Letteratura scientifica selezionata sul tema "Mating type gene"

When protoplasts carrying metalaxyl-resistant (Mr) nuclei from the A1 isolate of Phytophthora parasitica were fused with protoplasts carrying chloroneb-resistant (Cnr) nuclei from the A2 isolate of the same species, fusion products carrying Mr nuclei were either the A2 or A1A2 type, while those carrying Cnr nuclei were the A1, A2, or A1A2 type. Fusion products carrying Mr and Cnr nuclei also behaved as the A1, A2, or A1A2 type. The result refutes the hypothesis that mating types in Phytophthora are controlled by nuclear genes. When nuclei from the A1 isolate of P. parasitica were fused with protoplasts from the A2 isolate of the same species and vice versa, all of the nuclear hybrids expressed the mating type characteristics of the protoplast parent. The same was true when the nuclei from the A1 isolate of P. parasitica were fused with the protoplasts from the A0 isolate of Phytophthora capsici and vice versa. These results confirm the observation that mating type genes are not located in the nuclei and suggest the presence of mating type genes in the cytoplasms of the recipient protoplasts. When mitochondria from the A1 isolate of P. parasitica were fused with protoplasts from the A2 isolate of the same species, the mating type of three out of five regenerated protoplasts was changed to the A1 type. The result demonstrated the decisive effect of mitochondrial donor sexuality on mating type characteristics of mitochondrial hybrids and suggested the presence of mating type genes in mitochondria. All of the mitochondrial hybrids resulting from the transfer of mitochondria from the A0 isolate of P. capsici into protoplasts from the A1 isolate of P. parasitica were all of the A0 type. The result supports the hypothesis of the presence of mating type genes in mitochondria in Phytophthora.Key words: mating type, mitochondrial gene, Phytophthora parasitica, Phytophthora capsici.

In heterothallic basidiomycete fungi, sexual compatibility is restricted by mating types, typically controlled by two loci: PR, encoding pheromone precursors and pheromone receptors, and HD, encoding two types of homeodomain transcription factors. We analysed the single mating-type locus of the commercial button mushroom variety, Agaricus bisporus var. bisporus, and of the related variety burnettii. We identified the location of the mating-type locus using genetic map and genome information, corresponding to the HD locus, the PR locus having lost its mating-type role. We found the mip1 and β-fg genes flanking the HD genes as in several Agaricomycetes, two copies of the β-fg gene, an additional HD2 copy in the reference genome of A. bisporus var. bisporus and an additional HD1 copy in the reference genome of A. bisporus var. burnettii. We detected a 140 kb-long inversion between mating types in an A. bisporus var. burnettii heterokaryon, trapping the HD genes, the mip1 gene and fragments of additional genes. The two varieties had islands of transposable elements at the mating-type locus, spanning 35 kb in the A. bisporus var. burnettii reference genome. Linkage analyses showed a region with low recombination in the mating-type locus region in the A. bisporus var. burnettii variety. We found high differentiation between β-fg alleles in both varieties, indicating an ancient event of recombination suppression, followed more recently by a suppression of recombination at the mip1 gene through the inversion in A. bisporus var. burnettii and a suppression of recombination across whole chromosomes in A. bisporus var. bisporus, constituting stepwise recombination suppression as in many other mating-type chromosomes and sex chromosomes.

Transposable elements (TEs) have had immense impact on the structure, function and evolution of eukaryotic genomes. The work in this thesis identified Kat1, a novel domesticated DNA transposase of the hAT family in the yeast Kluyveromyces lactis. Kat1 triggers a genome rearrangement that results in a switch of mating type from MATa to MATα. Furthermore, Kat1 acts on sequences that presumably are ancient remnants of a long-lost transposable element. Therefore, Kat1 provides a remarkable example of the intricate relationship between transposable elements and their hosts. We showed that Kat1 generates two DNA double strand breaks (DSBs) in MATa and that the DDE motif and several other conserved amino acid residues are important for Kat1 cleavage activity. DNA hairpins were formed on one end of the DSBs whereas the DNA between the DSBs was joined into a circle. Kat1 was transcriptionally activated by nutrient limitation through the transcription factor Mts1 and negatively regulated by translational frameshifting. In conclusion, Kat1 is a highly regulated domesticated transposase that induces sexual differentiation.  In another study, we developed an assay to measure switching rates in K. lactis and found that the switching rate was ~6x10-4 events/generation. In a genetic screen for mutations that increased mating-type switching, we found mutations in the RAS1 gene. The small GTPase Ras1 regulates cellular cyclic AMP levels and we demonstrated that Mts1 transcription is regulated by the RAS/cAMP pathway and the transcription factor Msn2. Since Ras activity is regulated by nutrient availability, these data likely explains why nutrient limitation induces mating-type switching.

