Effects of a nursery CO2 enriched atmosphere on the germination and seedling morphology of two Mediterranean oaks with contrasting leaf habit

Abstract: The use of an enriched CO2 atmosphere in tree nurseries has been envisaged as a promising technique to increase productivity and to obtain seedlings with a higher root/shoot ratio, an essential trait to respond to water stress in Mediterranean-type ecosystems. In that framework, we have analyzed the effects of three levels of atmospheric CO2 concentration (350, 500 and 700 ppm) on the germination rate, growth and morphology of seedlings of two Mediterranean oaks used in reforestation programs: the evergreen Quercus ilex L. and the deciduous Quercus cerrioides Wilk. et Costa. CO2 enrichment increased the germination rate of Q. cerrioides (from 70±7 to 81±3 %) while it decreased that of Q. ilex (from 71±10 to 41±12 %). Seedlings of both species increased approximately 60% their total biomass in response to CO2 enrichment but at two different CO2 concentrations: 500 ppm for Q. cerrioides and 700 ppm for Q. ilex. This increase in seedlings biomass was entirely due to an augmentation of root biomass. Considering germination and biomass partitioning, an enriched CO2 atmosphere might not be appropriate for growing Mediterranean evergreen oaks, such as Q. ilex, since it reduces acorn germination and the only gains in root biomass occur at a high concentration (700 ppm). On the other hand, a moderate CO2 enrichment (500 ppm) appears as a promising nursery technique to stimulate the germination, growth and root/shoot ratio of deciduous oaks, such as Q. cerrioides. Resumen: El uso de una atmósfera enriquecida en CO2 durante la fase de vivero puede contribuir a aumentar la producción viverística, a la vez que ayudar a conseguir plántulas con una mayor relación biomasa subterránea/biomasa aérea, más adecuadas para hacer frente al severo estrés hídrico que generalmente limita el éxito de las repoblaciones en el clima Mediterráneo. En este estudio hemos analizado el efecto de tres niveles de abonado carbónico atmosférico (350, 500 y 750 ppm) en la germinación y morfología de plántulas de encina (Quercus ilex) y roble cerrioide (Quercus cerrioides). Una atmósfera enriquecida en CO2 incrementó la germinación de Q. cerrioides (de 70±7 a 81±3 %) mientras que disminuyó la de Q. ilex (de 71±10 a 41±12 %). Las plántulas de ambas especies incrementaron aproximadamente un 60% su biomasa en respuesta a una mayor concentración de CO2, aunque esta respuesta se produjo a diferentes dosis: 500 ppm en Q. cerrioides y 700 ppm en Q. ilex. El aumento en la biomasa total de las plántulas se debió enteramente a un mayor desarrollo de su sistema radical, Considerando tanto la germinación como los efectos sobre la relación biomasa subterránea/biomasa aérea, una atmósfera enriquecida en CO2 no parece ser un tratamiento adecuado para la producción en vivero de plántulas de Q.ilex, puesto que diminuye su germinación y solo aumenta su sistema radicular a dosis muy elevadas (700 ppm). Por el contrario, un aumento moderado en la concentración de CO2 (500 ppm) aparece como una técnica interesante para estimular el crecimiento y obtener plántulas de Q. cerrioides con un sistema radical más desarrollado.

Biomphalaria Tenagophila Guaibensis ssp. n. from Southern Brazil and Uruguay (pulmonata: Planorbidae). I. Morphology

A new subspecies of planorbid snail, biomphalaria tenagophila guaibensis, is described. It has been found along the coastal belt of the Brazilian state of rio grande do Sul and the middle part of Uruguay, from Porto Alegre to Mercedes. It differs from the nominate subspecies, Biomphalaria tenagophila tenagophila, in the appearance of the penial complex (prepuce longer and proportionally slenderer in B. t. guaibensis, shorter and proportionally stouter in b. t. tenagophila), in the ratio between the lengths of the penial sheath and the prepuce, in the ratio between the lengths of the uterine complex and the penial complex, and in a coefficient of difference of 2.44 for the ratio between the penis sheath and prepuce and of 2.02 for the ratio between the uterine complex and penial complex. The shell and the other organs of the genital system are similar in both subspecies. B. t. guaibensis is very similar to Biomphalaria occidentalis Paraense, 1981, but is readily separated from it by the presence of a vaginal pouch, which is lacking in the latter, besides showing highly significant difference in the penis sheath: prepuce and uterine complex: penial complex ratios. Crossbreeding experiments which lend additional support to the recognition of B. t. guaibensis as a subspecies will be reported elsewhere.

