Heated fiber optic distributed temperature sensing for measuring soil volumetric heat capacity and water content: A dual probe heat-pulse approach

The first feasibility study of using dual-probe heated fiber optics with distributed temperature sensing to measure soil volumetric heat capacity and soil water content is presented. Although results using different combinations of cables demonstrate feasibility, further work is needed to gain accuracy, including a model to account for the finite dimension and the thermal influence of the probes. Implementation of the dual-probe heat-pulse (DPHP) approach for measurement of volumetric heat capacity (C) and water content (θ) with distributed temperature sensing heated fiber optic (FO) systems presents an unprecedented opportunity for environmental monitoring (e.g., simultaneous measurement at thousands of points). We applied uniform heat pulses along a FO cable and monitored the thermal response at adjacent cables. We tested the DPHP method in the laboratory using multiple FO cables at a range of spacings. The amplitude and phase shift in the heat signal with distance was found to be a function of the soil volumetric heat capacity. Estimations of C at a range of moisture contents (θ = 0.09– 0.34 m3 m−3) suggest the feasibility of measurement via responsiveness to the changes in θ, although we observed error with decreasing soil water contents (up to 26% at θ = 0.09 m3 m−3). Optimization will require further models to account for the finite radius and thermal influence of the FO cables. Although the results indicate that the method shows great promise, further study is needed to quantify the effects of soil type, cable spacing, and jacket configurations on accuracy.

Mapping variability of soil water content and flux across 1¿1000 m scales using the actively heated fiber optic method

The Actively Heated Fiber Optic (AHFO) method is shown to be capable of measuring soil water content several times per hour at 0.25 m spacing along cables of multiple kilometers in length. AHFO is based on distributed temperature sensing (DTS) observation of the heating and cooling of a buried fiber-optic cable resulting from an electrical impulse of energy delivered from the steel cable jacket. The results presented were collected from 750 m of cable buried in three 240 m colocated transects at 30, 60, and 90 cm depths in an agricultural field under center pivot irrigation. The calibration curve relating soil water content to the thermal response of the soil to a heat pulse of 10 W m−1 for 1 min duration was developed in the lab. This calibration was found applicable to the 30 and 60 cm depth cables, while the 90 cm depth cable illustrated the challenges presented by soil heterogeneity for this technique. This method was used to map with high resolution the variability of soil water content and fluxes induced by the nonuniformity of water application at the surface.

Methodology of image analysis for study of the vertisols moisture content

The main problem to study vertical drainage from the moisture distribution, on a vertisol profile, is searching for suitable methods using these procedures. Our aim was to design a digital image processing methodology and its analysis to characterize the moisture content distribution of a vertisol profile. In this research, twelve soil pits were excavated on a ba re Mazic Pellic Vertisols ix of them in May 13/2011 and the rest in May 19 /2011 after a moderate rainfall event. Digital RGB images were taken from each vertisol pit using a Kodak? camera selecting a size of 1600x945 pixels. Each soil image was processed to homogenized brightness and then a spatial filter with several window sizes was applied to select the optimum one. The RGB image obtained were divided in each matrix color selecting the best thresholds for each one, maximum and minimum, to be applied and get a digital binary pattern. This one was analyzed by estimating two fractal scaling exponents box counting dimension D BC) and interface fractal dimension (D) In addition, three pre-fractal scaling coefficients were determinate at maximum resolution: total number of boxes intercepting the foreground pattern (A), fractal lacunarity (?1) and Shannon entropy S1). For all the images processed the spatial filter 9x9 was the optimum based on entropy, cluster and histogram criteria. Thresholds for each color were selected based on bimodal histograms.

Is the polarity of content producers strongly influenced by the results of the event?

