In vitro displacement by rat serum of adsorbed radiolabeled poloxamer and poloxamine copolymers from model and biodegradable nanospheres

Poloxamer 407 and poloxamine 908 have been used by many research groups to modify the surface of both model latex and biodegradable nanospheres, thereby producing nanospheres that have shown reduced protein adsorption in vitro and extended circulation times in vivo. A potential limitation of such systems is the desorption of the copolymer coating layer. We describe a two-stage process to radiolabel poloxamer 407 and poloxamine 908 that has facilitated an investigation into this potential desorption, in vitro. The first stage of the labeling procedure involved the substitution of the terminal hydroxyl groups in each poly(ethylene oxide) (PEO) chain of poloxamer 407 and poloxamine 908 with an amino group. The aminated copolymers were then radiolabeled with 125Iodine Bolton-Hunter reagent. The efficiency of labeling was calculated to be approximately 20% for the tetramine poloxamine 908 and approximately 33% for the diamine poloxamer 407. Remaining free amino groups were then either acetylated, using acetic anhydride, or left in the free amino form. Covalent linkage of the radiolabel to the copolymer was confirmed by nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. The stability of the link between radiolabel and copolymer to hydrolysis was also confirmed; <4% loss of radiolabel occurred from poloxamine 908 after incubation in phosphate-buffered saline (PBS) at 37 degrees C for 8 days. The radiolabeled copolymers (with the free amino groups acetylated) were then used in experiments that have given the first direct evidence that adsorbed copolymers can be displaced by serum proteins in significant amounts from the surface of model and biodegradable nanospheres. The displacement was highly dependent on copolymer-nanosphere compatibility, with up to 78% of 125I tetramine poloxamine 908 being displaced from poly(lactide-co-glycolide) (PLGA) nanospheres in 24 h, compared with 20% displacement of 125I tetramine poloxamine 908 in 24 h from polystyrene nanospheres. These results have direct implication for the future design of drug delivery systems based on coated nanospheres.     

Analysis and neural network modeling of the nonlinear correlates of habituation in the vestibulo-ocular reflex

Through the process of habituation, continued exposure to low-frequency (0.01 Hz) rotation in the dark produced suppression of the low-frequency response of the vestibulo-ocular reflex (VOR) in goldfish. The response did not decay gradually, as might be expected from an error-driven learning process, but displayed several nonlinear and nonstationary features. They included asymmetrical response suppression, magnitude-dependent suppression for lower- but not higher-magnitude head rotations, and abrupt-onset suppressions suggestive of a switching mechanism. Microinjection of lidocaine into the vestibulocerebellum of habituated goldfish resulted in a temporary dishabituation. This suggests that the vestibulocerebellum mediates habituation, presumably through Purkinje cell inhibition of vestibular nuclei neurons. The habituated VOR data were simulated with a feed-forward, nonlinear neural network model of the VOR in which only Purkinje cell inhibition of vestibular nuclei neurons was varied. The model suggests that Purkinje cell inhibition may switch in to introduce nonstationarities, and cause asymmetry and magnitude-dependency in the VOR to emerge from the essential nonlinearity of vestibular nuclei neurons.     

A urine patch framework to simulate nitrogen leaching on New Zealand dairy farms

On New Zealand dairy farms, it is the nitrogen excreted directly onto pasture, particularly urine, that drives nitrogen (N) leaching from the farm. A new framework (UPF: Urine Patch Framework) is presented that post-processes the results of a whole farm model and runs a mechanistic soil model to simulate the urine patches. Two alternative methods to simulate the spatial distribution of urine patches were implemented and compared (Grid: spatially explicit, and Probabilistic: based on the probability of different temporal urination patterns). This paper describes the implementation of these two methods in connection with a Whole Farm Model; and compares the N leaching predictions with observed data. Two examples are provided, one analyzing the impact of urine patch overlap and another, the relative risk of N leaching at different times of urinary N deposition. The model showed good correlation and predictive ability between simulated annual N leaching results and observed data [R² = 94 %, mean relative prediction error (MRPE) = 10 % for Grid and R² = 72 %, MRPE = 20 % for Probabilistic]. The two methods produced similar results across an 8-year period for monthly and annual N leaching (R² = 96 %, MRPE = 10 % and R² = 86 %, MRPE = 8 %; respectively). Only 8 % of the paddock area was covered with multiple urinations during 1 year, but as much as 39 % of the total urine volume was deposited on overlapped patches. Systematically removing all urinary N for 1 month in either May or June reduced N leaching by approximately 20 %. Avoiding urinary N deposition during autumn or early winter could be highly effective in mitigating N leached during the following winter.     

