Dystroglycan receptor is involved in integrin activation in intestinal epithelia

The dystroglycans (alpha-DG and beta-DG), which play important roles in the formation of basement membranes, have been well studied in skeletal muscle and nerve, but their expression and localization in intestinal epithelial cells has not been previously investigated. Here, we demonstrated that the DG complex, composed of alpha-DG, beta-DG, and utrophin, is specifically expressed in the basolateral membrane of the Caco-2-BBE monolayer. The DG complex coprecipitated with beta(1)-integrin, suggesting a possible interaction among these proteins. In addition, we observed that activation of DG receptors by laminin-1 enhanced the interaction between beta(1)-integrin and laminin-1, whereas activation of DG receptors by laminin-2 reduced the interaction between beta(1)-integrin and laminin-2. Finally, we demonstrated that the intracellular COOH-terminal tail of beta-DG and its binding to the DG binding domain of utrophin are crucial for the interactions between laminin-1/-2 and beta(1)-integrin. Collectively, these novel results indicate that dystroglycans play important roles in the regulation of interactions between intestinal epithelial cells and the extracellular matrix.     

Melatonin ingestion after exhaustive late-evening exercise improves sleep quality and quantity,and short-term performances in teenage athletes

ABSTRACT

The present study aimed to explore the effects of a single 10-mg dose of melatonin (MEL) administration after exhaustive late-evening exercise on sleep quality and quantity, and short-term physical and cognitive performances in healthy teenagers. Ten male adolescent athletes (mean ± SD, age = 15.4 ± 0.3 years, body-mass = 60.68 ± 5.7 kg, height = 167.9 ± 6.9 cm and BMI = 21.21 ± 2.5) performed two test sessions separated by at least one week. During each session, participants completed the Yo-Yo intermittent-recovery-test level-1 (YYIRT-1) at ~20:00 h. Then, sleep polysomnography was recorded from 22:15 min to 07:00 h, after a double blind randomized order administration of a single 10-mg tablet of MEL (MEL-10 mg) or Placebo (PLA). The following morning, Hooper wellness index was administered and the participants performed the Choice Reaction Time (CRT) test, the Zazzo test and some short-term physical exercises (YYIRT-1, vertical and horizontal Jumps (VJ; HJ), Hand grip strength (HG), and five-jump test (5-JT)). Evening total distance covered in the YYIRT-1 did not change during the two conditions (p > 0.05). Total sleep time (Δ = 24.55 mn; p < 0.001), sleep efficiency (Δ = 4.47%; p < 0.001), stage-3 sleep (N3 sleep) (Δ = 1.73%; p < 0.05) and rapid-eye-movement sleep (Δ = 2.15%; p < 0.001) were significantly higher with MEL in comparison with PLA. Moreover, sleep-onset-latency (Δ = –8.45mn; p < 0.001), total time of nocturnal awakenings after sleep-onset (NA) (Δ = –11 mn; p < 0.001), stage-1 sleep (N1 sleep) (Δ = –1.7%; p < 0.001) and stage-2 sleep (N2 sleep) (Δ = ?1.9%; p < 0.05) durations were lower with MEL. The Hooper index showed a better subjective sleep quality, a decrease of the subjective perception of fatigue and a reduced level of muscle soreness with MEL. Moreover, MEL improved speed and performance but not inaccuracy during the Zazzo test. CRT was faster with MEL. Morning YYIRT-1 (Δ = 82 m; p < 0.001) and 5-JT (Δ = 0.08 m; p < 0.05) performances were significantly higher with MEL in comparison with PLA. In contrast, HG, VJ and HJ performances did not change during the two conditions (p > 0.05). The administration of a single dose of MEL-10 mg after strenuous late-evening exercise improved sleep quality and quantity, selective attention, subjective assessment of the general wellness state, and some short-term physical performances the following morning in healthy teenagers.     

