Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.

Budget-aware role based access control

The suitability of Role Based Access Control (RBAC) is being challenged in dynamic environments like healthcare. In an RBAC system, a user's legitimate access may be denied if their need has not been anticipated by the security administrator at the time of policy specification. Alternatively, even when the policy is correctly specified an authorised user may accidentally or intentionally misuse the granted permission. The heart of the challenge is the intrinsic unpredictability of users' operational needs as well as their incentives to misuse permissions. In this paper we propose a novel Budget-aware Role Based Access Control (B-RBAC) model that extends RBAC with the explicit notion of budget and cost, where users are assigned a limited budget through which they pay for the cost of permissions they need. We propose a model where the value of resources are explicitly defined and an RBAC policy is used as a reference point to discriminate the price of access permissions, as opposed to representing hard and fast rules for making access decisions. This approach has several desirable properties. It enables users to acquire unassigned permissions if they deem them necessary. However, users misuse capability is always bounded by their allocated budget and is further adjustable through the discrimination of permission prices. Finally, it provides a uniform mechanism for the detection and prevention of misuses.

Collective dynamics and control of a 3-D small-world network with time delays

The paper presents a detailed analysis on the collective dynamics and delayed state feedback control of a three-dimensional delayed small-world network. The trivial equilibrium of the model is first investigated, showing that the uncontrolled model exhibits complicated unbounded behavior. Then three control strategies, namely a position feedback control, a velocity feedback control, and a hybrid control combined velocity with acceleration feedback, are then introduced to stabilize this unstable system. It is shown in these three control schemes that only the hybrid control can easily stabilize the 3-D network system. And with properly chosen delay and gain in the delayed feedback path, the hybrid controlled model may have stable equilibrium, or periodic solutions resulting from the Hopf bifurcation, or complex stranger attractor from the period-doubling bifurcation. Moreover, the direction of Hopf bifurcation and stability of the bifurcation periodic solutions are analyzed. The results are further extended to any "d" dimensional network. It shows that to stabilize a "d" dimensional delayed small-world network, at least a "d – 1" order completed differential feedback is needed. This work provides a constructive suggestion for the high dimensional delayed systems.

Protein monoubiquitination and polyubiquitination generate structural diversity to control distinct biological processes

Ubiquitination involves the attachment of ubiquitin (Ub) to lysine residues on substrate proteins or itself, which can result in protein monoubiquitination or polyubiquitination. Polyubiquitination through different lysines (seven) or the N-terminus of Ub can generate different protein-Ub structures. These include monoubiquitinated proteins, polyubiqutinated proteins with homotypic chains through a particular lysine on Ub or mixed polyubiquitin chains generated by polymerization through different Ub lysines. The ability of the ubiquitination pathway to generate different protein-Ub structures provides versatility of this pathway to target proteins to different fates. Protein ubiquitination is catalyzed by Ub-conjugating and Ub-ligase enzymes, with different combinations of these enzymes specifying the type of Ub modification on protein substrates. How Ub-conjugating and Ub-ligase enzymes generate this structural diversity is not clearly understood. In the current review, we discuss mechanisms utilized by the Ub-conjugating and Ub-ligase enzymes to generate structural diversity during protein ubiquitination, with a focus on recent mechanistic insights into protein monoubiquitination and polyubiquitination.

INT6/EIF3E interacts with ATM and is required for proper execution of the DNA damage response in human cells

Altered expression of the INT6 gene, encoding the e subunit of the translational initiation factor eIF3, occurs in human breast cancers, but how INT6 relates to carcinogenesis remains unestablished. Here, we show that INT6 is involved in the DNA damage response. INT6 was required for cell survival following γ-irradiation and G(2)-M checkpoint control. RNA interference-mediated silencing of INT6 reduced phosphorylation of the checkpoint kinases CHK1 and CHK2 after DNA damage. In addition, INT6 silencing prevented sustained accumulation of ataxia telangiectasia mutated (ATM) at DNA damage sites in cells treated with γ-radiation or the radiomimetic drug neocarzinostatin. Mechanistically, this result could be explained by interaction of INT6 with ATM, which together with INT6 was recruited to the sites of DNA damage. Finally, INT6 silencing also reduced ubiquitylation events that promote retention of repair proteins at DNA lesions. Accordingly, accumulation of the repair factor BRCA1 was defective in the absence of INT6. Our findings reveal unexpected and striking connections of INT6 with ATM and BRCA1 and suggest that the protective action of INT6 in the onset of breast cancers relies on its involvement in the DNA damage response.