The aim of this thesis was to study the evolution of reproductive systems and reproductive traits in the fungal genus Neurospora. More specifically, I have investigated the evolutionary forces shaping the genes involved in sexual reproduction, focusing on mating-type (mat) and pheromone receptor (pre) genes. Neurospora contains species exhibiting three different mating systems, i.e., heterothallism (self-incompatibility), homothallism (self-compatibility) and pseudohomothallism (partial self-incompatibility). First, a robust phylogeny of Neurospora was established. The phylogenetic analyses revealed multiple independent transitions in reproductive life style during the evolutionary history of the genus. We argued for a heterothallic ancestor of the genus, although our subsequent ancestral reconstruction analyses favored a homothallic ancestor. To be able to settle the ancestral mating system, we zoomed in on the structural architecture of the mat-locus in four homothallic species of Neurospora, thought to have arisen from independent transitions. Our results led us to suggest two different genetic mechanisms (translocation and unequal crossover) to explain the transitions in mating system from heterothallism to homothallism. We pointed out that the mating-system transitions in Neurospora are unidirectional, and suggested that transposable elements might be driving the transitions. In conclusion, we suggest a heterothallic ancestor for Neurospora, and that at least six transitions to homothallism and two transitions to pseudohomothallism have occurred in its evolutionary history. Further, we used the phylogeny of Neurospora as a framework to test if the evolution of pre-genes (pre-1 and pre-2) in hetero- and homothallic Neurospora is dependent on mating systems and/or even the homothallic clades themselves (i.e., mating-system and/or switch-dependent). The molecular evolution results suggest that pre-1 and pre-2 are overall functional in both homothallic and heterothallic Neurospora. The molecular evolution of pre-1 seems to be independent of mating-system or homothallic clade, and we detected signs for positive selection in the C-terminal tail. For pre-2 we found no support for mating-system dependent evolution, but indications for switch-dependent evolution. In this study we also included expression analyses of both pre- as well as mat-genes, with the prospect to assess functionality and regulation. During this thesis work, we also performed a phylogenetic study were we found that reproductive genes might be more permeable to introgression than other genes, which is in contrast to theoretical expectations. In the last study, we confirmed the co-existence of two alternative splice variants of the pheromone receptor gene pre-1 in Neurospora crassa, and performed expression profiles studies using quantitative RT-PCR. I hope this thesis work will further strengthen Neurospora as a model for research in evolutionary genetics.

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Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG
Rice is a worldwide cultivated and consumed grain, playing an important role on the diet of half of the world’s population. Several losses in production and grain quality have been reported due to biotic factors, such as rice blast, caused by Magnaporthe oryzae, which is the major disease in rice crops. One of the most effective ways to control this disease is the use of resistant cultivars. However, the high genetic variability of the pathogen results in a rapid resistance loss. The discovery of highly fertile, hermaphrodites individuals outside of the rice center of origin, suggests that sexual reproduction may contribute to this genetic variability, which influences the appropriate control strategies. M. oryzae reproduction studies begins with the determination of mating types, controlled by two idiomorphic genes (MAT1-1 e MAT1-2), along with the sexuality (hermaphrodite, female or male) and fertility (number of perithecia). Another important approach under investigation for this crop is the detection of avirulence genes from M. oryzae, to understand the pathogen variability. Our study focused on the investigation of MAT1-1 or MAT1-2 genes, and the presence of the avirulence gene AVR1-CO39in field isolates collected from all rice production regions from Brazil. Sexuality and fertility were also characterized. 208 selected isolates were cultivated in PDA medium and the fungus mycelia were used for DNA extraction and PCR detection of the above-mentioned genes. For the sexual characterization, 106 field isolates were paired in Petri dishes containing rice bran medium with two reference isolates: KA-3 (MAT1-1) and GUY11 (MAT1-2), known worldwide for their mating type and high fertility. The AVR1-CO39 gene was only detected in two field isolates. One of them was able to infect the rice cultivar CO39, which has the resistance gene Pi-CO39(t). A mutation on AVR1-CO39 gene could impair the recognition of its effector by Pi-CO39(t) protein. Only one mating type (MAT1-2) was observed on the 208 field isolates. It was also observed that, among the 106 analyzed isolates, one (0,94%) was identified as a female; three (2,8%) as hermaphrodite, 62 (57,9%) as male; and 41 (38,3%) were not determined, considered infertile. We also observed the formation of perithecia inside of rice leaves. Despite the predominance of one mating type among rice field isolates, there is a possibility that sexual reproduction may occur as the other idiomorphic gene (MAT1-1) is present on field isolates collected from other Poaceae. The identification of highly fertile hermaphrodites and fertile-female individuals in this study also highlight this possibility.
