Markovian models for the developmental study of social behavior

Behavioral development involves changes in the probabilities of both social and nonsocial activities and the sequential pattern of activities over time. A number of methods have been offered for the analysis of these patterns of behavioral sequences. However, there continue to be problematic issues, including the analysis of nonstationary data; accommodation of changes in patterns within an observation period, or over repeated observations or age; and identification of differences in pattern changes between individuals or groups, and the factors responsible for these differences. In this work, we analyze data from 15 young monkeys (Macaca nemestrina) using classification and Markovian methods, including a new approach to nonstationary data called the double-chain Markov model (DCCM). These methods allowed us to identify differences in behavior patterns that differentiate between normal subjects and those presenting developmental anomalies.     

Cysteine modification of a putative pore residue in ClC-0: implication for the pore stoichiometry of ClC chloride channels

The ClC channel family consists of chloride channels important for various physiological functions. Two members in this family, ClC-0 and ClC-1, share approximately 50-60% amino acid identity and show similar gating behaviors. Although they both contain two subunits, the number of pores present in the homodimeric channel is controversial. The double-barrel model proposed for ClC-0 was recently challenged by a one-pore model partly based on experiments with ClC-1 exploiting cysteine mutagenesis followed by modification with methanethiosulfonate (MTS) reagents. To investigate the pore stoichiometry of ClC-0 more rigorously, we applied a similar strategy of MTS modification in an inactivation-suppressed mutant (C212S) of ClC-0. Mutation of lysine 165 to cysteine (K165C) rendered the channel nonfunctional, but modification of the introduced cysteine by 2-aminoethyl MTS (MTSEA) recovered functional channels with altered properties of gating-permeation coupling. The fast gate of the MTSEA-modified K165C homodimer responded to external Cl(-) less effectively, so the P(o)-V curve was shifted to a more depolarized potential by approximately 45 mV. The K165C-K165 heterodimer showed double-barrel-like channel activity after MTSEA modification, with the fast-gating behaviors mimicking a combination of those of the mutant and the wild-type pore, as expected for the two-pore model. Without MTSEA modification, the heterodimer showed only one pore, and was easier to inactivate than the two-pore channel. These results showed that K165 is important for both the fast and slow gating of ClC-0. Therefore, the effects of MTS reagents on channel gating need to be carefully considered when interpreting the apparent modification rate.     

Kinetic characterization of the double mutant R148A/E182S of glycogen phosphorylase kinase catalytic subunit: the role of the activation loop

Many protein kinases are activated by phosphorylation in a highly conserved region of their catalytic subunit, termed activation loop. Phosphorylase kinase is constitutively active without the requirement for phosphorylation of residues in the activation loop. The residue which plays an analogous role to the phosphorylatable residues in other protein kinases is Glu182, which makes contacts to a highly conserved Arg148. In turn, Arg148 adjacent to the catalytic Asp149, enabling information to be transmitted from the activation loop to the catalytic machinery. The double mutant R148A/E182S has been kinetically characterized. The mutation resulted in an approximate 16- to 22-fold decrease in the k _cat/K _m value of the enzyme. The kinetic data, discussed in the light of the structural data from previously determined complexes of the enzyme, lead to the suggestion that the activation loop has a major role in substrate binding but also in correct orientation of the groups participating in catalysis.     

Reducing Meloidogyne incognita Injury to Cucumber in a Tomato-Cucumber Double-Cropping System

The effects of a root-knot nematode-resistant tomato cultivar and application of the nematicide ethoprop on root-knot nematode injury to cucumber were compared in a tomato-cucumber double-cropping system. A root-knot nematode-resistant tomato cultivar, Celebrity, and a susceptible cultivar, Heatwave, were grown in rotation with cucumber in 1995 and 1996. Celebrity suppressed populations of Meloidogyne incognita in the soil and resulted in a low root-gall rating on the subsequent cucumber crop. Nematode population densities were significantly lower at the termination of the cucumber crop in plots following Celebrity than in plots following Heatwave. Premium and marketable yields of cucumbers were higher in plots following Celebrity than in plots following Heatwave. Application of ethoprop through drip irrigation at 4.6 kg a.i./ha reduced root galling on the cucumber crop but had no effect on the nematode population density in the soil at crop termination. Ethoprop did not affect cucumber yield. These results indicate that planting a resistant tomato cultivar in a tomato-cucumber double-cropping system is more effective than applying ethoprop for managing M. incognita.     

新生鼠大脑皮质和中脑多巴胺神经元原代培养中NGF诱导c-jun mRNA表达的研究

用神经生长因子（nervegrowthfactor,NGF）分别处理原代培养的新生大鼠大脑皮质和中脑腹侧部神经元,应用免疫组织化学和原位杂交双重标记方法,观察不同时间NGF处理的神经元表达原癌基因c－junmRNA的情况。结果发现,神经特异性烯醇化酶（neuronspecificenolase,NSE）阳性的大脑皮层神经细胞在NGF处理15分钟即可表达c－junmRNA,2小时达高峰,4小时后开始下降,到8小时后基本消失．未经NGF处理的大鼠大脑皮层神经细胞不表达c－junmRNA;酪氨酸羟化酶（tyrosinehydroxylase,TH）阳性的中脑多巴胺能（Dopaminergic,DA）神经元经NGF处理也不表达c－jun基因。提示NGF与其受体结合可以激活神经细胞快速,短暂的一过性表达c-jun基因,作为第三信使,调节从细胞质膜到核的信号传递,同时也间接证明了新生大鼠大脑皮层神经细胞膜上存在神经生长因子受体（nervegrowthfactorreceptor,NGFR）,而中脑DA神经细胞对NGF无应答反应。     