The impact of anxiety on processing efficiency : implications for the attentional control theory

The Attentional Control Theory (ACT) proposes that high-anxious individuals maintain performance effectiveness (accuracy) at the expense of processing efficiency (response time), in particular, the two central executive functions of inhibition and shifting. In contrast, research has generally failed to consider the third executive function which relates to the function of updating. In the current study, seventy-five participants completed the Parametric Go/No-Go and n-back tasks, as well as the State-Trait Anxiety Inventory in order to explore the effects of anxiety on attention. Results indicated that anxiety lead to decay in processing efficiency, but not in performance effectiveness, across all three Central Executive functions (inhibition, set-shifting and updating). Interestingly, participants with high levels of trait anxiety also exhibited impaired performance effectiveness on the n-back task designed to measure the updating function. Findings are discussed in relation to developing a new model of ACT that also includes the role of preattentive processes and dual-task coordination when exploring the effects of anxiety on task performance.

ASAP ECMO : antibiotic, sedative and analgesic pharmacokinetics during extracorporeal membrane oxygenation : a multi-centre study to optimise drug therapy during ECMO

BACKGROUND: Given the expanding scope of extracorporeal membrane oxygenation (ECMO) and its variable impact on drug pharmacokinetics as observed in neonatal studies, it is imperative that the effects of the device on the drugs commonly prescribed in the intensive care unit (ICU) are further investigated. Currently, there are no data to confirm the appropriateness of standard drug dosing in adult patients on ECMO. Ineffective drug regimens in these critically ill patients can seriously worsen patient outcomes. This study was designed to describe the pharmacokinetics of the commonly used antibiotic, analgesic and sedative drugs in adult patients receiving ECMO. METHODS: This is a multi-centre, open-label, descriptive pharmacokinetic (PK) study. Eligible patients will be adults treated with ECMO for severe cardiac and/or respiratory failure at five Intensive Care Units in Australia and New Zealand. Patients will receive the study drugs as part of their routine management. Blood samples will be taken from indwelling catheters to investigate plasma concentrations of several antibiotics (ceftriaxone, meropenem, vancomycin, ciprofloxacin, gentamicin, piperacillin-tazobactum, ticarcillin-clavulunate, linezolid, fluconazole, voriconazole, caspofungin, oseltamivir), sedatives and analgesics (midazolam, morphine, fentanyl, propofol, dexmedetomidine, thiopentone). The PK of each drug will be characterised to determine the variability of PK in these patients and to develop dosing guidelines for prescription during ECMO. DISCUSSION: The evidence-based dosing algorithms generated from this analysis can be evaluated in later clinical studies. This knowledge is vitally important for optimising pharmacotherapy in these most severely ill patients to maximise the opportunity for therapeutic success and minimise the risk of therapeutic failure

An analytical method for assessing stage-specific drug activity in Plasmodium vivax malaria : implications for ex vivo drug susceptibility testing

The emergence of highly chloroquine (CQ) resistant P. vivax in Southeast Asia has created an urgent need for an improved understanding of the mechanisms of drug resistance in these parasites, the development of robust tools for defining the spread of resistance, and the discovery of new antimalarial agents. The ex vivo Schizont Maturation Test (SMT), originally developed for the study of P. falciparum, has been modified for P. vivax. We retrospectively analysed the results from 760 parasite isolates assessed by the modified SMT to investigate the relationship between parasite growth dynamics and parasite susceptibility to antimalarial drugs. Previous observations of the stage-specific activity of CQ against P. vivax were confirmed, and shown to have profound consequences for interpretation of the assay. Using a nonlinear model we show increased duration of the assay and a higher proportion of ring stages in the initial blood sample were associated with decreased effective concentration (EC50) values of CQ, and identify a threshold where these associations no longer hold. Thus, starting composition of parasites in the SMT and duration of the assay can have a profound effect on the calculated EC50 for CQ. Our findings indicate that EC50 values from assays with a duration less than 34 hours do not truly reflect the sensitivity of the parasite to CQ, nor an assay where the proportion of ring stage parasites at the start of the assay does not exceed 66%. Application of this threshold modelling approach suggests that similar issues may occur for susceptibility testing of amodiaquine and mefloquine. The statistical methodology which has been developed also provides a novel means of detecting stage-specific drug activity for new antimalarials.

