Mus in Morocco: a Quaternary sequence of intraspecific evolution

North Africa is an intricate biogeographical region at the crossroads of immigration waves from tropical Africa and Asia. Species confined between various barriers (Atlas Mountains, arid environments such as the Sahara in the south, water masses such as the Mediterranean Sea in the north, and the Atlantic Ocean in the west) were generally forced to adapt locally to environmental changes instead of tracking their habitat by shifting their distribution area. The present study aims at providing first insight into the evolution of the genus Mus, and more specifically of the western Mediterranean species Mus spretus in this area. The study relies on the abundant Late Pleistocene and Middle Holocene fossil assemblage from the El Harhoura 2 cave (Rabat‐Témara, Morocco). This exceptional record was studied using geometric morphometrics applied to first upper and lower molars, constituting the most informative and best preserved fossil remains for such small rodents. Two main issues were addressed. (1) Geometric morphometrics was used to clarify taxonomic status and phylogenetic relationships among fossil and modern species in this area. Morphometric analysis revealed good discrimination of most modern and fossil species but failed to document intermediate forms tracing anagenetic evolution. Not mutually exclusive, the occurrence of complex processes of morphological evolution in this genus such as parallel evolution and the action of stabilizing selection may make it difficult to translate patterns of morphological evolution into phylogenetic conclusions. (2) The record was shown to document a sequence of intraspecific evolution of M. spretus. The morphology of the molars through the fossil record of El Harhoura 2 was surprisingly stable despite extensive modern variation. The limited temporal variation largely failed to correlate to palaeoenvironmental proxies. The mouse fossil record at El Harhoura 2 thus presents an intriguing case of morphological stasis despite extensive environmental changes. This long‐term stability may have been recently perturbed by anthropogenic factors including landscape changes and introduction of various competitors and predators, leading to a size reduction. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 599–621.     

Comparing methods for metabolic network analysis and an application to metabolic engineering

Bioinformatics tools have facilitated the reconstruction and analysis of cellular metabolism of various organisms based on information encoded in their genomes. Characterization of cellular metabolism is useful to understand the phenotypic capabilities of these organisms. It has been done quantitatively through the analysis of pathway operations. There are several in silico approaches for analyzing metabolic networks, including structural and stoichiometric analysis, metabolic flux analysis, metabolic control analysis, and several kinetic modeling based analyses. They can serve as a virtual laboratory to give insights into basic principles of cellular functions. This article summarizes the progress and advances in software and algorithm development for metabolic network analysis, along with their applications relevant to cellular physiology, and metabolic engineering with an emphasis on microbial strain optimization. Moreover, it provides a detailed comparative analysis of existing approaches under different categories.     

Liebenberg syndrome is caused by a deletion upstream to the PITX1 gene resulting in transformation of the upper limbs to reflect lower limb characteristics

Liebenberg syndrome (MIM 186550) is a very rare autosomal dominant condition characterized by three main features: dysplasia of all of the bony components of the elbow joint, abnormalities in the shape of carpal bones, and brachydactyly. In this paper, we report a Saudi Arabian family with Liebenberg syndrome. Comparative genomic hybridization (CGH) revealed a 275-kb deletion within the cytogenetic band 5q31.1 which contains the H2AFY gene and 190,428 bp of its downstream region. The deleted region is upstream to the PITX1 gene. The radiological features in the upper limbs of all affected members of the family were almost identical to the phenotype in the mouse model with ectopic expression of Pitx1 in the forelimbs. We therefore re-define the phenotype of Liebenberg syndrome as a transformation of the upper limbs to reflect lower limb characteristics and speculate that the area of deletion contains a regulatory sequence that suppresses the expression of PITX1 in the upper limb buds.     

祁连山东段高寒植被群落特征及其与地形气候因子关系研究

地形气候因子与物种分布的关系影响着高寒植被群落的演替,同时对山地水源涵养林功能和结构的维持具有重要的意义。以祁连山东段水源涵养林为研究对象,基于野外植物群落物种组成及地形气候因子调查数据,运用数量分类与排序等方法,探究了高寒植物群落特征及其与地形气候因子的关系。结果表明:65个调查样方中出现181个植物种,隶属40科,124属。科的物种组成及占总物种数比例分别为菊科30个种,占16.57%;蔷薇科17个,占9.44%;禾本科13个种,占7.22%;豆科11个种,占6.11%。毛茛科10个种,占5.56%。单属种82个,占总属数的66.13%。群落层片由乔木层、灌木层和草本层组成,乔木8种,灌木25种,草本148种。乔木层优势种有青海云杉、祁连圆柏、红桦。灌木层优势种有金露梅、山生柳、匙叶小檗、高山绣线菊。草本层优势种有甘肃薹草、珠芽蓼、早熟禾、唐松草、甘青蒿。TWINSPAN将高寒植被群落划分为7个群丛类型:群丛I红桦-红花蔷薇-甘肃薹草B.albosinensis-Rosa moyesii-C.kansuensis,群丛II青海云杉-匙叶小檗-甘肃薹草P.crassifolia-B.vernae-C.kansuensis,群丛III祁连圆柏-高山绣线菊-珠芽蓼Sabina chinensis-Spiraea alpine-P.viviparum,群丛IV高山绣线菊+鬼箭锦鸡儿-珠芽蓼S.alpine+Caragana jubata-P.viviparum,群丛V沙棘+甘青蒿+鼠掌老鹳草Hippophae rhamnoides+A.tangutica+Geranium sibiricum,群丛VI沙棘+苦荬菜+苦荞麦H.rhamnoides+Ixeris polycephala+Fagopyrum tataricum,群丛VII沙棘-冰草+西北沼委陵菜H.rhamnoides-Agropyron cristatum+Comarum salesovianum。7个群丛在DCA排序图上聚集分布,反映了较好的环境梯度。CCA排序结果表明,海拔是祁连山高寒植被群落植物种分布的最重要环境因子,其次是降水、温度、坡向、坡度。     