Effects of worker size on the dynamics of fire ant tunnel construction

Social insects work together to complete tasks. However, different individuals within a colony may vary in task proficiency. We investigated if fire ant (Solenopsis invicta) worker body size influenced the ability to construct tunnels—a key component of subterranean nests. We monitored excavation by worker groups in a substrate of small wetted glass particles in quasi-two-dimensional arenas. Morphological and network features of the tunnel system were measured. Total tunnel area did not differ significantly between groups of large and small workers, although the tunnel area of control sized workers was significantly larger than that of large workers. Moreover, large workers created wider but shorter tunnels, with slower growth rate of tunnel number. However, edge–vertex scaling and degree distribution of the tunnel network were similar across all treatments. In all cases, the amount of excavated material was correlated with the number of active workers. Our study reveals that morphological features of excavated tunnels show modest variation when constructed by workers of varying sizes, but topological features associated with the tunnel network are conserved. These results suggest that important behavioural aspects of tunnel construction—and thus nest building—are similar among morphologically distinct members of fire ant societies.     

Entropic Contributions to Protein Stability

Thermodynamic stability is an important property of proteins that is linked to many of the trade-offs that characterize a protein molecule and therefore its function. Designing a protein with a desired stability is a complicated task given the intrinsic trade-off between enthalpy and entropy which applies for both the folded and unfolded states. Traditionally, protein stability is manipulated by point mutations which regulate the folded state enthalpy. In some cases, the entropy of the unfolded state has also been manipulated by means that drastically restrict its conformational dynamics such as engineering disulfide bonds. In this mini-review, we survey various approaches to modify protein stability by manipulating the entropy of either the unfolded or the folded states. We show that point mutations that involve elimination of long-range contacts may have a greater destabilization effect than mutations that eliminate shorter-range contacts. Protein conjugation can also affect the entropy of the unfolded state and thus the overall stability. In addition, we show that entropy can contribute to shape the folded state and yield greater protein stabilization. Hence, we argue that the entropy component can be practically manipulated both in the folded and unfolded state to modify protein stability.     

可回收式锥形肝静脉支架的动物实验研究

【摘要】 目的 通过动物实验评估可回收式锥形肝静脉支架的可行性、安全性及稳定性,探讨支架回收最佳时间。方法 健康家猪24头,随机分为4组,每组6头。全身麻醉下经右颈内静脉入路接受肝中静脉可回收式锥形支架置入术,术后3、7、11、15 d分别接受支架回收。采用t检验对比各组支架置入后、回收前所测得的支架内血流压力变化差异。采集支架中段相邻血管组织标本作病理学检查,测量计算新生血管内膜厚度,单因素方差分析对比各组间新生内膜厚度差异。结果 24枚支架均成功置入,技术成功率100%;支架回收技术成功率为50%（12/24）。肝中静脉内压力测量显示,3 d组支架置入后与回收前比较,差异无统计学意义（P＞0.05）;7 d组、11 d组、15 d组支架置入后与回收前比较,差异有统计学意义（P＜0.05）。各组间支架中段相邻组织新生内膜平均厚度两两比较,差异均有显著统计学意义（F＝338.29,P＜0.001）。结论 可回收式锥形支架置入家猪肝静脉安全、可行,其最佳安全回收时间为3 d,最长不宜超过7 d。

     

Soft-Chemistry Synthesis, Characterization, and Stabilization of CGO/Al2O3/Pt Nanostructured Composite Powders