Late Carboniferous-Early Permian Tetrapod Ichnofauna from the Khenifra Basin, Central Morocco

In terms of cumulative thickness and areal extent, the Khenifra Basin is one of the most important outcrops of Late Palaeozoic red-beds in central Morocco. Macro- and microfloral remains near the centre of the 1800 m-thick succession of interbedded conglomerates, sandstones, and mudstones are considered to be of middle to late Early Permian age. Here we give the first comprehensive analysis of the vertebrate ichnofossil record from the study area, based on 17 specimens of isolated footprints and incomplete step cycles collected at three localities that are lithostratigraphically equivalent to the plant-bearing horizons. The tetrapod ichnofauna comprises tracks of the plexus Batrachichnus Woodworth - Limnopus Marsh, Ichniotherium sphaerodactylum (Pabst), Dimetropus Romer and Price, and Dromopus Marsh which can be referred to temnospondyl, diadectomorph, synapsid (“pelycosaurian”) and early sauropsid trackmakers. This clearly Euramerican footprint assemblage, including the first occurrences of Ichniotherium and Dimetropus from outside Europe and North America, indicates a Late Carboniferous to Early Permian age of the fossiliferous strata. Judging from the relatively diverse ichnofauna and flora, the Khenifra Basin must have represented a well-established terrestrial ecosystem during that period. Its habitat could be specially important for the understanding of the phylogeny and dispersal of early tetrapods, inasmuch as we are able to report on an extremely rare type of diadectomorph footprint hitherto known only from the Early Permian of central Germany.     

New insights in the removal of the hydantoins, oxidation product of pyrimidines, via the base excision and nucleotide incision repair pathways

Background

Oxidative damage to DNA, if not repaired, can be both miscoding and blocking. These genetic alterations can lead to mutations and/or cell death, which in turn cause cancer and aging. Oxidized DNA bases are substrates for two overlapping repair pathways: base excision (BER) and nucleotide incision repair (NIR). Hydantoin derivatives such as 5-hydroxyhydantoin (5OH-Hyd) and 5-methyl-5-hydroxyhydantoin (5OH-5Me-Hyd), major products of cytosine and thymine oxidative degradation pathways, respectively, have been detected in cancer cells and ancient DNA. Hydantoins are blocking lesions for DNA polymerases and excised by bacterial and yeast DNA glycosylases in the BER pathway. However little is known about repair of pyrimidine-derived hydantoins in human cells.

Methodology/Principal Findings

Here, using both denaturing PAGE and MALDI-TOF MS analyses we report that the bacterial, yeast and human AP endonucleases can incise duplex DNA 5′ next to 5OH-Hyd and 5OH-5Me-Hyd thus initiating the NIR pathway. We have fully reconstituted the NIR pathway for these lesions in vitro using purified human proteins. Depletion of Nfo in E. coli and APE1 in HeLa cells abolishes the NIR activity in cell-free extracts. Importantly, a number of redundant DNA glycosylase activities can excise hydantoin residues, including human NTH1, NEIL1 and NEIL2 and the former protein being a major DNA glycosylase activity in HeLa cells extracts.

Conclusions/Significance

This study demonstrates that both BER and NIR pathways can compete and/or back-up each other to remove hydantoin DNA lesions in vivo.     

Jurassic reef events in the Moroccan Atlas: A review

In the Moroccan Atlas, sedimentary deposits provide important data on reef events that characterize the Jurassic period. Recent work allows us to enhance knowledge of the Jurassic reefs in the Atlas, in particular their age, character and palaeogeographic distribution. Numerous localities with sponge-microbial mud mounds, coral reefs, and lithiotid bioherms are recorded from the Middle and High Atlas regions. These different biogenic constructions occupy different palaeogeographic settings: on the top of tilted blocks within the basin center; in slightly deeper positions, at the basin platform junction; and on adjacent platforms in the middle of the coastal area. The main episodes of reef building span nearly 30 million years, as follows: (1) Sinemurian, (2) early Pliensbachian, (3) late Toarcian, (4) Aalenian–early Bajocian (pars), and (5) late Bajocian. These five distinct reef events can be linked with general fluctuations of sea level and tectonism, and have palaeoclimatic implications.     

