Monitoring and isolation of blood dendritic cells from apheresis products in healthy individuals: a platform for cancer immunotherapy

The fundamental role of dendritic cells (DC in initiating and directing the primary immune response is well established. Furthermore, it is now accepted that DC may be useful in new vaccination strategies for preventing certain malignant and infectious diseases. As blood DC (BDC physiology differs from that of the DC homologues generated in vitro from monocyte precursors, it is becoming more relevant to consider BDC for therapeutic interventions. Until recently, protocols for the isolation of BDC were laborious and inefficient; therefore, their use for investigative cancer immunotherapy is not widespread. In this study, we carefully documented BDC counts, yields and subsets during apheresis (Cobe Spectra), the initial and essential procedure in creating a BDC isolation platform for cancer immunotherapy. We established that an automated software package (Version 6,0 AutoPBPC) provides an operator-independent reliable source of motionuclear cells (MNC for BDC preparation. Further, we observed that BDC might be recovered in high yields, often greater than 100% relative to the number of circulating BDC predicted by blood volume. An average of 66 million (range, 17-179) BDC per 10-1 procedure were obtained, largely satisfying the needs for immunization. Higher yields were possible on total processed blood volumes of 151. BDC were not activated by the isolation procedure and, more importantly, both BDC subsets (CD11c(+)CD123(low) and CD11c(-)CD123(high)) were equally represented. Finally, we established that the apheresis product could be used for antibody-based BDC immunoselection and demonstrated that fully functional BDC can be obtained by this procedure. (C) 2002 Published by Elsevier Science B.V.

Molecular assays for detection of human metapneumovirus

The recent description of the respiratory pathogen human metapneumovirus (hMPV) has highlighted a deficiency in current diagnostic techniques for viral agents associated with acute lower respiratory tract infections. We describe two novel approaches to the detection of viral RNA by use of reverse transcriptase PCR (RT-PCR). The PCR products were identified after capture onto a solid-phase medium by hybridization with a sequence-specific, biotinylated oligonucleotide probe. The assay was applied to the screening of 329 nasopharyngeal aspirates sampled from patients suffering from respiratory tract disease. These samples were negative for other common microbial causes of respiratory tract disease. We were able to detect hMPV sequences in 32 (9.7%) samples collected from Australian patients during 2001. To further reduce result turnaround times we designed a fluorogenic TaqMan oligoprobe and combined it with the existing primers for use on the LightCycler platform. The real-time RT-PCR proved to be highly reproducible and detected hMPV in an additional 6 out of 62 samples (9.6%) tested during the comparison of the two diagnostic approaches. We found the real-time RT-PCR to be the test of choice for future investigation of samples for hMPV due to its speed, reproducibility, specificity, and sensitivity.

Lipid rafts and caveolae as portals for endocytosis: New insights and common mechanisms

Clathrin-coated pits and caveolae are two of the most recognizable features of the plasma membrane of mammalian cells. While our understanding of the machinery regulating and driving clathrin-coated pit-mediated endocytosis has progressed dramatically, including the elucidation of the structure of individual components and partial in vitro reconstitution, the role of caveolae as alternative endocytic carriers still remains elusive 50 years after their discovery. However, recent work has started to provide new insights into endocytosis by caveolae and into apparently related pathways involving lipid raft domains. These pathways, distinguished by their exquisite sensitivity to cholesterol-sequestering agents, can involve caveolae but also exist in cells devoid of caveolins and caveolae. This review examines the current evidence for the involvement of rafts and caveolae in endocytosis and the molecular players involved in their regulation.

The formative platform of the Congress of Panama (1810-1826): the Pan-American conjecture revisited

This article examines the formative platform of the Congress of Panama of 1826. It seeks to support the hypothesis that the nature and scope of the first test of integration in the Western Hemisphere depended critically on the platform created by Simón Bolívar and other Latin American Independence heroes from the Declaration of Independence of Venezuela in 1810 until the last bilateral agreement of 1826. In that respect, it corroborates the Latin American Identity of the initiative.

