Chemical inhibitors: a tool for plant cell cycle studies

AbstractSynchrony provides a large number of cells at defined points of the cell cycle. Highly synchronised cells are powerful and effective tools for molecular analyses and for studying the biochemical events of the cell cycle in plants. Usually, plant cell suspensions can be synchronised by chemical agents, which arrest the cell cycle by acting on the driving forces of the cell cycle engine such as cyclin-dependent kinase activity, enzymes involved in DNA synthesis or proteolysis of cell cycle regulators or by acting on the cell cycle apparatus (mitotic spindle). The specificity, reversibility and efficiency of each type of cell cycle inhibitor are described and related to their mode of action

Paleogeographic Interpretation of a Peat Layer at Torson di Sotto (Lagoon of Venice, Italy)

Ein 3,3 m langer Kern aus dem zentralen Bereich der Lagune von Venedig enthielt eine Torflage zwischen 1,3 und 2,0 m unter dem mittleren Meeresspiegel. Pollenanalysen zeigen ein Vorherrschen von Quercus und Alnus und eine transgressive Phase an: der untere Teil des Torfes (nach 14C-Daten 1730±80 v.h.) ist in einem Süßwasser-Environment gebildet worden, der obere Teil (nach 14C-Daten 1140 + 80 v.h.) in einem brackischen. Foraminiferenanalysen zeigen an, daß der Transgression eine Regression vorangegangen ist. Die gesamte Abfolge enthält von unten nach oben (Abb. 3): (1) ein oligohalines bis mesohalines Environment (von —3,3 bis —2,5 m) mit sehr zahlreichen Foraminiferen; (2) ein Süßwasser-Environment (von —2,5 bis —2,0 m); (3) die 0,7 m mächtige Torflage, die fortschreitend brackisch wird; (4) ein mesohalines Environment (oberhalb —1,3 m).Korrelationen mit benachbarten Lokalitäten (Abb. 1) weisen darauf hin, daß vergleichbare Erscheinungen zu gleicher Zeit im Zentralteil der Lagune, aber nicht in ihrem Nordteil, aufgetreten sind. Diese Regressions-Transgressions-Abfolge wird auf eine Periode erhöhter Niederschläge um das Ende der Römerzeit zurückgeführt, die zur Überflutung und zu einer Verlagerung des Laufes zahlreicher Flüsse geführt hat; dabei ist das ökologische Gleichgewicht dieses Marschengebietes zeitweilig verändert worden. Im Verlauf der vergangenen 100 Jahre hat sich die Transgression beschleunigt, überwiegend infolge anthropogener Eingriffe. Die „Barene" werden jetzt erodiert und verschwinden rasch (Abb. 2). Sie werden durch weite Meerwasserbecken eingenommen.researc

Trajectory tracking and time delay management of 4-mecanum wheeled mobile robots (4-MWMR)

International audienceNowadays, wheeled mobile robots have a very important role in industrial applications, namely in transportation tasks thanks to their accuracy and rapidity. However, meeting obstacles while executing a mission can cause an important time delay, which is not appreciable in industry where production must be optimal. This paper deals with the time delay management, the trajectory generation and the tracking problem applied on four wheeled omnidirectional mobile robots. A strategy is proposed to minimize or compensate the time delay caused by obstacles. The approach is done by updating the reference trajectory. This update helps to track the trajectory in real time, a new control law based on the feedback linearization control theory is synthesized to track perfectly generated or updated trajectories

Detection & isolation of sensor and actuator additive faults in a 4-mecanum wheeled mobile robot (4-MWMR)

International audienceIn this paper, the fault detection and isolation problem regarding actuation and sensing of a 4-mecanum wheeled mobile robot (4-MWMR) is studied. The challenge with respect to the current state of the art lies in detecting and distinguishing wheel sensor from wheel actuator additive faults for this kind of robots. An approach based on generating residuals is proposed. Sensor faults isolation is based on simply analyzing residual signatures which are different under each sensor fault. Due to omni-move properties, actuator faults are, however, more difficult to be isolated. More residual characteristics must be taken into consideration to achieve the isolation

Gut barrier-microbiota imbalances in early life lead to higher sensitivity to inflammation in a murine model of C-section delivery

