943 resultados para Biotechnology laboratories
Resumo:
A straightforward procedure for the acid digestion of geological samples with SiO2 concentrations ranging between about 40 to 80%, is described. A powdered sample (200 mesh) of 500 mg was used and fused with 1000 mg spectroflux at about 1000 degreesC in a platinum crucible. The molten was subsequently digested in an aqueous solution of HNO3 at 100 degreesC. Several systematic digestion procedures were followed using various concentrations of HNO3. It was found that a relationship could be established between the dissolution-time and acid concentration. For an acid concentration of 15% an optimum dissolution-time of under 4 min was recorded. To verify that the dissolutions were complete, they were subjected to rigorous quality control tests. The turbidity and viscosity were examined at different intervals and the results were compared with that of deionised water. No significant change in either parameter was observed. The shelf-life of each solution lasted for several months, after which time polymeric silicic acid formed in some solutions, resulting in the presence of a gelatinous solid. The method is cost effective and is clearly well suited for routine applications on a small scale, especially in laboratories in developing countries. ICP-MS was applied to the determination of 13 Rare Earth Elements and Hf in a set of 107 archaeological samples subjected to the above digestion procedure. The distribution of these elements was examined and the possibility of using the REE's for provenance studies is discussed.
Resumo:
Background & Research Focus Managing knowledge for innovation and organisational benefit has been extensively investigated in studies of large firms (Smith, Collins & Clark, 2005; Zucker, et al., 2007) and to a large extent there is limited research into studies of small- and medium- sized enterprises (SMEs). There are some investigations in knowledge management research on SMEs, but what remains to be seen in particular is the question of where are the potential challenges for managing knowledge more effectively within these firms? Effective knowledge management (KM) processes and systems lead to improved performance in pursuing distinct capabilities that contribute to firm-level innovation (Nassim 2009; Zucker et al. 2007; Verona and Ravasi 2003). Managing internal and external knowledge in a way that links it closely to the innovation process can assist the creation and implementation of new products and services. KM is particularly important in knowledge intensive firms where the knowledge requirements are highly specialized, diverse and often emergent. However, to a large extent the KM processes of small firms that are often the source of new knowledge and an important element of the value networks of larger companies have not been closely studied. To address this gap which is of increasing importance with the growing number of small firms, we need to further investigate knowledge management processes and the ways that firms find, capture, apply and integrate knowledge from multiple sources for their innovation process. This study builds on the previous literature and applies existing frameworks and takes the process and activity view of knowledge management as a starting point of departure (see among others Kraaijenbrink, Wijnhoven & Groen, 2007; Enberg, Lindkvist, & Tell, 2006; Lu, Wang & Mao, 2007). In this paper, it is attempted to develop a better understanding of the challenges of knowledge management within the innovation process in small knowledge-oriented firms. The paper aims to explore knowledge management processes and practices in firms that are engaged in the new product/service development programs. Consistent with the exploratory character of the study, the research question is: How is knowledge integrated, sourced and recombined from internal and external sources for innovation and new product development? Research Method The research took an exploratory case study approach and developed a theoretical framework to investigate the knowledge situation of knowledge-intensive firms. Equipped with the conceptual foundation, the research adopted a multiple case study method investigating four diverse Australian knowledge-intensive firms from IT, biotechnology, nanotechnology and biochemistry industries. The multiple case study method allowed us to document in some depth the knowledge management experience of the theses firms. Case study data were collected through a review of company published data and semi-structured interviews with managers using an interview guide to ensure uniform coverage of the research themes. This interview guide was developed following development of the framework and a review of the methodologies and issues covered by similar studies in other countries and used some questions common to these studies. It was framed to gather data around knowledge management activity within the business, focusing on the identification, acquisition and utilisation of knowledge, but collecting a range of information about subject as well. The focus of the case studies was on the use of external and internal knowledge to support their knowledge intensive products and services. Key Findings Firstly a conceptual and strategic knowledge management framework has been developed. The knowledge determinants are related to the nature of knowledge, organisational context, and mechanism of the linkages between internal and external knowledge. Overall, a number of key observations derived from this study, which demonstrated the challenges of managing knowledge and how important KM is as a management tool for innovation process in knowledge-oriented firms. To summarise, findings