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Labor Intensive (labor + intensive)
Selected AbstractsNovel application of flow cytometry: Determination of muscle fiber types and protein levels in whole murine skeletal muscles and heartCYTOSKELETON, Issue 12 2007Connie Jackaman Abstract Conventional methods for measuring proteins within muscle samples such as immunohistochemistry and western blot analysis can be time consuming, labor intensive and subject to sampling errors. We have developed flow cytometry techniques to detect proteins in whole murine heart and skeletal muscle. Flow cytometry and immunohistochemistry were performed on quadriceps and soleus muscles from male C57BL/6J, BALB/c, CBA and mdx mice. Proteins including actins, myosins, tropomyosin and ,-actinin were detected via single staining flow cytometric analysis. This correlated with immunohistochemistry using the same antibodies. Muscle fiber types could be determined by dual labeled flow cytometry for skeletal muscle actin and different myosins. This showed similar results to immunohistochemistry for I, IIA and IIB myosins. Flow cytometry of heart samples from C57BL/6J and BALB/c mice dual labeled with cardiac and skeletal muscle actin antibodies demonstrated the known increase in skeletal actin protein in BALB/c hearts. The membrane-associated proteins ,-sarcoglycan and dystrophin could be detected in C57BL/6J mice, but were decreased or absent in mdx mice. With the ability to label whole muscle samples simultaneously with multiple antibodies, flow cytometry may have advantages over conventional methods for certain applications, including assessing the efficacy of potential therapies for muscle diseases. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Early pyrotechnology in the Near East: Experimental lime-plaster production at the Pre-Pottery Neolithic B site of Kfar HaHoresh, IsraelGEOARCHAEOLOGY: AN INTERNATIONAL JOURNAL, Issue 6 2008Y. Goren A characteristic hallmark of the Pre-Pottery Neolithic B (PPNB) in the southern Levant was the extensive use of lime plaster for architectural and other purposes. Yet no obvious kilns have been identified in archaeological contexts. Here we present details of an experimental pit-kiln modeling lime-plaster production based on observed burnt stone accumulations in pits at the PPNB site of Kfar HaHoresh in the lower Galilee. The experimental kiln was loaded in layers with ,500 kg of limestone (pebbles and stones) and ,1000 kg of fuel (branches and dung). Fired for 24 hours, and reaching a maximum 870°C, the kiln yielded almost 250 kg of quicklime (calcium oxide, CaO). Micromorphological samples, general observations, and scaled plan view drawings made immediately following and nine years after ignition demonstrate that the original shape of the kiln and residual quicklime within and around it rapidly dissipated through bioturbation, trampling by animals, erosion, rain, and exposure to the elements. This could account for the seeming absence of kilns within sites, although they were probably located close to where lime-plaster was applied, given the unstable nature and toxic effects of handling quicklime. Calculations of the manpower and fuel involved indicate that PPNB lime-plaster production may have been less labor intensive and less detrimental to the environment than previously portrayed. © 2008 Wiley Periodicals, Inc. [source] ATM mutations on distinct SNP and STR haplotypes in ataxia-telangiectasia patients of differing ethnicities reveal ancestral founder effects,HUMAN MUTATION, Issue 1 2003Catarina Campbell Abstract Due to the large size (150 kb) of the ataxia-telangiectasia mutated (ATM) gene and the existence of over 400 mutations, identifying mutations in patients with ataxia-telangiectasia (A-T) is labor intensive. We compared the SNP and STR haplotypes of A-T patients from varying ethnicities who were carrying common ATM mutations. We used SSCP to determine SNP haplotypes. To our surprise, all of the most common ATM mutations in our large multiethnic cohort were associated with specific SNP haplotypes, whereas the STR haplotypes varied, suggesting that ATM mutations predated STR haplotypes but not SNP haplotypes. We conclude that these frequently observed ATM mutations are not hot spots, but have occurred only once and spread with time to different ethnic populations. More generally, a combination of SNP and STR haplotyping could be used as a screening strategy for identifying mutations in other large genes by first determining the ancestral SNP and STR haplotypes in order to identify specific founder mutations. We estimate this approach will identify approximately 30% of mutations in A-T patients across all ethnic groups. Hum Mutat 21:80,85, 2002. © 2002 Wiley-Liss, Inc. [source] The chick chorioallantoic membrane as a novel in vivo model for the testing of biomaterialsJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2002T.I. Valdes Abstract Current in vivo models for testing biomaterials are time and labor intensive as well as expensive. This article describes a new approach for testing biomaterials in vivo using the chorioallantoic membrane (CAM) of the developing chicken embryo, as an alternative to the traditional mammalian models. Fertilized chicken eggs were incubated for 4 days, at which time a small window was cut in the shell of the egg. After 1 week of incubation, the CAM received several test materials, including the endotoxin LPS, a cotton thread and a Silastic tubing. One day and 1 week later, the tissue response to the test materials was assessed using gross, histological, and scanning electron microscope evaluations. The inflammatory response of the chorioallantoic membrane to biomaterials was fully characterized and found to be similar to that of the mammalian response and was also seen to vary according to test materials. We also found that the structure and geometry of the test materials greatly influenced the incorporation of the samples in the CAM. The similarity of the tissue response of the CAM with the mammalian models, plus the low cost, simplicity, and possibility to continuously visualize the test site through the shell window make this animal model particularly attractive for the rapid in vivo screening of biomaterials. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 62: 273,282, 2002 [source] Validating the use of temperature data loggers to measure survival of songbird nestsJOURNAL OF FIELD ORNITHOLOGY, Issue 4 2006Karel Weidinger ABSTRACT Accurate determination of nest fates and nest predators is possible through continuous video monitoring, but such monitoring is relatively expensive and labor intensive. If documenting of the timing of nest termination events is sufficient, then data loggers (DL) may allow more extensive sampling and may represent a viable alternative. I validated temperature DL records of nest survival time by simultaneous videotaping and compared results derived from DL records with those obtained by regular nest visits by an observer. I estimated the fate of 937 nests of nine species of open cup-nesting songbirds, including 673 nests monitored using DL, 165 monitored using video cameras, 33 validation nests monitored simultaneously using both DL and video cameras, and 132 control nests monitored only by observer visits. Deployment of DL did not negatively influence nest survival rate. DL reliably recorded survival time and allowed classification of nest fates based on the potential fledging age, regardless of the frequency of nest visits by an observer. The true fate of nests that survived beyond the potential fledging age can not be safely determined from time of failure, except for nocturnal events that suggest partial predation. Video revealed frequent partial or complete predation on nests with old nestlings that would have been categorized as successful by other methods. I conclude that temperature DL are efficient, reliable, and relatively inexpensive tools for recording exact nest survival times and classification of nest fates, with implications for nest survival modeling and discriminating between diurnal and nocturnal predation. SINOPSIS Es posible determinar con precisión la sobrevicencia en nidos y la depredación en estos mediante el uso de videos contínuos. Pero dicho monitoreo es relativamente costoso y requiere mucho trabajo. Si el documentar el momento en que se termina el anidamiento es suficiente para obtener la información, previamente mencionada, el uso de bitácoras electrónicas de temperatura (loggers) pudieran permitir el tomar muestras más amplias y por ende, representar una alternativa viable. Validé la toma de temperaturas con bitácoras electrónicas para determinar la sobrevivencia en nidos con la toma simultánea de videos y comparé los resultados obtenidos (con la bitácora) con los datos tomados por un observador que visitó regularmente los nidos. El estudio se hizo con 937 nidos de nueve especies de aves canoras cuyo nido es en forma de copa. De estos 673 se monitorearon utilizando bitácoras electrónicas de temperatura, 165 con cámaras de video, 33 con monitoreo simultáneo de bitacora y video y 132, como control, monitoreados mediante observación directa. El uso de las bitácoras no influyó negativamente en la tasa de sobrevivencia. La bitácora grabó el tiempo de sobrervivencia y permitió la clasificación de los nidos (exitoso o no exitoso) basado en el tiempo potencial de la edad de dejar el nido, sin importar la frecuencia de visita a los nidos por parte de observadores. La verdadera finalidad de los nidos que sobreviven, más alla de la edad potencial de dejar el nido los pichones, no puede ser determinado con exactitud, excepto en eventos nocturnos que surgieren depredación. El uso de videos permitió determinar la depredación parcial o completa en nidos, particularmente de pichones que se tardaron más que el tiempo promedio en dejar el nido y que en estudios se asume que sobrevivieron. Puedo concluir que las bitácoras electrónicas de temperatura son eficientes, confiables, de bajo costo y permiten determinar con precisión la sobrevivencia en nidos y la clasificación de estos entre exitosos y no exitosos, con implicaciones para construir modelos de sobrevivencia y discriminar entre depredores diurnos y nocturnos. [source] Perspective on Bifurcation PCIJOURNAL OF INTERVENTIONAL CARDIOLOGY, Issue 2 2009SHAO-LIANG CHEN M.D. Coronary bifurcation lesion is a complex lesion with suboptimal angiographic and clinical results. There has been no satisfactory classification of the lesion that can guide selection of strategies and predict short- and long-term outcomes. The difference between left main (LM) bifurcation lesions and non-LM bifurcation is striking. So many stenting strategies have been proposed and tried in trials. They include the V, T, Y, one-stent, two-stent, crush, mini-crush, DK, and SKS techniques. However, because these techniques are time and labor intensive, dedicated bifurcated stents have been invented and trialed in humans. This review presents a historical perspective of interventions in bifurcated lesions, with the strengths and weaknesses of the major strategies and of the new dedicated stents. [source] Technical note: An R program for automating bone cross section reconstructionAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 4 2010Adam D. Sylvester Abstract Many recent studies have used long bone cross-sectional geometric properties in various comparative analyses. Methods have been described for reconstructing diaphyseal cross sections from external molds and biplanar radiographs that produce accurate results (within 5% of true values on average). The manual image processing required, however, is both time and labor intensive. A new freely available program developed here for the computational freeware, R, automates much of the process. This study