Generalized factor models: a bayesian approach

There is recent interest in the generalization of classical factor models in which the idiosyncratic factors are assumed to be orthogonal and there are identification restrictions on cross-sectional and time dimensions. In this study, we describe and implement a Bayesian approach to generalized factor models. A flexible framework is developed to determine the variations attributed to common and idiosyncratic factors. We also propose a unique methodology to select the (generalized) factor model that best fits a given set of data. Applying the proposed methodology to the simulated data and the foreign exchange rate data, we provide a comparative analysis between the classical and generalized factor models. We find that when there is a shift from classical to generalized, there are significant changes in the estimates of the structures of the covariance and correlation matrices while there are less dramatic changes in the estimates of the factor loadings and the variation attributed to common factors.

Nuclear factor I-C links platelet-derived growth factor and transforming growth factor beta1 signaling to skin wound healing progression.

Transforming growth factor beta (TGF-beta) and platelet-derived growth factor A (PDGFAlpha) play a central role in tissue morphogenesis and repair, but their interplay remain poorly understood. The nuclear factor I C (NFI-C) transcription factor has been implicated in TGF-beta signaling, extracellular matrix deposition, and skin appendage pathologies, but a potential role in skin morphogenesis or healing had not been assessed. To evaluate this possibility, we performed a global gene expression analysis in NFI-C(-/-) and wild-type embryonic primary murine fibroblasts. This indicated that NFI-C acts mostly to repress gene expression in response to TGF-beta1. Misregulated genes were prominently overrepresented by regulators of connective tissue inflammation and repair. In vivo skin healing revealed a faster inflammatory stage and wound closure in NFI-C(-/-) mice. Expression of PDGFA and PDGF-receptor alpha were increased in wounds of NFI-C(-/-) mice, explaining the early recruitment of macrophages and fibroblasts. Differentiation of fibroblasts to contractile myofibroblasts was also elevated, providing a rationale for faster wound closure. Taken together with the role of TGF-beta in myofibroblast differentiation, our results imply a central role of NFI-C in the interplay of the two signaling pathways and in regulation of the progression of tissue regeneration.

Ectodysplasin A1 promotes placodal cell fate during early morphogenesis of ectodermal appendages.

Organs developing as appendages of the ectoderm are initiated from epithelial thickenings called placodes. Their formation is regulated by interactions between the ectoderm and underlying mesenchyme, and several signalling molecules have been implicated as activators or inhibitors of placode formation. Ectodysplasin (Eda) is a unique signalling molecule in the tumour necrosis factor family that, together with its receptor Edar, is necessary for normal development of ectodermal organs both in humans and mice. We have shown previously that overexpression of the Eda-A1 isoform in transgenic mice stimulates the formation of several ectodermal organs. In the present study, we have analysed the formation and morphology of placodes using in vivo and in vitro models in which both the timing and amount of Eda-A1 applied could be varied. The hair and tooth placodes of K14-Eda-A1 transgenic embryos were enlarged, and extra placodes developed from the dental lamina and mammary line. Exposure of embryonic skin to Eda-A1 recombinant protein in vitro stimulated the growth and fusion of placodes. However, it did not accelerate the initiation of the first wave of hair follicles giving rise to the guard hairs. Hence, the function of Eda-A1 appears to be downstream of the primary inductive signal required for placode initiation during skin patterning. Analysis of BrdU incorporation indicated that the formation of the epithelial thickening in early placodes does not involve increased cell proliferation and also that the positive effect of Eda-A1 on placode expansion is not a result of increased cell proliferation. Taken together, our results suggest that Eda-A1 signalling promotes placodal cell fate during early development of ectodermal organs.

Insulin secretion is regulated by the glucose-dependent production of islet beta cell macrophage migration inhibitory factor.

Macrophage migration inhibitory factor (MIF), originally identified as a cytokine secreted by T lymphocytes, was found recently to be both a pituitary hormone and a mediator released by immune cells in response to glucocorticoid stimulation. We report here that the insulin-secreting beta cell of the islets of Langerhans expresses MIF and that its production is regulated by glucose in a time- and concentration-dependent manner. MIF and insulin colocalize by immunocytochemistry within the secretory granules of the pancreatic islet beta cells, and once released, MIF appears to regulate insulin release in an autocrine fashion. In perifusion studies performed with isolated rat islets, immunoneutralization of MIF reduced the first and second phase of the glucose-induced insulin secretion response by 39% and 31%, respectively. Conversely, exogenously added recombinant MIF was found to potentiate insulin release. Constitutive expression of MIF antisense RNA in the insulin-secreting INS-1 cell line inhibited MIF protein synthesis and decreased significantly glucose-induced insulin release. MIF is therefore a glucose-dependent, islet cell product that regulates insulin secretion in a positive manner and may play an important role in carbohydrate metabolism.