suggest that knowledge management process in these firms is very much project focused and not embedded within the overall organisational routines and mainly based on ad hoc and informal processes. Our findings highlighted lack of formal knowledge management process within our sampled firms. This point to the need for more specialised capabilities in knowledge management for these firms. We observed a need for an effective knowledge transfer support system which is required to facilitate knowledge sharing and particularly capturing and transferring tacit knowledge from one team members to another. In sum, our findings indicate that building effective and adaptive IT systems to manage and share knowledge in the firm is one of the biggest challenges for these small firms. Also, there is little explicit strategy in small knowledge-intensive firms that is targeted at systematic KM either at the strategic or operational level. Therefore, a strategic approach to managing knowledge for innovation as well as leadership and management are essential to achieving effective KM. In particular, research findings demonstrate that gathering tacit knowledge, internal and external to the organization, and applying processes to ensure the availability of knowledge for innovation teams, drives down the risks and cost of innovation. KM activities and tools, such as KM systems, environmental scanning, benchmarking, intranets, firm-wide databases and communities of practice to acquire knowledge and to make it accessible, were elements of KM. Practical Implications The case study method that used in this study provides practical insight into the knowledge management process within Australian knowledge-intensive firms. It also provides useful lessons which can be used by other firms in managing the knowledge more effectively in the innovation process. The findings would be helpful for small firms that may be searching for a practical method for managing and integrating their specialised knowledge. Using the results of this exploratory study and to address the challenges of knowledge management, this study proposes five practices that are discussed in the paper for managing knowledge more efficiently to improve innovation: (1) Knowledge-based firms must be strategic in knowledge management processes for innovation, (2) Leadership and management should encourage various practices for knowledge management, (3) Capturing and sharing tacit knowledge is critical and should be managed, (4)Team knowledge integration practices should be developed, (5) Knowledge management and integration through communication networks, and technology systems should be encouraged and strengthen. In sum, the main managerial contribution of the paper is the recognition of knowledge determinants and processes, and their effects on the effective knowledge management within firm. This may serve as a useful benchmark in the strategic planning of the firm as it utilises new and specialised knowledge.
Resumo:
Human saliva harbours proteins of clinical relevance and about 30% of blood proteins are also present in saliva. This highlights that saliva can be used for clinical applications just as urine or blood. However, the translation of salivary biomarker discoveries into clinical settings is hampered by the dynamics and complexity of the salivary proteome. This review focuses on the current status of technological developments and achievements relating to approaches for unravelling the human salivary proteome. We discuss the dynamics of the salivary proteome, as well as the importance of sample preparation and processing techniques and their influence on downstream protein applications; post-translational modifications of salivary proteome and protein: protein interactions. In addition, we describe possible enrichment strategies for discerning post-translational modifications of salivary proteins, the potential utility of selected-reaction-monitoring techniques for biomarker discovery and validation, limitations to proteomics and the biomarker challenge and future perspectives. In summary, we provide recommendations for practical saliva sampling, processing and storage conditions to increase the quality of future studies in an emerging field of saliva clinical proteomics. We propose that the advent of technologies allowing sensitive and high throughput proteome-wide analyses, coupled to well-controlled study design, will allow saliva to enter clinical practice as an alternative to blood-based methods due to its simplistic nature of sampling, non-invasiveness, easy of collection and multiple collections by untrained professionals and cost-effective advantages.
Resumo:
The measurements of plasma natriuretic peptides (NT-proBNP, proBNP and BNP) are used to diagnose heart failure but these are expensive to produce. We describe a rapid, cheap and facile production of proteins for immunoassays of heart failure. DNA encoding N-terminally His-tagged NT-proBNP and proBNP were cloned into the pJexpress404 vector. ProBNP and NT-proBNP peptides were expressed in Escherichia coli, purified and refolded in vitro. The analytical performance of these peptides were comparable with commercial analytes (NT-proBNP EC50 for the recombinant is 2.6 ng/ml and for the commercial material is 5.3 ng/ml) and the EC50 for recombinant and commercial proBNP, are 3.6 and 5.7 ng/ml respectively). Total yield of purified refolded NT-proBNP peptide was 1.75 mg/l and proBNP was 0.088 mg/l. This approach may also be useful in expressing other protein analytes for immunoassay applications. To develop a cost effective protein expression method in E. coli to obtain high yields of NT-proBNP (1.75 mg/l) and proBNP (0.088 mg/l) peptides for immunoassay use.
