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Metabolic engineering of microorganisms aims to design strains capable of producing valuable compounds under relevant industrial conditions and in an economically competitive manner. From this perspective, and beyond the need for a catalyst, biomass is essentially a cost-intensive, abundant by-product of a microbial conversion. Yet, few broadly applicable strategies focus on the optimal balance between product and biomass formation. Here, we present a genetic control module that can be used to precisely modulate growth of the industrial bacterial chassis Pseudomonas putida KT2440. The strategy is based on the controllable expression of the key metabolic enzyme complex pyruvate dehydrogenase (PDH) which functions as a metabolic valve. By tuning the PDH activity, we accurately controlled biomass formation, resulting in six distinct growth rates with parallel overproduction of excess pyruvate. We deployed this strategy to identify optimal growth patterns that improved the production yield of 2-ketoisovalerate and lycopene by 2.5- and 1.38-fold, respectively. This ability to dynamically steer fluxes to balance growth and production substantially enhances the potential of this remarkable microbial chassis for a wide range of industrial applications.

Many cellular activities in bacteria are organized according to their growth rate. The notion that ppGpp measures the cell’s growth rate is well accepted in the field of bacterial physiology. However, despite decades of interrogation and the identification of multiple molecular interactions that connects ppGpp to some aspects of cell growth, we lack a system-level, quantitative picture of how this alleged “measurement” is performed. Through quantitative experiments, we show that the ppGpp pool responds inversely to the rate of translational elongation in Escherichia coli. Together with its roles in inhibiting ribosome biogenesis and activity, ppGpp closes a key regulatory circuit that enables the cell to perceive and control the rate of its growth across conditions. The celebrated linear growth law relating the ribosome content and growth rate emerges as a consequence of keeping a supply of ribosome reserves while maintaining elongation rate in slow growth conditions. Further analysis suggests the elongation rate itself is detected by sensing the ratio of dwelling and translocating ribosomes, a strategy employed to collapse the complex, high-dimensional dynamics of the molecular processes underlying cell growth to perceive the physiological state of the whole.

Many cellular activities in bacteria are organized according to their growth rate. The notion that ppGpp measures the cell’s growth rate is well accepted in the field of bacterial physiology. However, despite decades of interrogation and the identification of multiple molecular interactions that connects ppGpp to some aspects of cell growth, we lack a system-level, quantitative picture of how this alleged “measurement” is performed. Through quantitative experiments, we show that the ppGpp pool responds inversely to the rate of translational elongation in Escherichia coli. Together with its roles in inhibiting ribosome biogenesis and activity, ppGpp closes a key regulatory circuit that enables the cell to perceive and control the rate of its growth across conditions. The celebrated linear growth law relating the ribosome content and growth rate emerges as a consequence of keeping a supply of ribosome reserves while maintaining elongation rate in slow growth conditions. Further analysis suggests the elongation rate itself is detected by sensing the ratio of dwelling and translocating ribosomes, a strategy employed to collapse the complex, high-dimensional dynamics of the molecular processes underlying cell growth to perceive the physiological state of the whole.

During development, multicellular organisms undergo stereotypical patterns of tissue growth in space and time. How developmental growth is orchestrated remains unclear, largely due to the difficulty of observing and quantitating this process in a living organism. Drosophila histoblast nests are small clusters of progenitor epithelial cells that undergo extensive growth to give rise to the adult abdominal epidermis and are amenable to live imaging. Our quantitative analysis of histoblast proliferation and tissue mechanics reveals that tissue growth is driven by cell divisions initiated through basal extracellular matrix degradation by matrix metalloproteases secreted by the neighboring larval epidermal cells. Laser ablations and computational simulations show that tissue mechanical tension does not decrease as the histoblasts fill the abdominal epidermal surface. During tissue growth, the histoblasts display oscillatory cell division rates until growth termination occurs through the rapid emergence of G0/G1 arrested cells, rather than a gradual increase in cell-cycle time as observed in other systems such as the Drosophila wing and mouse postnatal epidermis. Different developing tissues can therefore achieve their final size using distinct growth termination strategies. Thus, adult abdominal epidermal development is characterized by changes in the tissue microenvironment and a rapid exit from the cell cycle.

Microbial co-cultures and consortia are of interest in cell-based molecular production and even as “smart” therapeutics in that one can take advantage of division of labor and specialization to expand both the range of available functions and mechanisms for control. The development of tools that enable coordination and modulation of consortia will be crucial for future application of multi-population cultures. In particular, these systems would benefit from an expanded toolset that enables orthogonal inter-strain communication. We created a co-culture for the synthesis of a redox-active phenazine signaling molecule, pyocyanin (PYO), by dividing its synthesis into the generation of its intermediate, phenazine carboxylic acid (PCA) from the first strain, followed by consumption of PCA and generation of PYO in a second strain. Interestingly, both PCA and PYO can be used to actuate gene expression in cells engineered with the soxRS oxidative stress regulon, although importantly this signaling activity was found to depend on growth media. That is, like other signaling motifs in bacterial systems, the signaling activity is context dependent. We then used this co-culture’s phenazine signals in a tri-culture to modulate gene expression and production of three model products: quorum sensing molecule autoinducer-1 and two fluorescent marker proteins, eGFP and DsRed. We also showed how these redox-based signals could be intermingled with other quorum-sensing (QS) signals which are more commonly used in synthetic biology, to control complex behaviors. To provide control over product synthesis in the tri-cultures, we also showed how a QS-induced growth control module could guide metabolic flux in one population and at the same time guide overall tri-culture function. Specifically, we showed that phenazine signal recognition, enabled through the oxidative stress response regulon soxRS, was dependent on media composition such that signal propagation within our parsed synthetic sys

Gradients of decapentaplegic (Dpp) pattern Drosophila wing imaginal discs, establishing gene expression boundaries at specific locations. As discs grow, Dpp gradients expand, keeping relative boundary positions approximately stationary. Such scaling fails in mutants for Pentagone (pent), a gene repressed by Dpp that encodes a diffusible protein that expands Dpp gradients. Although these properties fit a recent mathematical model of automatic gradient scaling, that model requires an expander that spreads with minimal loss throughout a morphogen field. Here, we show that Pent's actions are confined to within just a few cell diameters of its site of synthesis and can be phenocopied by manipulating non-diffusible Pent targets strictly within the Pent expression domain. Using genetics and mathematical modeling, we develop an alternative model of scaling driven by feedback downregulation of Dpp receptors and co-receptors. Among the model's predictions is a size beyond which scaling fails-something we observe directly in wing discs.

