The role of the c-myc gene in pigment cell development and homeostasis

Résumé La dérégulation de c-Myc est un événement fréquent de la transformation cellulaire. Une régulation positive de cette oncoprotéine a été démontrée dans divers mélanomes cutanés primaires et métastatiques et est associée à un pronostic défavorable (Grover et al., 1996; Zhuang et al., 2008). c-Myc est considéré comme une molécule centrale impliquée dans plusieurs processus de l'homéostasie cellulaire. En raison de sa contribution importante dans la progression tumorale, la fonction de c-Myc a été étudiée intensément. Cependant nous connaissons peu le rôle de ce facteur de transcription dans l'embryogenèse et dans la spécification tissulaire. Un déficit total de c-Myc pendant l'embryogenèse conduit à la mort embryonnaire avant 10.5 jours de gestation. Cette mort est causée par de multiples imperfections du développement touchant la taille de l'embryon, le coeur, le péricarde, le tube neural et les cellules sanguines (Davis et al., 1993; Trumpp et al., 2001). Récemment, il a été montré que la plupart de ces anomalies sont secondaires et résultent d'une insuffisance du placenta dans les embryons c-myc-/- (Dubois et al., 2008). Sachant que c-Myc est important dans la maintenance des lignées de la crête neurale (Wei et al., 2007), nous nous sommes intéressés au rôle de c-Myc dans le développement des cellules pigmentaires et à leur homéostasie après la naissance. Un allèle floxé de c-myc (Trumpp et al., 2001) a été utilisé pour supprimer ce gène spécifiquement dans la lignée mélanocytaire à l'aide d'une souris transgénique Tyr::Cre (Delmas et al., 2003). L'ablation des deux allèles de c-myc dans les mélanocytes des souris c-myccKO conduit au phénotype de grisonnement des poils, observé directement après la naissance et associé à une diminution du nombre de mélanocytes dans le bulbe des follicules pileux. Les cellules pigmentaires restantes expriment les marqueurs mélanogéniques (Tyr, TRP-1, Dct and MITF) et semblent être fonctionnelles puisqu'elles peuvent produire et transférer la mélanine. De plus, la capacité de prolifération des mélanocytes déficients en c-Myc dans le bulbe des follicules pileux ne semble pas être affectée chez les nouveaux-nés. Les cellules souches mélanocytaires sont présentes, mais en nombre réduit, dans le bulge des follicules pileux à la fin de la morphogenèse chez les souris c-myccKO âgées de huit jours. Ces cellules sont maintenues sans changement durant le premier cycle pileux (vérifié à l'âge de trente jours), ce qui sous-entend que la fonction de c-Myc n'est pas nécessaire pour ce processus. Ceci explique pourquoi, en supposant que des cellules souches mélanocytaires fonctionnelles sont présentes dans la peau, nous n'observons pas de dilution de couleur de la robe liée à l'âge. Cependant, la présence de ces cellules souches mélanocytaires dans la peau c-myccKO ne suffit pas à assurer une quantité normale de mélanocytes différenciés dans le bulbe des follicules pileux. Cette population de cellules pigmentaires matures est sévèrement affectée par la suppression de c-Myc, ce qui contribue amplement au phénotype de grisonnement des poils. De plus, c-Myc paraît être important pour le développement des mélanocytes. Ainsi, le nombre de mélanoblastes diminue dans les embryons c-myccKO à partir du douzième jour de gestation. A treize jours de gestation, au stade où les mélanoblastes pénètrent dans l'épiderme et prolifèrent, les mélanoblastes déficients en c-Myc ne s'adaptent pas aux signaux de prolifération et se retrouvent en nombre réduit dans l'épiderme. Finalement, nous nous sommes intéressés, au rôle de N-Myc, un homologue proche de c-Myc, dans la lignée mélanocytaire. Nos expériences ont montré que. N-Myc était superflu pour le développement et l'homéostasie des mélanocytes, une seule copie du gène c-myc étant suffisante pour maintenir une pigmentation normale de la robe des souris c-mycc-myccKO/+~N_ myccKO/KO. Cependant, le