This paper presents an approach to compare two types of data, subjective data (Polarity of Pan American Games 2011 event by country) and objective data (the number of medals won by each participating country), based on the Pearson corre- lation. When dealing with events described by people, knowledge acquisition is difficult because their structure is heterogeneous and subjective. A first step towards knowing the polarity of the information provided by people consists in automatically classifying the posts into clusters according to their polarity. The authors carried out a set of experiments using a corpus that consists of 5600 posts extracted from 168 Internet resources related to a specific event: the 2011 Pan American games. The approach is based on four components: a crawler, a filter, a synthesizer and a polarity analyzer. The PanAmerican approach automatically classifies the polarity of the event into clusters with the following results: 588 positive, 336 neutral, and 76 negative. Our work found out that the polarity of the content produced was strongly influenced by the results of the event with a correlation of .74. Thus, it is possible to conclude that the polarity of content is strongly affected by the results of the event. Finally, the accuracy of the PanAmerican approach is: .87, .90, and .80 according to the precision of the three classes of polarity evaluated.

Effect of content characteristics on quality of experience of adaptive streaming

The growing popularity of adaptive streaming-based video delivery nowadays has raised the interest about the user's perception when experiencing quality adaptation. The impact of the video content characteristics on user's perceptual quality has already become evident. The aim of this study is to investigate the influence of this factor on the quality of experience of adaptive streaming scenarios. Our results show that the perceptual quality of adaptation strategies applied on videos with high spatial and low temporal amount of activity is significantly lower compared to the other content types.

Effects of nutrition on digestión efficicency and gaseous emissions from slurry in growing-finishing pigs. I. Influence of the inclusión of two levels of orange pulp and carb meal in isofibrous diets

An experiment was conducted to investigate the effects of increasing the level of two sources of fibrous by-products, orange pulp (OP) and carob meal (CM), in iso-NDF growing-finishing pig diets on nutrient balance, slurry composition and potential ammonia (NH3) and methane (CH4) emissions. Thirty pigs (85.4 ± 12.3 kg) were fed five iso-nutritive diets: a commercial control wheat/barley (C) and four experimental diets including two sources of fibrous by-products (OP and CM) and two dietary levels (75 and 150 g/kg) in a 2 × 2 factorial arrangement. After a 14-day adaptation period, faeces and urine were collected separately for 7 days to measure nutrient digestibility and the excretory patterns of N from pigs (6 replicates per diet) housed individually in metabolic pens. For each animal, the derived NH3 and CH4 emissions were measured in samples of slurry over an 11- and 100-day storage periods, respectively. Source and level of the fibrous by-products affected digestion efficiency in a different way as the coefficients of total tract apparent digestibility (CTTAD) for dry matter (DM), organic matter (OM), fibre fractions and gross energy increased with OP but decreased with CM (P < 0.05). Crude protein CTTAD decreased with the inclusion of both sources of fibre, being lower at the highest dietary level. Faecal concentration of fibre fractions increased (P < 0.05) with the level of inclusion of CM but decreased with that of OP (P < 0.01). High dietary level for both sources of fibre increased (P < 0.02) CP faecal content but urine N content decreased (from 205 to 168 g/kg DM, P < 0.05) in all the fibre-supplemented compared to C diet. Additionally, the proportions of undigested dietary, water soluble, and bacterial and endogenous debris of faecal N excretion were not affected by treatments. The initial slurry characteristics did not differ among different fibre sources and dietary levels, except pH, which decreased at the highest by-product inclusion levels. Ammonia emission per kg of slurry was lower in all the fibre-supplemented diets than in C diet (from 2.44 to 1.81 g, P < 0.05). Additionally, slurries from the highest dietary level of by-products tended (P < 0.06) to emit less NH3 per kg of initial total Kjeldahl N and showed a lower B0, independently of the fibre source. Thus, the fibre sources and their dietary levels affected pig nutrient digestion and composition of urine and faeces, showing potential to decrease NH3 and CH4 emissions at high levels of inclusion, independently of type of fibre.

The effect of moisture content on nondestructive probing measurements

When assessing existing timber structures it is not possible to obtain density as the ratio mass/volume, so nondestructive probing methods are used to predict density. As in other nondestructive techniques, moisture content influences measurements. The goal of this paper is to study the influence of timber moisture content on two nondestructive probing techniques (penetration resistance and pullout resistance). 25 large cross section specimens of laricio pine from Spain were measured. The moisture content ranged from 65.1% to 8.3%. Penetration depth decreases and screw withdrawal strength increases when moisture content decreases below the fiber saturation point. There are lineal tendencies in both techniques. No moisture content influence was found in measures above fiber saturation point.