Sensor-driven agenda for intelligent home care of the elderly

The exponential increase of home-bound persons who live alone and are in need of continuous monitoring requires new solutions to current problems. Most of these cases present illnesses such as motor or psychological disabilities that deprive of a normal living. Common events such as forgetfulness or falls are quite common and have to be prevented or dealt with. This paper introduces a platform to guide and assist these persons (mostly elderly people) by providing multisensory monitoring and intelligent assistance. The platform operates at three levels. The lower level, denominated ‘‘Data acquisition and processing’’performs the usual tasks of a monitoring system, collecting and processing data from the sensors for the purpose of detecting and tracking humans. The aim is to identify their activities in an intermediate level called ‘‘activity detection’’. The upper level, ‘‘Scheduling and decision-making’’, consists of a scheduler which provides warnings, schedules events in an intelligent manner and serves as an interface to the rest of the platform. The idea is to use mobile and static sensors performing constant monitoring of the user and his/her environment, providing a safe environment and an immediate response to severe problems. A case study on elderly fall detection in a nursery home bedroom demonstrates the usefulness of the proposal.

A Platform to Emulate Ambient Assisted Living Environments

According to the opinion of clinicians, emerging medical conditions can be timely detected by observing changes in the activities of daily living and/or in the physiological signals of a person. To accomplish such purpose, it is necessary to properly monitor both the person’s physiological signals as well as the home environment with sensing technology. Wireless sensor networks (WSNs) are a promising technology for this support. After receiving the data from the sensor nodes, a computer processes the data and extracts information to detect any abnormality. The computer runs algorithms that should have been previously developed and tested in real homes or in living-labs. However, these installations (and volunteers) may not be easily available. In order to get around that difficulty, this paper suggests the making of a physical model to emulate basic actions of a user at home, thus giving autonomy to researchers wanting to test the performance of their algorithms. This paper also studies some data communication issues in mobile WSNs namely how the orientation of the sensor nodes in the body affects the received signal strength, as well as retransmission aspects of a TDMA-based MAC protocol in the data recovery process.

A Wireless EEG Acquisition Platform based on Embedded Systems

This paper proposes a wireless EEG acquisition platform based on Open Multimedia Architecture Platform (OMAP) embedded system. A high-impedance active dry electrode was tested for improving the scalp- electrode interface. It was used the sigma-delta ADS1298 analog-to-digital converter, and developed a “kernelspace” character driver to manage the communications between the converter unit and the OMAP’s ARM core. The acquired EEG signal data is processed by a “userspace” application, which accesses the driver’s memory, saves the data to a SD-card and transmits them through a wireless TCP/IP-socket to a PC. The electrodes were tested through the alpha wave replacement phenomenon. The experimental results presented the expected alpha rhythm (8-13 Hz) reactiveness to the eyes opening task. The driver spends about 725 μs to acquire and store the data samples. The application takes about 244 μs to get the data from the driver and 1.4 ms to save it in the SD-card. A WiFi throughput of 12.8Mbps was measured which results in a transmission time of 5 ms for 512 kb of data. The embedded system consumes about 200 mAh when wireless off and 400 mAh when it is on. The system exhibits a reliable performance to record EEG signals and transmit them wirelessly. Besides the microcontroller-based architectures, the proposed platform demonstrates that powerful ARM processors running embedded operating systems can be programmed with real-time constrains at the kernel level in order to control hardware, while maintaining their parallel processing abilities in high level software applications.

Molybdenum mixed with glyphosate and alone via foliar spray in no-tillage common bean grown on corn stover

The effect of molybdenum (Mo) on common bean grown in desiccated corn stover in a no-tillage system was evaluated under two application modes: Mo mixed with the desiccant glyphosate and Mo direct spray to the bean leaves. The treatments (four replicates) were assigned to a completely randomized block design in a split-plot arrangement with the application of Mo (0, 100, 200, 400 and 800 g ha-1) mixed with glyphosate in the main plots and Mo foliar spray (0 and 100 g ha-1) in the sub-plots. The field experiments were carried out in 2009 and 2010 in the municipality of Coimbra, Minas Gerais State, with the common bean cultivar Ouro Vermelho. Mo mixed with glyphosate had neither an effect on common bean yield nor on the Mo and N contents in leaves, however it increased the Mo and N contents in seeds. Application of Mo via foliar spray increased Mo content in leaves and Mo and N contents in seeds. The reapplication of molybdenum with glyphosate for desiccation in subsequent crops caused a cumulative effect of Mo content in bean seeds.