rôle essentiel de N-Myc dans la maintenance des cellules mélanocytaires précurseurs apparaît lorsque c-Myc est absent, puisque la suppression simultanée des deux Myc résulte en une perte complète de la coloration de la robe. Ceci implique la présence d'un mécanisme compensatoire entre c- et N-Myc dans la lignée mélanocytaire, avec un rôle prédominant de c-Myc. Summary Deregulation of c-Myc is known to be a common event in cellular transformation. Upregulation of this oncoprotein was shown in a variety of primary and metastatic cutaneous melanomas and has been associated with a poor prognosis (Grover et al., 1996; Zhuang et al., 2008). c-myc is seen as a central molecule involved in many aspects of cellular homeostasis. c-Myc function has been intensively studied mostly because of its significant contribution to tumour progression. However little is known on the role of this transcription factor in embryogenesis and tissue specification. Complete loss of c-Myc during embryogenesis results in embryonic death before E10.5 due to multiple developmental defects including embryonic size, heart, pericardium, neural tube and blood cells (Davis et al., 1993; Trumpp et al., 2001). Recently it was discovered that most of these abnormalities are secondary and results of placental insufficiency in c-Myc-/- embryos (Dubois et al., 2008). Here, we focused on the role of c-Myc in pigment cell development and homeostasis after birth, knowing that c-Myc is important in the maintenance of neural crest lineages (Wei et al., 2007). A floxed allele of c-Myc (Trumpp et al., 2001) was used to specifically delete this gene in the melanocyte lineage using Tyr::Cre transgenic mice (Delmas et al., 2003). Removal of both c-Myc alleles in melanocytes of c-MyccKO mouse led to the grey hair phenotype which is seen directly after birth and was associated with a decrease in the melanocyte number in the bulb of the hair follicle. The remaining population of pigment cells express melanogenic markers (Tyr, TRP-1, Dct and MITF) and seem functionally normal since they can produce and transfer melanin. Furthermore proliferation capacity of c-Myc deficient melanocytes in the bulb of hair follicle seems not to be affected in newborn animals. Melanocyte stem cells (MSCs) are present but reduced in numbers in the bulge of the hair follicle at the end of morphogenesis in 8 days old c-MyccKO mice. These cells are maintained through the first hair cycle (as verified at P30) without any further changes, suggesting that c-Myc function is not required for this process. This explains why we did not detect any agerelated coat color dilution, assuming a presence of functional MSCs in the skin. Importantly, presence of MSCs in c-MyccKO skin was not sufficient for assuring a normal number of differentiated melanocytes in the bulb of the hair follicle. This population of mature pigmented cells is severely affected upon c-myc deletion thus largely contributing to the grey hair phenotype. Moreover, c-Myc appears to be important for melanocyte development. Thus, melanoblast number is affected in c-MyccKO embryos day 12 of gestation onwards. At E13.5, when melanoblasts enter the epidermis and proliferate, c-myc deficient melanoblasts failed to adapt to proliferation signals and are therefore reduced in number in the epidermis. Finally, we addressed the role of N-Myc, a closest homologue of c-Myc, in the melanocyte lineage. In these experiments, N-Myc was dispensable for melanocyte development and homeostasis, and even one copy of the c-myc gene was sufficient to maintain normal coat color pigmentation in c-mycc-mycCKO/+ ,N-myccKO/KO mice. However the crucial role of N-Myc in maintenance of melanocyte precursor cells became apparent when c-myc is eliminated since simultaneous deletion of both Myc results in complete loss of coat color pigmentation. This suggests compensatory mechanisms between c- and N-Myc with a predominant role of c-Myc in melanocyte lineage.