A Novel Approach to Context-Aware Content-Based Middleware

La computación ubicua está extendiendo su aplicación desde entornos específicos hacia el uso cotidiano; el Internet de las cosas (IoT, en inglés) es el ejemplo más brillante de su aplicación y de la complejidad intrínseca que tiene, en comparación con el clásico desarrollo de aplicaciones. La principal característica que diferencia la computación ubicua de los otros tipos está en como se emplea la información de contexto. Las aplicaciones clásicas no usan en absoluto la información de contexto o usan sólo una pequeña parte de ella, integrándola de una forma ad hoc con una implementación específica para la aplicación. La motivación de este tratamiento particular se tiene que buscar en la dificultad de compartir el contexto con otras aplicaciones. En realidad lo que es información de contexto depende del tipo de aplicación: por poner un ejemplo, para un editor de imágenes, la imagen es la información y sus metadatos, tales como la hora de grabación o los ajustes de la cámara, son el contexto, mientras que para el sistema de ficheros la imagen junto con los ajustes de cámara son la información, y el contexto es representado por los metadatos externos al fichero como la fecha de modificación o la de último acceso. Esto significa que es difícil compartir la información de contexto, y la presencia de un middleware de comunicación que soporte el contexto de forma explícita simplifica el desarrollo de aplicaciones para computación ubicua. Al mismo tiempo el uso del contexto no tiene que ser obligatorio, porque si no se perdería la compatibilidad con las aplicaciones que no lo usan, convirtiendo así dicho middleware en un middleware de contexto. SilboPS, que es nuestra implementación de un sistema publicador/subscriptor basado en contenido e inspirado en SIENA [11, 9], resuelve dicho problema extendiendo el paradigma con dos elementos: el Contexto y la Función de Contexto. El contexto representa la información contextual propiamente dicha del mensaje por enviar o aquella requerida por el subscriptor para recibir notificaciones, mientras la función de contexto se evalúa usando el contexto del publicador y del subscriptor. Esto permite desacoplar la lógica de gestión del contexto de aquella de la función de contexto, incrementando de esta forma la flexibilidad de la comunicación entre varias aplicaciones. De hecho, al utilizar por defecto un contexto vacío, las aplicaciones clásicas y las que manejan el contexto pueden usar el mismo SilboPS, resolviendo de esta forma la incompatibilidad entre las dos categorías. En cualquier caso la posible incompatibilidad semántica sigue existiendo ya que depende de la interpretación que cada aplicación hace de los datos y no puede ser solucionada por una tercera parte agnóstica. El entorno IoT conlleva retos no sólo de contexto, sino también de escalabilidad. La cantidad de sensores, el volumen de datos que producen y la cantidad de aplicaciones que podrían estar interesadas en manipular esos datos está en continuo aumento. Hoy en día la respuesta a esa necesidad es la computación en la nube, pero requiere que las aplicaciones sean no sólo capaces de escalar, sino de hacerlo de forma elástica [22]. Desgraciadamente no hay ninguna primitiva de sistema distribuido de slicing que soporte un particionamiento del estado interno [33] junto con un cambio en caliente, además de que los sistemas cloud actuales como OpenStack u OpenNebula no ofrecen directamente una monitorización elástica. Esto implica que hay un problema bilateral: cómo puede una aplicación escalar de forma elástica y cómo monitorizar esa aplicación para saber cuándo escalarla horizontalmente. E-SilboPS es la versión elástica de SilboPS y se adapta perfectamente como solución para el problema de monitorización, gracias al paradigma publicador/subscriptor basado en contenido y, a diferencia de otras soluciones [5], permite escalar eficientemente, para cumplir con la carga de trabajo sin sobre-provisionar o sub-provisionar recursos. Además está basado en un algoritmo recientemente diseñado que muestra como añadir elasticidad a una aplicación con distintas restricciones sobre el estado: sin estado, estado aislado con coordinación externa y estado compartido con coordinación general. Su evaluación enseña como se pueden conseguir notables speedups, siendo el nivel de red el principal factor limitante: de hecho la eficiencia calculada (ver Figura 5.8) demuestra cómo se comporta cada configuración en comparación con las adyacentes. Esto permite conocer la tendencia actual de todo el sistema, para saber si la siguiente configuración compensará el coste que tiene con la ganancia que lleva en el throughput de notificaciones. Se tiene que prestar especial atención en la evaluación de los despliegues con igual coste, para ver cuál es la mejor solución en relación a una carga de trabajo dada. Como último análisis se ha estimado el overhead introducido por las distintas configuraciones a fin de identificar el principal factor limitante del throughput. Esto ayuda a determinar la parte secuencial y el overhead de base [26] en un despliegue óptimo en comparación con uno subóptimo. Efectivamente, según el tipo de carga de trabajo, la estimación puede ser tan baja como el 10 % para un óptimo local o tan alta como el 60 %: esto ocurre cuando se despliega una configuración sobredimensionada para la carga de