The mechanism underlying control of organ size is not only a fascinating topic in biology, but also has profound significance in health care and disease prevention. The question can be considered at the level of cells that make the organs. In the field of developmental biology, the concept of cell lineages is widely used. A feedback control model has been proposed to address the problem that how cell lineages use short-range signals to control the growth of organs on a large scale. However, the detailed strategies used in the feedback controls that balance cell proliferation and cell differentiation in this process are little known. In this research, counting methods have been explored to estimate the rates of cell divisions in mouse olfactory epithelium (OE). New markers for cell lineage stages in OE have also been tested. The results can serve as a foundation for further detailed analysis of the cell lineage in OE and its regulation.

The mechanism underlying control of organ size is not only a fascinating topic in biology, but also has profound significance in health care and disease prevention. The question can be considered at the level of cells that make the organs. In the field of developmental biology, the concept of cell lineages is widely used. A feedback control model has been proposed to address the problem that how cell lineages use short-range signals to control the growth of organs on a large scale. However, the detailed strategies used in the feedback controls that balance cell proliferation and cell differentiation in this process are little known. In this research, counting methods have been explored to estimate the rates of cell divisions in mouse olfactory epithelium (OE). New markers for cell lineage stages in OE have also been tested. The results can serve as a foundation for further detailed analysis of the cell lineage in OE and its regulation.

Gradients of decapentaplegic (Dpp) pattern Drosophila wing imaginal discs, establishing gene expression boundaries at specific locations. As discs grow, Dpp gradients expand, keeping relative boundary positions approximately stationary. Such scaling fails in mutants for Pentagone (pent), a gene repressed by Dpp that encodes a diffusible protein that expands Dpp gradients. Although these properties fit a recent mathematical model of automatic gradient scaling, that model requires an expander that spreads with minimal loss throughout a morphogen field. Here, we show that Pent's actions are confined to within just a few cell diameters of its site of synthesis and can be phenocopied by manipulating non-diffusible Pent targets strictly within the Pent expression domain. Using genetics and mathematical modeling, we develop an alternative model of scaling driven by feedback downregulation of Dpp receptors and co-receptors. Among the model's predictions is a size beyond which scaling fails-something we observe directly in wing discs.

The evolutionarily conserved Target of Rapamycin (TOR) complex-2 (TORC2) is an essential regulator of plasma membrane homeostasis in budding yeast (Saccharomyces cerevisiae). In this yeast, TORC2 phosphorylates and activates the effector protein kinase Ypk1 and its paralog Ypk2. These protein kinases, in turn, carry out all the crucial functions of TORC2 by phosphorylating and thereby controlling the activity of at least a dozen downstream substrates. A previously uncharacterized interplay between the Rab5 GTPases and TORC2 signaling was uncovered through analysis of a newly suspected Ypk1 target. Muk1, one of two guanine nucleotide exchange factors for the Rab5 GTPases, was found to be a physiologically relevant Ypk1 substrate; and, genetic analysis indicates that Ypk1-mediated phosphorylation activates the guanine nucleotide exchange activity of Muk1. Second, it was demonstrated both in vivo and in vitro that the GTP-bound state of the Rab5 GTPase Vps21/Ypt51 physically associates with TORC2 and acts as a direct positive effector required for full TORC2 activity. These interrelationships provide a self-reinforcing control circuit for sustained up-regulation of TORC2-Ypk1 signaling. In this overview, we summarize the experimental basis of these findings, their implications, and speculate as to the molecular basis for Rab5-mediated TORC2 activation.

Introduction. The vanilla production in Mexico has decreased by 70 % in the last fifteen years, due to the fall of its fruits. This occurs in extreme climatic conditions when the temperature is higher than 32 °C and the relative humidity is less than 80 %. Objective. The objective was to determine the effect of some commercial growth regulators (GR) on the retention of vanilla (Vanilla planifolia Jacks ex Andrews) fruits Mansa cultivar. Materials and methods. The study was carried out in 2016 in the Tecolutla Municipality, Veracruz, Mexico. The treatments evaluated were: aminoethoxyvinylglycine hydrochloride (AVG; 250 ppm), 1-naphthaleneacetic acid (ANA; 27 ppm), benzyl-adenine (BA; 150 ppm), forchlorfenuron (CPPU; 5 ml l-1 of water), 2,4-Dichlorophenoxyacetic acid (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 of water) and control, which consisted of applying only one acidifier (8 ml in 10 l of water). Treatments were applied three times to the same fruits, at 6, 20 and 31 days after the last flowers pollinated by bunch. Results. All GR retained more labelled fruits than the control. Treatments 2,4-D and 2,4-D + CPPU retained more fruits per raceme and produced significantly longer fruits than the control. In addition, the 2,4-D + CPPU mixture produced fruits with larger diameter than the control. The growth regulators AVG, ANA BA and CPPU did not differ in the number of labelled fruits, fruits per raceme, fruit length, and fruit diameter. Conclusion. GR had a positive effect on vanilla fruit retention, mainly the 2,4-D + CPPU mixture with which more labelled fruits and per bunch were obtained compared to the control. The use of GR was effective in reducing vanilla abortion., Introducción. La producción de vainilla en México ha disminuido en un 70 % en los últimos quince años, debido a la caída de sus frutos que ocurre en condiciones climáticas extremas, cuando la temperatura es mayor a 32 °C y la humedad relativa es menor a 80 %. Objetivo. El objetivo fue determinar el efecto de algunos reguladores de crecimiento (RC) comerciales sobre la retención de frutos de vainilla (Vanilla planifolia Jacks ex Andrews) cultivar Mansa. Materiales y métodos. El estudio se realizó en el 2016 en el municipio de Tecolutla, Veracruz, México. Los tratamientos evaluados fueron: clorhidrato de aminoetoxivinilglicina (AVG; 250 ppm), 1-ácido naftalenacético (ANA; 27 ppm), Benciladenina (BA; 150 ppm), Forclorfenurón (CPPU; 5 ml l-1 de agua), Ácido 2,4-diclorofenoxiacético (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 de agua) y testigo, que consistió en aplicar solamente un acidificante (8 ml en 10 l de agua). Los tratamientos se aplicaron en tres ocasiones en los mismos frutos a los 6, 20 y 31 días después de las últimas flores polinizadas por racimo. Resultados. Todos los RC retuvieron más frutos etiquetados que el testigo. Los tratamientos 2,4-D y 2,4-D + CPPU retuvieron más frutos por racimo y produjeron frutos significativamente más largos que el testigo. Además, la mezcla 2,4-D + CPPU produjo frutos con mayor diámetro que el testigo. Los reguladores de crecimiento AVG, ANA BA y CPPU no difirieron en el número de frutos etiquetados, frutos por racimo, longitud del fruto y diámetro del fruto. Conclusión. Los RC tuvieron un efecto positivo en la retención de frutos de vainilla, principalmente la mezcla 2,4-D + CPPU, con la cual se obtuvieron más frutos etiquetados y por racimo, con respecto al testigo. El uso de RC fue efectivo para reducir el aborto en vainilla.