Tempestitic shell beds formed by a new serpulid polychaete from the Bajocian (Middle Jurassic) of the Central High Atlas (Morocco)

Serpulid-dominated shell beds from the Lower Bajocian (Middle Jurassic) of the Central High Atlas (Morocco) are primarily formed by quadrangular tubes of the serpulid polychaete Nogrobs moroccensis sp. nov. The transverse tube ornament consisting predominately of wrinkles, the lack of narrow spirals and the shortness or absence of attached posterior tube portion represent the diagnostic features of this new species. Tubes of Nogrobs moroccensis co-occur with gastropods and oyster–crinoidal debris. We suggest that these serpulids were free-lying in their adult stage and formed benthic meadows on soft-bottom habitats. They frequently show parallel alignment in plane views in both graded and non-graded, few-cm-thick shell beds, corresponding to distal tempestites deposited close to and below storm wave base in an outer-ramp environment. The excellent preservation of serpulids implies short residence time on the seafloor, and the lack of compositional mixing with other communities and their restriction to outer-ramp habitats indicate that they represent parautochthonous assemblages. Analyses of serpulid orientation on the bottom bedding planes of shell beds revealed weak unidirectional arrangement of tubes, whereas orientation on the top bedding planes revealed strong unidirectional arrangement of tubes, with significantly different orientation relative to the bottom planes. This change could suggest a shift from predominantly a current component of the combined storm flow (with uneven seafloor surface and adhesion of skeletal grains to muddy substrate) to significant oscillatory component of the waning storm, as also suggested by bidirectional orientation of gastropods.     

The role of sucrose during maturation of black spruce (Picea mariana) and white spruce (Picea glauca) somatic embryos

The present study was conducted to understand the role of sucrose in the medium on the maturation of black spruce and white spruce somatic embryos. A maturation medium containing 6% sucrose, which hydrolyzed into glucose and fructose, gave significantly more embryos than a medium containing 3.16% of each glucose and fructose. Preventing the complete sucrose hydrolysis by a daily transfer of the tissues onto fresh medium significantly decreased the yield of somatic embryos compared to when sucrose was allowed to complete its hydrolysis. This reduction was not due to the manipulation of the tissues during the transfer, since a daily in situ transfer did not affect embryo production. To verify if the better embryo production observed on a medium containing 6% sucrose was due to the increasing osmotic pressure of the medium, this increasing osmotic pressure was simulated with a sequence of media containing different concentrations of glucose and fructose. Unexpectedly and for both species, this simulation did not improve somatic embryo production, which stayed similar to the one obtained on constant osmotic pressure. To understand these results, embryos produced on the different treatments were analyzed in terms of sucrose, glucose, fructose and starch levels and protein contents. The embryo carbohydrate content was independent from the carbohydrate used in the maturation medium. However, embryos matured on 6% sucrose allowed to hydrolyze during the maturation period contained significantly more soluble and insoluble proteins than embryos matured on any other treatment. Furthermore, embryos with a higher protein content also exhibited a higher epicotyl appearance frequency. The role of sucrose as a regulatory factor during the maturation of spruce somatic embryos is discussed.     

Distinguishing the optimal binding mechanism of an E3 ubiquitin ligase: Covalent versus noncovalent inhibition

The development of covalent drugs, specifically in cancer therapeutics, has recently sparked interest among the pharmaceutical research community. While representing a significant fraction of the drugs in the market, very few have been deliberately designed to interact covalently with their biological target. One of the enzymes that have been both covalently and noncovalently targeted is the Neural Precursor Cell Expressed Developmentally Downregulated gene 4-1 (Nedd4-1). This enzyme has been found to have multiple physiological implications, including its involvement in cancer invasion. A critical gap still remains in the molecular understanding of the structural mechanism upon the covalent and noncovalent binding to Nedd4-1. In this study, we explore the most optimal binding mechanism in the inhibition of the catalytic site of the Nedd4-1. Our results exhibited a greater stability in the covalent complex compared with the noncovalent complex. This was supported by the secondary structure elements that were more dominant in the covalently inhibited complex. This complex disclosed an optimal free binding energy landscape, induced by the catalytic site energy contributions that showed to be more favorable. The insights demonstrating the above binding mechanism of Nedd4-1 establishes covalent inhibition as the preferred method of inhibition of the enzyme. This investigation aids in the understanding of the structural mechanism of Nedd4-1 inhibition and would assist in the design of more potent covalent inhibitors at the catalytic site of Nedd4-1.