trabajo. Esta estimación de la métrica de Karp-Flatt es importante para el sistema de gestión porque le permite conocer en que dirección (ampliar o reducir) es necesario cambiar el despliegue para mejorar sus prestaciones, en lugar que usar simplemente una política de ampliación. ABSTRACT The application of pervasive computing is extending from field-specific to everyday use. The Internet of Things (IoT) is the shiniest example of its application and of its intrinsic complexity compared with classical application development. The main characteristic that differentiates pervasive from other forms of computing lies in the use of contextual information. Some classical applications do not use any contextual information whatsoever. Others, on the other hand, use only part of the contextual information, which is integrated in an ad hoc fashion using an application-specific implementation. This information is handled in a one-off manner because of the difficulty of sharing context across applications. As a matter of fact, the application type determines what the contextual information is. For instance, for an imaging editor, the image is the information and its meta-data, like the time of the shot or camera settings, are the context, whereas, for a file-system application, the image, including its camera settings, is the information and the meta-data external to the file, like the modification date or the last accessed timestamps, constitute the context. This means that contextual information is hard to share. A communication middleware that supports context decidedly eases application development in pervasive computing. However, the use of context should not be mandatory; otherwise, the communication middleware would be reduced to a context middleware and no longer be compatible with non-context-aware applications. SilboPS, our implementation of content-based publish/subscribe inspired by SIENA [11, 9], solves this problem by adding two new elements to the paradigm: the context and the context function. Context represents the actual contextual information specific to the message to be sent or that needs to be notified to the subscriber, whereas the context function is evaluated using the publisher’s context and the subscriber’s context to decide whether the current message and context are useful for the subscriber. In this manner, context logic management is decoupled from context management, increasing the flexibility of communication and usage across different applications. Since the default context is empty, context-aware and classical applications can use the same SilboPS, resolving the syntactic mismatch that there is between the two categories. In any case, the possible semantic mismatch is still present because it depends on how each application interprets the data, and it cannot be resolved by an agnostic third party. The IoT environment introduces not only context but scaling challenges too. The number of sensors, the volume of the data that they produce and the number of applications that could be interested in harvesting such data are growing all the time. Today’s response to the above need is cloud computing. However, cloud computing applications need to be able to scale elastically [22]. Unfortunately there is no slicing, as distributed system primitives that support internal state partitioning [33] and hot swapping and current cloud systems like OpenStack or OpenNebula do not provide elastic monitoring out of the box. This means there is a two-sided problem: 1) how to scale an application elastically and 2) how to monitor the application and know when it should scale in or out. E-SilboPS is the elastic version of SilboPS. I t is the solution for the monitoring problem thanks to its content-based publish/subscribe nature and, unlike other solutions [5], it scales efficiently so as to meet workload demand without overprovisioning or underprovisioning. Additionally, it is based on a newly designed algorithm that shows how to add elasticity in an application with different state constraints: stateless, isolated stateful with external coordination and shared stateful with general coordination. Its evaluation shows that it is able to achieve remarkable speedups where the network layer is the main limiting factor: the calculated efficiency (see Figure 5.8) shows how each configuration performs with respect to adjacent configurations. This provides insight into the actual trending of the whole system in order to predict if the next configuration would offset its cost against the resulting gain in notification throughput. Particular attention has been paid to the evaluation of same-cost deployments in order to find out which one is the best for the given workload demand. Finally, the overhead introduced by the different configurations has been estimated to identify the primary limiting factor for throughput. This helps to determine the intrinsic sequential part and base overhead [26] of an optimal versus a suboptimal deployment. Depending on the type of workload, this can be as low as 10% in a local optimum or as high as 60% when an overprovisioned configuration is deployed for a given workload demand. This Karp-Flatt metric estimation is important for system management because it indicates the direction (scale in or out) in which the deployment has to be changed in order to improve its performance instead of simply using a scale-out policy.

