Robust Proximal Femur Segmentation in Conventional X-Ray Images via Random Forest Regression on Multi-resolution Gradient Features

In this paper, we propose a fully automatic, robust approach for segmenting proximal femur in conventional X-ray images. Our method is based on hierarchical landmark detection by random forest regression, where the detection results of 22 global landmarks are used to do the spatial normalization, and the detection results of the 59 local landmarks serve as the image cue for instantiation of a statistical shape model of the proximal femur. To detect landmarks in both levels, we use multi-resolution HoG (Histogram of Oriented Gradients) as features which can achieve better accuracy and robustness. The efficacy of the present method is demonstrated by experiments conducted on 150 clinical x-ray images. It was found that the present method could achieve an average point-to-curve error of 2.0 mm and that the present method was robust to low image contrast, noise and occlusions caused by implants.

Design, synthesis and biological evaluation of 5$\sp\prime$-mononucleotide prodrugs: Strategies to introduce nucleotides into cells

The neutral bis ((pivaloyloxy)methyl) (PIV$\sb2\rbrack$ derivatives of FdUMP, ddUMP, and AZTMP were synthesized as potential membrane-permeable prodrugs of FdUMP, ddUMP, and AZTMP. These compounds were designed to enter cells by passive diffusion and revert to the parent nucleotides after removal of the PIV groups by hydrolytic enzymes. These prodrugs were prepared by condensation of FUdR, ddU, and AZT with PIV$\sb2$ phosphate in the presence of triphenylphosphine and diethyl azodicarboxylate (the Mitsunobo reagent). PIV$\sb2$-FdUMP, PIV$\sb2$-ddUMP, and PIV$\sb2$-AZTMP were stable in the pH range 1.0-4.0 (t$\sb{1/2} = {>}$100 h). They were also fairly stable at pH 7.4 (t$\sb{1/2} = {>}$40 h). In 0.05 M NaOH solution, however, they were rapidly degraded (t$\sb{1/2} < 2$ min). In the presence hog liver carboxylate esterase, they were converted quantitatively to the corresponding phosphodiesters, PIV$\sb1$-FdUMP, PIV$\sb1$-ddUMP, and PIV$\sb1$-AZTMP; after 24 h incubation, only trace amounts of FdUMP, ddUMP, and AZTMP (1-5%) were observed indicating that the PIV$\sb1$ compounds were poor substrates for the enzyme. In human plasma, the PIV$\sb2$ compounds were rapidly degraded with half-lives of less than 5 min. The rate of degradation of the PIV$\sb2$ compounds in the presence of phosphodiesterase I was the same as that in buffer controls, indicating that they were not substrates for this enzyme. In the presence of phosphodiesterase I, PIV$\sb1$-FdUMP, PIV$\sb1$-ddUMP, and PIV$\sb1$-AZTMP were converted quantitatively to FdUMP, ddUMP, and AZTMP.^ PIV$\sb2$-ddUMP and PIV$\sb2$-AZTMP were effective at controlling HIV type 1 infection in MT-4 and CEM tk$\sp-$ cells in culture. Mechanistic studies demonstrated that PIV$\sb2$-ddUMP and PIV$\sb2$-AZTMP were taken up by the cells and converted to ddUTP and AZTTP, both potent inhibitors of HIV reverse transcriptase. However, a potential shortcoming of PIV$\sb2$-ddUMP and PIV$\sb2$-AZTMP as clinical therapeutic agents is that they are rapidly degraded (t$\sb{1/2}$ = approx. 4 minutes) in human plasma by carboxylate esterases. To circumvent this limitation, chemically-labile nucleotide prodrugs and liposome-encapsulated nucleotide prodrugs were investigated. In the former approach, the protective groups bis(N, N-(dimethyl)carbamoyloxymethyl) (DM$\sb2$) and bis (N-(piperidino)carbamoyloxymethyl) (DP$\sb2$) were used to synthesize DM$\sb2$-ddUMP and DP$\sb2$-ddUMP, respectively. In aqueous buffers (pH range 1.0-9.0) these compounds were degraded with half-lives of 3 to 4 h. They had similar half-lives in human plasma demonstrating that they were resistant to esterase-mediated cleavage. However, neither compound gave rise to significant concentrations of ddUMP in CEM or CEM tk$\sp-$ cells. In the liposome-encapsulated nucleotide prodrug approach, three different liposomal formulations of PIV$\sb2$-ddUMP (L-PIV$\sb2$-ddUMP) were investigated. The half-lifes of these L-PIV$\sb2$-ddUMP preparations in human plasma were 2 h compared with 4 min for the free drug. The preparations were more effective at controlling HIV-1 infection than free PIV$\sb2$-ddUMP in human T cells in culture. Collectively, these data indicate that PIV$\sb2$-FdUMP, PIV$\sb2$-ddUMP, and PIV$\sb2$-AZTMP are effective membrane-permeable prodrugs of FdUMP, ddUMP, and AZTMP. ^