Soft-chemistry routes were used to synthesize Ce_0.9Gd_0.1O_1.95-based powders with attractive and stable structural, morphological, and textural properties. In the intermediate temperature range between 500° and 700°C, the average Gd-doped CeO₂ (CGO) crystallite size is in the range 9–22 nm and the specific surface area varies from 43.4 to 8 m²/g. Above 700°C, a phase separation occurs between ceria and gadolinium oxide. Addition of alumina was found to be useful in stabilizing the CGO nanocrystallites at a high temperature and to avoid phase separation. A homogeneous dispersion of Pt nanoparticles (<10 nm at 1000°C) in the CGO materials was found to be possible by post-impregnation, although direct insertion of the Pt precursors during the synthesis led to aggregated particles, with less potential for catalytic applications.     

高精度伺服系统基于Pade近似的数字前馈控制器

针对指令超前信息未知的情况,提出一种基于Pade近似原理的数字前馈控制器。以某电机伺服系统为例进行仿真,结果表明了本控制器的有效性。     

Neurodevelopmental Disorders Associated with PSD-95 and Its Interaction Partners

The postsynaptic density (PSD) is a massive protein complex, critical for synaptic strength and plasticity in excitatory neurons. Here, the scaffolding protein PSD-95 plays a crucial role as it organizes key PSD components essential for synaptic signaling, development, and survival. Recently, variants in DLG4 encoding PSD-95 were found to cause a neurodevelopmental disorder with a variety of clinical features including intellectual disability, developmental delay, and epilepsy. Genetic variants in several of the interaction partners of PSD-95 are associated with similar phenotypes, suggesting that deficient PSD-95 may affect the interaction partners, explaining the overlapping symptoms. Here, we review the transmembrane interaction partners of PSD-95 and their association with neurodevelopmental disorders. We assess how the structural changes induced by DLG4 missense variants may disrupt or alter such protein–protein interactions, and we argue that the pathological effect of DLG4 variants is, at least partly, exerted indirectly through interaction partners of PSD-95. This review presents a direction for functional studies to elucidate the pathogenic mechanism of deficient PSD-95, providing clues for therapeutic strategies.     

SUPERCRITICAL EXTRACTION OF ISRAELI MISHOR ROTEM OIL SHALE 1. BATCH EXTRACTION WITH TOLUENE

ABSTRACT

Recovery of organic material from Israeli Mishor Rotem oil shale with toluene under supercritical conditions was investigated. The rate of solubilization, change of structural aspects and molecular weights of solubilized products with time were investigated. Experiments to recover organic material from shale were performed in a stainless steel high pressure autoclave. Shale Sample was charged in a sintered glass crucible suspended from the cap of the autoclave; toluene was the solvent in all experiments. Solvent/shale ratio was 20; experimental temperature was 34O^°C. The gaseous products were analyzed by gas chromatography. Molecular weights of the organic material recovered was measured by gel permeation chromatography technique. FTIR spectra of the organic material recovered and fractions isolated by extractive and chromatographical methods were measured. The amount of kerogen remaining in the spent shale was determined by oxidative derivative thermal gravimetry. A steady state in the production of solubles was reached within 60 minutes at 340^°C with a yield of 60 percent. After this time no further amounts of organic material was recovered. The molecular weights of the recovered organic material decreased at extended times after the steady state was reached. It appeared that the structure of the organic material recovered remained unchanged until the steady state condition is reached whereupon some structural changes occurred. At extended times the organic material was converted into more aromatic and less hydroxyl containing structures of lower molecular weight. The organic material recovered upon reaching steady state was fractionated into 63 percent oils (pentane solubles) and 32 percent asphaltenes (toluene solubles). The oile contained aliphatics and monoaromatic structures and the asphaltenes contained polyaromatic polar structures. Gases which constituted 4 percent of the initial kerogen were produced during the heating period to 340^°C. The amount of carbon monoxide produced remained constant and amounts of hydrogen, carbon dioxide and methane decreased after supercritical conditions were attained.