Observations on the morphology of Pomacea lineata (Spix, 1827) (Mollusca, Ampullariidae)

This paper deals with the morpholgy of Pomacea lineata (Spix, 1827) collected at its type locality. The shell is globose, moderately heavy, horn-colored with brown spiral bands; apex subelevated; 4 - 5 rounded whorls increasing in diameter rather rapidly, separated by deep suture. Aperture large and ovoid; outer lip sharp; umbilicus narrow and deep; operculum concentric, corneous. Ratios: shell width/shell length = 0.74 - 0.83 (mean 0.78); spire length/shell length = 0.10 - 0.18 (mean 0.13); aperture length/shell length = 0.70 - 0.77 (mean 0.73). The animal is longisiphonate. Renal organ brownish with marked invagination at its right edge. Ureter elongated with its long axis transverse to the main axis of the kidney. The radula is taenioglossate (2.1.1.1.2) and has on average 35 transverse rows of teeth. The form and arrangement of the radula teeth are nearly the same as in other Ampullariidae. The testis is cream-colored and lies in the first three whorls of the spire. Spermiduct uniformly narrow, running to the base of the spire. Seminal vesicle whitish, slightly pressed dorsoventrally. Prostate cylindric and thick, similar in color to the testis. Penis whiplike, with a closed circular spermiduct. Penis pouch ovoid completely envelping the penis. Penis sheath elongated, broad prosimally, tapering distally. Its inner surface shows a longitudinal channel along its proximal half and two glands, one on the middle and the other apical. Ovary composed of branched whitish tubules situated on the surface of the digestive gland. Oviduct slender running along the columellar axis toward the base of the spire. Seminal receptalble tubiform, thick-walled and rounded proximally. Albumen gland large, pink, enclosing the receptacle and the spiral capsule gland. Vestigial male copulatory apparatus (penis and its sheath) present in all females examined.

Regulació del factor de transcripció NFAT5 per les quinases WNK1, SPAK i OSR1 i cerca de quinases activadores de la pròpia WNK1 en resposta a estrés osmòtic

Estudi elaborat a partir d’una estada a la School of Life Sciences de la University of Dundee, Gran Bretanya, entre gener i març del 2007.L'estrès osmòtic causa rà pidament l'activació de la quinasa WNK1, que fosforila i activa a continuació les quinases SPAK i OSR1, que alhora regulen canals i transportadors d’ions preexistents a la membrana cel•lular. El factor de transcripció NFAT5 és el principal regulador de la resposta cel•lular transcripcional secundà ria a hipertonicitat i s’ha descrit que les quinases p38, Fyn, PKA, ERK/MEK i ATM estan involucrades en la seva regulació post-traduccional. No obstant, com que la funció d’aquestes quinases no explica totalment els mecanismes d'activació de NFAT5, s’ha estudiat si l’activitat transcripcional de NFAT5 pot estar regulada per WNK1, SPAK o OSR1. Així doncs, es va observar que l’activitat d’un reporter dependent de NFAT5 no es veu afectada per la presència de cap de les quinases anteriors, en la seva forma wild-type o dominant negatiu. D’altra banda, es va estudiar quin domini de WNK1 és necessari per a que pugui respondre a hipertonicitat i quines quinases poden estar involucrades en la fosforilació de la serina 382 de WNK1. En conclusió, les dades obtingudes apunten que l’activació de WNK1 en resposta a estrès osmòtic requereix la seva fosforilació en la serina 382 per quinases upstream com PAK2 o RSK i que també és necessari un dels seus dominis coiled-coil, almenys els aminoà cids 558 i 561. Aquests processos, però, semblen ser independents de l’activació de NFAT5 en resposta a hipertonicitat.   

Array de DNA per identificar espècies de Fusarium: ampliació d'un array existent per incloure espècies del Sud d'Europa

