Digestibility, nutrient balance and urinary purine derivative excretion in dry yak cows fed oat hay at different levels of intake

A feeding trial A as conducted at the farm of Qinghai Academy of Animal and Veterinary Science, Xining, China during 1996 - 1997 with three dry yak cows (initial body weight 163 - 197 kg, age 5 - 6 years) by using 3 x 3 Latin Square Design to determine the effect of levels of feed intake on digestion, nitrogen balance and purine derivative excretion in urine of yak cows. The animals were fed oat hay (nitrogen 13.5 g/kg dry matter (DM), metabolisable energy 8.3 MJ/kg DM), i.e., 0.3, 0.6 and 0.9 of voluntary intake (VI). Each intake treatment lasted for 17 days and the samples (feeds, faeces and urine) were collected during last 7 days of each period. The results indicate that digestibility of dietary DM, OM, NDF and ash declined when intake levels increased from 0.3 to 0.9 VI [DM, from 66.1% to 59.1% (P < 0.05); OM, from 68.1% to 59.9% (P < 0.05); NDF, from 62.1% to 54.3% (P < 0.05); and ash, from 33.9% to 11.8% (P < 0.05)]. Around 0.10 g N/kg W-0.75 was deficient daily in yak cows at 0.3 VI, and positive N balances were observed at 0.6 and 0.9 VI. Intake levels significantly (P < 0.05) affected total PD excretion in yak urine. The proportion of allantoin increased (P < 0.05) and uric acid decreased (P < 0.05) as intake level of feed increased. (C) 2004 Elsevier B.V. All rights reserved.

生产线监控与管理系统的设计

介绍在Labview的开发环境下,装配检测生产线监控与管理系统的开发方法。该系统实现了检测数据的人机界面显示,查询打印,统计过程控制(SPC)以及每月的不合格工件计数。统计过程控制(SPC)实现了对生产线的实时监控及质量管理,其他三个软件模块实现了对生产线的自动化生产管理。这种模块化的生产线管理系统对设计同类软件具有借鉴意义。

牡丹江碰撞带的组成及演化

Jiamusi Massif is an important tectonic unit in Northeast China. It’s significant for understanding the evolution of Paleo-Asian Ocean and reconstruction of the tectonic framework of Northeast China. Mudanjiang area is located in the southern margin of Jiamusi Massif and is the key to understand the evolution of Jiamusi Massif. However, the detailed geological research for Mudanjiang area has long been deficient in many important problems, such as the tectonic components of the Mudanjiang collision zone (MCZ), the age of collisional complexes and the scenario of tectonic evolution. Based on the lithology, geochemistry and the SHRIMP zircon U-Pb geochronology in Mudanjiang area, our new data and results come to some constraints for the tectonic reconstruction of MCZ as follows: 1） It is identified that the former suggestion, which the so-called “Heilongjiang Group” in Mudanjiang area is the vestige of oceanic crust, is correct. The oceanic relics represent the Neo-Proterozoic-Early Paleozoic oceanic basins based on the SHRIMP zircon U-Pb geochronology. 2） One sheet of gabbroic complex with oceanic island-type geochemical signature was discovered by this work in Mudanjiang area. 3） It is verified that the Proterozoic concordant U-Pb ages of the migmatites developed along the southern margin of Jiamusi massif, which represent the events of magmatic intrusion, as the direct evidence for the existence of the Proterozoic crystalline basements of the Jiamusi Massif. Based on geochronology, we suggest that the migmatization and coeval S-type granite magmatism of the southern margin of Jiamusi Massif took place about 490Ma. 4） The island arc complex has been found in the Heilongjiang Group, and the oceanic relics was found distributing on both sides, as provided important constraint for the tectonic reconstruction of the MCZ. 5） ~440Ma metamorphic event and coeval post-collisional granite magmatism have been firmly identified in the MCZ and its southern neighboring area. Together with previous data obtained by other researchers, our conclusions on the reconstruction of the tectonic architecture and evolution of the MCZ as follows: 1） The orogenic assemblages developed in the Mudanjiang collisional zone are featured by a sequence of ancient active continental margins and ensuing orogenic processing. The Mashan Group is the reworking basement of Jiamusi Massif, whereas the Heilongjiang Group represents arc and oceanic complexes characterized by imbricate deep-seated sliced and slivering sheets due to multi-phases of thrusting and nappe stacking. 2） The northern sub-belt of MCZ is probably the arc-continent collisional boundary related to the closure of main oceanic basin. The collisional age can be constrained by the events of syn-orogenic migmatization of migmatite, coeval S-type granite magmatism and the related granulite-facies metamorphism. Therefore, we suggested the collisional age of northern sub-belt is probably Cambrian-Early Ordovician. The extensive granulite-facies metamorphism of the Mashan Group in Jiamusi Massif, as affirmed by former works, was probably related with the collisional event. 3） The southern sub-belt of the MCZ was possibly related with the closure of back-arc basin. We presumed that the collisional age of southern sub-belt is about Ordovician-Early Silurian according to the ~440Ma extensive metamorphism and the occurrence of coeval post-collisional granite magmatism. 4） The extant structural architecture of the MCZ is related to the multi-phases of intra-continental superimposition, which is characterized by the Mesozoic nappe structure.