The evolutionarily conserved Target of Rapamycin (TOR) complex-2 (TORC2) is an essential regulator of plasma membrane homeostasis in budding yeast (Saccharomyces cerevisiae). In this yeast, TORC2 phosphorylates and activates the effector protein kinase Ypk1 and its paralog Ypk2. These protein kinases, in turn, carry out all the crucial functions of TORC2 by phosphorylating and thereby controlling the activity of at least a dozen downstream substrates. A previously uncharacterized interplay between the Rab5 GTPases and TORC2 signaling was uncovered through analysis of a newly suspected Ypk1 target. Muk1, one of two guanine nucleotide exchange factors for the Rab5 GTPases, was found to be a physiologically relevant Ypk1 substrate; and, genetic analysis indicates that Ypk1-mediated phosphorylation activates the guanine nucleotide exchange activity of Muk1. Second, it was demonstrated both in vivo and in vitro that the GTP-bound state of the Rab5 GTPase Vps21/Ypt51 physically associates with TORC2 and acts as a direct positive effector required for full TORC2 activity. These interrelationships provide a self-reinforcing control circuit for sustained up-regulation of TORC2-Ypk1 signaling. In this overview, we summarize the experimental basis of these findings, their implications, and speculate as to the molecular basis for Rab5-mediated TORC2 activation.

The evolutionarily conserved Target of Rapamycin (TOR) complex-2 (TORC2) is an essential regulator of plasma membrane homeostasis in budding yeast (Saccharomyces cerevisiae). In this yeast, TORC2 phosphorylates and activates the effector protein kinase Ypk1 and its paralog Ypk2. These protein kinases, in turn, carry out all the crucial functions of TORC2 by phosphorylating and thereby controlling the activity of at least a dozen downstream substrates. A previously uncharacterized interplay between the Rab5 GTPases and TORC2 signaling was uncovered through analysis of a newly suspected Ypk1 target. Muk1, one of two guanine nucleotide exchange factors for the Rab5 GTPases, was found to be a physiologically relevant Ypk1 substrate; and, genetic analysis indicates that Ypk1-mediated phosphorylation activates the guanine nucleotide exchange activity of Muk1. Second, it was demonstrated both in vivo and in vitro that the GTP-bound state of the Rab5 GTPase Vps21/Ypt51 physically associates with TORC2 and acts as a direct positive effector required for full TORC2 activity. These interrelationships provide a self-reinforcing control circuit for sustained up-regulation of TORC2-Ypk1 signaling. In this overview, we summarize the experimental basis of these findings, their implications, and speculate as to the molecular basis for Rab5-mediated TORC2 activation.

Introduction. The vanilla production in Mexico has decreased by 70 % in the last fifteen years, due to the fall of its fruits. This occurs in extreme climatic conditions when the temperature is higher than 32 °C and the relative humidity is less than 80 %. Objective. The objective was to determine the effect of some commercial growth regulators (GR) on the retention of vanilla (Vanilla planifolia Jacks ex Andrews) fruits Mansa cultivar. Materials and methods. The study was carried out in 2016 in the Tecolutla Municipality, Veracruz, Mexico. The treatments evaluated were: aminoethoxyvinylglycine hydrochloride (AVG; 250 ppm), 1-naphthaleneacetic acid (ANA; 27 ppm), benzyl-adenine (BA; 150 ppm), forchlorfenuron (CPPU; 5 ml l-1 of water), 2,4-Dichlorophenoxyacetic acid (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 of water) and control, which consisted of applying only one acidifier (8 ml in 10 l of water). Treatments were applied three times to the same fruits, at 6, 20 and 31 days after the last flowers pollinated by bunch. Results. All GR retained more labelled fruits than the control. Treatments 2,4-D and 2,4-D + CPPU retained more fruits per raceme and produced significantly longer fruits than the control. In addition, the 2,4-D + CPPU mixture produced fruits with larger diameter than the control. The growth regulators AVG, ANA BA and CPPU did not differ in the number of labelled fruits, fruits per raceme, fruit length, and fruit diameter. Conclusion. GR had a positive effect on vanilla fruit retention, mainly the 2,4-D + CPPU mixture with which more labelled fruits and per bunch were obtained compared to the control. The use of GR was effective in reducing vanilla abortion., Introducción. La producción de vainilla en México ha disminuido en un 70 % en los últimos quince años, debido a la caída de sus frutos que ocurre en condiciones climáticas extremas, cuando la temperatura es mayor a 32 °C y la humedad relativa es menor a 80 %. Objetivo. El objetivo fue determinar el efecto de algunos reguladores de crecimiento (RC) comerciales sobre la retención de frutos de vainilla (Vanilla planifolia Jacks ex Andrews) cultivar Mansa. Materiales y métodos. El estudio se realizó en el 2016 en el municipio de Tecolutla, Veracruz, México. Los tratamientos evaluados fueron: clorhidrato de aminoetoxivinilglicina (AVG; 250 ppm), 1-ácido naftalenacético (ANA; 27 ppm), Benciladenina (BA; 150 ppm), Forclorfenurón (CPPU; 5 ml l-1 de agua), Ácido 2,4-diclorofenoxiacético (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 de agua) y testigo, que consistió en aplicar solamente un acidificante (8 ml en 10 l de agua). Los tratamientos se aplicaron en tres ocasiones en los mismos frutos a los 6, 20 y 31 días después de las últimas flores polinizadas por racimo. Resultados. Todos los RC retuvieron más frutos etiquetados que el testigo. Los tratamientos 2,4-D y 2,4-D + CPPU retuvieron más frutos por racimo y produjeron frutos significativamente más largos que el testigo. Además, la mezcla 2,4-D + CPPU produjo frutos con mayor diámetro que el testigo. Los reguladores de crecimiento AVG, ANA BA y CPPU no difirieron en el número de frutos etiquetados, frutos por racimo, longitud del fruto y diámetro del fruto. Conclusión. Los RC tuvieron un efecto positivo en la retención de frutos de vainilla, principalmente la mezcla 2,4-D + CPPU, con la cual se obtuvieron más frutos etiquetados y por racimo, con respecto al testigo. El uso de RC fue efectivo para reducir el aborto en vainilla.