O arroz é cultivado e consumido em todos os continentes, desempenhando um importante papel na dieta de mais da metade da população mundial. O seu cultivo vem sofrendo perdas na produção e na qualidade de grãos, devido a fatores bióticos como a brusone, causada pelo fungo Magnaporthe oryzae, que é a principal doença da cultura do arroz, representando uma ameaça à segurança alimentar mundial. O uso de cultivares resistentes é considerado o método mais efetivo para o controle da doença, porém, a alta variabilidade do patógeno resulta em uma rápida suplantação da resistência. Com a descoberta de isolados de alta fertilidade, hermafroditas, fora do centro de origem do arroz, sugere-se que a reprodução sexuada possa estar contribuindo para esta variabilidade genética, o que consequentemente influencia as estratégias apropriadas de controle. O estudo da reprodução sexuada em M. oryzae inicia-se com a definição dos tipos compatíveis, característica controlada pelo gene mating type com dois idiomorfos (MAT1-1 e MAT1-2); além das características como sexualidade (hermafrodita, fêmea ou macho) e fertilidade (número de peritécios). Outra abordagem de grande importância para a cultura é a detecção de genes de avirulência de M. oryzae, visando estudos de sua variabilidade. Dessa forma, o objetivo desta pesquisa foi investigar a presença dos genes MAT1-1 ou MAT1-2, e do gene de avirulência AVR1-CO39, em isolados coletados em todas as regiões produtoras de arroz do Brasil, além de caracterizá-los quanto à sexualidade e fertilidade. Foram selecionados 208 isolados que forma cultivados em BDA e seus micélios utilizados para extração de DNA e detecção dos genes citados. Para a caracterização sexual, 106 isolados de campo foram pareados em placa de Petri, contendo meio de farelo de arroz, com dois isolados: KA-3 (MAT1-1) e GUY11 (MAT1-2), os quais apresentam mating types e alta fertilidade, conhecidos mundialmente. O gene AVR1-CO39 foi detectado em apenas dois isolados, e um deles é patogênico à cultivar CO39, portadora do gene de resistência Pi-CO39(t), levantando a possibilidade de que uma mutação possa ter ocorrido, como deleção, o que impossibilita o reconhecimento do efetor pela proteína do gene Pi-CO39(t). Apenas um tipo compatível (MAT1-2) foi observado nos 208 isolados de campo. Foi observado também que, entre os 106 isolados analisados, um (0,94%), foi identificado como fêmea; três (2,8%), como hermafroditas; 62 (57,9%), como machos; e 41 (38,3%), como não determinados, sendo considerados inférteis. Observou-se também a formação de peritécios no interior da folha de arroz. Apesar do predomínio de um mating type, entre os isolados do arroz, há a possibilidade de ocorrência da reprodução sexuada, devido à presença do outro idiomorfo (MAT1-1) em isolados coletados de outras gramíneas, juntamente com a presença de hermafroditas e da fêmea-fértil, com alta fertilidade, de isolados MAT1-2 identificados nesse trabalho.

This work is concerned with the molecular analysis of ascomycete mating type genes of various Sordaria species. Work previously published has reported the cloning and characterisation of mating type genes from several Neurospora species. In heterothallic species the genotype at the mating type locus (mtA or mta) determines the mating type. Homothallic species, which proceed through the sexual cycle without the need to mate, have no obvious mating types but molecular analysis has been used to demonstrate the presence of mating type genes in species with this life cycle. Neurospora species and Sordaria species both belong to the Sordariaceae and are closely related. Several λ clones containing putative Sordaria mating type genes from heterothallic and homothallic species had been isolated previously using N. crassa mtA and mta probes. In this study the mtA-1 gene of the heterothallic species S. sclerogenia was subcloned from a λ clone and sequenced. The equivalent gene from S. equina (a homothallic species containing only the mtA sequence) was also subcloned and sequenced. A λ clone for the species S. fimicola was found to hybridise with both the mtA and mta probes. S. fimicola is a homothallic species containing mtA and mta in the same nucleus. On using the lambda clone it was found that the mtA and mta genes are linked in this species. All the Sordaria mtA-1 genes contained putative DNA binding domains, α domains. The mta-1 gene sequenced from s. fimicola contained a putative HMG box. The S. equina mtA-1 gene was expressed in a sterile N. crassa mta mutant and was found to restore mating type function to the mutant. The mtA-1 gene did not however confer homothallic behaviour on the recipient mutant. S. equina and S. sclerogenia contain a 59bp common region following on from the mtA-1 gene which is conserved in both these species and in Neurospora species. A variable region continues on from the common region in S. equina and S. sclerogenia and in Neuropora species.