太阳辐射对热层和电离层变化性的影响

Solar ultraviolet (UV) radiation at wavelengths less than 400 nm is an important source of energy for aeronomic processes throughout the solar system. Solar UV photons are absorbed in planetary atmospheres, as well as throughout the heliosphere, via photodissociation of molecules, photoionization of molecules and atoms, and photoexcitation toexcitation including resonance scattering. In this paper, the solar irradiances data measured by TIMED SEE, as well as the solar proxies such as F10.7 and Mg II, thermosphere neutral density of CHAMP measurements and topside ionospheric plasmas densities from DMSP, are used to analyze solar irradiance effects on the variabilities of the thermosphere and the ionosphere. First, thermosphere densities near 410 km altitude are analyzed for solar irradiance variability effects during the period 2002-2004. Correlations between the densities and the solar irradiances for different spectral lines and wavelength ranges reveal significantly different characteristics. The density correlates remarkably well with all the selected solar irradiances except the lower chromospheric O I (130.4 nm) emission. Among the chosen solar proxies, the Mg II core-to-wing ratio index, EUV (30-120 nm) and F10.7 show the highest correlations with the density for short-term (< ~27 days) variations. For both long- (> ~27 days) and short-term variations, linear correlation coefficients exhibit a decreasing trend from low latitudes towards high latitudes. The density variability can be effectively modeled (capturing 71% of the variance) using multiple solar irradiance indices, including F10.7, SEUV (the EUV 30-120 nm index), and SFUV (the FUV 120-193 nm index), in which a lag time of 1 day was used for both F10.7 and SEUV, and 5 days for SFUV. In our regression formulation SEUV has the largest contribution to the density variation (40%), with the F10.7 having the next largest contribution (32%) and SFUV accounting for the rest (28%). Furthermore, a pronounced period of about 27.2 days (mean period of the Sun's rotation) is present in both density and solar irradiance data of 2003 and 2004, and a pronounced period of about 54.4 days (doubled period of the solar rotation) is also revealed in 2004. However, soft X-ray and FUV irradiances did not present a pronounced 54.4 day period in 2004, in spite of their high correlation with the densities. The Ap index also shows 54-day periodicities in 2004, and magnetic activity, together with solar irradiance, affects the 54-day variation in density significantly. In addition, NRLMSISE00, DTM-2000 and JB2006 model predictions are compared with density measurements from CHAMP to assess their accuracy, and the results show that these models underestimate the response of the thermosphere to variations induced by solar rotation. Next, the equatorial topside ionospheric plasmas densities Ni are analyzed for solar irradiance variability effects during the period 2002-2005. Linear correlations between Ni and the solar irradiances for different wavelength ranges reveal significantly different characteristics. XUV (0-35 nm) and EUV (115-130 nm) show higher correlation with Ni for the long-term variations, whereas EUV (35-115 nm) show higher correlation for the short-term variations. Moreover, partial correlation analysis shows that the long-term variations of Ni are affected by both XUV (0-35 nm) and EUV (35-115 nm), whereas XUV (0-35 nm) play a more important role; the short-term variations of Ni are mostly affected by EUV (35-115 nm). Furthermore, a pronounced period of about 27 days is present in both Ni and solar irradiance data of 2003 and 2004, and a pronounced period of about 54 days is also revealed in 2004. Finally, prompted by previous studies that have suggested solar EUV radiation as a means of driving the semiannual variation, we investigate the intra-annual variation in thermosphere neutral density near 400 km during 2002-2005. The intra-annual variation, commonly referred to as the ‘semiannual variation’, is characterized by significant latitude structure, hemispheric asymmetries, and inter-annual variability. The magnitude of the maximum yearly difference, from the yearly minimum to the yearly maximum, varies by as much as 60% from year to year, and the phases of the minima and maxima also change by 20-40 days from year to year. Each annual harmonic of the intra-annual variation, namely, annual, semiannual, ter-annual and quatra-annual, exhibits a decreasing trend from 2002 through 2005 that is correlated with the decline in solar activity. In addition, some variations in these harmonics are correlated with geomagnetic activity, as represented by the daily mean value of Kp. Recent empirical models of the thermosphere are found to be deficient in capturing most of the latitude dependencies discovered in our data. In addition, the solar flux and geomagnetic activity proxies that we have employed do not capture some latitude and inter-annual variations detected in our data. It is possible that these variations are partly due to other effects, such as seasonal-latitudinal variations in turbopause altitude (and hence O/N2 composition) and ionosphere coupling processes that remain to be discovered in the context of influencing the intra-annual variations depicted here. Our results provide a new dataset to challenge and validate thermosphere-ionosphere general circulation models that seek to delineate the thermosphere intra-annual variation and to understand the various competing mechanisms that may contribute to its existence and variability. We furthermore suggest that the term “intra-annual” variation be adopted to describe the variability in thermosphere and ionosphere parameters that is well-captured through a superposition of annual, semiannual, ter-annual, and quatra-annual harmonic terms, and that “semiannual’ be used strictly in reference to a pure 6-monthly sinusoidal variation. Moreover, we propose the term “intra-seasonal” to refer to those shorter-term variations that arise as residuals from the above Fourier representation.