Fully automatic segmentation of AP pelvis X-rays via random forest regression with efficient feature selection and hierarchical sparse shape composition

In clinical practice, traditional X-ray radiography is widely used, and knowledge of landmarks and contours in anteroposterior (AP) pelvis X-rays is invaluable for computer aided diagnosis, hip surgery planning and image-guided interventions. This paper presents a fully automatic approach for landmark detection and shape segmentation of both pelvis and femur in conventional AP X-ray images. Our approach is based on the framework of landmark detection via Random Forest (RF) regression and shape regularization via hierarchical sparse shape composition. We propose a visual feature FL-HoG (Flexible- Level Histogram of Oriented Gradients) and a feature selection algorithm based on trace radio optimization to improve the robustness and the efficacy of RF-based landmark detection. The landmark detection result is then used in a hierarchical sparse shape composition framework for shape regularization. Finally, the extracted shape contour is fine-tuned by a post-processing step based on low level image features. The experimental results demonstrate that our feature selection algorithm reduces the feature dimension in a factor of 40 and improves both training and test efficiency. Further experiments conducted on 436 clinical AP pelvis X-rays show that our approach achieves an average point-to-curve error around 1.2 mm for femur and 1.9 mm for pelvis.

Efficacy of a live attenuated vaccine in classical swine fever virus postnatally persistently infected pigs

Classical swine fever (CSF) causes major losses in pig farming, with various degrees of disease severity. Efficient live attenuated vaccines against classical swine fever virus (CSFV) are used routinely in endemic countries. However, despite intensive vaccination programs in these areas for more than 20 years, CSF has not been eradicated. Molecular epidemiology studies in these regions suggests that the virus circulating in the field has evolved under the positive selection pressure exerted by the immune response to the vaccine, leading to new attenuated viral variants. Recent work by our group demonstrated that a high proportion of persistently infected piglets can be generated by early postnatal infection with low and moderately virulent CSFV strains. Here, we studied the immune response to a hog cholera lapinised virus vaccine (HCLV), C-strain, in six-week-old persistently infected pigs following post-natal infection. CSFV-negative pigs were vaccinated as controls. The humoral and interferon gamma responses as well as the CSFV RNA loads were monitored for 21 days post-vaccination. No vaccine viral RNA was detected in the serum samples and tonsils from CSFV postnatally persistently infected pigs for 21 days post-vaccination. Furthermore, no E2-specific antibody response or neutralising antibody titres were shown in CSFV persistently infected vaccinated animals. Likewise, no of IFN-gamma producing cell response against CSFV or PHA was observed. To our knowledge, this is the first report demonstrating the absence of a response to vaccination in CSFV persistently infected pigs.