Le champignon filamenteux, Podospora anserina, possède deux types sexuels, mat+ et mat-, caractérisés chacun par une séquence spécifique. La séquence mat+ contient un seul gène FPR1; la séquence mat- contient trois gènes : FMR1, SMR1 et SMR2. La fonction moléculaire de SMR1 est inconnue, les autres gènes codent des facteurs de transcription qui contrôlent la fécondation (reconnaissance intercellulaire), et le passage d'un syncytium à un hyphe spécialisé binucléé contenant un noyau mat+ et un noyau mat- (reconnaissance internucléaire). Il n'y a pas eu d'analyse exhaustive des gènes impliqués dans la reconnaissance intercellulaire et le mécanisme de la reconnaissance internucléaire est encore inconnu. Afin de déterminer les cibles de FPR1 et FMR1, et les différents mécanismes impliqués, nous avons utilisé une approche microarray. Le profil transcriptomique des souches mat+ et mat- compétentes pour la fécondation a permis d'identifier 157 gènes cibles, et l'analyse transcriptomique des souches mutantes fpr1- et fmr1- a révélé que ces cibles peuvent être soit réprimées, soit activées par FMR1 ou FPR1, ou être sous le contrôle de ces deux facteurs. Ces expériences ont aussi détecté l'existence de 10 gènes activés ou réprimés au même niveau dans mat+ et mat-. La délétion de 32 gènes choisis parmi ces 167 gènes cibles n'a permis de mettre en évidence que deux gènes impliqués dans la fécondation. Les comparaisons des gènes cibles des facteurs de transcription MAT de Gibberella moniliformis et Sordaria macrospora avec ceux de P. anserina révèlent un nombre significatif de gènes cibles communs entre ces espèces, mais ces gènes ont des profils transcriptomiques différents, soulevant la question du rôle de ces gènes cibles. La recherche des gènes cibles de FPR1, FMR1 et SMR2 impliqués dans la reconnaissance internuléaire a été effectuée en comparant le transcriptome des périthèces issus de deux croisements, l'un n'exprimant que les gènes spécifiques mat+, l'autre que les gènes spécifiques mat-. Les résultats ont été interprétés selon le modèle d'identité nucléaire et le modèle de ségrégation aléatoire. Le premier modèle a conduit à l'identification de 27 gènes cibles, tandis que 154 gènes cibles ont été identifiés en appliquant le deuxième modèle. Au total 46 souches mutantes ont été construites. Cependant aucune délétion n'a affecté le développement sexué. En parallèle de ces expériences transcriptomiques, nous avons invalidé tous les gènes à HMG-box de P. anserina. Les résultats montrent que ces derniers ont un rôle très important dans le développement sexué, particulièrement Pa_1_13940 qui code un régulateur des gènes des types sexuels, le premier identifié chez les Pezizomycotina.

Sporisorium scitamineum é um fungo basidiomiceto causador do carvão da cana-de-açúcar, uma doença com impacto negativo no cultivo da cana-de-açúcar, e com ocorrência em todos os países produtores. A manifestação da doença na cultura da cana depende da formação de uma hifa dicariótica a partir da anastomose de duas hifas haplóides compatíveis com relação ao tipo de reação sexual (mating-type). O controle do cruzamento sexuado (mating) é realizado pela expressão de um conjunto de genes presentes em dois loci, a e b. O locus a codifica um lipopeptídeo com função de feromônio e um receptor de feromônio, responsáveis pelo reconhecimento de células compatíveis e fusão de hifas, enquanto o locus b codifica fatores de transcrição que controlam a expressão de genes responsáveis pela manutenção das hifas dicarióticas durante o processo de infecção e crescimento do fungo dentro da planta. Apesar de desempenharem função essencial no processo de infecção e manutenção da doença em cana-de-açúcar, o conhecimento a respeito da organização genômica ou da função dos demais genes presentes nos loci a e b em S. scitamineum e em outros fungos causadores de carvão é ainda incipiente. Desta forma, o objetivo geral do presente trabalho foi isolar as regiões genômicas relacionadas aos genes de cruzamento em S. scitamineum e analisar comparativamente com regiões similares já descritas e depositadas em bancos de dados públicos. Para o isolamento destas regiões, foi construída uma biblioteca genômica em BAC de uma linhagem haplóide de S. scitamineum, a Ssc39 (+), isolada de uma variedade de cana-de-açúcar com sintomas de alta susceptibilidade. Foram selecionados 11 clones por PCR. Os insertos foram sequenciados e utilizados para confirmação da montagem dos loci no sequenciamento do genoma do fungo. Apesar do fungo S. scitamineum apresentar sistema bipolar de reação sexual assim como o fungo U. hordei, as análises comparativas de ambos os locus indicaram que S. scitamineum apresenta maior similaridade com o fungo S. reilianum principalmente com o alelo a1, no qual apresenta sistema tetrapolar de reação sexual. A anotação e caracterização dos genes do tipo de reação sexual (mating type) possibilitaram a comparação e melhor entendimento sobre esses genes de grande importância na patogenicidade e no ciclo de vida do fungo.
Sporisorium scitamineum is a basidiomycete fungus causing the smut disease in sugarcane, with a negative impact on the cultivation of sugarcane, and occurring in all producing countries. The manifestation of the disease in sugarcane crop depends on the formation of a dikaryotic hyphae originated of the anastomosis of two haploid mating type compatible cells. The control of the sexual crossing (mating) is performed by expression of a set of genes present in two loci, a and b. The locus a encodes a lipopeptide with the function of pheromone and pheromone membrane receptor responsible for cell recognition and compatible hyphal fusion, whereas the locus b encodes transcription factors that control the expression of genes responsible for the maintenance of the dikaryotic hyphal growth in plant. Although they play an essential role in the maintenance of infection and disease in sugarcane process, knowledge about the genomic organization and function of other genes in these two loci of S. scitamineum and other smut fungi is still incipient. Thus, the overall goal of this work was to isolate genomic regions related to the mating type in S. scitamineum and to perform a comparative analyze with similar regions described and deposited in public databases. For the isolation of these regions, we constructed a genomic BAC library of a haploid strain of S. scitamineum, the Ssc39 (+), isolated from a variety of sugarcane with symptoms of high susceptibility. Eleven clones were selected by PCR. The inserts were sequenced and used to confirm the assembly of both loci in the genome sequencing of the fungus. Although S. scitamineum belongs to the class of bipolar system of sexual response as well as the fungus U. hordei , the comparative analysis of both loci indicated that S. scitamineum shows greater similarity to the S. reilianum mainly with A1 allele, which has a tetrapolar system sexual response. The annotation of the genes and characterization mating type genes enabled the comparison and better understanding of the importance of these genes in the life cycle of the fungus.