Resumo:
This article considers the extent to which a claimed process must be repeatable or reproducible in order to be patentable according to Australian patent law. It asks whether a process must yield identical or near-identical results each time the process is invoked, or if not, what degree of repeatability is required. The question is relevant when considering, among other things, the patentability of some methods of medical treatment and diagnosis, biotechnology inventions and business methods.
Resumo:
The biological impact of Rho depends critically on the precise subcellular localization of its active, GTP-loaded form. This can potentially be determined by the balance between molecules that promote nucleotide exchange or GTP hydrolysis. However, how these activities may be coordinated is poorly understood. We now report a molecular pathway that achieves exactly this coordination at the epithelial zonula adherens. We identify an extramitotic activity of the centralspindlin complex, better understood as a cytokinetic regulator, which localizes to the interphase zonula adherens by interacting with the cadherin-associated protein, α-catenin. Centralspindlin recruits the RhoGEF, ECT2, to activate Rho and support junctional integrity through myosin IIA. Centralspindlin also inhibits the junctional localization of p190 B RhoGAP, which can inactivate Rho. Thus, a conserved molecular ensemble that governs Rho activation during cytokinesis is used in interphase cells to control the Rho GTPase cycle at the zonula adherens
Resumo:
Directional cell migration requires force generation that relies on the coordinated remodeling of interactions with the extracellular matrix (ECM), which is mediated by integrin-based focal adhesions (FAs). Normal FA turnover requires dynamic microtubules, and three members of the diverse group of microtubule plus-end-tracking proteins are principally involved in mediating microtubule interactions with FAs. Microtubules also alter the assembly state of FAs by modulating Rho GTPase signaling, and recent evidence suggests that microtubule-mediated clathrin-dependent and -independent endocytosis regulates FA dynamics. In addition, FA-associated microtubules may provide a polarized microtubule track for localized secretion of matrix metalloproteases (MMPs). Thus, different aspects of the molecular mechanisms by which microtubules control FA turnover in migrating cells are beginning to emerge.
Resumo:
The palette of fluorescent proteins (FPs) has grown exponentially over the past decade, and as a result, live imaging of cells expressing fluorescently tagged proteins is becoming more and more mainstream. Spinning disk confocal (SDC) microscopy is a high-speed optical sectioning technique and a method of choice to observe and analyze intracellular FP dynamics at high spatial and temporal resolution. In an SDC system, a rapidly rotating pinhole disk generates thousands of points of light that scan the specimen simultaneously, which allows direct capture of the confocal image with low-noise scientific grade-cooled charge-coupled device cameras, and can achieve frame rates of up to 1000 frames per second. In this chapter, we describe important components of a state-of-the-art spinning disk system optimized for live cell microscopy and provide a rationale for specific design choices. We also give guidelines of how other imaging techniques such as total internal reflection microscopy or spatially controlled photoactivation can be coupled with SDC imaging and provide a short protocol on how to generate cell lines stably expressing fluorescently tagged proteins by lentivirus-mediated transduction.
Resumo:
Cross-talk between microtubule networks and sites of cell-matrix and cell-cell adhesion has profound impact on these structures and is essential for proper cell organization, polarization and motility. Components of adhesion sites can interact directly with microtubules or with proteins that specifically associate with microtubule plus ends and minus ends and in this way capture, stabilize or destabilize microtubules. In their turn, microtubules can serve as routes for delivery of structural and regulatory factors that control adhesion site turnover. In addition, the microtubule lattice or growing microtubule plus ends can serve as diffusional sinks that accumulate and scaffold regulatory molecules, thereby affecting their activity in the vicinity of adhesions. Combination of these mechanisms underlies the functional co-operation between microtubules and adhesion sites and defines their dynamic behavior.
Resumo:
Classical cadherins are fundamental determinants of tissue organization both in health and disease. It has long been recognized that cadherins function in close cooperation with the cytoskeleton, particularly with actin. Less appreciated is the capacity for cadherins to also interact functionally and biochemically with microtubules and their associated proteins. In this review, we aim to highlight the potential for cooperativity between cadherins and microtubules. Cadherins can regulate the organization and dynamics of microtubules through mechanisms such as anchorage of minus ends and cortical capture of plus ends. Such cadherin-induced reorganization of microtubules may then affect cadherin biology by diverse processes that include directed vesicular traffic by microtubule-based motors and regulation of cortical signaling and organization. Ultimately, we hope this will stimulate fresh interest and research to understand a neglected partnership.
