Molecular and cellular basis of ornithine δ-aminotransferase deficiency caused by the V332M mutation associated with gyrate atrophy of the choroid and retina

Gyrate atrophy (GA) is a rare recessive disorder characterized by progressive blindness, chorioretinal degeneration and systemic hyperornithinemia. GA is caused by point mutations in the gene encoding ornithine δ-aminotransferase (OAT), a tetrameric pyridoxal 5′-phosphate-dependent enzyme catalysing the transamination of l-ornithine and α-ketoglutarate to glutamic–γ-semialdehyde and l-glutamate in mitochondria. More than 50 OAT variants have been identified, but their molecular and cellular properties are mostly unknown. A subset of patients is responsive to pyridoxine administration, although the mechanisms underlying responsiveness have not been clarified. Herein, we studied the effects of the V332M mutation identified in pyridoxine-responsive patients. The Val332-to-Met substitution does not significantly affect the spectroscopic and kinetic properties of OAT, but during catalysis it makes the protein prone to convert into the apo-form, which undergoes unfolding and aggregation under physiological conditions. By using the CRISPR/Cas9 technology we generated a new cellular model of GA based on HEK293 cells knock-out for the OAT gene (HEK-OAT_KO). When overexpressed in HEK-OAT_KO cells, the V332M variant is present in an inactive apodimeric form, but partly shifts to the catalytically-competent holotetrameric form in the presence of exogenous PLP, thus explaining the responsiveness of these patients to pyridoxine administration. Overall, our data represent the first integrated molecular and cellular analysis of the effects of a pathogenic mutation in OAT. In addition, we validated a novel cellular model for the disease that could prove instrumental to define the molecular defect of other GA-causing variants, as well as their responsiveness to pyridoxine and other putative drugs.     

MSTARA and MSTARI: interactive PC algorithms for simple, model independent evaluation of sedimentation equilibrium data

This paper describes a program available for PC's for the evaluation of molecular weights from sedimentation equilibrium. This program, in its two forms – MSTARA for absorption optical records and MSTARI for interference optical records – requires no prior assumption of the nature of the system (ideal, non-ideal, monodisperse, polydisperse, self-associating etc.) and takes into consideration the whole solute distribution (i.e. from solution meniscus to cell base) in the ultracentrifuge cell rather than just a selected data-set. MSTARA or MSTARI are therefore recommended as a first analysis programme of sedimentation equilibrium data coming off an absorption or interference based analytical ultracentrifuge. These programmes are therefore particularly well suited if heterogeneity (polydispersity or interaction phenomena) or non-ideality is suspected. Their use is demonstrated for a series of data-set types (ideal, non-ideal, polydisperse and self-associating). Although MSTARA and MSTARI are model independent, they provide the basis for more detailed analysis of interactions, polydisperse distributions or non-ideality via easy export of ASCII datafiles to model dependent routines. Received: 4 November 1996 / Accepted: 15 November 1996     

单侧前庭神经切断后猫运动方式的改变和补偿

本工作首次将一种运动分析系统,即视觉自动电视摄相信息处理系统（ｏｐｔｉｃａｌａｕｔｏｍａｔｉｃＴＶ－ｉｍａｇｅｐｒｏｃｅｓｓｏｒ,Ｅ．Ｌ．Ｉ．Ｔ．Ｅｓｙｓｔｅｍ）用于动物运动方式的定量分析,比较了猫一侧前庭神经切断前后在跨越转动横梁测试中运动方式的变化。结果如下：在横梁静止条件下,猫述后行走的初始身高明显下降,步距变短,步 频降低,呈缓慢的蹒中山步态,继而各参量逐渐增加,分别在术后４５和７０ｄ恢复     

Development of a boron buffered solution culture system for controlled studies of plant boron nutrition

Chelated-buffered nutrient solutions are used for studies on micronutrient metals but so far no equivalent system exists for boron nutrition studies: the present investigation was initiated with that intention. From a literature review, it was noted that a range of substances form chelates with boron including polyhydric alcohols, sugars and phenolic compounds. However, none apart from hydrofluoric acid formed chelates with formation constants comparable to those of micronutrient metal chelates like diethylenetriaminepentaacetic acid (DTPA). Moreover, most chelating substances had deleterious side effects which reduced their possible use in water culture: many of the compounds are substrates for bacterial growth, some are harmful to handle, and others are toxic to plants or humans. Borosilicate glass; was tested in a laboratory experiment but found to release boron too slowly into solution to maintain constant boron concentration in solution even when very finely ground. Current investigations centre around the use of a boron-specific resin, which strongly complexes H₃BO₃ on its N-methyl glucamine functional groups. The boron sorption capacity of the resin varied from 2.2 to 5.0 mg B g^-1 resin. Boron saturated resin maintained an equilibrium solution boron concentration of 46 t M when added at the rate of 2 g of resin to 1 L of boron free triple deionised water. Plants grown in complete nutrient solution with boron saturated resin added at 1 g per litre of nutrient solution grew as well as plants grown in conventional nutrient solution containing 9.2 t M boron and their shoots contained adequate boron concentrations for growth. There was no evidence that the resin had effects on plant growth other than in releasing and equilibrating boron concentration in the nutrient solution.     