改造型盆地油气成藏机理与油气预测－以银根－额济纳旗盆地为例

Reformed basin is a basin that underwent multiple immense reformation after the sedimentary stage, the major geologic elements of the petroleum system in the prototyped basin are destroyed to a certain extent, and their petroleum system has been reconstructed. This type of basin is frequently found in the course of exploration both home and abroad. In China, especially in the western and southern part of China, the basins in which oil explorations have been conducted are mostly reformed basins. The reformed basins from Paleozoic, Late Mesozoic to Cenozoic are widely distributed in West and South China. They are, and moreover, will be a challenge for oil and gas exploration. The conventional investigation and exploration techniques used in the slightly reconstructed basin just don't work well when facing the reformed basin. Therefore, the study on the reformed basin, especially the study on the pool-forming mechanism and reservoir prediction becomes a focus and one of difficulties for the geologists overseas and domestic. Yingen-Ejinaqi Basin is a typical case of the Late Mesozoic and Cenozoic reformed basins in China. It locates in West China and is a exploration frontier with difficulties and no break through is made for years. A comprehensive research on it will be of significance for oil and gas exploration in similar basins of China. The late research for reformed basin in China now is mainly concentrated on basin classification, formation mechanism, geologic features, and survey technique, distribution regularity of oil accumulation and its dominating factors, assessment of oil exploration prospect and target zones, etc. On the other hand, the study on the pool-forming mechanism and reservoir prediction seems insufficient in systematization, and the research is deficient in methodology and combination of qualitative and quantitative studies, as well as the application of the new theory and techniques. The current efforts are mainly directed to structures (faults), sedimentation, the relationship between reservoir evolution and oil accumulation, and some other relevant fields. However, the application of the new theory and techniques seems to be insufficient such as petroleum system, pool-forming dynamics, fluid pressure compartment, and basin simulation, etc. So is the dynamic and integrated research. As a result, incomplete knowledge and understandings derived from the research on pool-forming mechanism and reservoir prediction often do not accord with rea-lity of the basin. The study and exploration under the guidance of this knowledge will inevitably lead to errors and failure. This paper, based on the previous study of the other geologists on reformed basins, with emphasis on "wholeness or systematic, dynamic and integrated" research, presents a reverse thinking of beginning from conserved units in the basin and the combination of qualitative and quantitative study with new theory and technique by building a geological model. The paper also puts forward a new thought for studying the oil & gas accumulation and reservoir prediction , and establishes a new research system for reformed basin. It is verified by the known reservoir and oil accumulation area in the basin and has a practical value for use and reference. The new ideas and achievements in this research are as following: 1.This is the first time that the system for studying the reformed basin and its pool-forming mechanism and reservoir prediction is presented. A reverse thinking and combination of qualitative & quantitative are applied here with emphasis on "wholeness or systematic, dynamic and integrated" research, new theory, techniques & methods comprehensive use and geologic models building. 2. Identifying criterion and methods, classifying schemes, and denominating principles for the conserved units of reformed basins are presented in this paper. The geologic model of conserved units of Yingen-Ejinaqi Basin has been built. It is a practical method when combined with the traditional way for basin survey and the conserved units study. 3.The dynamic sources of basin deformation are believed to be stress, gravity and thermodynamics. The stress and gravity are key factors in basin deformation and pool forming, especially stress. Scientific proof is provided by classifying the functional type, style and range of the stress, gravity and thermodynamics. 4.The pool forming and reservoir distribution of Yingen-Ejinaqi Basin are controlled by multiple factors or geologic conditions or/and co-controlled by both of them. The qualitative and quantitative research on petroleum system and basin modeling will help us understand and determine the pool-forming period of the conserved unit (timing), the oil migrating direction (orientation), the oil accumulating region (location), the oil distributing border (bordering) and the size of oil accumulation (quantification). Thus the pool-forming and distribution zones can be predicted. 5.Three generating modes (reform-succession type, reform-destroyed type and reform-regenerating type or reform-newborn type) of pool forming for reformed basins are presented here, together with the inner relationships between basin deformation type, overlapping style and pool-forming modes. The pool-forming modes are determined by deformation type and overlapping style. Reservoir distribution will be predicted based on the modes and other concrete pool-forming conditions. 6.The evaluation methods of the conserved units and zones and the parameter selection are reliable in optimal selecting of target zones. The technical terms, new ideas and methods for the study of reformed basins, the pool-forming mechanism and reservoir prediction are presented in this paper. The concepts and terms, the identifying criterion, the denominating principles, the generating modes for pool forming, the methods of reservoir prediction, and the evaluation techniques for conserved units and zones can be used for reference in studies on the petroleum exploration of reformed basins in China and abroad. It serves as a typical example for further research of the reformed basins and the geologic regulations of oil accumulation. It has a practical value of use and reference. The future research in the field of pool-forming mechanism of the reformed basins may well be on the process simulation of pool-forming dynamics of the reformed basins. Experimental work has been conducted to simulate the processes by using quantitative and qualitative methods combined. The further study in this field calls for more efforts.

