Characterization of the C584R variant in the mtDNA depletion syndrome gene FBXL4, reveals a novel role for FBXL4 as a regulator of mitochondrial fusion

Mutations in FBXL4 (F-Box and Leucine rich repeat protein 4), a nuclear-encoded mitochondrial protein with an unknown function, cause mitochondrial DNA depletion syndrome. We report two siblings, from consanguineous parents, harbouring a previously uncharacterized homozygous variant in FBXL4 (c.1750 T > C; p.Cys584Arg). Both patients presented with encephalomyopathy, lactic acidosis and cardiac hypertrophy, which are reported features of FBXL4 impairment. Remarkably, dichloroacetate (DCA) administration to the younger sibling improved metabolic acidosis and reversed cardiac hypertrophy. Characterization of FBXL4 patient fibroblasts revealed severe bioenergetic defects, mtDNA depletion, fragmentation of mitochondrial networks, and abnormalities in mtDNA nucleoids. These phenotypes, observed with other pathogenic FBXL4 variants, confirm the pathogenicity of the p.Cys584Arg variant. Although treating FBXL4 fibroblasts with DCA improved extracellular acidification, in line with reduced lactate levels in patients, DCA treatment did not improve any of the other mitochondrial functions. Nonetheless, we highlight DCA as a potentially effective drug for the management of elevated lactate and cardiomyopathy in patients with pathogenic FBXL4 variants. Finally, as the exact mechanism through which FBXL4 mutations lead to mtDNA depletion was unknown, we tested the hypothesis that FBXL4 promotes mitochondrial fusion. Using a photo-activatable GFP fusion assay, we found reduced mitochondrial fusion rates in cells harbouring a pathogenic FBXL4 variant. Meanwhile, overexpression of wildtype FBXL4, but not the p.Cys584Arg variant, promoted mitochondrial hyperfusion. Thus, we have uncovered a novel function for FBXL4 in promoting mitochondrial fusion, providing important mechanistic insights into the pathogenic mechanism underlying FBXL4 dysfunction.     

老年患者脓毒性休克液体复苏速度与预后的关系

目的探讨老年患者脓毒性休克液体复苏过程中输液速度与预后的相关性。方法回顾性分析92例严重感染病人液体复苏效果。按液体复苏期间的输液速度分为慢速组（〈500ml/h）和快速组（≥500ml/h）,并对两组患者的心率、有创及无创血压、血氧饱和度、呼吸、中心静脉压（CVP）、每小时尿量、尿比重、血乳酸、ScVO2及有无发生肺水肿、急性心功能不全、急性肾功能不全、MODS和7天死亡率、28天死亡率及入住ICU时间、住院时间等指标进行分析比较。结果两组患者在液体复苏过程中CVP和MAP两项指标比较无统计学意义。慢速组的尿量达标时间耗时较长（P〈0.05）;ScVO2及乳酸清除率比较无明显差异。快速组对液体的需求量多,而应用血管活性药物的病例数则少于慢速组;发生肺水肿、急性心功能不全的比例高于慢速组（P〈0.05）。两组急性肾功能不全的发生率相近,但慢速组出现MODS的例数少（P〈0.05）。7天死亡率和28天死亡率比较,慢速组低于快速组,但无统计学意义。两组患者住院时间无显著统计学意义,入住ICU时间慢速组明显短于快速组（P〈0.05）。结论在脓毒性休克复苏过程中不可一味靠加快输液速度来保证组织灌注,尤其对于老年患者,更应控制单位时间内的输液量,避免给储备能力较差的心功能造成更大的负担。在保证主要脏器灌注的情况下可以适当应用血管活性药物。     

Exact and user-friendly kinetic analysis of the two-step rapid equilibrium Michaelis-Menten mechanism

Most enzyme kinetic experiments are carried out under pseudo-first-order conditions, that is, when one of the reactant species (the enzyme or the substrate) is in a large excess of the other species. More accurate kinetic information about the system can be gained without the restrictions of the pseudo-first-order conditions. We present a practical and general method of analysis of the common two-step rapid equilibrium Michaelis-Menten mechanism. The formalism is exact in that it does not involve any other approximations such as the steady-state, limitations on the reactant concentrations or on reaction times. We apply this method to the global analysis of kinetic progress curves for bovine alkaline phosphatase assays carried out under both pseudo-first-order and pseudo-second-order conditions.     

Survival and fidelity of an enclosed white-tailed deer population using capture–recapture-reporting data

The number of animal populations enclosed by impermeable fences has increased, which poses issues related to the behavior of individuals and populations. Despite the increased number of fenced enclosures, there is a paucity of survival and fidelity data on white-tailed deer (Odocoileus virginianus) from fenced enclosures. Therefore, we examined marked deer recaptures and resightings over 13 years for an enclosed population of white-tailed deer in Oklahoma, USA, to estimate survival and fidelity parameters. We found that a step model was the best model of survival for both sexes. Survival of females and males was greater after hunting was suspended. Average female survival was 77% before hunting was suspended but increased to 98% after. Male survival was also greater after (99%) hunting was suspended compared to before (58%). Females exhibited greater site fidelity (84–94%) than males for all age groups except old individuals, which showed similar site fidelities for males and females. Fidelity was highest for old males (85%), followed by adult (74%), fawn (61%), and yearling males (56%). Our high-tensile electric fence allowed management goals to be achieved through increased survival while potentially maintaining genetic diversity through the exchange of limited numbers of individuals from surrounding areas. However, if the goal is to confine deer to limit disease spread or protect sensitive areas (e.g., airports) from deer encroachment, then other fence designs may be necessary, because our fence was not completely effective at controlling deer movements for these purposes.     