Reproductive isolation is an essential stage in speciation. In Ascomycetes, the ubiquitous distribution of many species suggests that sympatric speciation through assertive mating should be an important factor. The MAT locus and the pheromone/receptor system could both potentially contribute to the development of such sexual isolation. Alterations at the MAT loci could lead to distinct reproductive habits or a change in mating system, both of which can reduce gene-flow between species. However, if deliberate pre-mating sexual preferences exist, they are more likely to be determined by the pheromone/receptor system. This study of Fusarium reproductive genes, and comparisons with other Ascomycetes, has yielded numerous interesting findings regarding the evolution of these mate-recognition mechanisms and the implications thereof. The G. fujikuroi and F. graminearum species complexes have offered an interesting comparison between heterothallic and homothallic MAT locus evolution. The value of comparative sequence analysis has been demonstrated in the discovery of a previously unknown gene, MAT1-2-3, which may be specific to members of the Order Hypocreales. While all MAT genes share similar regulatory elements, this is the first report of evidence that a transition to homothallism can be accompanied by the recruitment of distinct elements that could facilitate alternate expression of MAT genes. The MAT genes are also highly divergent between Fusarium spp., largely due to relaxed selective constraint, particularly in homothallic species. However, inter-specific gene-flow could curb MAT gene divergence among homothallic species. Despite strong reproductive barriers in the G. fujikuroi complex, the F. sacchari MAT1-1 sequence appears to have been acquired through lateral transfer from a distant relative. Analytical analysis of the MAT locus novelties reported here, including the new MAT gene, will be necessary to determine their biological significance. To investigate the extent of pheromone diversity in the Ascomycetes, and to gain clues as to its biological importance, pheromone peptides from seventy ascomycete species were compared. A number of reproductively incompatible species, such as those in the G. fujikuroi complex, share identical pheromones; which implies that another mechanism must be responsible for the observed reproductive barriers. However, on the whole, pheromones are highly divergent among species. Both adaptive and non-adaptive evolution could have contributed to this pattern. In fact the structure of the á-class pheromone precursor gene, which consists of multiple repeats of the pheromone module, could facilitate rapid diversification through “birth-and-death” evolution. Within species, selection maintains pheromone peptides, implying that much of the inter-specific variation is functionally relevant. This further suggests that pheromone evolution could contribute to the generation of reproductive isolation between species. The most general trend in the findings of this study is that ascomycete reproductive genes are highly divergent. This is in agreement with findings in other Kingdoms. A number of evolutionary forces are probably involved but weaker selective constraint, resulting from the fact that reproduction is not essential in these fungi, appears to be a common factor. This reproductive gene variability could be directly linked to speciation and, therefore, the great diversity in Ascomycetes. Additional information on the appendices is available on a CD, stored at the Merensky Library on Level 3
Dissertation (MSc)--University of Pretoria, 2011.
Genetics
Unrestricted

Bones are multifunctional passive organs of movement that supports soft tissue and directly attached muscles. They also protect internal organs and are a reserve of calcium, phosphorus and magnesium. Each bone is covered with periosteum, and the adjacent bone surfaces are covered by articular cartilage. Histologically, the bone is an organ composed of many different tissues. The main component is bone tissue (cortical and spongy) composed of a set of bone cells and intercellular substance (mineral and organic), it also contains fat, hematopoietic (bone marrow) and cartilaginous tissue. Bones are a tissue that even in adult life retains the ability to change shape and structure depending on changes in their mechanical and hormonal environment, as well as self-renewal and repair capabilities. This process is called bone turnover. The basic processes of bone turnover are: • bone modeling (incessantly changes in bone shape during individual growth) following resorption and tissue formation at various locations (e.g. bone marrow formation) to increase mass and skeletal morphology. This process occurs in the bones of growing individuals and stops after reaching puberty • bone remodeling (processes involve in maintaining bone tissue by resorbing and replacing old bone tissue with new tissue in the same place, e.g. repairing micro fractures). It is a process involving the removal and internal remodeling of existing bone and is responsible for maintaining tissue mass and architecture of mature bones. Bone turnover is regulated by two types of transformation: • osteoclastogenesis, i.e. formation of cells responsible for bone resorption • osteoblastogenesis, i.e. formation of cells responsible for bone formation (bone matrix synthesis and mineralization) Bone maturity can be defined as the completion of basic structural development and mineralization leading to maximum mass and optimal mechanical strength. The highest rate of increase in pig bone mass is observed in the first twelve weeks after birth. This period of growth is considered crucial for optimizing the growth of the skeleton of pigs, because the degree of bone mineralization in later life stages (adulthood) depends largely on the amount of bone minerals