Selection of common bean lines with high grain yield and high grain calcium and iron concentrations

Genetic improvement of common bean nutritional quality has advantages in marketing and can contribute to society as a food source. The objective of this study was to evaluate the genetic variability for grain yield, calcium and iron concentrations in grains of inbred common bean lines obtained by different breeding methods. For this, 136 F7 inbred lines were obtained using the Pedigree method and 136 F7 inbred lines were obtained using the Single-Seed Descent (SSD) method. The lines showed genetic variability for grain yield, and concentrations of calcium and iron independently of the method of advancing segregating populations. The Pedigree method allows obtaining a greater number of lines with high grain yield. Selection using the SSD method allows the identification of a larger number of lines with high concentrations of calcium and iron in grains. Weak negative correlations were found between grain yield and calcium concentration (r = -0.0994) and grain yield and iron concentration (r = -0.3926). Several lines show genetic superiority for grain yield and concentrations of calcium and iron in grains and their selection can result in new common bean cultivars with high nutritional quality.

A Caregiver Support Platform within the Scope of an Ambient Assisted Living Ecosystem

The Ambient Assisted Living (AAL) area is in constant evolution, providing new technologies to users and enhancing the level of security and comfort that is ensured by house platforms. The Ambient Assisted Living for All (AAL4ALL) project aims to develop a new AAL concept, supported on a unified ecosystem and certification process that enables a heterogeneous environment. The concepts of Intelligent Environments, Ambient Intelligence, and the foundations of the Ambient Assisted Living are all presented in the framework of this project. In this work, we consider a specific platform developed in the scope of AAL4ALL, called UserAccess. The architecture of the platform and its role within the overall AAL4ALL concept, the implementation of the platform, and the available interfaces are presented. In addition, its feasibility is validated through a series of tests.

Selection of anthracnose resistant common beans using detached leaves in partially controlled environment

The objectives of this study were to evaluate the possibility of selecting anthracnose resistant common bean plants using detached primary leaves in partially controlled environment of a greenhouse and identify differences in the reaction of genotypes to anthracnose. The common bean cultivars Ouro Negro, OuroVermelho, ManteigãoFosco 11, Rudá, Rudá-R, VP8, BRSMG Madrepérola, Pérola, MeiaNoite and BRSMG Talismãwere characterizedfor resistance to the races 65, 81 and 453 of Colletotrichum lindemuthianum and the method of detached primary leaves was compared to the method with the traditional inoculation of plants at the phenological stage V2. The lines Rudá, Rudá-R and Pérola were inoculated with the races 65 and 453 of C. lindemuthianum, aiming to assess the rate of coincidence of anthracnose severity by both inoculation methods. In general, the two methods presented similar results for the reaction of the cultivars. The use of detached primary leaves of common bean plants in the partially controlled environment was feasible for selection of plants resistant to anthracnose and has the advantages of low-needed infrastructure and reduction of resources, space and time.

Agronomic performance of common bean in straw mulch systems and topdressing nitrogen rates in no-tillage

ABSTRACTIn no-tillage systems, straw coverage on soil surface is the key to success, and the choice of crops for rotation is crucial to achieve the sustainability and quality that conservation agriculture requires. The objective of this study was to evaluate the agronomic performance of the common bean cultivar IAC Formoso sown in succession to three straw mulch systems (corn alone, corn/Urochloa ruziziensisintercrop and U. ruziziensisalone) and topdress nitrogen rates (0; 40; 80; 120 and 160 kg ha-1N), at the four-leaf stage, three years after the implementation of no-tillage. The experiment was arranged in a randomized block split plot design, with three replications. Common bean highest yields were achieved in succession to U. ruziziensisalone and intercropped with corn. The corn/U. ruziziensisintercrop provided both straw and seed production, allowing for quality no-tillage. Topdressed nitrogen influenced the common bean yield when in succession to corn alone, U. ruziziensisalone and corn/U. ruziziensisintercrop in no-tillage.