电阻率测井反演方法应用研究

Formation resistivity is one of the most important parameters to be evaluated in the evaluation of reservoir. In order to acquire the true value of virginal formation, various types of resistivity logging tools have been developed. However, with the increment of the proved reserves, the thickness of interest pay zone is becoming thinner and thinner, especially in the terrestrial deposit oilfield, so that electrical logging tools, limited by the contradictory requirements of resolution and investigation depth of this kinds of tools, can not provide the true value of the formation resistivity. Therefore, resitivity inversion techniques have been popular in the determination of true formation resistivity based on the improving logging data from new tools. In geophysical inverse problems, non-unique solution is inevitable due to the noisy data and deficient measurement information. I address this problem in my dissertation from three aspects, data acquisition, data processing/inversion and applications of the results/ uncertainty evaluation of the non-unique solution. Some other problems in the traditional inversion methods such as slowness speed of the convergence and the initial-correlation results. Firstly, I deal with the uncertainties in the data to be processed. The combination of micro-spherically focused log (MSFL) and dual laterolog(DLL) is the standard program to determine formation resistivity. During the inversion, the readings of MSFL are regarded as the resistivity of invasion zone of the formation after being corrected. However, the errors can be as large as 30 percent due to mud cake influence even if the rugose borehole effects on the readings of MSFL can be ignored. Furthermore, there still are argues about whether the two logs can be quantitatively used to determine formation resisitivities due to the different measurement principles. Thus, anew type of laterolog tool is designed theoretically. The new tool can provide three curves with different investigation depths and the nearly same resolution. The resolution is about 0.4meter. Secondly, because the popular iterative inversion method based on the least-square estimation can not solve problems more than two parameters simultaneously and the new laterolog logging tool is not applied to practice, my work is focused on two parameters inversion (radius of the invasion and the resistivty of virgin information ) of traditional dual laterolog logging data. An unequal weighted damp factors- revised method is developed to instead of the parameter-revised techniques used in the traditional inversion method. In this new method, the parameter is revised not only dependency on the damp its self but also dependency on the difference between the measurement data and the fitting data in different layers. At least 2 iterative numbers are reduced than the older method, the computation cost of inversion is reduced. The damp least-squares inversion method is the realization of Tikhonov's tradeoff theory on the smooth solution and stability of inversion process. This method is realized through linearity of non-linear inversion problem which must lead to the dependency of solution on the initial value of parameters. Thus, severe debates on efficiency of this kinds of methods are getting popular with the developments of non-linear processing methods. The artificial neural net method is proposed in this dissertation. The database of tool's response to formation parameters is built through the modeling of the laterolog tool and then is used to training the neural nets. A unit model is put