Retrieving Cloud Characteristics from Ground-Based Daytime Color All-Sky Images

A discussion is presented of daytime sky imaging and techniques that may be applied to the analysis of full-color sky images to infer cloud macrophysical properties. Descriptions of two different types of skyimaging systems developed by the authors are presented, one of which has been developed into a commercially available instrument. Retrievals of fractional sky cover from automated processing methods are compared to human retrievals, both from direct observations and visual analyses of sky images. Although some uncertainty exists in fractional sky cover retrievals from sky images, this uncertainty is no greater than that attached to human observations for the commercially available sky-imager retrievals. Thus, the application of automatic digital image processing techniques on sky images is a useful method to complement, or even replace, traditional human observations of sky cover and, potentially, cloud type. Additionally, the possibilities for inferring other cloud parameters such as cloud brokenness and solar obstruction further enhance the usefulness of sky imagers

Guidelines for Removal of Traffic Control Devices in Rural Areas, TR-527, 2005

It is commonly regarded that the overuse of traffic control devices desensitizes drivers and leads to disrespect, especially for low-volume secondary roads with limited enforcement. The maintenance of traffic signs is also a tort liability concern, exacerbated by unnecessary signs. The Federal Highway Administration’s (FHWA) Manual on Uniform Traffic Control Devices (MUTCD) and the Institute of Transportation Engineer’s (ITE) Traffic Control Devices Handbook provide guidance for the implementation of STOP signs based on expected compliance with right-of-way rules, provision of through traffic flow, context (proximity to other controlled intersections), speed, sight distance, and crash history. The approach(es) to stop is left to engineering judgment and is usually dependent on traffic volume or functional class/continuity of system. Although presently being considered by the National Committee on Traffic Control Devices, traffic volume itself is not given as a criterion for implementation in the MUTCD. STOP signs have been installed at many locations for various reasons which no longer (or perhaps never) met engineering needs. If in fact the presence of STOP signs does not increase safety, removal should be considered. To date, however, no guidance exists for the removal of STOP signs at two-way stop-controlled intersections. The scope of this research is ultra-low-volume (< 150 daily entering vehicles) unpaved intersections in rural agricultural areas of Iowa, where each of the 99 counties may have as many as 300 or more STOP sign pairs. Overall safety performance is examined as a function of a county excessive use factor, developed specifically for this study and based on various volume ranges and terrain as a proxy for sight distance. Four conclusions are supported: (1) there is no statistical difference in the safety performance of ultra-low-volume stop-controlled and uncontrolled intersections for all drivers or for younger and older drivers (although interestingly, older drivers are underrepresented at both types of intersections); (2) compliance with stop control (as indicated by crash performance) does not appear to be affected by the use or excessive use of STOP signs, even when adjusted for volume and a sight distance proxy; (3) crash performance does not appear to be improved by the liberal use of stop control; (4) safety performance of uncontrolled intersections appears to decline relative to stop-controlled intersections above about 150 daily entering vehicles. Subject to adequate sight distance, traffic professionals may wish to consider removal of control below this threshold. The report concludes with a section on methods and legal considerations for safe removal of stop control.

Sequentially aerated membrane biofilm reactors for autotrophic nitrogen removal: microbial community composition and dynamics

Membrane-aerated biofilm reactors performing autotrophic nitrogen removal can be successfully applied to treat concentrated nitrogen streams. However, their process performance is seriously hampered by the growth of nitrite oxidizing bacteria (NOB). In this work we document how sequential aeration can bring the rapid and long-term suppression of NOB and the onset of the activity of anaerobic ammonium oxidizing bacteria (AnAOB). Real-time quantitative polymerase chain reaction analyses confirmed that such shift in performance was mirrored by a change in population densities, with a very drastic reduction of the NOB Nitrospira and Nitrobacter and a 10-fold increase in AnAOB numbers. The study of biofilm sections with relevant 16S rRNA fluorescent probes revealed strongly stratified biofilm structures fostering aerobic ammonium oxidizing bacteria (AOB) in biofilm areas close to the membrane surface (rich in oxygen) and AnAOB in regions neighbouring the liquid phase. Both communities were separated by a transition region potentially populated by denitrifying heterotrophic bacteria. AOB and AnAOB bacterial groups were more abundant and diverse than NOB, and dominated by the r-strategists Nitrosomonas europaea and Ca. Brocadia anammoxidans, respectively. Taken together, the present work presents tools to better engineer, monitor and control the microbial communities that support robust, sustainable and efficient nitrogen removal

Potassium permanganate effects in postharvest conservation of the papaya cultivar Sunrise Golden

The objective of this work was to evaluate the effects of KMnO4 on the extension of postharvest life of 'Sunrise Golden' papaya, stored under modified atmosphere and refrigeration. Fruit with up to 10% yellow peel were harvested in a commercial orchard in Linhares, state of Espírito Santo, Brazil. Sets of three fruit (unit mass of 289.9±18.5 g) were wrapped in low-density polyethylene films (28 ¼m thick) containing sachets of KMnO4 at 0, 0.5, 1, 1.5, and 2 g per bag. The bags were sealed and stored at 10.4±0.9°C and 90±5% relative humidity for 25 days. After this period, the fruit were removed from the bags and maintained at 21±0.8°C and 90±5% relative humidity until complete ripening. Four days after bag sealing, CO2 concentration stabilized in all treatments, and was higher in bags without KMnO4. In all treatments, fruit reached the climacteric respiratory peak on the third day after bag removal, coinciding with peel color index of 3.5. Increasing the KMnO4 dose reduced the losses in fruit fresh matter, consistency and pulp electrolyte leakage. Potassium permanganate was effective in maintaining the fruit at the pre-climacteric stage during the 25-day storage, and did not interfere with normal ripening after bag removal.