Effect of the season on the free phytoprostanes content in Cornicabra extra virgin olive oil from deficit irrigated olive trees

BACKGROUND: The effect of regulated deficit irrigation (RDI) on the phytoprostane (PhytoP) content in extra virgin olive (Olea europaea L., cv. Cornicabra) oil (EVOO) was studied. During the 2012 and 2013 seasons, T0 plants were irrigated at 100% ETc, while T1 and T2 plants were irrigated avoiding water deficit during phases I and III of fruit growth and saving water during the non-critical phenological period of pit hardening (phase II), developing amore severewater deficit in T2 plants. In 2013, a fourth treatment (T3) was also performed, which was similar to T2 except that water saving was from the beginning of phase II to 15 days after the end of phase II. RESULTS: 9-F1t-PhytoP, 9-epi-9-F1t-PhytoP, 9-epi-9-D1t-PhytoP, 9-D1t-PhytoP, 16-B1-PhytoP and 9-L1-PhytoP were present in Cornicabra EVOO, and their contents increased in the EVOO fromRDI plants. CONCLUSION: Deficit irrigation during pit hardening or for a further period of 2 weeks thereafter to increase irrigation water saving is clearly critical for EVOO composition because of the enhancement of free PhytoPs, which have potential beneficial effects on human health. The response of individual free PhytoPs to changes in plant water status was not as perceptible as expected, preventing their use as biomarkers of water stress.

Face, content, and construct validity of the EndoViS training system for objective assessment of psychomotor skills of laparoscopic surgeons

Background The aim of this study is to present face, content, and constructs validity of the endoscopic orthogonal video system (EndoViS) training system and determines its efficiency as a training and objective assessment tool of the surgeons’ psychomotor skills. Methods Thirty-five surgeons and medical students participated in this study: 11 medical students, 19 residents, and 5 experts. All participants performed four basic skill tasks using conventional laparoscopic instruments and EndoViS training system. Subsequently, participants filled out a questionnaire regarding the design, realism, overall functionality, and its capabilities to train hand–eye coordination and depth perception, rated on a 5-point Likert scale. Motion data of the instruments were obtained by means of two webcams built into a laparoscopic physical trainer. To identify the surgical instruments in the images, colored markers were placed in each instrument. Thirteen motion-related metrics were used to assess laparoscopic performance of the participants. Statistical analysis of performance was made between novice, intermediate, and expert groups. Internal consistency of all metrics was analyzed with Cronbach’s α test. Results Overall scores about features of the EndoViS system were positives. Participants agreed with the usefulness of tasks and the training capacities of EndoViS system (score >4). Results presented significant differences in the execution of three skill tasks performed by participants. Seven metrics showed construct validity for assessment of performance with high consistency levels. Conclusions EndoViS training system has been successfully validated. Results showed that EndoViS was able to differentiate between participants of varying laparoscopic experience. This simulator is a useful and effective tool to objectively assess laparoscopic psychomotor skills of the surgeons.