The Cenomanian/Turronian Boundary Event at ODP Hole 103-641A

Drilling at ODP Site 641 (on the western margin of Galicia Bank, off northwestern Spain) revealed a thin, but pronounced, interval of black shale and gray-green claystone. Our high-resolution study combines the sedimentology, micropaleontology (palynomorphs and others), organic and inorganic geochemistry, and isotopic values of this layer to demonstrate the distinct nature of the sediment and prove that the sequence represents the local sedimentary expression of the global Cenomanian/Turonian Oceanic Anoxic Event (OAE) of Schlanger and Jenkyns (1976), Arthur and Schlanger (1979), and Jenkyns (1980), also called the Cenomanian/Turonian Boundary Event (CTBE). The most striking evidence is that the strong positive d13C excursion characterizing the CTBE sequences in shallow areas can be traced into a pronounced deep-sea expression, thus providing a good stratigraphic marker for the CTBE in various paleosettings. The isotopic excursion at Site 641 coincides with an extremely enriched trace metal content, with values that were previously unknown for the Cretaceous Atlantic. Similar to other CTBE occurrences, the organic carbon content is high (up to 11%) and the organic matter is of dominantly marine origin (kerogen type II). The bulk mineralogy of the CTBE sediments does not differ significantly from the general trend of Cretaceous North Atlantic sediments (dominance of smectite and zeolite with minor amounts of illite and scattered palygorskite, kaolinite, and chlorite); thus, no evidence for either increased volcanic activity nor a drastic climatic change in the borderlands was found. Results from Site 641 are compared with the CTBE section found at Site 398, DSDP Leg 47B (Vigo Seamount at the southern end of the Galicia Bank).

Pedogeomorphology, geochronology and paleobotany of soil profiles obtained in the Nagqu area, central Tibet

Vast areas on the Tibetan Plateau are covered by alpine sedge mats consisting of different species of the genus Kobresia. These mats have topsoil horizons rich in rhizogenic organic matter which creates turfs. As the turfs have recently been affected by a complex destruction process, knowledge concerning their soil properties, age and pedogenesis are needed. In the core area of Kobresia pygmaea mats around Nagqu (central Tibetan Plateau, ca. 4500 m a.s.l.), four profiles were subjected to pedological, paleobotanical and geochronological analyses concentrating on soil properties, phytogenic composition and dating of the turf. The turf of both dry K. pygmaea sites and wet Kobresia schoenoides sites is characterised by an enrichment of living (dominant portion) and dead root biomass. In terms of humus forms, K. pygmaea turfs can be classified as Rhizomulls mainly developed from Cambisols. Wet-site K. schoenoides turfs, however, can be classified as Rhizo-Hydromors developed from Histic Gleysols. At the dry sites studied, the turnover of soil organic matter is controlled by a non-permafrost cold thermal regime. Below-ground remains from sedges are the most frequent macroremains in the turf. Only a few pollen types of vascular plants occur, predominantly originating from sedges and grasses. Large amounts of microscopic charcoal (indeterminate) are present. Macroremains and pollen extracted from the turfs predominantly have negative AMS 14C ages, giving evidence of a modern turf genesis. Bulk-soil datings from the lowermost part of the turfs have a Late Holocene age comprising the last ca. 2000 years. The development of K. pygmaea turfs was most probably caused by an anthropo(zoo)-genetically initiated growth of sedge mats replacing former grass-dominated vegetation ('steppe'). Thus the turfs result from the transformation of pre-existing topsoils comprising a secondary penetration and accumulation of roots. K. schoenoides turfs, however, are characterised by a combined process of peat formation and penetration/accumulation of roots probably representing a (quasi) natural wetland vegetation.

9-28 d of exposure to elevated pCO2 reduces avoidance of predator odour but had no effect on behavioural lateralization or swimming activity in a temperate wrasse(Ctenolabrus rupestris)