forward to simplify the dada space and an additional physical limitation is applied to optimize the net after the cross-validation method is done. Results show that the neural net inversion method could replace the traditional inversion method in a single formation and can be used a method to determine the initial value of the traditional method. No matter what method is developed, the non-uniqueness and uncertainties of the solution could be inevitable. Thus, it is wise to evaluate the non-uniqueness and uncertainties of the solution in the application of inversion results. Bayes theorem provides a way to solve such problems. This method is illustrately discussed in a single formation and achieve plausible results. In the end, the traditional least squares inversion method is used to process raw logging data, the calculated oil saturation increased 20 percent than that not be proceed compared to core analysis.

Strong effect of transitional metals on the sulfur resistance of Pd/HY-Al2O3 catalysts for aromatic hydrogenation

In order to improve the sulfur resistance of noble metal catalysts in the aromatic hydrogenation of diesel fuel, the alloying effect of non-noble metals with Pd was studied. Toluene hydrogenation over Pd and Pd-M bimetallic catalysts (M = Cr, W,La, Mn, Mo, Ag) on a mixed HY-Al2O3 support was investigated in the presence of 3000 ppm sulfur as thiophene in the feedstock. The results showed that the addition of the second metals strongly affected the activity of toluene hydrogenation, which suggests that the sulfur resistibility of Pd-M bimetallic catalysts is much different from single Pd. La, Mn, Mo and Ag decreased the sulfur resistance of the palladium catalysts. For example, the toluene conversion at 553 K was observed to decrease sharply from 39.4 wt.% on Pd to 1.6 wt.% on Pd-Ag, which is by a factor of 25. One of the important findings in this article is that Cr and W increase hydrogenation activity of Pd catalysts. The reactions occurring on these catalysts include hydrogenation, isomerization and hydrocracking, The addition of the second metals has no noticeable effects on the hydrogenation and isomerization selectivity, but it slightly suppresses hydrocracking reactions. The four typical catalysts, Pd-Cr, Pd-W, Pd-Ag and Pd were characterized by infrared (IR) spectroscopy of pyridine and CO. LR spectra of CO revealed the strong interaction between Pd and the second metal as Cr, W and Ag (or their oxide), indicating that the improvement in sulfur resistance originates from electron-deficient Pd with the addition of second metals. (C) 2001 Elsevier Science B.V. All rights reserved.

The EXACT description of biomedical protocols

Soldatova, L. N., Aubrey, W., King, R. D., Clare, A. J. (2008). The EXACT description of biomedical protocols. Bioinformatics, 24 (13), i295-i303 Sponsorship: BBSRC / RAEng / EPSRC specialissue: ISMB

Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana

Clarke, S. M., Mur, L. A. J., Wood, J. E., & Scott, I. M. (2004). Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana. The Plant Journal, 38(3), 432-447. Sponsorship: BBSRC RAE2008

Salicylate accumulation inhibits growth at chilling temperature in Arabidopsis

Ian M. Scott, Shannon M. Clarke, Jacqueline E. Wood and Luis A.J. Mur (2004). Salicylate accumulation inhibits growth at chilling temperature in Arabidopsis. Plant Physiology, 135(2), 1040-1049. RAE2008

Struktura fizyczna gruntu, zawartość substancji organicznej oraz skład chemiczny gleb w podłożach 21 stanowisk zieleni miejskiej na terenie Poznania. Część I. Struktura fizyczna gruntu