基于不同分析和取样方法研究褪黑激素对育成期母貂行为分配的影响

本文基于不同分析和取样方法研究褪黑激素对育成期母貂行为分配的影响。从貂场中随机选取当年出生的6只未埋植褪黑激素、6只埋植褪黑激素的美国短毛黑母貂。采用高清摄像头分别在埋植褪黑激素后的第30天（Ⅰ期）、第60天（Ⅱ期）、第75天（Ⅲ期）记录所有水貂的行为。埋植褪黑激素对水貂行为分配的影响,混合线性模型（mixed linear model,MLM）和广义线性混合模型（generalized linear mixed model,GLMM）对于水貂的位置、采食、修饰、异常和饮水行为的分析结果不同,广义线性混合模型的分析结果更符合褪黑激素已知的作用效果;瞬时取样间隔≤1.5 min时,水貂的所有行为比例与连续取样无显著差异（P>0.05）。由此可以推断,广义线性混合模型更适合无法正态化的实验数据的分析;采用瞬时扫描取样法研究水貂行为分配,扫描时间间隔以1.5 min为宜。     

研究生基因克隆与功能分析课程教学方式的探索与实践

基因工程是现代生物技术的重要核心。基因克隆与功能分析是生物化学与分子生物学专业硕士研究生的专业主干课,在生物技术人才的培养过程中起着重要的作用。时代的发展对研究生基因克隆与功能分析课程的教学模式提出了新的要求。本文探讨了翻转课堂教学法、研讨式教学法、案例式教学法、问题导向学习教学法、任务驱动教学法在研究生基因克隆与功能分析教学中的综合运用,以期提高人才培养质量。     

Development of a novel,high resolution melting analysis based genotyping method for Cryptosporidium parvum

This study employed the post-real-time PCR application, high resolution melting (HRM) analysis, in order to differentiate between characterised clinical and reference Cryptosporidium parvum samples obtained from Cork University Hospital (Cork, Ireland) and the Cryptosporidium Reference Unit (Swansea, Wales). A sample set composed of 18 distinct C. parvum gp60-subtypes of the IIa gp60-subtype family (an allele family accounting for over 80% of all cryptosporidiosis cases in Ireland) was employed. HRM analysis-based interrogation of the gp60, MM5 and MS9-Mallon tandem repeat loci was found to completely differentiate between 10 of the 18 studied gp60-subtypes. The remaining eight gp60-subtypes were differentiated into three distinct groupings, with the designations within these groupings resolved to two to three potential gp60-subtypes.The current study aimed to develop a novel, reproducible, real-time PCR based multi-locus genotyping method to distinguish between C. parvum gp60-subtypes. These preliminary results support the further expansion of the multi-locus panel in order to increase the discriminatory capabilities of this novel method.     

An ion-insensitive cAMP biosensor for long term quantitative ratiometric fluorescence resonance energy transfer (FRET) measurements under variable physiological conditions

Ratiometric measurements with FRET-based biosensors in living cells using a single fluorescence excitation wavelength are often affected by a significant ion sensitivity and the aggregation behavior of the FRET pair. This is an important problem for quantitative approaches. Here we report on the influence of physiological ion concentration changes on quantitative ratiometric measurements by comparing different FRET pairs for a cAMP-detecting biosensor. We exchanged the enhanced CFP/enhanced YFP FRET pair of an established Epac1-based biosensor by the fluorophores mCerulean/mCitrine. In the case of enhanced CFP/enhanced YFP, we showed that changes in proton, and (to a lesser extent) chloride ion concentrations result in incorrect ratiometric FRET signals, which may exceed the dynamic range of the biosensor. Calcium ions have no direct, but an indirect pH-driven effect by mobilizing protons. These ion dependences were greatly eliminated when mCerulean/mCitrine fluorophores were used. For such advanced FRET pairs the biosensor is less sensitive to changes in ion concentration and allows consistent cAMP concentration measurements under different physiological conditions, as occur in metabolically active cells. In addition, we verified that the described FRET pair exchange increased the dynamic range of the FRET efficiency response. The time window for stable experimental conditions was also prolonged by a faster biosensor expression rate in transfected cells and a greatly reduced tendency to aggregate, which reduces cytotoxicity. These properties were verified in functional tests in single cells co-expressing the biosensor and the 5-HT(1A) receptor.