Evolving and expanding : the scope of practice of the infection control practitioner

Infection control practitioners (ICPs) work across the full spectrum of health care settings and carry out a broad range of practice activities. Whilst several studies have reported on the role of the ICP, there has been little investigation of the scope of infection control practice. This knowledge is essential to inform the professional, legal, educational and financial implications of this specialist role. One hundred and thirteen ICPs from a range of health care settings across Queensland were surveyed. Respondents were asked to rate the extent to which they were and should be engaging in the range of practices identified by Gardner, Jones & Olesen (1999). Significant differences were evident between what ICPs said was their actual practice versus what they thought they should be doing. Overall, the respondents consistently reported that they should be engaging in more of the range of infection control activities than they were, particularly with regard to management practices. A number of differences were found according to the context in which the practitioners worked, such as the type and size of facility and their employment status. The results of this study indicate that the scope of infection control practice has clearly moved beyond those practices that are confined by the hospital wall and defined by surveillance activities.

Use of focus group technique to investigate infection control practice

Use of focus groups as a technique of inquiry is gaining attention in the area of health-care research. This paper will report on the technique of focus group interviewing to investigate the role of the infection control practitioner. Infection control is examined as a specialty area of health-care practice that has received little research attention to date. Additionally, it is an area of practice that is expanding in response to social, economic and microbiological forces. The focus group technique in this study helped a group of infection control practitioners from urban, regional and rural areas throughout Queensland identify and categorise their daily work activities. The outcomes of this process were then analysed to identify the growth in breadth and complexity of the role of the infection control practitioner in the contemporary health-care environment. Findings indicate that the role of the infection control practitioner in Australia has undergone changes consistent with and reflecting changing models of health-care delivery.

Credentialing the infection control practitioner

The following proposal is submitted by the AICA's credentialling and certification subcommittee for your consideration. It outlines a process and procedure for interim credentialling of infection control practitioners. In submitting this proposal, the subcommittee acknowledges that, while competency-based education is the preferred process for credentialling, there are clinicians who, in the absence of educational opportunities, have developed a specialist level of competency in infection control practice through self education and experience. Committee members also recognise the need for self-regulation of accrediting processes, to maintain standards in practice and support members in their clinical roles. We ask you to review the following proposal and invite your comments and critique, to be received by the last week in January 1998.

Key priorities for Australian infection control : summary of findings from the launch of the Centre for Research Excellence in Reducing Healthcare Associated Infections

The launch of the Centre of Research Excellence in Reducing Healthcare Associated Infection (CRE-RHAI) took place in Sydney on Friday 12 October 2012. The mission of the CRE-RHAI is to generate new knowledge about strategies to reduce healthcare associated infections and to provide data on the cost-effectiveness of infection control programs. As well as launching the CRE-RHAI, an important part of this event was a stakeholder Consultation Workshop, which brought together several experts in the Australian infection control community. The aims of this workshop were to establish the research and clinical priorities in Australian infection control, assess the importance of various multi-resistant organisms, and to gather information about decision making in infection control. We present here a summary and discussion of the responses we received.

Effects of type 2 diabetes behavioural telehealth interventions on glycaemic control and adherence : a systematic review

We reviewed the effect of behavioural telehealth interventions on glycaemic control and diabetes self-management in patients with type 2 diabetes. The databases CINAHL, Medline and psychINFO were searched in August 2012. Journal articles were selected that had been published in English with a randomized controlled trial design using a usual care comparison group, and in which the primary intervention component was delivered by telehealth. Relevant outcome measures were glycaemic control and one or more of the following diabetes self-care areas: diet, physical activity, blood glucose self-monitoring (BGSM) or medication adherence. Interventions were excluded if they were primarily based on telemonitoring. The search retrieved 1027 articles, from which 49 were selected based on their title and abstract. Fourteen articles (reporting 13 studies) met the eligibility criteria for inclusion. Four studies reported significant improvements in glycaemic control. Five of eight studies on dietary adherence reported significant treatment effects, as did five of eight on physical activity, four of nine on blood glucose self-monitoring, and three of eight on medication adherence. Overall, behavioural telehealth interventions show promise in improving the diabetes self-care and glycaemic control of people with type 2 diabetes.

Identification of evidence-based biospecimen quality-control tools

Control of biospecimen quality that is linked to processing is one of the goals of biospecimen science. Consensus is lacking, however, regarding optimal sample quality-control (QC) tools (ie, markers and assays). The aim of this review was to identify QC tools, both for fluid and solid-tissue samples, based on a comprehensive and critical literature review. The most readily applicable tools are those with a known threshold for the preanalytical variation and a known reference range for the QC analyte. Only a few meaningful markers were identified that meet these criteria, such as CD40L for assessing serum exposure at high temperatures and VEGF for assessing serum freeze-thawing. To fully assess biospecimen quality, multiple QC markers are needed. Here we present the most promising biospecimen QC tools that were identified.