Most studies on the impact of near-future levels of carbon dioxide on fish behaviour report behavioural alterations, wherefore abnormal behaviour has been suggested to be a potential consequence of future ocean acidification and therefore a threat to ocean ecosystems. However, an increasing number of studies show tolerance of fish to increased levels of carbon dioxide. This variation among studies in susceptibility highlights the importance of continued investigation of the possible effects of elevated pCO2. Here, we investigated the impacts of increased levels of carbon dioxide on behaviour using the goldsinny wrasse (Ctenolabrus rupestris), which is a common species in European coastal waters and widely used as cleaner fish to control sea lice infestation in commercial fish farming in Europe. The wrasses were exposed to control water conditions (370 µatm) or elevated pCO2 (995 µatm) for 1 month, during which time behavioural trials were performed. We investigated the possible effects of CO2 on behavioural lateralization, swimming activity, and prey and predator olfactory preferences, all behaviours where disturbances have previously been reported in other fish species after exposure to elevated CO2. Interestingly, we failed to detect effects of carbon dioxide for most behaviours investigated, excluding predator olfactory cue avoidance, where control fish initially avoided predator cue while the high CO2 group was indifferent. The present study therefore shows behavioural tolerance to increased levels of carbon dioxide in the goldsinny wrasse. We also highlight that individual fish can show disturbance in specific behaviours while being apparently unaffected by elevated pCO2 in other behavioural tests. However, using experiments with exposure times measured in weeks to predict possible effects of long-term drivers, such as ocean acidification, has limitations, and the behavioural effects from elevated pCO2 in this experiment cannot be viewed as proof that these fish would show the same reaction after decades of evolution.

Problemática estructural y de funcionamiento de la cadena de valor del jamón ibérico de bellota: el caso de Guijuelo en España