In the present study, Chafino et al. identify the upregulation of E93 expression as the trigger for metamorphosis in the holometabolous insect Tribolium castaneum. Depletion of E93 prevents pupa formation, whereas precocious E93 upregulation, by either JH depletion or nutrition restriction, induces premature onset of metamorphosis.Body size in holometabolous insects is determined by the size at which the juvenile larva undergoes metamorphosis to the pupal stage. To undergo larva-pupa transition, larva must reach a critical developmental checkpoint, the threshold size (TS); however, the molecular mechanisms through which the TS cues this transition remain to be fully characterized. Here, we use the flour beetle Tribolium castaneum to characterize the molecular mechanisms underlying entry into metamorphosis. We found that T. castaneum reaches a TS at the beginning of the last larval instar, which is associated with the downregulation of TcKr-h1 and the upregulation of TcE93 and TcBr-C. Unexpectedly, we found that while there is an association between TS and TcE93 upregulation, it is the latter that constitutes the molecular trigger for metamorphosis initiation. In light of our results, we evaluate the interactions that control the larva-pupa transition and suggest alternative models.

Zentrales Thema dieser Dissertation ist die Untersuchung anorganisch/organischer Hybridsysteme (HIOS) mit besonderem Fokus auf den speziellen Prozessen an der Grenzfläche beider Materialklassen. Organische Moleküle, in Verbindung mit anorganischen Halbleitern haben ein großes Potenzial für Anwendungen in zukünftigen optoelektronischen Hybridbauteilen, indem sie Vorteile zweier unterschiedlicher Welten kombinieren. Entscheidend für die Herstellung von hybriden Strukturen ist das Verständnis der Wechselwirkungen an der Grenzfläche zwischen organischem und anorganischem Material. In dieser Arbeit werden diese Wechselwirkungen analysiert, um eine Wachstumskontrolle an der Grenzfläche zwischen konjugierten organischen Molekül und anorganischem Halbleiter zu ermöglichen. Hierfür werden unterschiedliche Ansätze verfolgt: Im ersten Teil der Arbeit wird die Wechselwirkung des Modellsystems Sexiphenyl (6P) an der Grenzfläche zu ZnO untersucht, sowie das Wachstum des Moleküls mittels verschiedener Methoden kontrolliert. Das daraus gewonnene Wissen kann im zweiten Teil dazu verwendet werden einen hybriden ZnO/6P/ZnO-Stapel zu realisieren, bei dem die organische Schicht ohne Beeinträchtigung der Kristallstruktur, mit definierten Grenzflächen bis hin zur atomaren/molekularen Ebene, überwachsen werden kann. Der letzte Teil der Arbeit befasst sich mit der optischen Echtzeit-Beobachtung während des organischen Wachstums verschiedener Moleküle. Dadurch ist es möglich Veränderungen von Struktureigenschaften und Wechselwirkungen zwischen Molekülen und dem Substrat zerstörungsfrei zu bestimmen, während diese aufgewachsen werden. Hierdurch können schlussendlich mögliche Mechanismen aufgezeigt werden, um elektronische und optische Wechselwirkung an der Grenzfläche zwischen organischem Molekül und anorganischen Halbleitern zu analysieren, sowie Wachstumsprozesse weiter zu verstehen und kontrollieren., The central subject of this thesis are hybrid inorganic/organic systems (HIOS) with a focus on the specific processes at the interface between the two material classes. Organic molecules used together with inorganic semiconductors, have a great potential for future optoelectronic applications in hybrid components, by combining the advantages of two dissimilar worlds. Decisive for the production of hybrid structures is the understanding of the interactions at the interface between organic and inorganic material. In this thesis, the interactions are analyzed to enable growth control at the interface between conjugated organic molecules and inorganic semiconductors. In the first part of the thesis, the interaction of the model system sexiphenyl (6P) at the interface with ZnO, as well as approaches to control the growth of the molecule are being investigated. The knowledge gained here is used in the second part to realize a hybrid ZnO/6P/ ZnO stack, in which the organic layer can be overgrown without affecting the crystal structure, exhibiting defined interfaces down to the atomic/molecular level. The last part of the thesis deals with real time optical observation during organic growth of different molecules. By this changes in structural properties and interactions between molecules and the substrate can be non-destructively determined as they are growing. Ultimately, a comprehensive insight into the optical and electronic interactions at the interface between organic molecules and inorganic semiconductors can be gained and possible control mechanisms are shown.

Zentrales Thema dieser Dissertation ist die Untersuchung anorganisch/organischer Hybridsysteme (HIOS) mit besonderem Fokus auf den speziellen Prozessen an der Grenzfläche beider Materialklassen. Organische Moleküle, in Verbindung mit anorganischen Halbleitern haben ein großes Potenzial für Anwendungen in zukünftigen optoelektronischen Hybridbauteilen, indem sie Vorteile zweier unterschiedlicher Welten kombinieren. Entscheidend für die Herstellung von hybriden Strukturen ist das Verständnis der Wechselwirkungen an der Grenzfläche zwischen organischem und anorganischem Material. In dieser Arbeit werden diese Wechselwirkungen analysiert, um eine Wachstumskontrolle an der Grenzfläche zwischen konjugierten organischen Molekül und anorganischem Halbleiter zu ermöglichen. Hierfür werden unterschiedliche Ansätze verfolgt: Im ersten Teil der Arbeit wird die Wechselwirkung des Modellsystems Sexiphenyl (6P) an der Grenzfläche zu ZnO untersucht, sowie das Wachstum des Moleküls mittels verschiedener Methoden kontrolliert. Das daraus gewonnene Wissen kann im zweiten Teil dazu verwendet werden einen hybriden ZnO/6P/ZnO-Stapel zu realisieren, bei dem die organische Schicht ohne Beeinträchtigung der Kristallstruktur, mit definierten Grenzflächen bis hin zur atomaren/molekularen Ebene, überwachsen werden kann. Der letzte Teil der Arbeit befasst sich mit der optischen Echtzeit-Beobachtung während des organischen Wachstums verschiedener Moleküle. Dadurch ist es möglich Veränderungen von Struktureigenschaften und Wechselwirkungen zwischen Molekülen und dem Substrat zerstörungsfrei zu bestimmen, während diese aufgewachsen werden. Hierdurch können schlussendlich mögliche Mechanismen aufgezeigt werden, um elektronische und optische Wechselwirkung an der Grenzfläche zwischen organischem Molekül und anorganischen Halbleitern zu analysieren, sowie Wachstumsprozesse weiter zu verstehen und kontrollieren., The central subject of this thesis are hybrid inorganic/organic systems (HIOS) with a focus on the specific processes at the interface between the two material classes. Organic molecules used together with inorganic semiconductors, have a great potential for future optoelectronic applications in hybrid components, by combining the advantages of two dissimilar worlds. Decisive for the production of hybrid structures is the understanding of the interactions at the interface between organic and inorganic material. In this thesis, the interactions are analyzed to enable growth control at the interface between conjugated organic molecules and inorganic semiconductors. In the first part of the thesis, the interaction of the model system sexiphenyl (6P) at the interface with ZnO, as well as approaches to control the growth of the molecule are being investigated. The knowledge gained here is used in the second part to realize a hybrid ZnO/6P/ ZnO stack, in which the organic layer can be overgrown without affecting the crystal structure, exhibiting defined interfaces down to the atomic/molecular level. The last part of the thesis deals with real time optical observation during organic growth of different molecules. By this changes in structural properties and interactions between molecules and the substrate can be non-destructively determined as they are growing. Ultimately, a comprehensive insight into the optical and electronic interactions at the interface between organic molecules and inorganic semiconductors can be gained and possible control mechanisms are shown.

