MAN1B1 Deficiency: An Unexpected CDG-II

Congenital disorders of glycosylation (CDG) are a group of rare metabolic diseases, due to impaired protein and lipid glycosylation. In the present study, exome sequencing was used to identify MAN1B1 as the culprit gene in an unsolved CDG-II patient. Subsequently, 6 additional cases with MAN1B1-CDG were found. All individuals presented slight facial dysmorphism, psychomotor retardation and truncal obesity. Generally, MAN1B1 is believed to be an ER resident alpha-1,2-mannosidase acting as a key factor in glycoprotein quality control by targeting misfolded proteins for ER-associated degradation (ERAD). However, recent studies indicated a Golgi localization of the endogenous MAN1B1, suggesting a more complex role for MAN1B1 in quality control. We were able to confirm that MAN1B1 is indeed localized to the Golgi complex instead of the ER. Furthermore, we observed an altered Golgi morphology in all patients'' cells, with marked dilatation and fragmentation. We hypothesize that part of the phenotype is associated to this Golgi disruption. In conclusion, we linked mutations in MAN1B1 to a Golgi glycosylation disorder. Additionally, our results support the recent findings on MAN1B1 localization. However, more work is needed to pinpoint the exact function of MAN1B1 in glycoprotein quality control, and to understand the pathophysiology of its deficiency.     

Phagocyte migration: an overview

Phagocytes are the first line of host defense thanks to their capacity to infiltrate infected and wounded tissues, where they exert their bactericidal and tissue repair functions. However, tissue infiltration of phagocytes also stimulates the progression of pathologies such as cancer and chronic inflammatory diseases. It is therefore necessary to identify the molecular and cellular mechanisms that control this process to identify new therapeutic targets. Phagocytes leave the blood stream by crossing the vascular wall and infiltrate interstitial tissues, a three-dimensional environment. A state-of-the-art of the different steps of phagocyte tissue recruitment in vivo and of the different in vitro models is developed in this synthesis. We focus on recent data concerning the migration of phagocytes in three-dimensional environments. The use of two different migration modes, amoeboid and mesenchymal, by macrophages and the role of podosomes and proteases in the mesenchymal migration are discussed.     

Organelle trafficking of chimeric ribozymes and genetic manipulation of mitochondria

With the expansion of the RNA world, antisense strategies have become widespread to manipulate nuclear gene expression but organelle genetic systems have remained aside. The present work opens the field to mitochondria. We demonstrate that customized RNAs expressed from a nuclear transgene and driven by a transfer RNA-like (tRNA-like) moiety are taken up by mitochondria in plant cells. The process appears to follow the natural tRNA import specificity, suggesting that translocation indeed occurs through the regular tRNA uptake pathway. Upon validation of the strategy with a reporter sequence, we developed a chimeric catalytic RNA composed of a specially designed trans-cleaving hammerhead ribozyme and a tRNA mimic. Organelle import of the chimeric ribozyme and specific target cleavage within mitochondria were demonstrated in transgenic tobacco cell cultures and Arabidopsis thaliana plants, providing the first directed knockdown of a mitochondrial RNA in a multicellular eukaryote. Further observations point to mitochondrial messenger RNA control mechanisms related to the plant developmental stage and culture conditions. Transformation of mitochondria is only accessible in yeast and in the unicellular alga Chlamydomonas. Based on the widespread tRNA import pathway, our data thus make a breakthrough for direct investigation and manipulation of mitochondrial genetics.     

Effects of playing surface (hard and clay courts) on heart rate and blood lactate during tennis matches played by high-level players

The aim of this study was to compare tennis matches played on clay (CL) and resin (R) courts. Six matches were played (3 on CL courts and 3 on R courts) by 6 high-level players. Heart rate (HR) was monitored continuously while running time (4.66 m), and blood lactate concentration ([La]) were measured every 4 games. Mean duration of points and effective playing time (EPT) were measured for each match. Mean HR (154 ± 12 vs. 141 ± 9 b · min(-1)) and [La] values (5.7 ± 1.8 vs. 3.6 ± 1.2 mmol · L(-1)) were significantly higher on CL (p < 0.05). The [La] increased significantly during the match on CL court. Mean duration of rallies (8.5 ± 0.2 vs. 5.9 ± 0.5 seconds) and EPT (26.2 ± 1.9 vs. 19.5 ± 2.0%) were significantly longer (p < 0.05) on CL. Running time values in speed tests were not significantly different between CL and R. Running time performance was not significantly decreased during the match, whatever the playing surface. This study shows that the court surface influences the characteristics of the match and the player's physiological responses. The court surface should be a key factor for consideration when coaches determine specific training programs for high-level tennis players.     