compares cross-sectional properties calculated using the new R program to those from peripheral quantitative CT (pQCT) and the original manual method. We find that the R program works aswell as the original manual image processing for most cross sections eliminates the chance for entry errors at several steps and greatly speeds up data collection. Am J Phys Anthropol 142:665,669, 2010. © 2010 Wiley-Liss, Inc. [source] Large-Scale Zostera marina (eelgrass) Restoration in Chesapeake Bay, Maryland, USA.RESTORATION ECOLOGY, Issue 4 2010Associated Costs, Part I: A Comparison of Techniques The Chesapeake Bay, like many other temperate estuaries, has exhibited dramatic declines in the abundance of submerged aquatic vegetation (SAV) during the later half of the twentieth century. Because of the functions SAV serve in maintaining a healthy estuarine ecosystem, SAV restoration has become an important component of Chesapeake Bay restoration. Specifically, recent water quality improvements in areas from which populations of Zostera marina (eelgrass) have been extirpated have suggested that Z. marina restoration could succeed. Early restoration efforts involved transplanting Z. marina plants from healthy source beds to restoration locations, but this was labor intensive, time consuming, expensive, and potentially detrimental to donor beds. This multi-year project investigated new techniques for large-scale Z. marina seed collection and processing and compared two seed dispersal methods to evaluate cost effectiveness. Tens of millions of mature Z. marina seeds were collected through snorkeling, SCUBA, or with a mechanical harvester. Seed storage conditions and processing techniques were manipulated in order to maximize seed yield. Seeds were dispersed using two methods: spring seed buoys and fall seed broadcasts. Our costs for planting 1 ha of bottom with Z. marina seeds ranged from $6,674 to $165,699 depending on seeding density and dispersal method used. The average cost per Z. marina seed was $0.17. Interannual variations in seed collection yield and seed viability after summer storage had great impact on final costs. Our results suggest that the use of seeds for large-scale Z. marina restoration offers a competitive advantage to more traditional transplanting methods. [source] In this issue: Biotechnology Journal 11/2009BIOTECHNOLOGY JOURNAL, Issue 11 2009Article first published online: 13 NOV 200 Forensic identification on chips Choi and Seo et al., Biotechnol. J. 2009, 4, 1530,1541 Short tandem repeat (STR) analysis can be used for genetic fingerprinting of individuals as it is done for forensic human identification. However, the current state-of-the-art STR genotyping processes and instruments are labor intensive, expensive, time consuming, and lack portability. Micro-total-analysis systems or lab-on-a-chip platforms based on microfabrication technologies have the capability to miniaturize and integrate bioanalysis steps in a single format and have already been successfully applied for forensic STR typing. Researchers from Daejeon, Korea, highlight up-to-date work on advanced microdevices for high-throughput STR genotyping, and a portable integrated microsystem for on-site forensic DNA analysis. Surface plasmon resonance on chips Maynard et al., Biotechnol. J. 2009, 4, 1542,1558 Technologies based on surface plasmon resonance (SPR) have allowed rapid, label-free characterization of protein-protein and protein-small molecule interactions. SPR has become the gold standard in industrial and academic settings, in which the interaction between a pair of soluble binding partners is characterized in detail or a library of molecules is screened for binding against a single soluble protein. In spite of these successes, SPR is only beginning to be adapted to the needs of membrane-bound proteins which are promising targets for drug and biomarker development. This team of authors from Austin, Minneapolis and Rochester (all USA) describe current SPR instrumentation and the potential for SPR nanopore arrays to enable quantitative, high-throughput screening of G-protein coupled receptor ligands and applications in cellular biology. Nucleotide immobilization on chips Sethi et al., Biotechnol. J. 2009, 4, 1513,1529 The development of oligonucleotide-based microarrays (biochips) is of major interest in science and biotechnology industry and has applications in a wide range of research areas including genomics, proteomics, computational biology and pharmaceuticals. Especially microarrays have proven to be a unique method for time and cost efficient analysis of thousands of genes at one. Authors from Delhi and Lucknow, India discuss currently used chemical strategies for immobilization of oligonucleotides and put a special emphasis on post-synthetic immobilization on glass surfaces. Recent advances on these synthesis pathways are presented in detail. [source] An integrated microdevice for high-performance short tandem repeat genotypingBIOTECHNOLOGY JOURNAL, Issue 11 2009Jong Young Choi Abstract Short tandem repeat (STR) analysis provides genetic fingerprinting of individuals, and is considered as a powerful and indispensable technique for forensic human identification. However, the current state-of-the-art STR genotyping processes and instruments are labor intensive, expensive, time consuming, and lack portability. Micro-total-analysis systems or lab-on-a-chip platforms based on microfabrication technologies have the capability to miniaturize and integrate bioanalysis steps in a single format. Recent progress in microsystems has demonstrated their successful performance for the forensic STR typing with a reduced cost, high speed, and improved high throughput. The purpose of this review article is to highlight up-to-date work on advanced microdevices for high-throughput STR genotyping, and a portable integrated microsystem for on-site forensic DNA analysis. [source] | ||||||||||