This article is the first one in the series of four publications describing the condition of soil environment in the area of broadly understood centre of Poznań. The paper presents the results of studies into a ph ph ph ysical structure of the soil collected around horse chestnuts (Aesculus hippocastanum), growing in various habitats in 21 locations in the area of Poznań. In the majority of cases, 75 per cent of them, the obtained data point to a deficient ph ph ysical structure of subsoils. Only in five cases, out of a general number of 21 locations, the roots of the trees developed in soil conditions that were favourable for the plants, and which enabled them the retention of precipitation water, oxygention of rhizosph ph ere and accumulation of humus.

Nutrição Artificial nos Serviços Farmacêuticos Hospitalares

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Variations in total and differential milk somatic cell counts and plasmin levels and their role in proteolysis and quality of milk and cheese

Increased plasmin and plasminogen levels and elevated somatic cell counts (SCC) and polymorphonuclear leucocyte levels (PMN) were evident in late lactation milk. Compositional changes in these milks were associated with increased SCC. The quality of late lactation milks was related to nutritional status of herds, with milks from herds on a high plane of nutrition having composition and clotting properties similar to, or superior to, early-mid lactation milks. Nutritionally-deficient cows had elevated numbers of polymorphonuclear leucocytes (PMNs) in their milk, elevated plasmin levels and increased overall proteolytic activity. The dominant effect of plasmin on proteolysis in milks of low SCC was established. When present in elevated numbers, somatic cells and PMNs in particular had a more significant influence on the proteolysis of both raw and pasteurised milks than plasmin. PMN protease action on the caseins showed proteolysis products of two specific enzymes, cathepsin B and elastase, which were also shown in high SCC milk. Crude extracts of somatic cells had a high specificity on αs1-casein. Cheeses made from late lactation milks had increased breakdown of αs1-casein, suggestive of the action of somatic cell proteinases, which may be linked to textural defects in cheese. Late lactation cheeses also showed decreased production of small peptides and amino acids, the reason for which is unknown. Plasmin, which is elevated in activity in late lactation milk, accelerated the ripening of Gouda-type cheese, but was not associated with defects of texture or flavour. The retention of somatic cell enzymes in cheese curd was confirmed, and a potential role in production of bitter peptides identified. Cheeses made from milks containing high levels of PMNs had accelerated αs1-casein breakdown relative to cheeses made from low PMN milk of the same total SCC, consistent with the demonstrated action of PMN proteinases. The two types of cheese were determined significantly different by blind triangle testing.