accumulated in the early stages of their growth. The development of the technique allows to determine the condition of the skeletal system (or individual bones) in living animals by methods used in human medicine, or after their slaughter. For in vivo determination of bone properties, Abstract 10 double energy X-ray absorptiometry or computed tomography scanning techniques are used. Both methods allow the quantification of mineral content and bone mineral density. The most important property from a practical point of view is the bone’s bending strength, which is directly determined by the maximum bending force. The most important factors affecting bone strength are: • age (growth period), • gender and the associated hormonal balance, • genotype and modification of genes responsible for bone growth • chemical composition of the body (protein and fat content, and the proportion between these components), • physical activity and related bone load, • nutritional factors: – protein intake influencing synthesis of organic matrix of bone, – content of minerals in the feed (CA, P, Zn, Ca/P, Mg, Mn, Na, Cl, K, Cu ratio) influencing synthesis of the inorganic matrix of bone, – mineral/protein ratio in the diet (Ca/protein, P/protein, Zn/protein) – feed energy concentration, – energy source (content of saturated fatty acids - SFA, content of polyun saturated fatty acids - PUFA, in particular ALA, EPA, DPA, DHA), – feed additives, in particular: enzymes (e.g. phytase releasing of minerals bounded in phytin complexes), probiotics and prebiotics (e.g. inulin improving the function of the digestive tract by increasing absorption of nutrients), – vitamin content that regulate metabolism and biochemical changes occurring in bone tissue (e.g. vitamin D3, B6, C and K). This study was based on the results of research experiments from available literature, and studies on growing pigs carried out at the Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences. The tests were performed in total on 300 pigs of Duroc, Pietrain, Puławska breeds, line 990 and hybrids (Great White × Duroc, Great White × Landrace), PIC pigs, slaughtered at different body weight during the growth period from 15 to 130 kg. Bones for biomechanical tests were collected after slaughter from each pig. Their length, mass and volume were determined. Based on these measurements, the specific weight (density, g/cm3) was calculated. Then each bone was cut in the middle of the shaft and the outer and inner diameters were measured both horizontally and vertically. Based on these measurements, the following indicators were calculated: • cortical thickness, • cortical surface, • cortical index. Abstract 11 Bone strength was tested by a three-point bending test. The obtained data enabled the determination of: • bending force (the magnitude of the maximum force at which disintegration and disruption of bone structure occurs), • strength (the amount of maximum force needed to break/crack of bone), • stiffness (quotient of the force acting on the bone and the amount of displacement occurring under the influence of this force). Investigation of changes in physical and biomechanical features of bones during growth was performed on pigs of the synthetic 990 line growing from 15 to 130 kg body weight. The animals were slaughtered successively at a body weight of 15, 30, 40, 50, 70, 90, 110 and 130 kg. After slaughter, the following bones were separated from the right half-carcass: humerus, 3rd and 4th metatarsal bone, femur, tibia and fibula as well as 3rd and 4th metatarsal bone. The features of bones were determined using methods described in the methodology. Describing bone growth with the Gompertz equation, it was found that the earliest slowdown of bone growth curve was observed for metacarpal and metatarsal bones. This means that these bones matured the most quickly. The established data also indicate that the rib is the slowest maturing bone. The femur, humerus, tibia and fibula were between the values of these features for the metatarsal, metacarpal and rib bones. The rate of increase in bone mass and length differed significantly between the examined bones, but in all cases it was lower (coefficient b <1) than the growth rate of the whole body of the animal. The fastest growth rate was estimated for the rib mass (coefficient b = 0.93). Among the long bones, the humerus (coefficient b = 0.81) was characterized by the fastest rate of weight gain, however femur the smallest (coefficient b = 0.71). The lowest rate of bone mass increase was observed in the foot bones, with the metacarpal bones having a slightly higher value of coefficient b than the metatarsal bones (0.67 vs 0.62). The third bone had a lower growth rate than the fourth bone, regardless of whether they were metatarsal or metacarpal. The value of the bending force increased as the animals grew. Regardless of the growth point tested, the highest values were observed for the humerus, tibia and femur, smaller for the metatarsal and metacarpal bone, and the lowest for the fibula and rib. The rate of change in the value of this indicator increased at a similar rate as the body weight changes of the animals in the case of the fibula and the fourth metacarpal bone (b value = 0.98), and more slowly in the case of the metatarsal bone, the third metacarpal bone, and the tibia bone (values of the b ratio 0.81–0.85), and the slowest femur, humerus and rib (value of b = 0.60–0.66). Bone stiffness increased as animals grew. Regardless of the growth point tested, the highest values were observed for the humerus, tibia and femur, smaller for the metatarsal and metacarpal bone, and the lowest for the fibula and rib. Abstract 12 The rate of change in the value of this indicator changed at a faster rate than the increase in weight of pigs in the case of metacarpal and metatarsal bones (coefficient b = 