Coordination of soccer players during preseason training

This study aimed to verify whether coordination improves as a result of a preseason soccer training. During 5 experimental sessions (days 1, 6, 11, 15, and 19), 16 semiprofessional male soccer players (22.0 ± 3.6 years) were administered 3 specific soccer tests (speed dribbling, shooting a dead ball, and shooting from a pass) and an interlimb coordination test (total duration of a trial: 60 seconds), consisting of isodirectional and nonisodirectional synchronized (1:1 ratio) hand and foot flexions and extensions at an increasing velocity of execution (80, 120, and 180 b·min(-1)). Furthermore, subjective ratings were monitored to assess the recovery state (RestQ) of the players, their perceived exertion (rating of perceived exertion [RPE]) for the whole body, and the perceived muscle pain (rating of muscle pain [RMP]) for the lower limbs and the internal training load by means of the session-RPE method. The ratios between post and pretraining RPE and RMP increased only during the first 2 experimental sessions and decreased after the second week of the training camp (p = 0.001). The Rest-Q showed increases (p < 0.05) for general stress, conflict/pressure, social recovery, and being in shape dimensions. Conversely, decreases (p < 0.05) were observed for social stress, fatigue, physical complaints dimensions. Throughout the preseason, the players improved their speed dribbling (p = 0.03), Shooting from a Pass (p = 0.02), and interlimb coordination (p < 0.0001) performances. These coordination tests succeeded in discriminating coordination in soccer players and could integrate field test batteries during the whole soccer season, because they were easily and inexpensively administrable by coaches.     

GENETIC COMPATIBILITY AFFECTS DIVISION OF LABOR IN THE ARGENTINE ANT LINEPITHEMA HUMILE

Division of labor is central to the organization of insect societies. Within‐colony comparisons between subfamilies of workers (patrilines or matrilines) revealed genetic effects on division of labor in many social insect species. Although this has been taken as evidence for additive genetic effects on division of labor, it has never been experimentally tested. To determine the relative roles of additive and nonadditive genetic effects (e.g., genetic compatibility, epistasis, and parent‐of‐origin imprinting effects) on worker behavior, we performed controlled crosses using the Argentine ant Linepithema humile. Three of the measured behaviors (the efficiency to collect pupae, the foraging propensity, and the distance between non‐brood‐tenders and brood) were affected by the maternal genetic background and the two others (the efficiency to feed larvae and the distance between brood‐tenders and brood) by the paternal genetic background. Moreover, there were significant interactions between the maternal and paternal genetic backgrounds for three of the five behaviors. These results are most consistent with parent‐of‐origin and genetic compatibility effects on division of labor. The finding of nonadditive genetic effects is in strong contrast with the current view and has important consequences for our understanding of division of labor in insect societies.     

A method to estimate phenological variation using data from large‐scale abundance monitoring programmes

Capsule Large‐scale abundance monitoring programmes can be used to estimate annual phenological shifts.

Aims Phenology refers to the timing of any annually repeated biological event. The method developed here aims at measuring phenological variation in an indirect way by modelling seasonal abundance variations. Thus, it provides the opportunity to use a large number of datasets which have rarely been used in phenological studies. Phenological variations computed using this standardized method are comparable between species.

Methods The data used for the development of this method originates from the French Breeding Bird Survey, a large‐scale abundance monitoring programme launched in 2001. For each species, the phenological shift between two seasonal abundance trends is computed using maximum likelihood.

Results Phenological shifts relative to the year 2005 (reference year) were estimated for 46 species over a 5‐year period (2001–6). The standard deviations of the shifts do not differ significantly between species with different migratory status. Moreover, at the species level, the computed phenological shifts relate to the shifts of the mean date weighted by abundance. However, mean date, cannot be used in studies incorporating species with different migratory status (e.g. trans‐Saharan migrant, sedentary) because of ambiguous changes for the same biological shift in timing.

Conclusions The method described here is of particular value in determining how the phenology of common bird species changes in relation to climate. It offers the opportunity to increase the spatial scale of phenological studies and to include multi‐species analyses. This method could be applied to any abundance or constant effort site programme to study the timing of any biological process for which a seasonal distribution is available.     

Iron-mediated effect of alcohol on hepatocyte differentiation in HepaRG cells

The development of alcoholic liver diseases depends on the ability of hepatocyte to proliferate and differentiate in the case of alcohol-induced injury. Our previous work showed an inhibitory effect of alcohol on hepatocyte proliferation. However, the effect of alcohol on hepatocyte differentiation has not yet been precisely characterized. In the present study, we evaluated the effect of alcohol on hepatocyte differentiation in relationship with changes of iron metabolism in HepaRG cells. This unique bipotent human cell line can differentiate into hepatocytes and biliary epithelial cells, paralleling liver development. Results showed that alcohol reduced cell viability, total protein level and enhanced hepatic enzymes leakage in differentiated HepaRG cells. Moreover, it caused cell enlargement, decreased number of hepatocyte and expression of C/EBPα as well as bile canaliculi F-actin. Alcohol increased expression of hepatic cell-specific markers and alcohol-metabolizing enzymes (ADH2, CYP2E1). This was associated with a lipid peroxidation and an iron excess expressed by an increase in total iron content, ferritin level, iron uptake as well as an overexpression of genes involved in iron transport and storage. Alcohol-induced hepatoxicity was amplified by exogenous iron via exceeding iron overload. Taken together, our data demonstrate that in differentiated hepatocytes, alcohol reduces proliferation while increasing expression of hepatic cell-specific markers. Moreover, iron overload could be one of the underlying mechanisms of effect of alcohol on the whole differentiation process of hepatocytes.