Investigating the function of PINK1 in cancer development and pathogenesis

PTEN‐induced kinase 1 (PINK1) was identified initially in cancer cells as a gene up‐regulated by overexpression of the central tumour suppressor, PTEN. Loss‐of‐function mutations in PINK1 were discovered subsequently to cause autosomal recessive Parkinsonʹs disease (ARPD). Despite much research focusing on the proposed mechanism(s) through which loss of PINKI function causes neurodegeneration, few studies have focused on a direct role for this serine/threonine kinase in cancer biology. The focus of this thesis was to examine a direct role for PINK1 function in tumourigenesis. Initial studies showed that loss of PINK1 reduces tumour‐associated phenotypes including cell growth, colony formation and invasiveness, in several cell types in vitro, indicating a pro‐tumourigenic role for PINK1 in cancer. Furthermore, results revealed for the first time that PINK1 deletion, examined in mouse embryonic fibroblasts (MEFS) from PINK1 knock‐out animals, causes cell cycle defects, whereby cells arrest at in cytokinesis, giving rise to a highly significant increase in the number of multinucleated cells. This results in several key changes in the expression profile of cell cycle associated protein. In addition, PINK1‐deficient MEFs were found to resist cell cycle exit, with a proportion of cells remaining in proliferative phases upon removal of serum. The ability of cells to progress through mitosis conferred by PINK1 expression was independent of its kinase activity, while the cell cycle exit following serum withdrawal was kinase dependent. Investigations into the mechanism through which loss of PINK1 function gives rise to cell cycle defects revealed that dynamin related protein 1 (Drp1)‐mediated mitochondrial fission is enhanced in PINK1‐ deficient MEFs, and that increased expression of Drp1 on mitochondria and activation of Drp1 is highly significant in PINK1‐deficient multinucleated cells. Deregulated and increased levels and activation of mitochondrial fission via Drp1 was shown to be a major feature of cell cycle defects caused by PINK1 deletion, both during progression through G2/M and cell cycle exit following serum removal. Altered PINK1 localisation was also observed during progression of mitosis, and upon serum deprivation. Thus, PINK1 dissociated from the mitochondria during the mitotic phases and localised to mitochondria upon serum withdrawal. During serum withdrawal deletion of PINK1 disabled the ability of MEFs to increase mitochondrial membrane potential (ΔΨm), and increase autophagy. This was co‐incident with increased mitochondrial fission, and increased localisation of Drp1 to mitochondria following serum deprivation. Together, this indicates an inability of PINK1‐negative cells to respond protectively to this stress‐induced state, primarily via impaired mitochondrial function. In contrast, PINK1 overexpression was found to protect cells from DNA damage following treatment with oxidants. In addition, deletion of PINK1 blocked the ability of cells to re‐enter the cell cycle in response to insulin‐like growth factor‐1 (IGF‐1), a major cancer promoting agonistwhich acts primarily via PI3‐kinase/Akt activation. Furthermore, PINK1 mRNA expression was significantly increased following serum deprivation of MCF‐7 cells, and this was rendered more significant upon additional inhibition of PI3‐kinase. Conversely, IGF‐1 activation of PI3‐kinase/Akt causes a time‐dependent and significant reduction of PINK1 mRNA expression that was PI3‐kinase dependent. Together these results indicate that PINK1 expression is necessary for IGF‐1 signalling and is regulated reciprocally in the absence and presence of IGF‐1, via PI3‐kinase/Akt, a signalling system which has major tumour‐promoting capacity in cancer cell biology. The results of this thesis indicate PINK1 is a candidate tumour-promoting gene which has a significant function in the regulation of the cell cycle, and growth factor responses, at key cell cycle checkpoints, namely, during progression through G2/M and during exit of the cell cycle following removal of serum. Furthermore, the results reveal that the regulation of mitochondrial fission and Drp1 function is mechanistically important in the regulation of cell cycle control by PINK1. As deregulation of the cell cycle is linked to both tumourigenesis and neurodegeneration, the findings of this thesis are of importance not just for understanding cancer biology, but also in the context of PINK1‐associated neurodegeneration.

Investigation of toll-like receptor tolerance in the regulation of inflammation

Inflammation is a complex and highly organised immune response to microbes and tissue injury. Recognition of noxious stimuli by pathogen recognition receptor families including Toll-like receptors results in the expression of hundreds of genes that encode cytokines, chemokines, antimicrobials and regulators of inflammation. Regulation of TLR activation responses is controlled by TLR tolerance which induces a global change in the cellular transcriptional expression profile resulting in gene specific suppression and induction of transcription. In this thesis the plasticity of TLR receptor tolerance is investigated using an in vivo, transcriptomics and functional approach to determine the plasticity of TLR tolerance in the regulation of inflammation. Firstly, using mice deficient in the negative regulator of TLR gene transcription, Bcl-3 (Bcl-3-/-) in a model of intestinal inflammation, we investigated the role of Bcl-3 in the regulation of intestinal inflammatory responses. Our data revealed a novel role for Bcl-3 in the regulation of epithelial cell proliferation and regeneration during intestinal inflammation. Furthermore this data revealed that increased Bcl-3 expression contributes to the development of inflammatory bowel disease (IBD). Secondly, we demonstrate that lipopolysaccharide tolerance is transient and recovery from LPS tolerance results in polarisation of macrophages to a previously un-described hybrid state (RM). In addition, we identified that RM cells have a unique transcriptional profile with suppression and induction of genes specific to this polarisation state. Furthermore, using a functional approach to characterise the outcomes of TLR tolerance plasticity, we demonstrate that cytokine transcription is uncoupled from cytokine secretion in macrophages following recovery from LPS tolerance. Here we demonstrate a novel mechanism of regulation of TLR tolerance through suppression of cytokine secretion in macrophages. We show that TNF-α is alternatively trafficked towards a degradative intracellular compartment. These studies demonstrate that TLR tolerance is a complex immunological response with the plasticity of this state playing an important role in the regulation of inflammation.