1.01–1.22), slightly slower in the case of fibula (coefficient b = 0.92), definitely slower in the case of the tibia (b = 0.73), ribs (b = 0.66), femur (b = 0.59) and humerus (b = 0.50). Bone strength increased as animals grew. Regardless of the growth point tested, bone strength was as follows femur > tibia > humerus > 4 metacarpal> 3 metacarpal> 3 metatarsal > 4 metatarsal > rib> fibula. The rate of increase in strength of all examined bones was greater than the rate of weight gain of pigs (value of the coefficient b = 2.04–3.26). As the animals grew, the bone density increased. However, the growth rate of this indicator for the majority of bones was slower than the rate of weight gain (the value of the coefficient b ranged from 0.37 – humerus to 0.84 – fibula). The exception was the rib, whose density increased at a similar pace increasing the body weight of animals (value of the coefficient b = 0.97). The study on the influence of the breed and the feeding intensity on bone characteristics (physical and biomechanical) was performed on pigs of the breeds Duroc, Pietrain, and synthetic 990 during a growth period of 15 to 70 kg body weight. Animals were fed ad libitum or dosed system. After slaughter at a body weight of 70 kg, three bones were taken from the right half-carcass: femur, three metatarsal, and three metacarpal and subjected to the determinations described in the methodology. The weight of bones of animals fed aa libitum was significantly lower than in pigs fed restrictively All bones of Duroc breed were significantly heavier and longer than Pietrain and 990 pig bones. The average values of bending force for the examined bones took the following order: III metatarsal bone (63.5 kg) <III metacarpal bone (77.9 kg) <femur (271.5 kg). The feeding system and breed of pigs had no significant effect on the value of this indicator. The average values of the bones strength took the following order: III metatarsal bone (92.6 kg) <III metacarpal (107.2 kg) <femur (353.1 kg). Feeding intensity and breed of animals had no significant effect on the value of this feature of the bones tested. The average bone density took the following order: femur (1.23 g/cm3) <III metatarsal bone (1.26 g/cm3) <III metacarpal bone (1.34 g / cm3). The density of bones of animals fed aa libitum was higher (P<0.01) than in animals fed with a dosing system. The density of examined bones within the breeds took the following order: Pietrain race> line 990> Duroc race. The differences between the “extreme” breeds were: 7.2% (III metatarsal bone), 8.3% (III metacarpal bone), 8.4% (femur). Abstract 13 The average bone stiffness took the following order: III metatarsal bone (35.1 kg/mm) <III metacarpus (41.5 kg/mm) <femur (60.5 kg/mm). This indicator did not differ between the groups of pigs fed at different intensity, except for the metacarpal bone, which was more stiffer in pigs fed aa libitum (P<0.05). The femur of animals fed ad libitum showed a tendency (P<0.09) to be more stiffer and a force of 4.5 kg required for its displacement by 1 mm. Breed differences in stiffness were found for the femur (P <0.05) and III metacarpal bone (P <0.05). For femur, the highest value of this indicator was found in Pietrain pigs (64.5 kg/mm), lower in pigs of 990 line (61.6 kg/mm) and the lowest in Duroc pigs (55.3 kg/mm). In turn, the 3rd metacarpal bone of Duroc and Pietrain pigs had similar stiffness (39.0 and 40.0 kg/mm respectively) and was smaller than that of line 990 pigs (45.4 kg/mm). The thickness of the cortical bone layer took the following order: III metatarsal bone (2.25 mm) <III metacarpal bone (2.41 mm) <femur (5.12 mm). The feeding system did not affect this indicator. Breed differences (P <0.05) for this trait were found only for the femur bone: Duroc (5.42 mm)> line 990 (5.13 mm)> Pietrain (4.81 mm). The cross sectional area of the examined bones was arranged in the following order: III metatarsal bone (84 mm2) <III metacarpal bone (90 mm2) <femur (286 mm2). The feeding system had no effect on the value of this bone trait, with the exception of the femur, which in animals fed the dosing system was 4.7% higher (P<0.05) than in pigs fed ad libitum. Breed differences (P<0.01) in the coross sectional area were found only in femur and III metatarsal bone. The value of this indicator was the highest in Duroc pigs, lower in 990 animals and the lowest in Pietrain pigs. The cortical index of individual bones was in the following order: III metatarsal bone (31.86) <III metacarpal bone (33.86) <femur (44.75). However, its value did not significantly depend on the intensity of feeding or the breed of pigs.

The method presented in this note mimics two fundamental mechanisms from nature, growth, and development, for the synthesis of new three-dimensional structures. The structures were synthesized to support a load generated by a wind. Every structure grows from a single artificial cell following a set of genes, encoded in an artificial genome shared by all cells. Genes are a set of commands that control the growth process. Genes are regulated by interaction with the environment. The environment is both external and internal to the structure. The performance each structure is measured by its ability to hold the load and other additional engineering criteria. A population of structures is evolved using a genetic algorithm, which alters the genome of two mating individuals. We will present evolved phenotypes with high degrees of modularity and symmetry which evolved according to engineering criteria. Neither one of these two characteristics has been directly imposed as the fitness evaluation, but rather spontaneously emerge as a consequence of natural selection. We will argue that the types of rules we are using in this model are not biased toward any of these characteristics, but rather basic rules for growth and development.

