 0.301 g/m², respectively. Oligochaetes and chironomids constituted the major groups, and Limnodrilus hoffmeisteri, Procladius sp., Branchiura sowerbyi, Tubificidae sp1. and Polypedilum scalaenum group sp. were the common taxa in this reservoir system. Among them, L. hoffmeisteri and Procladius sp. were the dominant species, and contributed 37.0% and 28.3% of the total density, respectively. The standing crop of zoobenthos in Xiangxi Bay was relative low, compared with other similar reservoirs. Our studies demonstrate that the standing crop increased gradually with time. The total density and biomass in August 2004 were 8.8 and 14.3 times those of the initial impoundment, separately. Oligochaetes did not become the sole dominant group, indicating that the sediment loading in Xiangxi Bay may be not serious. __________ Translated from Acta Hydrobiologica Sinica, 2006, 30(1): 64–69 [译自: 水生生物学报]     

医学超声治疗原理及其临床应用研究

本文从超声波对人体特有的生物效应出发,用生物医学工程的观点,阐述了现代超声治疗技术基本原理及最新临床应用成果,特别是在超声外科治疗技术中的最新发展,展望了超声治疗技术的应用发展前景。     

人血清蛋白质组学方法的比较和优化

目的:对人未做处理的血清以及去除白蛋白和免疫球蛋白G(IgG)血清的蛋白质组学方法进行比较和优化。方法:应用双向电泳(2-DE)方法分离了未做处理的以及去除白蛋白和免疫球蛋白G(IgG)的血清,比较优化了高温变性、水化液成份组成及泡胀方式等影响血清2-DE分离效果的因素,并用质谱分析鉴定未做处理和已处理血清的2-DE谱图中部分差异蛋白点。结果:得到了分辨率和重复性较好的2-DE谱图,未做处理的血清、去除白蛋白及IgG血清的平均蛋白质点分别为(482±18)个和(523±29)个,质谱分析了9个差异蛋白点,鉴定为8种蛋白质,其中7种为功能蛋白质。仅出现在未做处理血清中的蛋白有4种,分别是维生素A结合蛋白、可溶性尿激酶血纤维蛋白溶酶原激活剂受体、蛋白激酶1抗原、血清白蛋白。4种蛋白仅出现在去除白蛋白和IgG的血清中,分别是NADH脱氢酶辅酶β亚基、肌动蛋白结合蛋白M1、T细胞活性受体β链、血小板生长因子C。结论:去除高丰度蛋白可增加一些低丰度蛋白质的检出,但非特异性吸附会导致部分功能蛋白质的丢失。     

Vitellogenin cleavage products as indicators for toxic stress in zebra fish embryos: a proteomic approach

Vitellogenins (Vtgs) are the major yolk proteins in all oviparous animals. Systematic and regulated processing of these during embryogenesis is crucial for embryonic development. In the present study, toxicant-induced disturbance of Vtg degradation processes during Danio rerio (DR) embryogenesis was analysed to establish a sensitive tool for monitoring toxic stress at the molecular level. A 2-DE-based proteomic approach for whole DR embryos was established to study Vtg cleavage products (lipovitellin (Lv) derivatives). Ethanol was chosen as a positive control for a toxicity related change in the proteome of whole zebra fish embryos. Protein extracts from embryos treated with two ethanol concentrations, 0.5 and 2% v/v, showing either no or very strong visible effects, like absent heartbeat and blood circulation, were examined. Significant changes in the Lv pattern were detected for both conditions. The results are interpreted as scope for the use of the high abundant Lv derivatives as sensitive stress indicators in zebra fish embryos reflecting the overall fitness of the intact organisms.     

Partial characterization of the proteome of the mouse striatum

Many diseases of the mammalian CNS, including Parkinson's (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1-4]. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin-proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5-11]. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12]. Many studies have demonstrated that the both human LND patients and a mouse knockout model of HPRT deficiency have significantly reduced levels and uptake of dopamine in the striatum [4, 13-16] that is likely to be the principal cause of the CNS disorder. The precise molecular and cellular mechanisms that underlie this neurotransmitter defect are unknown.     

Exploring proteasome complexes by proteomic approaches

The ubiquitin proteasome system (UPS) represents a major pathway for intracellular protein degradation. Proteasome dependent protein quality control participates in cell cycle, immune response and apoptosis. Therefore, the UPS is in focus of therapeutic investigations and the development of pharmaceutical agents. Detailed analyses on proteasome structure and function are the foundation for drug development and clinical studies. Proteomic approaches contributed significantly to our current knowledge in proteasome research. In particular, 2-DE has been essential in facilitating the development of current models on molecular composition and assembly of proteasome complexes. Furthermore, developments in MS enabled identification of UPS proteins and their PTMs at high accuracy and high-throughput. First results on global characterization of the UPS are also available. Although the UPS has been intensively investigated within the last two decades, its functional significance and contribution to the regulation of cell and tissue phenotypes remain to be explored. This review recapitulates a variety of applied proteomic approaches in proteasome exploration, and presents an overview of current technologies and their potential in driving further investigations.