During embryonic development, Hox genes participate in the building of a functional digestive system in metazoans, and genetic conditions involving these genes lead to important, sometimes lethal, growth retardation. Recently, this phenotype was obtained after deletion of Haglr, the Hoxd antisense growth-associated long noncoding RNA (lncRNA) located between Hoxd1 and Hoxd3 In this study, we have analyzed the function of Hoxd genes in delayed growth trajectories by looking at several nested targeted deficiencies of the mouse HoxD cluster. Mutant pups were severely stunted during the suckling period, but many recovered after weaning. After comparing seven distinct HoxD alleles, including CRISPR/Cas9 deletions involving Haglr, we identified Hoxd3 as the critical component for the gut to maintain milk-digestive competence. This essential function could be abrogated by the dominant-negative effect of HOXD10 as shown by a genetic rescue approach, thus further illustrating the importance of posterior prevalence in Hox gene function. A role for the lncRNA Haglr in the control of postnatal growth could not be corroborated.

In dieser Arbeit werden neue Strategien zur Wachstumskontrolle funktionaler organischer Dünnschichtmaterialien aufgezeigt, mit denen sich die Eigenschaften molekularer Schichtsysteme gezielt beeinflussen lassen. Ein Fokus liegt dabei auf der Untersuchung des Einflusses von optischen Feldern auf das Wachstumsverhalten. Ein weiterer Schwerpunkt wird auf das Grundlagenverständnis von thermisch aktivierten, kinetischen Prozessen, die die Morphologie während und nach dem Schichtwachstum beeinflussen, gelegt. Zuerst wird am Beispiel des molekularen Halbleiters Sexithiophen (6T) gezeigt, wie sich Kontrolle über das Kristallphasengleichgewicht während des Wachstums auf Kaliumchlorid (KCl) erzielen lässt. Im zweiten Teil der Arbeit wird eine neue Herangehensweise zur direkten Ausrichtung von Molekülkristallen im optischen Feld während ihrer Entstehung studiert. Am Beispiel von Tetracene wird gezeigt wie sich so optische anisotrope Absorptionseigenschaften von Molekülen dazu nutzen lassen den Brechungsindex eines polykristallinen Films lokal durch ein photolitographisches Verfahren zu beeinflussen. Im dritten Teil wird der Einfluss von dynamisch variierenden Wachstumsbedingungen während des Schichtwachstums von PTCDI-C8 studiert. Es wird gezeigt, dass sich die Oberflächenrauigkeit stark reduzieren lässt, indem zu Beginn des Wachstums jeder individuellen molekularen Monolage die Nukleationsdichte stark erhöht und in den darauf folgenden Wachstumsphasen die Diffusivität gezielt gesteigert wird. Im vierten Teil wird das Diffusionsverhalten von n-Alkan Schichten unter thermischen Einflüssen betrachtet. Es wird gezeigt, dass die inhärente molekulare Anisotropie von C44H90 Molekülen (TTC) sowie die sehr geringe, stark anisotrope, Oberflächenenergie dieses Materials ein ungewöhnliches Diffusionsverhalten auslöst., The research performed in the framework of this thesis focuses on new strategies to effectively control the growth of crystalline thin films of functional organic molecules and attributes the quest for additional growth control parameters in organic molecular beam deposition (OMBD). First the influence of light on the growth process of the sexithiophene (6T) is studied. We find that 6T thin films deposited as conventional in dark environments on KCl exhibit a bimodal growth with phase coexistence of two crystal polymorphs. In contrast, films grown under illumination with 532 nm light show increased phase purity. Further, we establish light-directed molecular self-assembly (LDSA) to generate permanently aligned thin films of tetracene (C18H12) and demonstrate direct patterning with light. Polarized light illumination leads to azimuthally photoaligned films on isotropic, amorphous substrates. Thus, LDSA can be regarded as a new degree of freedom in the quest for control-parameters in organic thin film growth. Next the impact of dynamic temperature oscillations on the time scales of molecular monolayer growth during organic molecular beam deposition is discussed. We strongly increase the island density during nucleation and selectively increase interlayer diffusion at later stages of monolayer growth. We analyse the interplay between molecular interlayer transport and island sizes to understand kinetic processes during growth. In a fourth experiment we show how thermal annealing can be used to improve smoothness and to increase the lateral size of crystalline islands of n-alkane (TTC, C44H90) films. We employ real-time optical phase contrast microscopy to track the diffusion across monomolecular step edges which causes the unusual smoothing during annealing. We rationalise the smoothing behaviour with the highly anisotropic attachment energies and low surface energies of TTC.

During embryonic development, Hox genes participate in the building of a functional digestive system in metazoans, and genetic conditions involving these genes lead to important, sometimes lethal, growth retardation. Recently, this phenotype was obtained after deletion of Haglr, the Hoxd antisense growth-associated long noncoding RNA (lncRNA) located between Hoxd1 and Hoxd3 In this study, we have analyzed the function of Hoxd genes in delayed growth trajectories by looking at several nested targeted deficiencies of the mouse HoxD cluster. Mutant pups were severely stunted during the suckling period, but many recovered after weaning. After comparing seven distinct HoxD alleles, including CRISPR/Cas9 deletions involving Haglr, we identified Hoxd3 as the critical component for the gut to maintain milk-digestive competence. This essential function could be abrogated by the dominant-negative effect of HOXD10 as shown by a genetic rescue approach, thus further illustrating the importance of posterior prevalence in Hox gene function. A role for the lncRNA Haglr in the control of postnatal growth could not be corroborated.