Background Most interactions between the host and its microbiota occur at the gut barrier, and primary colonizers are essential in the gut barrier maturation in the early life. The mother–ofspring transmission of microorganisms is the most important factor infuencing microbial colonization in mammals, and C‑section delivery (CSD) is an impor‑ tant disruptive factor of this transfer. Recently, the deregulation of symbiotic host‑microbe interactions in early life has been shown to alter the maturation of the immune system, predisposing the host to gut barrier dysfunction and infammation. The main goal of this study is to decipher the role of the early‑life gut microbiota‑barrier alterations and its links with later‑life risks of intestinal infammation in a murine model of CSD. Results The higher sensitivity to chemically induced infammation in CSD mice is related to excessive exposure to a too diverse microbiota too early in life. This early microbial stimulus has short‑term consequences on the host homeo‑ stasis. It switches the pup’s immune response to an infammatory context and alters the epithelium structure and the mucus‑producing cells, disrupting gut homeostasis. This presence of a too diverse microbiota in the very early life involves a disproportionate short‑chain fatty acids ratio and an excessive antigen exposure across the vulnerable gut barrier in the frst days of life, before the gut closure. Besides, as shown by microbiota transfer experiments, the microbiota is causal in the high sensitivity of CSD mice to chemical‑induced colitis and in most of the phenotypical parameters found altered in early life. Finally, supplementation with lactobacilli, the main bacterial group impacted by CSD in mice, reverts the higher sensitivity to infammation in ex‑germ‑free mice colonized by CSD pups’ microbiota. Conclusions Early‑life gut microbiota‑host crosstalk alterations related to CSD could be the linchpin behind the phe‑ notypic efects that lead to increased susceptibility to an induced infammation later in life in mice. Keywords C‑section delivery, Microbiota, Primary colonization, Early life, Infammation, Gut barrier, Murine modelinfo:eu-repo/semantics/publishedVersio

Gut barrier-microbiota imbalances in early life lead to higher sensitivity to inflammation in a murine model of C-section delivery

Most interactions between the host and its microbiota occur at the gut barrier, and primary colonizers are essential in the gut barrier maturation in the early life. The mother-offspring transmission of microorganisms is the most important factor influencing microbial colonization in mammals, and C-section delivery (CSD) is an important disruptive factor of this transfer. Recently, the deregulation of symbiotic host-microbe interactions in early life has been shown to alter the maturation of the immune system, predisposing the host to gut barrier dysfunction and inflammation. The main goal of this study is to decipher the role of the early-life gut microbiota-barrier alterations and its links with later-life risks of intestinal inflammation in a murine model of CSD. The higher sensitivity to chemically induced inflammation in CSD mice is related to excessive exposure to a too diverse microbiota too early in life. This early microbial stimulus has short-term consequences on the host homeostasis. It switches the pup's immune response to an inflammatory context and alters the epithelium structure and the mucus-producing cells, disrupting gut homeostasis. This presence of a too diverse microbiota in the very early life involves a disproportionate short-chain fatty acids ratio and an excessive antigen exposure across the vulnerable gut barrier in the first days of life, before the gut closure. Besides, as shown by microbiota transfer experiments, the microbiota is causal in the high sensitivity of CSD mice to chemical-induced colitis and in most of the phenotypical parameters found altered in early life. Finally, supplementation with lactobacilli, the main bacterial group impacted by CSD in mice, reverts the higher sensitivity to inflammation in ex-germ-free mice colonized by CSD pups' microbiota. Early-life gut microbiota-host crosstalk alterations related to CSD could be the linchpin behind the phenotypic effects that lead to increased susceptibility to an induced inflammation later in life in mice

Pre-transplant CD45RC expression on blood T cells differentiates patients with cancer and rejection after kidney transplantation

Background Biological biomarkers to stratify cancer risk before kidney transplantation are lacking. Several data support that tumor development and growth is associated with a tolerant immune profile. T cells expressing low levels of CD45RC preferentially secrete regulatory cytokines and contain regulatory T cell subset. In contrast, T cells expressing high levels of CD45RC have been shown to secrete proinflammatory cytokines, to drive alloreactivity and to predict acute rejection (AR) in kidney transplant patients. In the present work, we evaluated whether pre-transplant CD45RClow T cell subset was predictive of post-transplant cancer occurrence. Methods We performed an observational cohort study of 89 consecutive first time kidney transplant patients whose CD45RC T cell expression was determined by flow cytometry before transplantation. Post-transplant events including cancer, AR, and death were assessed retrospectively. Results After a mean follow-up of 11.1±4.1 years, cancer occurred in 25 patients (28.1%) and was associated with a decreased pre-transplant proportion of CD4+CD45RChigh T cells, with a frequency below 51.9% conferring a 3.7-fold increased risk of post-transplant malignancy (HR 3.71 [1.24–11.1], p = 0.019). The sensibility, specificity, negative predictive and positive predictive values of CD4+CD45RChigh<51.9% were 84.0, 54.7, 89.8 and 42.0% respectively. Confirming our previous results, frequency of CD8+CD45RChigh T cells above 52.1% was associated with AR, conferring a 20-fold increased risk (HR 21.7 [2.67–176.2], p = 0.0004). The sensibility, specificity, negative predictive and positive predictive values of CD8+CD45RChigh>52.1% were 94.5, 68.0, 34.7 and 98.6% respectively. Frequency of CD4+CD45RChigh T cells was positively correlated with those of CD8+CD45RChigh (p<0.0001), suggesting that recipients with high AR risk display a low cancer risk. Conclusion High frequency of CD45RChigh T cells was associated with AR, while low frequency was associated with cancer. Thus, CD45RC expression on T cells appears as a double-edged sword biomarker of promising interest to assess both cancer and AR risk before kidney transplantation