Cambios de color y pH causados por el PVAc en bienes culturales

Este artículo presenta los primeros resultados del estudio "Identificación de patologías causadas por el PVAc en bienes culturales" que se está realizando en la Sección de Conservación-Restauración de la Facultad de Bellas Artes, Universidad de Barcelona. El trabajo se basa en el estudio de obras originales que fueron tratadas con PVAc en los años 70-80 y pretende identificar los problemas que genera el PVAc en materiales de archivo, arqueológicos, pintura sobre tela, sobre madera y pintura mural. Tras el análisis de las obras originales, se han preparado muestras probeta que reproducen sus características, así como de adhesivos de PVAc comerciales y de uso específico en restauración. Éstas han sido analizadas antes y después de someterlas a dos fases consecutivas de envejecimiento acelerado. También se han analizado obras originales con PVAc aplicado hace aproximadamente 30 años. El artículo presenta los resultados de las mediciones de color y pH en las muestras probeta antes y después de la primera fase de envejecimiento acelerado y, también, en las muestras envejecidas de forma natural durante 10 años.

Cambios de color y pH causados por el PVAc en bienes culturales

Este artículo presenta los primeros resultados del estudio "Identificación de patologías causadas por el PVAc en bienes culturales" que se está realizando en la Sección de Conservación-Restauración de la Facultad de Bellas Artes, Universidad de Barcelona. El trabajo se basa en el estudio de obras originales que fueron tratadas con PVAc en los años 70-80 y pretende identificar los problemas que genera el PVAc en materiales de archivo, arqueológicos, pintura sobre tela, sobre madera y pintura mural. Tras el análisis de las obras originales, se han preparado muestras probeta que reproducen sus características, así como de adhesivos de PVAc comerciales y de uso específico en restauración. Éstas han sido analizadas antes y después de someterlas a dos fases consecutivas de envejecimiento acelerado. También se han analizado obras originales con PVAc aplicado hace aproximadamente 30 años. El artículo presenta los resultados de las mediciones de color y pH en las muestras probeta antes y después de la primera fase de envejecimiento acelerado y, también, en las muestras envejecidas de forma natural durante 10 años.

Health consultation : Review of on-site air monitoring data during the removal action at Le Mars coal gas site, Le Mars coal gas plant Le Mars, Plymouth County, Iowa EPA facility ID: IA0001032556 (2005)

The Iowa Department of Public Health (IDPH), Hazardous Waste Site Health Assessment Program was asked by the US Environmental Protection Agency (EPA) to review a round of air sampling data. The air data was collected and analyzed during a removal action at the Le Mars Coal Gas Site in Le Mars, Iowa. EPA asked IDPH to determine from the air data if additional monitoring is necessary throughout the removal action to protect nearby residents from exposure.

Color of smoked loin from animals fed with bioactive compounds added to forage

The objective of this work was to evaluate the influence of the breed and of the addition of bioactive substances to forage on the color of smoked pork loin. Two pig breeds (Polish Landrace and the crossbreed Polish Landrace x Duroc), three types of bioactive components (organic selenium; 2% of canola oil and 1% of flaxseed oil; and 2% of flaxseed oil and 1% of canola oil), and a control treatment were evaluated. Computer image analysis included the color assessment of muscle, fat, connective tissues, and smoked loin surface. For Polish Landrace, selenium supplementation caused higher values of red, green, and blue color components of the muscle tissue, which were lower for the crossbreed. However, there was no difference in the color components of loin fat tissue of the Polish Landrace breed due to selenium supplementation. In the case of oil supplementation, values of the color components of the muscle tissue for the Polish Landrace x Duroc crossbreed were also lower. The color components of muscle, fat, connective tissues, and smoked loin surface depend on the pig breed and on the bioactive compounds added to the forage.

Proteomics of methylene blue photo-treated plasma before and after removal of the dye by an absorbent filter.