In dieser Arbeit werden neue Strategien zur Wachstumskontrolle funktionaler organischer Dünnschichtmaterialien aufgezeigt, mit denen sich die Eigenschaften molekularer Schichtsysteme gezielt beeinflussen lassen. Ein Fokus liegt dabei auf der Untersuchung des Einflusses von optischen Feldern auf das Wachstumsverhalten. Ein weiterer Schwerpunkt wird auf das Grundlagenverständnis von thermisch aktivierten, kinetischen Prozessen, die die Morphologie während und nach dem Schichtwachstum beeinflussen, gelegt. Zuerst wird am Beispiel des molekularen Halbleiters Sexithiophen (6T) gezeigt, wie sich Kontrolle über das Kristallphasengleichgewicht während des Wachstums auf Kaliumchlorid (KCl) erzielen lässt. Im zweiten Teil der Arbeit wird eine neue Herangehensweise zur direkten Ausrichtung von Molekülkristallen im optischen Feld während ihrer Entstehung studiert. Am Beispiel von Tetracene wird gezeigt wie sich so optische anisotrope Absorptionseigenschaften von Molekülen dazu nutzen lassen den Brechungsindex eines polykristallinen Films lokal durch ein photolitographisches Verfahren zu beeinflussen. Im dritten Teil wird der Einfluss von dynamisch variierenden Wachstumsbedingungen während des Schichtwachstums von PTCDI-C8 studiert. Es wird gezeigt, dass sich die Oberflächenrauigkeit stark reduzieren lässt, indem zu Beginn des Wachstums jeder individuellen molekularen Monolage die Nukleationsdichte stark erhöht und in den darauf folgenden Wachstumsphasen die Diffusivität gezielt gesteigert wird. Im vierten Teil wird das Diffusionsverhalten von n-Alkan Schichten unter thermischen Einflüssen betrachtet. Es wird gezeigt, dass die inhärente molekulare Anisotropie von C44H90 Molekülen (TTC) sowie die sehr geringe, stark anisotrope, Oberflächenenergie dieses Materials ein ungewöhnliches Diffusionsverhalten auslöst., The research performed in the framework of this thesis focuses on new strategies to effectively control the growth of crystalline thin films of functional organic molecules and attributes the quest for additional growth control parameters in organic molecular beam deposition (OMBD). First the influence of light on the growth process of the sexithiophene (6T) is studied. We find that 6T thin films deposited as conventional in dark environments on KCl exhibit a bimodal growth with phase coexistence of two crystal polymorphs. In contrast, films grown under illumination with 532 nm light show increased phase purity. Further, we establish light-directed molecular self-assembly (LDSA) to generate permanently aligned thin films of tetracene (C18H12) and demonstrate direct patterning with light. Polarized light illumination leads to azimuthally photoaligned films on isotropic, amorphous substrates. Thus, LDSA can be regarded as a new degree of freedom in the quest for control-parameters in organic thin film growth. Next the impact of dynamic temperature oscillations on the time scales of molecular monolayer growth during organic molecular beam deposition is discussed. We strongly increase the island density during nucleation and selectively increase interlayer diffusion at later stages of monolayer growth. We analyse the interplay between molecular interlayer transport and island sizes to understand kinetic processes during growth. In a fourth experiment we show how thermal annealing can be used to improve smoothness and to increase the lateral size of crystalline islands of n-alkane (TTC, C44H90) films. We employ real-time optical phase contrast microscopy to track the diffusion across monomolecular step edges which causes the unusual smoothing during annealing. We rationalise the smoothing behaviour with the highly anisotropic attachment energies and low surface energies of TTC.

The size and shape of organs are characteristic for each species. Even when organisms develop to different sizes due to varying environmental conditions, such as nutrition, organ size follows species-specific rules of proportionality to the rest of the body, a phenomenon referred to as allometry. Therefore, for a given environment, organs stop growth at a predictable size set by the species's genotype. How do organs stop growth? How can related species give rise to organs of strikingly different size? No definitive answer has been given to date. One of the major models for the studies of growth termination is the vinegar fly Drosophila melanogaster. Therefore, this review will focus mostly on work carried out in Drosophila to try to tease apart potential mechanisms and identify routes for further investigation. One general rule, found across the animal kingdom, is that the rate of growth declines with developmental time. Therefore, answers to the problem of growth termination should explain this seemingly universal fact. In addition, growth termination is intimately related to the problems of robustness (i.e. precision) and plasticity in organ size, symmetric and asymmetric organ development, and of how the 'target' size depends on extrinsic, environmental factors.

In eukaryotes, DNA polymerase I is the kilodalton enzyme responsible for ribosonal DNA (rDNA) transcription, leading to the synthesis of the ribosonal RNA precusor. Pol I's activity is crucial for ribosome biogenesis and therefore, its modulation influences cell growth. The intiation factor rDNA, conseved in eukaryotes, binds Pol I corresponds to an activated state of the enzyme, whose activity can be regulated by assembling or disrupting Pol I:Rm3. the cryo-electron microscopy (cryo-EM) structures of yeast free monomeric Pol I at 4.9 A resolution and Pol I in complex with Rm3 at 7.7 A are the major findings reported here. The derived pseudo-atomic models unveil the structural changes in the transition from inactive Pol I dimers. previously solved by X-ray crystalography, to free monovers and from these to the activated state bound to Rm.3 In addition, analytical ultracentrifugation suggets that yeast Rm3 dimers found in solution migth establish a dimer-monover equilibrium in vitro upon dilution. Also, electrophoretic mobility shift assays indicate yeast Rm3 could not bind DNA as previously described for the mammalian homolog. These resuts provide valuable information on Pol I and Rn3 changes required for enzyme activation.

BackgroundHow tissue and organ sizes are specified is one of the great unsolved mysteries in biology. Experiments and mathematical modeling implicate feedback control of cell lineage progression, but a broad understanding of what lineage feedback accomplishes is lacking.ResultsBy exploring the possible effects of various biologically relevant disturbances on the dynamic and steady state behaviors of stem cell lineages, we find that the simplest and most frequently studied form of lineage feedback - which we term renewal control - suffers from several serious drawbacks. These reflect fundamental performance limits dictated by universal conservation-type laws, and are independent of parameter choice. Here we show that introducing lineage branches can circumvent all such limitations, permitting effective attenuation of a wide range of perturbations. The type of feedback that achieves such performance - which we term fate control - involves promotion of lineage branching at the expense of both renewal and (primary) differentiation. We discuss the evidence that feedback of just this type occurs in vivo, and plays a role in tissue growth control.ConclusionsRegulated lineage branching is an effective strategy for dealing with disturbances in stem cell systems. The existence of this strategy provides a dynamics-based justification for feedback control of cell fate in vivo.