Interaction of detergents with bovine lens α-crystallin: evidence for an oligomeric structure based on amphiphilic interactions

We have studied the quaternary structure of α-crystallin in the presence of increasing concentrations of amphiphilic and neutral detergents using gel filtration, light-scattering, boundary and equilibrium sedimentation. We observed a continuous reduction of the molar mass of the polymeric α-crystallin on increasing the concentration of sodium dodecyl sulphate from 0.1 mM to 5 mM, ending up with the monomeric peptides. Dodecyltrimethylammonium bromide also disrupts the oligomeric structure of α-crystallin but the interaction appears to be cooperative: in the sharp transition region (for a 1 mg/ml protein solution) from 3 to 8 mM of the detergent, only the native protein and a mixture of monomeric and dimeric peptide-DTAB complexes can be observed. Concomitant studies of the circular dichroism in the far UV revealed a substantial decrease of the β-sheet and increase of the α-helix secondary structure. The latter can be related to the presence of amphiphilic polypeptide sequences in the constituent αA and αB peptides. These studies reveal for the first time a direct relation between changes in the secondary structure of the αA and αB peptides and the formation of the oligomeric α-crystallin structure: the binding of the amphiphilic detergent reduces the β-sheet content, induces the formation of α-helix secondary structure and reduces the tendency of the peptide to form large aggregates. The different mechanisms for reducing the oligomeric size by anionic and cationic detergents with identical apolar parts stresses the importance of charge interactions. Our findings support some aspects of the micelle model of α-crystallin and can be related to its chaperone activity. Accepted: 18 October 1996     

Seasonal Dynamics of Plant Diversity on lnselbergs in the Ivory Coast (West Africa)

Inselbergs are isolated monolithic outcrops which are characterized by large areas of exposed crystalline rock. Due to harsh edaphic and microclimatic conditions, inselbergs are completely differentiated from their surroundings. Consequently they host a very distinct vegetation which is being investigated on a global scale over a six year period. The seasonal dynamics of Selected plant communities (Afrotrilepis pilosa mat, shallow depression, ephemeral flush vegetation) on granitic inselbergs in the Comoe National Park (NE Ivory Coast) were studied during the rainy period from May to November 1991 by recording all vascular plant species at 12 intervals. For the habitats investigated, the seasonal vegetation dynamics were related to the rainfall pattern. Maximum values both in species diversity and richness were attained in the first third of the rainy period. Drought in August and September caused a decline in species number and diversity in the shallow depression and ephemeral flush vegetation, resulting in mortality of more than 20% of the species. The individual communities studied differed considerably in species diversity and richness. We conclude that ephemeral flush and shallow depression communities are more species rich than the mat community which is dominated by the highly competitive and specialized K-strategist Afrotrilepis pilosa (a poikilohydric Cyperaceae) due to stochastic climatic perturbations which allow the maintenance of species rich non-equilibrium assemblages with r-strategists as major components.     

一类具有稀疏效应的生态系统的极限环

研究如下一类具有稀疏效应的生态系统模型应用微分方程定性理论,得到了该系统极限环的存在性、唯一性及存在性的参数范围．     

Comparative Studies of the Indoleamine Dioxygenase-like Myoglobin from the Abalone Sulculus diversicolor

The abalone Sulculus diversicolor contains abundant myoglobin in its buccal mass. The myoglobin is homodimeric and the molecular mass of the constituent polypeptide chain is 41,000 Da. The amino acid sequence and gene structure are highly homologous with those of a vertebrate tryptophan-degrading enzyme, indoleamine dioxygenase (IDO). Thus Sulculus myoglobin evolved from an IDO gene, and represents a typical case of functional convergence. The oxygen equilibrium properties of Sulculus myoglobin were examined and compared with those of myoglobins from other sources. It binds oxygen reversibly, and the P₅₀ was determined to be 3.8 mmHg at 20°C and pH 7.4, showing that the oxygen affinity of Sulculus myoglobin is significantly lower than those of usual 16 kDa myoglobins. It also displays no cooperativity (n_max: 1.02–1.06) and no alkaline Bohr effect between pH 7.0 and 7.9. The cDNA-derived amino acid sequences of vertebrate IDOs, molluscan IDO-like myoglobins and a homolog in the yeast Saccharomyces were aligned, and several amino acid residues were proposed as candidates for key residues to control the function of IDO or myoglobin.     

Coevolutionary interactions in a host–parasite system

Interactions between parasitic cuckoos and their hosts represent a classic example of coevolution, where adaptations in the parasite to exploit the host select for defences, which in turn select for new parasite adaptations. Current interactions between the two parties may be at an evolutionary equilibrium or, alternatively, a coevolutionary arms race may be taking place. By taking into account the effect of gene flow in 15 European magpie ( Pica pica ) populations, we studied the coevolutionary interactions with its brood parasite, the great spotted cuckoo ( Clamator glandarius ). Our results suggest that, in Europe, magpies and cuckoos are engaged in an ongoing coevolutionary process because, despite controlling for the large amounts of gene flow among different magpie populations, we still found a positive relationship between host defence (i.e. foreign egg recognition and rejection) and parasite selection pressure.     

Disturbances prevent stem size-density distributions in natural forests from following scaling relationships

Enquist and Niklas propose that trees in natural forests have invariant size-density distributions (SDDs) that scale as a −2 power of stem diameter, although early studies described such distributions using negative exponential functions. Using New Zealand and ‘global’ data sets, we demonstrate that neither type of function accurately describes the SDD over the entire diameter range. Instead, scaling functions provide the best fit to smaller stems, while negative exponential functions provide the best fit to larger stems. We argue that these patterns are consistent with competition shaping the small-stem phase and exogenous disturbance shaping the large-stem phase. Mortality rates, estimated from repeat measurements on 1546 New Zealand plots, fell precipitously with stem size until 18 cm but remained constant after that, consistent with our arguments. Even in the small-stem phase, where SDDs were best described by scaling functions, the scaling exponents were not invariantly −2, but differed significantly from this value in both the ‘global’ and New Zealand data sets, and varied through time in the New Zealand data set.