Methylene blue (MB) and light are used for virus inactivation of plasma for transfusion. However, the presence of MB has been the subject of concern, and efforts have been made to efficiently remove the dye after photo-treatment. For this study, plasma was collected by apheresis from 10 donors (group A), then treated using the MacoPharma THERAFLEX procedure (MB; 1 microM, and light exposure; 180 J/cm(2)) (group B), and finally filtered in order to remove the dye (group C). Proteins were analyzed by two-dimensional electrophoresis, and peptides showing modifications were characterized by mass spectrometry. Clottable and antigenic fibrinogen levels, as well as fibrin polymerization time were measured. Analyses of the gels focused on a region corresponding to pI between 4.5 and 6.5, and M(r) from 7000 to 58 000. In this area, 387 +/- 47 spots matched, and four of these spots presented significant modifications. They corresponded to changes of the gamma-chain of fibrinogen, of transthyretin, and of apolipoprotein A-I, respectively. A decrease of clottable fibrinogen and a prolongation of fibrin polymerization time were observed in groups B and C. Removal of MB by filtration was not responsible for additional protein alterations. The effect of over-treatment of plasma by very high concentrations of MB (50 microM) in association with prolonged light exposure (3 h) was also analyzed, and showed complex alterations of most of the plasma proteins, including fibrinogen gamma-chain, transthyretin, and apolipoprotein A-I. Our data indicates that MB treatment at high concentration and prolonged illumination severely injure plasma proteins. By contrast, at the MB concentration used to inactivate viruses, damages are apparently very restricted.

Ethephon use and application timing of abscisic acid for improving color of 'Rubi' table grape

The objective of this work was to evaluate the effect of ethephon and of abscisic acid (ABA) application timing on the color of 'Rubi' Table grape. Eight treatments were evaluated: control, without application; ethephon 500 mg L‑1 applied seven days after veraison (7 DAV); and two concentrations of ABA (200 and 400 mg L‑1) arranged with three application timings at 7 DAV, at 15 days before harvest (DBH), and at 7 DAV + 15 DBH. ABA does not modify physical‑chemical characteristics of the cluster and improves the color of grapes, especially when applied twice (7 DAV + 15 DBH) at the concentration of 400 mg L‑1.

Removal of unusual, large high-velocity metallic maxillary sinus foreign bodies by a modified free bone flap technique.

Metallic foreign bodies are rarely found in the maxillary sinus, and usually they have a dental origin.Potential complications related to foreign bodies include recurrent sinusitis, rhinolith formation, cutaneous fistula,chemical poisoning, facial neuralgic pain and even malignancies.Two main surgical approaches are currently used for the removal of foreign bodies in the maxillary sinus: the bone flap and the endoscopic sinus techniques. We are reporting two unusual cases of large high-velocity foreign bodies removed by a modified maxillary lateral antrotomy,with free bone flap repositioning and fixation with a titanium miniplate.

Inheritance of seed coat color in sesame

The objective of this work was to determine the inheritance mode of seed coat color in sesame. Two crosses and their reciprocals were performed: UCLA37 x UCV3 and UCLA90 x UCV3, of which UCLA37 and UCLA90 are white seed, and UCV3 is brown seed. Results of reciprocal crosses within each cross were identical: F1 seeds had the same phenotype as the maternal parent, and F2 resulted in the phenotype brown color. These results are consistent only with the model in which the maternal effect is the responsible for this trait. This model was validated by recording the seed coat color of 100 F2 plants (F3 seeds) from each cross with its reciprocal, in which the 3:1 expected ratio for plants producing brown and white seeds was tested with the chi-square test. Sesame seed color is determined by the maternal genotype. Proposed names for the alleles participating in sesame seed coat color are: Sc1, for brown color; and Sc2, for white color; Sc1 is dominant over Sc2.

Color of berry and juice of 'Isabel' grape treated with abscisic acid in different ripening stages

Abstract : The objective of this work was to evaluate the effect of (S)-cis-abscisic acid (S-ABA) application at different ripening stages, in increasing phenolic compounds and color of berry and juice of 'Isabel' grape (Vitis labrusca). The evaluated treatments were: control, without S-ABA application; 400 mg L-1S-ABA applied 7 days before veraison (DBV) + 400 mg L-1S-ABA at 35 days after first application (DAFA); 400 mg L-1S-ABA applied at veraison (V) + 400 mg L-1S-ABA at 35 DAFA; and 400 mg L-1S-ABA applied 7 days after veraison (DAV) + 400 mg L-1S-ABA at 35 DAFA. There was no difference among treatments regarding the physical characteristics of berries and clusters, as well as total polyphenols in berry and juice. However, there was an increase in total anthocyanins in berry and juice with S-ABA application. Colorimetric variables indicated the increase in color of berry treated with S-ABA. Juices produced from grapes treated with S-ABA were more appreciated by tasters. The treatments with 400 mg L-1S-ABA applied 7 days before, during, or 7 days after veraison, combined with an additional application 35 days after the first one, increment total anthocyanin concentration and color of berry and juice of 'Isabel' grape, with better juice acceptance, without affecting total polyphenol concentration.