Immunodetection of retinoblastoma-related protein and its phosphorylated form in interphase and mitotic alfalfa cells

Plant retinoblastoma-related (RBR) proteins are primarily considered as key regulators of G1/S phase transition, with functional roles in a variety of cellular events during plant growth and organ development. Polyclonal antibody against the C-terminal region of the Arabidopsis RBR1 protein also specifically recognizes the alfalfa 115 kDa MsRBR protein, as shown by the antigen competition assay. The MsRBR protein was detected in all cell cycle phases, with a moderate increase in samples representing G2/M cells. Antibody against the human phospho-pRb peptide (Ser807/811) cross-reacted with the same 115 kDa MsRBR protein and with the in vitro phosphorylated MsRBR protein C-terminal fragment. Phospho-MsRBR protein was low in G1 cells. Its amount increased upon entry into the S phase and remained high during the G2/M phases. Roscovitine treatment abolished the activity of alfalfa MsCDKA1;1 and MsCDKB2;1, and the phospho-MsRBR protein level was significantly decreased in the treated cells. Colchicine block increased the detected levels of both forms of MsRBR protein. Reduced levels of the MsRBR protein in cells at stationary phase or grown in hormone-free medium can be a sign of the division-dependent presence of plant RBR proteins. Immunolocalization of the phospho-MsRBR protein indicated spots of variable number and size in the labelled interphase nuclei and high signal intensity of nuclear granules in prophase. Structures similar to phospho-MsRBR proteins cannot be recognized in later mitotic phases. Based on the presented western blot and immunolocalization data, the possible involvement of RBR proteins in G2/M phase regulation in plant cells is discussed

Arabidopsis A BOUT DE SOUFFLE is a putative mitochondrial transporter involved in photorespiratory metabolism and is required for meristem growth at ambient CO2 levels

Photorespiratory metabolism is essential in all oxygenic photosynthetic organisms. In plants, it is a highly compartmentalized pathway that involves chloroplasts, peroxisomes, mitochondria and the cytoplasm. The metabolic pathway itself is well characterized, and the enzymes required for its function have been identified. However, very little information is available on the transport proteins that catalyze the high metabolic flux between the involved compartments. Here we show that the A BOUT DE SOUFFLE (BOU) gene, which encodes a mitochondrial carrier, is involved in photorespiration in Arabidopsis. BOU was found to be co-expressed with photorespiratory genes in leaf tissues. The knockout mutant bou-2 showed the hallmarks of a photorespiratory growth phenotype, an elevated CO2 compensation point, and excessive accumulation of glycine. Furthermore, degradation of the P-protein, a subunit of glycine decarboxylase, was demonstrated for bou-2, and is reflected in strongly reduced glycine decarboxylase activity. The photorespiration defect in bou-2 has dramatic consequences early in the seedling stage, which are highlighted by transcriptome studies. In bou-2 seedlings, as in shm1, another photorespiratory mutant, the shoot apical meristem organization is severely compromised. Cell divisions are arrested, leading to growth arrest at ambient CO2. Although the specific substrate for the BOU transporter protein remains elusive, we show that it is essential for the function of the photorespiratory metabolism. We hypothesize that BOU function is linked with glycine decarboxylase activity, and is required for normal apical meristems functioning in seedlings

Predicting understory maximum shrubs cover using altitude and overstory basal area in different Mediterranean forests

In some areas of the Mediterranean basin where the understory stratum represents a critical fire hazard, managing the canopy cover to control the understory shrubby vegetation is an ecological alternative to the current mechanical management techniques. In this study, we determine the relationship between the overstory basal area and the cover of the understory shrubby vegetation for different dominant canopy species (Pinaceae and Fagaceae species) along a wide altitudinal gradient in the province of Catalonia (Spain). Analyses were conducted using data from the Spanish National Forest Inventory. At the regional scale, when all stands are analysed together, a strong negative relationship between mean shrub cover and site elevation was found. Among the Pinaceae species, we found fairly good relationships between stand basal area and the maximum development of the shrub stratum for species located at intermediate elevations (Pinus nigra, Pinus sylvestris). However, at the extremes of the elevationclimatic gradient (Pinus halepensis and Pinus uncinata stands), stand basal area explained very little of the shrub cover variation probably because microsite and topographic factors override its effect. Among the Fagaceae species, a negative relationship between basal area and the maximum development of the shrub stratum was found in Quercus humilis and Fagus sylvatica dominated stands but not in Quercus ilex. This can be due to the particular canopy structure and management history of Q. ilex stands. In conclusion, our study revealed a marked effect of the tree layer composition and the environment on the relationship between the development of the understory and overstory tree structure. More fine-grained studies are needed to provide forest managers with more detailed information about the relationship between these two forest strata