Both apolipoprotein E and A-I genes are present in a nonmammalian vertebrate and are highly expressed during embryonic development

Apolipoprotein E (apoE) is associated with several classes of plasma lipoproteins and mediates uptake of lipoproteins through its ability to interact with specific cell surface receptors. Besides its role in cardiovascular diseases, accumulating evidence has suggested that apoE could play a role in neurodegenerative diseases, such as Alzheimer disease. In vertebrates, apoA-I is the major protein of high-density lipoprotein. ApoA-I may play an important role in regulating the cholesterol content of peripheral tissues through the reverse cholesterol transport pathway. We have isolated cDNA clones that code for apoE and apoA-I from a zebrafish embryo library. Analysis of the deduced amino acid sequences showed the presence of a region enriched in basic amino acids in zebrafish apoE similar to the lipoprotein receptor-binding region of human apoE. We demonstrated by whole-mount in situ hybridization that apoE and apoA-I genes are highly expressed in the yolk syncytial layer, an extraembryonic structure implicated in embryonic and larval nutrition. ApoE transcripts were also observed in the deep cell layer during blastula stage, in numerous ectodermal derivatives after gastrulation, and after 3 days of development in a limited number of cells both in brain and in the eyes. Our data indicate that apoE can be found in a nonmammalian vertebrate and that the duplication events, from which apoE and apoA-I genes arose, occurred before the divergence of the tetrapod and teleost ancestors. Zebrafish can be used as a simple and useful model for studying the role of apolipoproteins in embryonic and larval nutrition and of apoE in brain morphogenesis and regeneration.

Insulin and insulin-like growth factor-I acutely inhibit surface translocation of growth hormone receptors in osteoblasts: A novel mechanism of growth hormone receptor regulation

We previously have demonstrated that insulin and insulin-like growth factor-I (IGF-I) down-regulate growth hormone (GH) binding in osteoblasts by reducing the number of surface GH receptors (GHRs). The present study was undertaken to investigate the mechanism of GHR down-regulation. Treatment with 5 nM insulin or IGF-I for 18 hr significantly decreased surface GH binding to 26.4 ± 2.9% and 23.0 ± 2.7% of control (mean ± SE; P < 0.05), respectively. No corresponding reductions in the mRNA level and total cellular content of GHR were found, nor was the rate of receptor internalization affected. The effects on GHR translocation were assessed by measuring the reappearance of GH binding of whole cells after trypsinization to remove the surface receptors. GH binding of control cultures significantly increased (P < 0.05) over 2 hr after trypsinization, whereas no recovery of binding activity was detected in insulin and IGF-I-treated cultures, indicating that GHR translocation was impaired. Studies on the time course of GHR down-regulation revealed that surface GH binding was reduced significantly by 3-hr treatment (P ≤ 0.0005), whereas GHR translocation was completely abolished by 75–90 min with insulin and IGF-I. The inhibition of receptor translocation by insulin, but not IGF-I, was attenuated by wortmannin. In conclusion, insulin and IGF-I down-regulated GH binding in osteoblasts by acutely impairing GHR translocation, with their effects exerted through distinct postreceptor signaling pathways.

Experimental hemochromatosis due to MHC class I HFE deficiency: Immune status and iron metabolism

The puzzling linkage between genetic hemochromatosis and histocompatibility loci became even more so when the gene involved, HFE, was identified. Indeed, within the well defined, mainly peptide-binding, MHC class I family of molecules, HFE seems to perform an unusual yet essential function. As yet, our understanding of HFE function in iron homeostasis is only partial; an even more open question is its possible role in the immune system. To advance on both of these avenues, we report the deletion of HFE α1 and α2 putative ligand binding domains in vivo. HFE-deficient animals were analyzed for a comprehensive set of metabolic and immune parameters. Faithfully mimicking human hemochromatosis, mice homozygous for this deletion develop iron overload, characterized by a higher plasma iron content and a raised transferrin saturation as well as an elevated hepatic iron load. The primary defect could, indeed, be traced to an augmented duodenal iron absorption. In parallel, measurement of the gut mucosal iron content as well as iron regulatory proteins allows a more informed evaluation of various hypotheses regarding the precise role of HFE in iron homeostasis. Finally, an extensive phenotyping of primary and secondary lymphoid organs including the gut provides no compelling evidence for an obvious immune-linked function for HFE.