El sector porcino ibérico es un sector típicamente español. La imagen que asocia el consumidor a los productos ibéricos es la correspondiente a los productos elaborados a partir de cerdos ibéricos en explotaciones extensivas, (dehesas), y cebados en montanera. Sin embargo, según los datos publicados por el Ministerio de Agricultura, Alimentación y Medio Ambiente, más de un 70 por ciento de los cerdos ibéricos que se producen en España son criados en granjas intensivas. El sector porcino ibérico está en pleno proceso de cambio, tanto por las variaciones que se derivan de la crisis que sufre el sector, como por las modificaciones que se esperan en la norma de calidad, recientemente acordadas. En este entorno en el que existe una reducción importante de los márgenes comerciales en todos los eslabones de la cadena de valor del jamón ibérico de bellota, adquiere importancia el estudio de su estructura y funcionamiento, la determinación de los aspectos que mejoran la eficiencia económica a lo largo de la misma y su concordancia a lo largo de ella. Así mismo, un sector de los consumidores comienza a buscar productos de calidad, se interesa por productos con determinados etiquetados que garantizan la calidad o unas determinadas prácticas de producción y elaboración, como sucede con los productos ecológicos. Existen numerosos estudios sobre distintos aspectos del sector porcino ibérico, realizados por distintas instituciones, como universidades, instituciones privadas y proyectos europeos. En general, se centran en la mejora de la producción de los cerdos ibéricos o en la mejora de la elaboración de los jamones. La novedad de la investigación realizada en la presente Tesis Doctoral reside en el estudio de los aspectos que determinan la mayor eficiencia económica de la cadena de valor del jamón ibérico de bellota de la denominación de origen de Guijuelo, y en las características que aporta al jamón ibérico de bellota la producción ecológica frente a la convencional. Resumen Los objetivos planteados en esta investigación son la mejora de la cadena de valor del jamón ibérico de bellota de la denominación de origen de Guijuelo, mediante el estudio de la problemática estructural y de funcionamiento de la misma. Como objetivos específicos se proponen la mejora de la eficiencia económica en las relaciones comerciales entre los distintos eslabones de la cadena de valor, y el impulso de la oferta de los productos ecológicos de ibérico, mediante el estudio comparativo de las producciones convencionales y ecológicas. Para alcanzar estos objetivos la metodología utilizada es la cadena de valor agroalimentaria, utilizando como herramientas de análisis el método Delphi y la realización de un análisis DAFO del sector porcino ibérico. Las fuentes de información primarias utilizadas proceden por una parte de la colaboración realizada en el proyecto europeo Q-Porkchains, coordinado por el Dr. Jacques H. Trienekens, y por otra parte de los cuestionarios especialmente diseñados para el análisis Delphi realizado. Las fuentes de información secundarias proceden de artículos académicos publicados, de artículos de revistas especializadas en el sector y de informes y estadísticas publicados por el Ministerio de Agricultura, Alimentación y Medio Ambiente. Las conclusiones de la investigación son las siguientes. Las relaciones comerciales de la cadena de valor estudiada están gestionadas, ya que se otorga gran valor a las relaciones comerciales basadas en la confianza, tanto en el aprovisionamiento de productos como en la venta de los mismos en cada eslabón. Sin embargo, no se gestiona adecuadamente la gama de productos que se oferta a lo largo de la cadena de valor, puesto que los eslabones de la misma obtienen diferente eficiencia económica para el mismo producto. Por otra parte, aunque la denominación de origen de Guijuelo es una marca conocida por el consumidor, el esfuerzo por tener una marca colectiva de calidad no ha sido totalmente satisfactorio, en especial para el eslabón de la distribución. Por esta razón, es importante resaltar que es recomendable la creación de una marca propia que cuide la calidad con especial atención, modificando incluso las empresas integrantes de la cadena de valor si el objetivo de calidad no coincide. Es importante garantizar la calidad del producto al consumidor, mediante la trasmisión la información generada por los informes de inspección y certificación realizados por las entidades correspondientes en relación a la norma de calidad de los productos de cerdo ibérico y por el Consejo Regulador de Guijuelo. Se requiere un estudio detallado de los posibles nichos de mercado para llegar a los clientes que buscan un producto de calidad diferenciada como es el jamón ibérico de denominación de origen de Guijuelo. Dentro de los posibles clientes debe tenerse en consideración al colectivo de personas alérgicas, ya que no se encuentran con facilidad en el mercado productos cárnicos de cerdo libres de alérgenos. Por último, la innovación es muy importante en este sector, y pueden ofertarse nuevos productos que ayuden al acortamiento de la cadena de valor, como podría ser la carne fresca de cerdo ibérico de bellota. La producción ecológica de porcino ibérico, en la actualidad no es significativa. El manejo que se realiza de los cerdos ibéricos convencionales de bellota es muy similar al manejo que se realiza de los cerdos ecológicos, salvo en la alimentación que reciben los cerdos ibéricos antes del cebo en montanera y en los medicamentos permitidos en ambas producciones. La producción de porcino ecológico de bellota puede ser interesante para la exportación. Los jamones ibéricos de bellota dispondrían de un sello que garantiza un manejo tradicional –como se realiza en muchas explotaciones extensivas de cerdos ibéricos de bellota-, y que es apreciado por los consumidores de otros países, dispuestos a pagar por productos de calidad diferenciada. Este tipo de producción quizá podría solventar la limitación al ámbito nacional de la norma de calidad de los productos de cerdo ibérico. La falta de claridad y unanimidad en el sector sobre la calidad de los