Projecte de recerca elaborat a partir d’una estada al Department for Feed and Food Hygiene del National Veterinary Institute, Noruega, entre novembre i desembre del 2006. Els grans de cereal poden estar contaminats amb diferents espècies de Fusarium capaces de produir metabolits secundaris altament tòxics com trichotecenes, fumonisines o moniliformines. La correcta identificació d’aquestes espècies és de gran importància per l’assegurament del risc en l’àmbit de la salut humana i animal. La identificació de Fusarium en base a la seva morfologia requereix coneixements taxonòmics i temps; la majoria dels mètodes moleculars permeten la identificació d’una única espècie diana. Per contra, la tecnologia de microarray ofereix l’anàlisi paral•lel d’un alt nombre de DNA dianes. En aquest treball, s’ha desenvolupat un array per a la identificació de les principals espècies de Fusarium toxigèniques del Nord i Sud d’Europa. S’ha ampliat un array ja existent, per a la detecció de les espècies de Fusarium productores de trichothecene i moniliformina (predominants al Nord d’Europa), amb l’addició de 18 sondes de DNA que permeten identificar les espècies toxigèniques més abundants al Sud d’Europa, les qual produeixen majoritàriament fumonisines. Les sondes de captura han estat dissenyades en base al factor d’elongació translació- 1 alpha (TEF-1alpha). L’anàlisi de les mostres es realitza mitjançant una única PCR que permet amplificar part del TEF-1alpha seguida de la hibridació al xip de Fusarium. Els resultats es visualitzen mitjançant un mètode de detecció colorimètric. El xip de Fusarium desenvolupat pot esdevenir una eina útil i de gran interès per a l’anàlisi de cereals presents en la cadena alimentària.

Epidermal growth factor receptor: a re-emerging target in glioblastoma.

PURPOSE OF REVIEW: Amplification and overexpression of the epidermal growth factor receptor (EGFR) gene are a hallmark of primary glioblastoma (45%), making it a prime target for therapy. In addition, these amplifications are frequently associated with oncogenic mutations in the extracellular domain. However, efforts at targeting the EGFR tyrosine kinase using small molecule inhibitors or antibodies have shown disappointing efficacy in clinical trials for newly diagnosed or recurrent glioblastoma. Here, we review recent insights into molecular mechanisms relevant for effective targeting of the EGFR pathway. RECENT FINDINGS: Molecular workup of glioblastoma tissue of patients under treatment with small molecule inhibitors has established drug concentrations in the tumor tissue, and has shed light on the effectiveness of target inhibition and respective effects on pathway signaling. Further, functional analyses of interaction of small molecule inhibitors with distinct properties to bind to the active or inactive form of EGFR have provided new insights that will impact the choice of drugs. Finally, vaccination approaches targeting the EGFRvIII mutant featuring a tumor-specific antigen have shown promising results that warrant larger controlled clinical trials. SUMMARY: A combination of preclinical and clinical studies at the molecular level has provided new insights that will allow refining strategies for targeting the EGFR pathway in glioblastoma.

Expression of Staphylococcus aureus clumping factor A in Lactococcus lactis subsp. cremoris using a new shuttle vector.

Staphylococcus aureus harbors redundant adhesins mediating tissue colonization and infection. To evaluate their intrinsic role outside of the staphylococcal background, a system was designed to express them in Lactococcus lactis subsp. cremoris 1363. This bacterium is devoid of virulence factors and has a known genetic background. A new Escherichia coli-L. lactis shuttle and expression vector was constructed for this purpose. First, the high-copy-number lactococcal plasmid pIL253 was equipped with the oriColE1 origin, generating pOri253 that could replicate in E. coli. Second, the lactococcal promoters P23 or P59 were inserted at one end of the pOri253 multicloning site. Gene expression was assessed by a luciferase reporter system. The plasmid carrying P23 (named pOri23) expressed luciferase constitutively at a level 10,000 times greater than did the P59-containing plasmid. Transcription was absent in E. coli. The staphylococcal clumping factor A (clfA) gene was cloned into pOri23 and used as a model system. Lactococci carrying pOri23-clfA produced an unaltered and functional 130-kDa ClfA protein attached to their cell walls. This was indicated both by the presence of the protein in Western blots of solubilized cell walls and by the ability of ClfA-positive lactococci to clump in the presence of plasma. ClfA-positive lactococci had clumping titers (titer of 4,112) similar to those of S. aureus Newman in soluble fibrinogen and bound equally well to solid-phase fibrinogen. These experiments provide a new way to study individual staphylococcal pathogenic factors and might complement both classical knockout mutagenesis and modern in vivo expression technology and signature tag mutagenesis.

Decay accelerating factor (DAF) as the host antigen with protective activity to complement killing of schistosomula

The acquisition of host antigens by Schistosoma mansoni was studied by evaluating the resistance of schistosomula to the complement attack mediated by lethal antibody. Schistosomula cultured for 24 hours with intact human erythrocytes (N-HuE) or ghosts of any type of ABO or Rh blood group, showed a marked resistance to complement damage. Sheep red blood cells, pronase-treated N-HuE or erythrocytes from patients with paroxysmal nocturnal hemoglobinuria, which are complement-sensitive cells, were unable to protect schistosomula. Schistosomula protected by N-HuE became again susceptible to complement killing after incubation with a monoclonal antibody anti-DAF. These results indicate that, in vitro, host DAF from N-HuE can be acquired by schistosomula surface in a biological active form that protects the parasite from the complement lesion.