Resumo:
The prospect of economically producing useful biologics in plants has greatly increased with the advent of viral vectors. The ability of viral vectors to amplify transgene expression has seen them develop into robust transient platforms for the high-level, rapid production of recombinant proteins. To adapt these systems to stably transformed plants, new ways of deconstructing the virus machinery and linking its expression and replication to chemically controlled promoters have been developed. The more advanced of these stable, inducible hyper-expression vectors provide both activated and amplified heterologous transgene expression. Such systems could be deployed in broad acre crops and provide a pathway to fully exploit the advantages of plants as a platform for the manufacture of a wide spectrum of products.
Resumo:
The transcriptome response of Atlantic salmon (Salmo salar) displaying advanced stages of amoebic gill disease (AGD) was investigated. Naïve smolt were challenged with AGD for 19 days, at which time all fish were euthanized and their severity of infection quantified through histopathological scoring. Gene expression profiles were compared between heavily infected and naïve individuals using a 17 K Atlantic salmon cDNA microarray with real-time quantitative RT-PCR (qPCR) verification. Expression profiles were examined in the gill, anterior kidney, and liver. Twenty-seven transcripts were significantly differentially expressed within the gill; 20 of these transcripts were down-regulated in the AGD-affected individuals compared with naïve individuals. In contrast, only nine transcripts were significantly differentially expressed within the anterior kidney and five within the liver. Again the majority of these transcripts were down-regulated within the diseased individuals. A down-regulation of transcripts involved in apoptosis (procathepsin L, cathepsin H precursor, and cystatin B) was observed in AGD-affected Atlantic salmon. Four transcripts encoding genes with antioxidant properties also were down-regulated in AGD-affected gill tissue according to qPCR analysis. The most up-regulated transcript within the gill was an unknown expressed sequence tag (EST) whose expression was 218-fold (± SE 66) higher within the AGD affected gill tissue. Our results suggest that Atlantic salmon experiencing advanced stages of AGD demonstrate general down-regulation of gene expression, which is most pronounced within the gill. We propose that this general gene suppression is parasite-mediated, thus allowing the parasite to withstand or ameliorate the host response. © 2008 Springer Science+Business Media, LLC.
Resumo:
When crystallization screening is conducted many outcomes are observed but typically the only trial recorded in the literature is the condition that yielded the crystal(s) used for subsequent diffraction studies. The initial hit that was optimized and the results of all the other trials are lost. These missing results contain information that would be useful for an improved general understanding of crystallization. This paper provides a report of a crystallization data exchange (XDX) workshop organized by several international large-scale crystallization screening laboratories to discuss how this information may be captured and utilized. A group that administers a significant fraction of the worlds crystallization screening results was convened, together with chemical and structural data informaticians and computational scientists who specialize in creating and analysing large disparate data sets. The development of a crystallization ontology for the crystallization community was proposed. This paper (by the attendees of the workshop) provides the thoughts and rationale leading to this conclusion. This is brought to the attention of the wider audience of crystallographers so that they are aware of these early efforts and can contribute to the process going forward. © 2012 International Union of Crystallography All rights reserved.
Resumo:
A laboratory with fully developed and accredited quality systems is highly desirable. In some cases it is mandatory. This is evidenced by the concerns raised when certain medical testing laboratories have not maintained appropriate standards to retain their accreditation with NATA. Whether or not third party accreditation is mandated, good laboratory practice based on sustainable systems is required. Internationally accepted standard ISO/IEC 17025 provides specific guidelines for operating a laboratory that may be accredited. To what extent should a university laboratory implement this standard? Is NATA accreditation necessary? As staffing and funding reduce is it possible to maintain high standard quality systems that are in reality truly required if assurance of results is due? Accreditation may be required where accreditation is due but is credit given by the institutions for the resources required to get there or stay there? These issues are considered in relation to university and commercial laboratories and specific case points are raised from experience with NATA accreditation in the School of Civil Engineering, QUT.