productos de ibéricos que se percibe en los cambios previstos en la legislación, y el interés del eslabón de la industria elaboradora de jamones en calidades no ligadas a producciones extensivas sino intensivas, por su mayor volumen de ventas y sus mejores rendimientos, mueven a los productores de “ibérico puro de bellota” a desmarcarse del sector “ibérico”, donde el peso del producto ibérico cruzado de pienso tiene mucho peso en las decisiones que se toman. The Iberian pork industry is a typically Spanish industry. The associated image by the consumer to these products corresponds to products made from Iberian pigs in extensive farms (meadows), and fattened in open range. However, according to data released by the Ministry of Agriculture, Food and Environment, more than 70 percent of the Iberian pigs produced in Spain are raised on factory farms. The Iberian pork industry is in process of change, both by modifications resulting from the crisis in the sector, and the expected ones in the quality standard, recently agreed. In this environment where there is a significant reduction in trade margins in all links of the value chain of Iberian ham, becomes important to study its structure and operation, identifying the aspects that improve economic efficiency along the value chain and their concordance along it. Likewise, part of the consumers begins searching for quality products, they are interested in certain products that guarantee some sort of quality or production and processing practices, like organic products. There are several studies on various aspects of the Iberian pork industry, made by different institutions, such as universities, private institutions and some European projects. In general, their goal is focused on improving the Iberian hog production or on improving the elaboration of hams. The novelty of the research conducted in this PhD thesis lies in the study of the aspects that determine the economic efficiency of the value chain of Iberian ham from Guijuelo designation of origin, and the comparison between the features that organic or conventional production of Iberian ham brings to it. The objectives propound in this research are the improvement of the value chain of Iberian ham from Guijuelo designation of origin, through the study of its structural and operational problematic. The proposed specific objectives are the improvement of the economic efficiency Resumen in trade relations between the different links in the value chain, and the promotion of the supply of Iberian organic pork products, through the comparative study of conventional and organic productions. In order to achieve these objectives, the used methodology is the agrifood value chain, using as analysis tools the Delphi method, and a SWOT analysis of the Iberian pork industry. The primary sources of information come partly from the collaboration in the European project Q-Porkchains, coordinated by Dr. Jacques Trienekens, and partly from the questionnaires specially designed for the Delphi analysis performed. The secondary sources come from published academic papers, specialized journal articles and reports published by the Ministry of Agriculture, Food and Environment. The research conclusions are as follows. Trade relations in the studied value chain are managed, as it is granted a great value to business relationships based on trust, both in the supply of products and in the sale in each link. However, the portfolio that is offered throughout the value chain is not properly managed, since the economic efficiency obtained for the same product is different in each link. Moreover, although the designation of origin of Guijuelo is a well-known brand, the effort made to acquire a collective quality trademark has not been entirely satisfactory, especially in the distribution link. Due to it, it is important to highlight that it is recommended to create a brand focused on quality, and indeed to change the firms integrating the vale chain, if the goal is not shared by all the companies. It is essential to ensure the quality of the product to the consumer, through the transmission of the available information related to the quality standard of Iberian pork products and of the Guijuelo Regulating Council. A detailed study of potential niche markets is required, in order to reach customers looking for a differentiated quality product as Iberian ham of Guijuelo designation of origin. It should be taken into consideration the group of allergy sufferers as potential customers, as they do not easily find pork products in the market free of allergens. Innovation is very important in this industry, and new products could be commercialized that help to shorten the value chain, as fresh acorn Iberian pork. Nowadays, the organic production of Iberian pork products is not significant. The operation is very similar in both conventional and organic production of Iberian pork products, except in the food received by Iberian hogs and the sort of medicinal treatments allowed in each production. The organic acorn Iberian pork production could be interesting for export. Acorn Iberian hams would have a well-known hallmark, which guarantees a traditional production, -as it is done in many extensive holdings of Iberian acorn-pigs-, and that is appreciated by consumers in other countries This niche of consumers could pay for differentiated quality products. Perhaps, this sort of production could solve the limitation to national scope of the quality standard of Iberian pork products. The lack of clarity and unanimity in the industry on the quality of Iberian pork products has been perceived in the agreed changes of the legislation as well as the interest of the processing industry in hams of quality categories which are not linked to extensive but intensive productions, due to their higher sales volume and better yields. This situation pushed "acorn pure Iberian " pigs farmers to split from the Iberian industry, where the importance of cross Iberian pig from intensive farms is growing so high that they can influence the negotiations and decisions taken inside and outside the industry.