A targeted mutation in the murine gene encoding the high density lipoprotein (HDL) receptor scavenger receptor class B type I reveals its key role in HDL metabolism

Plasma high density lipoprotein (HDL), which protects against atherosclerosis, is thought to remove cholesterol from peripheral tissues and to deliver cholesteryl esters via a selective uptake pathway to the liver (reverse cholesterol transport) and steroidogenic tissues (e.g., adrenal gland for storage and hormone synthesis). Despite its physiologic and pathophysiologic importance, the cellular metabolism of HDL has not been well defined. The class B, type I scavenger receptor (SR-BI) has been proposed to play an important role in HDL metabolism because (i) it is a cell surface HDL receptor which mediates selective cholesterol uptake in cultured cells, (ii) its physiologically regulated expression is most abundant in the liver and steroidogenic tissues, and (iii) hepatic overexpression dramatically lowers plasma HDL. To test directly the normal role of SR-BI in HDL metabolism, we generated mice with a targeted null mutation in the SR-BI gene. In heterozygous and homozygous mutants relative to wild-type controls, plasma cholesterol concentrations were increased by ≈31% and 125%, respectively, because of the formation of large, apolipoprotein A-I (apoA-I)-containing particles, and adrenal gland cholesterol content decreased by 42% and 72%, respectively. The plasma concentration of apoA-I, the major protein in HDL, was unchanged in the mutants. This, in conjunction with the increased lipoprotein size, suggests that the increased plasma cholesterol in the mutants was due to decreased selective cholesterol uptake. These results provide strong support for the proposal that in mice the gene encoding SR-BI plays a key role in determining the levels of plasma lipoprotein cholesterol (primarily HDL) and the accumulation of cholesterol stores in the adrenal gland. If it has a similar role in controlling plasma HDL in humans, SR-BI may influence the development and progression of atherosclerosis and may be an attractive candidate for therapeutic intervention in this disease.

Binding of Insulin-like Growth Factor (IGF)–Binding Protein-5 to Smooth-Muscle Cell Extracellular Matrix Is a Major Determinant of the Cellular Response to IGF-I

Insulin-like growth factor–binding protein-5 (IGFBP-5) has been shown to bind to fibroblast extracellular matrix (ECM). Extracellular matrix binding of IGFBP-5 leads to a decrease in its affinity for insulin-like growth factor-I (IGF-I), which allows IGF-I to better equilibrate with IGF receptors. When the amount of IGFBP-5 that is bound to ECM is increased by exogenous addition, IGF-I’s effect on fibroblast growth is enhanced. In this study we identified the specific basic residues in IGFBP-5 that mediate its binding to porcine smooth-muscle cell (pSMC) ECM. An IGFBP-5 mutant containing alterations of basic residues at positions 211, 214, 217, and 218 had the greatest reduction in ECM binding, although three other mutants, R214A, R207A/K211N, and K202A/R206N/R207A, also had major decreases. In contrast, three other mutants, R201A/K202N/R206N/R208A, and K217N/R218A and K211N, had only minimal reductions in ECM binding. This suggested that residues R207 and R214 were the most important for binding, whereas alterations in K211 and R218, which align near them, had minimal effects. To determine the effect of a reduction in ECM binding on the cellular replication response to IGF-I, pSMCs were transfected with the mutant cDNAs that encoded the forms of IGFBPs with the greatest changes in ECM binding. The ECM content of IGFBP-5 from cultures expressing the K211N, R214A, R217A/R218A, and K202A/R206N/R207A mutants was reduced by 79.6 and 71.7%, respectively, compared with cells expressing the wild-type protein. In contrast, abundance of the R201A/K202N/R206N/R208A mutant was reduced by only 14%. Cells expressing the two mutants with reduced ECM binding had decreased DNA synthesis responses to IGF-I, but the cells expressing the R201A/K202N/R206N/R208A mutant responded well to IGF-I. The findings suggest that specific basic amino acids at positions 207 and 214 mediate the binding of IGFBP-5 to pSMC/ECM. Smooth-muscle cells that constitutively express the mutants that bind weakly to ECM are less responsive to IGF-I, suggesting that ECM binding of IGFBP-5 is an important variable that determines cellular responsiveness.