The estimation of population size,migration rates and survival in a stratified population

Summary Estimates of survival, migration rates, and population size are developed for a triple catch marking experiment onn (n>-2) areas with migration among all areas and death in all areas occurring, but no recruitment (birth). This repressents the extension to three sampling times of the method ofChapman andJunge (1956) for estimates in a stratified population. The method is further extented to allow for ‘losses on capture’.     

Sex-biased movement in the guppy (<Emphasis Type="Italic">Poecilia reticulata</Emphasis>)

The movement strategies of birds and mammals are often closely linked to their mating system, but few studies have examined the relationship between mating systems and movement in fishes. We examined the movement patterns of the guppy (Poecilia reticulata) in the Arima river of Trinidad and predicted that sexual asymmetry in reproductive investment would result in male-biased movement. Since male guppies maximize their reproductive success by mating with as many different females as possible, there should be strong selection for males to move in search of mates. In agreement with our prediction, the percentage of fish that emigrated from release pools was higher for males than females (27.3% vs. 6.9%, respectively). Sex ratio was highly variable among pools and may influence a male's decision to emigrate or continue moving. We also detected a positive relationship between body length and the probability of emigration for males and a significant bias for upstream movement by males. Among the few females that did emigrate, a positive correlation was observed between body length and distance moved. Sex-biased movement appears to be related to mating systems in fishes, but the evidence is very limited. Given the implications for ecology, evolution, and conservation, future studies should explicitly address the influence of sex and mating systems on movement patterns.     

The relationship between relative growth rate and susceptibility to aphids in wild barley under different nutrient levels

The Resource Availability Hypothesis (RAH) states that plants with a low Relative Growth Rate (RGR) and high levels of defence against herbivores or pathogens are favoured in habitats with low resource availability, whereas plants with a high potential RGR and low levels of defence are favoured in environments with high resource availability. High levels of defence are expected to result in lower reproduction and/or growth of the herbivores or pathogens. To test this hypothesis, four accessions of each of nine natural Hordeum spontaneum (wild barley) populations were grown in a climate chamber under two levels of nutrient supply. Susceptibility to Schizaphis graminum (greenbug) was quantified by placing a single adult greenbug on each plant and measuring its realised fecundity after 8 days. Data on potential RGR were available from a previous experiment. No support for the RAH was found. The correlation between potential RGR and greenbug reproduction was not significant, neither at the high nor at the low level of nutrient supply. Furthermore, on average plants grown under high and low nutrients did not differ in susceptibility. However, accessions-within-populations differed in the way susceptibility was affected by nutrient supply, and most accessions had a higher susceptibility under nutrient-poor conditions. It could be that these accessions differed in the spectrum of secondary metabolites they produced. Whatever the cause, the genetic variation for the reaction in susceptibility to nutrient supply suggests that selection could act in favour of more or less plasticity in plants without any apparent change in potential RGR.Due to an error in the citation line, this revised PDF (published in December 2003) deviates from the printed version, and is the correct and authoritative version of the paper.     

Predominant role of basicity of leaving group in alpha-effect for nucleophilic ester cleavage

It has been found that alpha-effects in nucleophilic reactions, unexpectedly large nucleophilicity due to adjacent unpaired electrons, are strongly dependent on the structure of substrate. The nucleophilic cleavages of 4-nitrobenzoate esters and 4-methylbenzoate esters by HOO- have been systematically investigated in detail. When the leaving groups of substrates are sufficiently good (aryl, 2,2,2-trifluoroethyl, and 2,2-dichloroethyl esters), alpha-effect is evident. However, this effect drastically decreases as the leaving group gets poorer, and is only marginal for the cleavages of 2-fluoroethyl and methyl esters. In the nucleophilic cleavages by salicylaldoxime and acetohydroxamic acid, alpha-effect is also notable only for the esters having good leaving groups. These enormous dependences of alpha-effects on the substrate-structure have been interpreted in terms of the difference in the position of transition-state in the reaction coordinate.     

The rate of mitochondrial 12S rRNA gene evolution is similar in freshwater turtles and marsupials

Assertions that the ``conventional' rate of mitochondrial DNA (mtDNA) evolution is reduced in poikilotherms in general and turtles in particular were tested for side-necked turtles (Pleurodira: Chelidae). Homologous data sets of mitochondrial 12S rRNA gene sequences were used to compare the average divergence between the Australian and South American species for two Gondwanan groups: the chelid turtles and the marsupials. The mean nucleotide divergences between continental groups for both the turtles and the marsupials are remarkably similar. These data suggest that the rate of evolution of mitochondrial 12S rRNA gene is not substantially slower in turtles than in the homeothermic marsupials. Received: 24 February 1997 / Accepted: 30 June 1997     

Estimating the fraction of invariable codons with a capture-recapture method

Summary A codon-based approach to estimating the number of variable sites in a protein is presented. When first and second positions of codons are assumed to be replacement positions, a capture-recapture model can be used to estimate the number of variable codons from every pair of homologous and aligned sequences. The capture-recapture estimate is compared to a maximum likelihood estimate of the number of variable codons and to previous approaches that estimate the number of variable sites (not codons) in a sequence. Computer simulations are presented that show under which circumstances the capture-recapture estimate can be used to correct biases in distance matrices. Analysis of published sequences of two genes, calmodulin and serum albumin, shows that distance corrections that employ a capture-recapture estimate of the number of variable sites may be considerably different from corrections that assume that the number of variable sites is equal to the total number of positions in the sequence. Offprint requests to: A. Sidow