A study of feature combination for vehicle detection based on image processing

Video analytics play a critical role in most recent traffic monitoring and driver assistance systems. In this context, the correct detection and classification of surrounding vehicles through image analysis has been the focus of extensive research in the last years. Most of the pieces of work reported for image-based vehicle verification make use of supervised classification approaches and resort to techniques, such as histograms of oriented gradients (HOG), principal component analysis (PCA), and Gabor filters, among others. Unfortunately, existing approaches are lacking in two respects: first, comparison between methods using a common body of work has not been addressed; second, no study of the combination potentiality of popular features for vehicle classification has been reported. In this study the performance of the different techniques is first reviewed and compared using a common public database. Then, the combination capabilities of these techniques are explored and a methodology is presented for the fusion of classifiers built upon them, taking into account also the vehicle pose. The study unveils the limitations of single-feature based classification and makes clear that fusion of classifiers is highly beneficial for vehicle verification.

Histograms of oriented gradients for fast on-board vehicle verification

Histograms of Oriented Gradients (HoGs) provide excellent results in object detection and verification. However, their demanding processing requirements bound their applicability in some critical real-time scenarios, such as for video-based on-board vehicle detection systems. In this work, an efficient HOG configuration for pose-based on-board vehicle verification is proposed, which alleviates both the processing requirements and required feature vector length without reducing classification performance. The impact on classification of some critical configuration and processing parameters is in depth analyzed to propose a baseline efficient descriptor. Based on the analysis of its cells contribution to classification, new view-dependent cell-configuration patterns are proposed, resulting in reduced descriptors which provide an excellent balance between performance and computational requirements, rendering higher verification rates than other works in the literature.

Kinase Activity-dependent Nuclear Export Opposes Stress-induced Nuclear Accumulation and Retention of Hog1 Mitogen-activated Protein Kinase in the Budding Yeast Saccharomyces cerevisiae

Budding yeast adjusts to increases in external osmolarity via a specific mitogen-activated protein kinase signal pathway, the high-osmolarity glycerol response (HOG) pathway. Studies with a functional Hog1–green fluorescent protein (GFP) fusion reveal that even under nonstress conditions the mitogen-activated protein kinase Hog1 cycles between cytoplasmic and nuclear compartments. The basal distribution of the protein seems independent of its activator, Pbs2, and independent of its phosphorylation status. Upon osmotic challenge, the Hog1–GFP fusion becomes rapidly concentrated in the nucleus from which it is reexported after return to an iso-osmotic environment or after adaptation to high osmolarity. The preconditions and kinetics of increased nuclear localization correlate with those found for the dual phosphorylation of Hog1–GFP. The duration of Hog1 nuclear residence is modulated by the presence of the general stress activators Msn2 and Msn4. Reexport of Hog1 to the cytoplasm does not require de novo protein synthesis but depends on Hog1 kinase activity. Thus, at least three different mechanisms contribute to the intracellular distribution pattern of Hog1: phosphorylation-dependent nuclear accumulation, retention by nuclear targets, and a kinase-induced export.

Mitochondrial Inheritance Is Delayed in Saccharomyces cerevisiae Cells Lacking the Serine/Threonine Phosphatase PTC1

In wild-type yeast mitochondrial inheritance occurs early in the cell cycle concomitant with bud emergence. Cells lacking the PTC1 gene initially produce buds without a mitochondrial compartment; however, these buds later receive part of the mitochondrial network from the mother cell. Thus, the loss of PTC1 causes a delay, but not a complete block, in mitochondrial transport. PTC1 encodes a serine/threonine phosphatase in the high-osmolarity glycerol response (HOG) pathway. The mitochondrial inheritance delay in the ptc1 mutant is not attributable to changes in intracellular glycerol concentrations or defects in the organization of the actin cytoskeleton. Moreover, epistasis experiments with ptc1Δ and mutations in HOG pathway kinases reveal that PTC1 is not acting through the HOG pathway to control the timing of mitochondrial inheritance. Instead, PTC1 may be acting either directly or through a different signaling pathway to affect the mitochondrial transport machinery in the cell. These studies indicate that the timing of mitochondrial transport in wild-type cells is genetically controlled and provide new evidence that mitochondrial inheritance does not depend on a physical link between the mitochondrial network and the incipient bud site.