BackgroundHow tissue and organ sizes are specified is one of the great unsolved mysteries in biology. Experiments and mathematical modeling implicate feedback control of cell lineage progression, but a broad understanding of what lineage feedback accomplishes is lacking.ResultsBy exploring the possible effects of various biologically relevant disturbances on the dynamic and steady state behaviors of stem cell lineages, we find that the simplest and most frequently studied form of lineage feedback - which we term renewal control - suffers from several serious drawbacks. These reflect fundamental performance limits dictated by universal conservation-type laws, and are independent of parameter choice. Here we show that introducing lineage branches can circumvent all such limitations, permitting effective attenuation of a wide range of perturbations. The type of feedback that achieves such performance - which we term fate control - involves promotion of lineage branching at the expense of both renewal and (primary) differentiation. We discuss the evidence that feedback of just this type occurs in vivo, and plays a role in tissue growth control.ConclusionsRegulated lineage branching is an effective strategy for dealing with disturbances in stem cell systems. The existence of this strategy provides a dynamics-based justification for feedback control of cell fate in vivo.

Bacteria must constantly adapt their growth to changes in nutrient availability; yet despite large-scale changes in protein expression associated with sensing, adaptation, and processing different environmental nutrients, simple growth laws connect the ribosome abundance and the growth rate. Here, we investigate the origin of these growth laws by analyzing the features of ribosomal regulation that coordinate proteome-wide expression changes with cell growth in a variety of nutrient conditions in the model organism Escherichia coli. We identify supply-driven feedforward activation of ribosomal protein synthesis as the key regulatory motif maximizing amino acid flux, and autonomously guiding a cell to achieve optimal growth in different environments. The growth laws emerge naturally from the robust regulatory strategy underlying growth rate control, irrespective of the details of the molecular implementation. The study highlights the interplay between phenomenological modeling and molecular mechanisms in uncovering fundamental operating constraints, with implications for endogenous and synthetic design of microorganisms.

[EN] Cell expansion is an increase in cell size and thus plays an essential role in plant growth and development. Phytohormones and the primary plant cell wall play major roles in the complex process of cell expansion. In shoot tissues, cell expansion requires the auxin receptor AUXIN BINDING PROTEIN1 (ABP1), but the mechanism by which ABP1 affects expansion remains unknown. We analyzed the effect of functional inactivation of ABP1 on transcriptomic changes in dark-grown hypocotyls and investigated the consequences of gene expression on cell wall composition and cell expansion. Molecular and genetic evidence indicates that ABP1 affects the expression of a broad range of cell wall-related genes, especially cell wall remodeling genes, mainly via an SCFTIR/AFB-dependent pathway. ABP1 also functions in the modulation of hemicellulose xyloglucan structure. Furthermore, fucosidase-mediated defucosylation of xyloglucan, but not biosynthesis of nonfucosylated xyloglucan, rescued dark-grown hypocotyl lengthening of ABP1 knockdown seedlings. In muro remodeling of xyloglucan side chains via an ABP1-dependent pathway appears to be of critical importance for temporal and spatial control of cell expansion.

[EN] Cell expansion is an increase in cell size and thus plays an essential role in plant growth and development. Phytohormones and the primary plant cell wall play major roles in the complex process of cell expansion. In shoot tissues, cell expansion requires the auxin receptor AUXIN BINDING PROTEIN1 (ABP1), but the mechanism by which ABP1 affects expansion remains unknown. We analyzed the effect of functional inactivation of ABP1 on transcriptomic changes in dark-grown hypocotyls and investigated the consequences of gene expression on cell wall composition and cell expansion. Molecular and genetic evidence indicates that ABP1 affects the expression of a broad range of cell wall-related genes, especially cell wall remodeling genes, mainly via an SCFTIR/AFB-dependent pathway. ABP1 also functions in the modulation of hemicellulose xyloglucan structure. Furthermore, fucosidase-mediated defucosylation of xyloglucan, but not biosynthesis of nonfucosylated xyloglucan, rescued dark-grown hypocotyl lengthening of ABP1 knockdown seedlings. In muro remodeling of xyloglucan side chains via an ABP1-dependent pathway appears to be of critical importance for temporal and spatial control of cell expansion.

Bacteria must constantly adapt their growth to changes in nutrient availability; yet despite large-scale changes in protein expression associated with sensing, adaptation, and processing different environmental nutrients, simple growth laws connect the ribosome abundance and the growth rate. Here, we investigate the origin of these growth laws by analyzing the features of ribosomal regulation that coordinate proteome-wide expression changes with cell growth in a variety of nutrient conditions in the model organism Escherichia coli. We identify supply-driven feedforward activation of ribosomal protein synthesis as the key regulatory motif maximizing amino acid flux, and autonomously guiding a cell to achieve optimal growth in different environments. The growth laws emerge naturally from the robust regulatory strategy underlying growth rate control, irrespective of the details of the molecular implementation. The study highlights the interplay between phenomenological modeling and molecular mechanisms in uncovering fundamental operating constraints, with implications for endogenous and synthetic design of microorganisms.

Vegetative growth control is crucial for high fruit yield in pear orchards. The use of chemical growth retardants has been prohibited in Danish and European pear production due to increasing concerns about environment and chemical residues on fruits. Root pruning has been proven as a good alternative to control the vegetative growth of fruit trees substituting the use of chemical growth regulators. A field experiment was conducted in 2011 to investigate the effects of root pruning and different irrigation regimes on shoot growth, tree water status, leaf photosynthesis and fruit size and color in a pear orchard. The results showed that root pruning is very effective in controlling the vegetative growth of pear trees. In root pruned pear trees full irrigation and deficit irrigation did not stimulate additional shoot growth. Root pruning decreased the tree water status and stomatal conductance. However, leaf photosynthesis was not affected resulting in improved intrinsic water use efficiency. Root pruning resulted in lower fruit weight, size and less green color. Full irrigation and deficit irrigation tended to improve fruit weight and size in root pruned pear trees, and thus maybe used as an efficient tool to alleviate the negative effects of root pruning on fruit yield and quality.

Vegetative growth control is crucial for high fruit yield in pear orchards. The use of chemical growth retardants has been prohibited in Danish and European pear production due to increasing concerns about environment and chemical residues on fruits. Root pruning has been proven as a good alternative to control the vegetative growth of fruit trees substituting the use of chemical growth regulators. A field experiment was conducted in 2011 to investigate the effects of root pruning and different irrigation regimes on shoot growth, tree water status, leaf photosynthesis and fruit size and color in a pear orchard. The results showed that root pruning is very effective in controlling the vegetative growth of pear trees. In root pruned pear trees full irrigation and deficit irrigation did not stimulate additional shoot growth. Root pruning decreased the tree water status and stomatal conductance. However, leaf photosynthesis was not affected resulting in improved intrinsic water use efficiency. Root pruning resulted in lower fruit weight, size and less green color. Full irrigation and deficit irrigation tended to improve fruit weight and size in root pruned pear trees, and thus maybe used as an efficient tool to alleviate the negative effects of root pruning on fruit yield and quality.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Gibberellic acid (GA3) increases berry size of "Emperatriz" seedless grape, the response depending on the phenological stage of vine at treatment date and on the concentration applied. From berry fruit set to 21 days later, 80 mg/L GA3 increased commercial berry weight by 50%-90%, depending on the year, reaching similar size to that of "Aledo" seeded grape, used as comparison. This effect takes place through: a) a larger berry growth rate; b) an early glucose, fructose and sucrose uptake; c) an increase of absolute glucose and fructose content (mg/berry) of seedless berries up to similar values to those of seeded berries; and d) an increase of absolute berry water content but not of relative content to fresh weight, thus water potential and osmotic potential are not significantly modified by treatments. GA3 does not affect berry pericarp cell number but increases pericarp cell diameter.