基于叶绿体全基因组的云南樱花品种‘红霞''的种系分析

云南樱花品种‘红霞’(Cerasus‘Hongxia’)广泛种植于滇中地区，3月开花，半重瓣至重瓣，粉红色，观赏价值较高。‘红霞’曾被认为是红花高盆樱(C.cerasoides var.rubea)的园艺品种，而其分子标记结果却聚在钟花樱(C.campanulata)种系，可见‘红霞’的种系归属存疑，对今后的育种及栽培极为不利。为探索‘红霞’的种系问题，该研究以‘红霞’的叶片DNA为材料，应用Illumina软件对叶绿体基因组特征进行测序，通过SPAdes 3.13.0、Gapcloser 1.12和CPGAVAS2 2.0软件进行组装、注释，之后使用MISA和Geneious 10.0软件对叶绿体基因组特征进行分析，并使用RAxML 8.0软件构建Cerasus分支系统发育树，分析‘红霞’在樱属植物的系统关系。结果表明：(1)‘红霞’的叶绿体全基因组长度为157 832 bp,鸟嘌呤(Guanine)、胞嘧啶(Cytosine)含量为36.73%,包含一对反向重复(inverted repeat sequence, IR, 26 381 bp)区，由小单拷贝(short single...     

Dual‐Confined Flexible Sulfur Cathodes Encapsulated in Nitrogen‐Doped Double‐Shelled Hollow Carbon Spheres and Wrapped with Graphene for Li–S Batteries

Batteries with high energy and power densities along with long cycle life and acceptable safety at an affordable cost are critical for large‐scale applications such as electric vehicles and smart grids, but is challenging. Lithium–sulfur (Li‐S) batteries are attractive in this regard due to their high energy density and the abundance of sulfur, but several hurdles such as poor cycle life and inferior sulfur utilization need to be overcome for them to be commercially viable. Li–S cells with high capacity and long cycle life with a dual‐confined flexible cathode configuration by encapsulating sulfur in nitrogen‐doped double‐shelled hollow carbon spheres followed by graphene wrapping are presented here. Sulfur/polysulfides are effectively immobilized in the cathode through physical confinement by the hollow spheres with porous shells and graphene wrapping as well as chemical binding between heteronitrogen atoms and polysulfides. This rationally designed free‐standing nanostructured sulfur cathode provides a well‐built 3D carbon conductive network without requiring binders, enabling a high initial discharge capacity of 1360 mA h g^?1 at a current rate of C/5, excellent rate capability of 600 mA h g^?1 at 2 C rate, and sustainable cycling stability for 200 cycles with nearly 100% Coulombic efficiency, suggesting its great promise for advanced Li–S batteries.     

Essential gene disruptions reveal complex relationships between phenotypic robustness,pleiotropy, and fitness

The concept of robustness in biology has gained much attention recently, but a mechanistic understanding of how genetic networks regulate phenotypic variation has remained elusive. One approach to understand the genetic architecture of variability has been to analyze dispensable gene deletions in model organisms; however, the most important genes cannot be deleted. Here, we have utilized two systems in yeast whereby essential genes have been altered to reduce expression. Using high-throughput microscopy and image analysis, we have characterized a large number of morphological phenotypes, and their associated variation, for the majority of essential genes in yeast. Our results indicate that phenotypic robustness is more highly dependent upon the expression of essential genes than on the presence of dispensable genes. Morphological robustness appears to be a general property of a genotype that is closely related to pleiotropy. While the fitness profile across a range of expression levels is idiosyncratic to each gene, the global pattern indicates that there is a window in which phenotypic variation can be released before fitness effects are observable.     

Finite Element Method (FEM) Modeling of Freeze-drying: Monitoring Pharmaceutical Product Robustness During Lyophilization

Lyophilization is an approach commonly undertaken to formulate drugs that are unstable to be commercialized as ready to use (RTU) solutions. One of the important aspects of commercializing a lyophilized product is to transfer the process parameters that are developed in lab scale lyophilizer to commercial scale without a loss in product quality. This process is often accomplished by costly engineering runs or through an iterative process at the commercial scale. Here, we are highlighting a combination of computational and experimental approach to predict commercial process parameters for the primary drying phase of lyophilization. Heat and mass transfer coefficients are determined experimentally either by manometric temperature measurement (MTM) or sublimation tests and used as inputs for the finite element model (FEM)-based software called PASSAGE, which computes various primary drying parameters such as primary drying time and product temperature. The heat and mass transfer coefficients will vary at different lyophilization scales; hence, we present an approach to use appropriate factors while scaling-up from lab scale to commercial scale. As a result, one can predict commercial scale primary drying time based on these parameters. Additionally, the model-based approach presented in this study provides a process to monitor pharmaceutical product robustness and accidental process deviations during Lyophilization to support commercial supply chain continuity. The approach presented here provides a robust lyophilization scale-up strategy; and because of the simple and minimalistic approach, it will also be less capital intensive path with minimal use of expensive drug substance/active material.