Abstract The aim of this study is to explore the possibility of matching a cycle performance model to public data on a state-of-the-art commercial aircraft engine (GEnx-1B). The study is focused on obtaining valuable information on figure of merits for the technology level of the low-pressure system and associated uncertainties. It is therefore directed more specifically towards the fan and low-pressure turbine efficiencies, the Mach number at the fan-face, the distribution of power between the core and the bypass stream as well as the fan pressure ratio. Available cycle performance data have been extracted from the engine emission databank provided by the International Civil Aviation Organization (ICAO), type certificate datasheets from the European Union Aviation Safety Agency (EASA) and the Federal Aviation Administration (FAA), as well as publicly available data from engine manufacturer. Uncertainties in the available source data are estimated and randomly sampled to generate inputs for a model matching procedure. The results show that fuel performance can be estimated with some degree of confidence. However, the study also indicates that a high degree of uncertainty is expected in the prediction of key low-pressure system performance metrics, when relying solely on publicly available data. This outcome highlights the importance of statistic-based methods as a support tool for the inverse design procedures. It also provides a better understanding on the limitations of conventional thermodynamic matching procedures, and the need to complement with methods that take into account conceptual design, cost and fuel burn.

On the road transportation sector, considering its deep involvement with many social expectations, assumed such proportions to become one of the major source of air pollution, mainly in urban highly congested areas. The use of reciprocating internal combustion engines (ICE) dominates the sector and the environmental dimension of the problem is under a strong attention of Governments. European Community, for instance, through sequences of regulations (EURO) reduced the emission allowed of primary pollutants; more recently, the Community added limits to climate-altering gases which directly refer to fuel consumption reduction. These limits today appear the new driver of the future engine and vehicle technological evolution. Similar efforts are under commitment by other developed countries (USA, Japan, etc,…) as well as also by the other Countries whose economic importance will dominate the markets in a very near future (BRICS Countries). The need to fulfill these issues and to keep the traditional engine expectations (torque, speed, fun to drive, etc..) triggered, especially in recent decades, a virtuous cycle whose result will be a new engine and vehicle era. The evolution till had today has been driven by the EURO limits and it demonstrated surprisingly that emission reduction and engine performances can be matched without compromises in both sides. Today, adding severe limits on equivalent CO2, emissions, it appears very difficult to predict how future engines (and vehicles) will be improved; new technologies are entering to further improve the traditional thermal powertrain but the way to a massive and more convinced electrification seems to be definitely opened. The two aspects will match in the sector of energy recovery which appears one of the most powerful tools for fuel consumption saving and CO2 reduction. When the recovery is done on exhaust gases it has an additional interest, having a moderate cost per unit of CO2 saved. The potentiality of this recovery is huge: 30%–35% of the chemical energy provided by the fuel is lost with the flue gases. For different reasons engines for passengers cars or goods transportation (light and heavy unit engines) as well those used for electricity generation (gen-set) are interested to this recovery: the first sector for the CO2 reduction, the second for the increasing value of electrical energy on the market. This wide interest is increasing the probability to have in a near future a reliable technology, being different actors pushing in this direction. In recent years the literature focused the attention to this recovery through a working fluid (organic type) on which the thermal energy is recovered by increasing its enthalpy. Thanks to a sequence of thermodynamic transformations (Rankine or Hirn cycle), mechanical work is produced. Both concept (Organic working fluid used and Rankine Cycle) are addressed as ORC technology. This overall technology has an evident complexity and doesn’t match with the need to keep reduced costs: it needs an energy recovery system at the gas side, an expander, a condenser and a pump. The space required by these components represents a limiting aspect. The variation of the flow rate and temperature of the gas (typical in ICE), as well as that at the condenser, represents additional critical aspect and call for suitable control strategies not yet exploited. In this paper the Authors studied an energy recovery method integrated with the turbocharging system, which does not require a working fluid making the recovery directly on the gas leaving the cylinders. Considering that the enthalpy drop across the turbine is usually higher than that requested by the compressor to boost the intake air, the concept was to consider an additional turbine which operates in parallel to the existing one. Room for recovery is guaranteed if one considers that a correct matching between turbine and compressor is actually done bypassing part of the exhaust gas from the turbine (waste gate) or using a variable geometry turbine (VGT) which, in any case, represents an energy loss. An additional positive feature is that this recovery does not impact on engine performances and the main components which realizes the recovery (valves & turbine) are technologically proven. In order to evaluate the potentiality of such recovery, the Authors developed a theoretical activity which represents the matching between turbocharger and engine. Thanks to an experimental characterization done on an IVECO F1C 16v JTD engine, an overall virtual platform was set up. The result produced a very satisfactory representation of the cited engine in terms of mechanical engine performances, relevant engine flow rates, pressures and temperatures. The ECU functions were represented too, such as boost pressure, EGR rates, rack control of VGT, etc… Two new direct recovery configurations have been conceived and implemented in the engine virtual platform.