Three varieties of Kappaphycus alvarezii (Kapilaran, KAP), Tambalang purple (PUR), Adik-adik (AA), and one variety of Kappaphycus striatum var. sacol (green sacol (GS) were used to determine the efficiency of Acadian marine plant extract powder (AMPEP) as a culture medium at different concentrations, for the regeneration of young plants of Kappaphycus varieties, using tissue culture techniques for the production of seed stock for nursery and outplanting purposes for the commercial cultivation of carrageenophytes. A shorter duration for shoot formation was observed when the explant was treated with AMPEP + Plant Growth Regulator (PGR = PAA + zeatin at 1 mg L−1) compared to AMPEP when used singly. However, four explants responded differently to the number of days required for shoot formation. The KAP variety took 46 days to form shoots at 3–4 mg L−1 AMPEP + PGR; while PUR required 21 days at 3–5 mg L−1 AMPEP and 3–4 mg L−1 AMPEP + PGR. AA required 17 days at 3–5 mg L−1 AMPEP and AMPEP + PGR; and GS 25 days at 1 mg L−1 AMPEP + PGR. It was observed that among the four explants used, PUR and AA initiated shoot formation with the use of AMPEP only at higher concentrations (3–5 mg L−1) after a shorter period. Only PUR responded positively to ESS/2 for shoot initiation. The use of AMPEP alone and/or in combination with PGR as a culture medium in the propagation of microplantlets using tissue culture technique is highly encouraging.

We report here experiments aimed at understanding the connections between cell competition and growth in the Drosophila wing disc. The principal assay has been to generate discs containing marked cells that proliferate at different rates and to study their interactions and their contribution to the final structure. It is known that single clones of fast-dividing cells within a compartment may occupy the larger part of the compartment without affecting its size. This has suggested the existence of interactions involving cell competition between fast- and slow-dividing cells directed to accommodate the contribution of each cell to the final compartment. Here we show that indeed fast-dividing cells can outcompete slow-dividing ones in their proximity. However, we argue that this elimination is of little consequence because preventing apoptosis, and therefore cell competition, in those compartments does not affect the size of the clones or the size of the compartments. Our experiments indicate that cells within a compartment proliferate autonomously at their own rate. The contribution of each cell to the compartment is exclusively determined by its division rate within the frame of a size control mechanism that stops growth once the compartment has reached the final arresting size. This is supported by a computer simulation of the contribution of individual fast clones growing within a population of slower dividing cells and without interacting with them. The values predicted by the simulation are very close to those obtained experimentally.

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the

The mechanisms that control organ growth are among the least known in development. This is particularly the case for the process in which growth is arrested once final size is reached. We have studied this problem in the wing disc of Drosophila, the developmental and growth parameters of which are well known. We have devised a method to generate entire fast-growing Minute+ (M+) discs or compartments in slow developing Minute/+ (M/+) larvae. Under these conditions, a M+ wing disc gains at least 20 hours of additional development time. Yet it grows to the same size of Minute/+ discs developing in M/+ larvae. We have also generated wing discs in which all the cells in either the anterior (A) or the posterior (P) compartment are transformed from M/+ to M+. We find that the difference in the cell division rate of their cells is reflected in autonomous differences in the developmental progression of these compartments: each grows at its own rate and manifests autonomous regulation in the expression of the developmental genes wingless and vestigial. In spite of these differences, 'mosaic'discs comprising fast and slow compartments differentiate into adult wings of the correct size and shape. Our results demonstrate that imaginal discs possess an autonomous mechanism with which to arrest growth in anterior and posterior compartments, which behave as independent developmental units. We propose that this mechanism does not act by preventing cell divisions, but by lengthening the division cycle.

The Decapentaplegic (Dpp) morphogen gradient controls growth and patterning in the Drosophila appendages. There is recent evidence indicating that the Dpp gradient is converted into an inverse gradient of activity of the gene brinker (brk), which encodes a transcriptional repressor and is negatively regulated by the Dpp pathway. We have studied how alterations in the Brk gradient affect the growth of the wing disc. We find that there is a negative correlation between brk activity and growth of the disc: high levels of brk prevent or reduce growth, whereas loss of brk activity results in excessive growth. This effect is concentration dependent: different amounts of Brk produce distinct rates of growth. Furthermore, our results demonstrate that although brk is able to induce apoptosis where there is a sharp difference in Brk levels, its role as a growth repressor is not achieved by inducing apoptosis but by reducing cell proliferation. Brk appears to downregulate the activity of genes that control cell proliferation, such as bantam.

Outside the immediate visage of regional policy analysts and smart growth boosters, the local growth control & growth management (LGC&M) movement remains extremely active, especially at the ballot box and especially in California. Analyses show LGC&M activity to be strongly correlated with state and regional growth rates—rising during expansionary periods such as the late-1980s and late-1990s, and falling during slowdown periods such as the early-1990s. Although exact numbers are hard to come by, it is estimated that about three-quarters of California cities and two-thirds of California counties have adopted some form of LGC&M program since 1980.Their continuing popularity notwithstanding, many questions remain about the efficacy and effects of LGC&M programs. Four are of particular interest:(1) To what extent do different LGC&M approaches really restrict the amount, pace, or location of growth? (2) To what extent are the resulting supply restrictions reflected in local real estate prices, especially housing prices? (3) Which LGC&M programs and approaches yield their promised benefits and which do not? (4) Do LGC&M programs cause growth to be systematically displaced from more restrictive to less restrictive communities, leading to such negative outcomes as sprawl and wasteful commuting?Using California as its lens, this report focuses on these unresolved questions. In no other US state is growth as tightly managed at the local level as California. Likewise, in no other state is there such a diversity of local growth management approaches and experiences. From a research perspective, because California lacks a statewide growth management framework, the effectiveness or ineffectiveness of growth management can be traced back to its local implementation. The report is organized into six sections: (1) Recent theoretical and empirical research into the use and efficacy of LGC&M programs; (2) An updated taxonomy of growth management an