Functional Characterization of the Interaction of Ste50p with Ste11p MAPKKK in Saccharomyces cerevisiae

The Saccharomyces cerevisiae Ste11p protein kinase is a homologue of mammalian MAPK/extracellular signal-regulated protein kinase kinase kinases (MAPKKKs or MEKKs) as well as the Schizosaccharomyces pombe Byr2p kinase. Ste11p functions in several signaling pathways, including those for mating pheromone response and osmotic stress response. The Ste11p kinase has an N-terminal domain that interacts with other signaling molecules to regulate Ste11p function and direct its activity in these pathways. One of the Ste11p regulators is Ste50p, and Ste11p and Ste50p associate through their respective N-terminal domains. This interaction relieves a negative activity of the Ste11p N terminus, and removal of this negative function is required for Ste11p function in the high-osmolarity glycerol (HOG) pathway. The Ste50p/Ste11p interaction is also important (but not essential) for Ste11p function in the mating pathway; in this pathway binding of the Ste11p N terminus with both Ste50p and Ste5p is required, with the Ste5p association playing the major role in Ste11p function. In vitro, Ste50p disrupts an association between the catalytic C terminus and the regulatory N terminus of Ste11p. In addition, Ste50p appears to modulate Ste11p autophosphorylation and is itself a substrate of the Ste11p kinase. Therefore, both in vivo and in vitro data support a role for Ste50p in the regulation of Ste11p activity.

Rck2, a member of the calmodulin-protein kinase family, links protein synthesis to high osmolarity MAP kinase signaling in budding yeast

Rck2, a yeast Ser/Thr protein kinase homologous to mammalian calmodulin kinases, requires phosphorylation for activation. We provide evidence that in budding yeast, this step can be executed by the osmostress-activated mitogen-activated protein kinase Hog1. Rck2 phosphorylation was transiently increased during osmostress or in mutants with a hyperactive high osmolarity glycerol (HOG) pathway. This modification depended on catalytically active Hog1 kinase and two putative mitogen-activated protein kinase phosphorylation sites in Rck2. Immunokinase assays showed that Hog1 can directly phosphorylate Rck2 to stimulate its enzymatic activity toward translation elongation factor 2. We demonstrate that Hog1 and Rck2 are necessary for attenuation of protein synthesis in response to osmotic challenge and show that modification of elongation factor 2 induced by osmostress depends on Rck2 and Hog1 in vivo. Therefore, we propose that the transient down-regulation of protein synthesis after osmotic shock is a response not to damage but to an extracellular signal mediated by Hog1 and Rck2.

Human Motion Analysis Based on Sequential Modeling of Radar Signal and Stereo Image Features

Falls are one of the greatest threats to elderly health in their daily living routines and activities. Therefore, it is very important to detect falls of an elderly in a timely and accurate manner, so that immediate response and proper care can be provided, by sending fall alarms to caregivers. Radar is an effective non-intrusive sensing modality which is well suited for this purpose, which can detect human motions in all types of environments, penetrate walls and fabrics, preserve privacy, and is insensitive to lighting conditions. Micro-Doppler features are utilized in radar signal corresponding to human body motions and gait to detect falls using a narrowband pulse-Doppler radar. Human motions cause time-varying Doppler signatures, which are analyzed using time-frequency representations and matching pursuit decomposition (MPD) for feature extraction and fall detection. The extracted features include MPD features and the principal components of the time-frequency signal representations. To analyze the sequential characteristics of typical falls, the extracted features are used for training and testing hidden Markov models (HMM) in different falling scenarios. Experimental results demonstrate that the proposed algorithm and method achieve fast and accurate fall detections. The risk of falls increases sharply when the elderly or patients try to exit beds. Thus, if a bed exit can be detected at an early stage of this motion, the related injuries can be prevented with a high probability. To detect bed exit for fall prevention, the trajectory of head movements is used for recognize such human motion. A head detector is trained using the histogram of oriented gradient (HOG) features of the head and shoulder areas from recorded bed exit images. A data association algorithm is applied on the head detection results to eliminate head detection false alarms. Then the three dimensional (3D) head trajectories are constructed by matching scale-invariant feature transform (SIFT) keypoints in the detected head areas from both the left and right stereo images. The extracted 3D head trajectories are used for training and testing an HMM based classifier for recognizing bed exit activities. The results of the classifier are presented and discussed in the thesis, which demonstrates the effectiveness of the proposed stereo vision based bed exit detection approach.