Probiotics modify tight junction proteins in an animal model of non-alcoholic fatty liver disease

Objectives We have investigated the effects of a multi–species probiotic preparation containing a combination of probiotic bacterial genera that included Bifidobacteria, Lactobacilli and a Streptococcus in a mouse model of high fat diet/obesity induced liver steatosis. Methods Three groups of C57B1/6J mice were fed either a standard chow or a high fat diet for 20 weeks, while a third group was fed a high fat diet for 10 weeks and then concomitantly administered probiotics for a further 10 weeks. Serum, liver and large bowel samples were collected for analysis. Results The expression of the tight junction proteins ZO-1 and ZO-2 was reduced (p < 0.05) in high fat diet fed mice compared to chow fed mice. Probiotic supplementation helped to maintain tight ZO-1 and ZO-2 expression compared with the high fat diet group (p < 0.05), but did not restore ZO-1 or ZO-2 expression compared with chow fed mice. Mice fed a high fat diet ± probiotics had significant steatosis development compared to chow fed mice (p < 0.05); steatosis was less severe in the probiotics group compared to the high fat diet group. Hepatic triglycerides concentration was higher in mice fed a high fat diet ± probiotics compared to the chow group (p < 0.05), and was lower in the probiotics group compared to the high fat diet group (p < 0.05). Compared to chow fed mice, serum glucose and cholesterol concentrations, and the activity of alanine transaminase were higher (p < 0.05), whereas serum triglyceride concentration was lower (p < 0.05) in mice fed a high fat diet ± probiotics. Conclusions Supplementation with a multi-species probiotic formulation helped to maintain tight junction proteins ZO-1 and ZO-2, and reduced hepatic triglyceride concentrations compared with a HFD alone.

Towards a quantitative theory of epidermal calcium profile formation in unwounded skin

We propose and mathematically examine a theory of calcium profile formation in unwounded mammalian epidermis based on: changes in keratinocyte proliferation, fluid and calcium exchange with the extracellular fluid during these cells' passage through the epidermal sublayers, and the barrier functions of both the stratum corneum and tight junctions localised in the stratum granulosum. Using this theory, we develop a mathematical model that predicts epidermal sublayer transit times, partitioning of the epidermal calcium gradient between intracellular and extracellular domains, and the permeability of the tight junction barrier to calcium ions. Comparison of our model's predictions of epidermal transit times with experimental data indicates that keratinocytes lose at least 87% of their volume during their disintegration to become corneocytes. Intracellular calcium is suggested as the main contributor to the epidermal calcium gradient, with its distribution actively regulated by a phenotypic switch in calcium exchange between keratinocytes and extracellular fluid present at the boundary between the stratum spinosum and the stratum granulosum. Formation of the extracellular calcium distribution, which rises in concentration through the stratum granulosum towards the skin surface, is attributed to a tight junction barrier in this sublayer possessing permeability to calcium ions that is less than 15 nm/s in human epidermis and less than 37 nm/s in murine epidermis. Future experimental work may refine the presented theory and reduce the mathematical uncertainty present in the model predictions.

Laser capture microdissection and multiplex-tandem PCR analysis of proximal tubular epithelial cell signaling in human kidney disease

Interstitial fibrosis, a histological process common to many kidney diseases, is the precursor state to end stage kidney disease, a devastating and costly outcome for the patient and the health system. Fibrosis is historically associated with chronic kidney disease (CKD) but emerging evidence is now linking many forms of acute kidney disease (AKD) with the development of CKD. Indeed, we and others have observed at least some degree of fibrosis in up to 50% of clinically defined cases of AKD. Epithelial cells of the proximal tubule (PTEC) are central in the development of kidney interstitial fibrosis. We combine the novel techniques of laser capture microdissection and multiplex-tandem PCR to identify and quantitate “real time” gene transcription profiles of purified PTEC isolated from human kidney biopsies that describe signaling pathways associated with this pathological fibrotic process. Our results: (i) confirm previous in-vitro and animal model studies; kidney injury molecule-1 is up-regulated in patients with acute tubular injury, inflammation, neutrophil infiltration and a range of chronic disease diagnoses, (ii) provide data to inform treatment; complement component 3 expression correlates with inflammation and acute tubular injury, (iii) identify potential new biomarkers; proline 4-hydroxylase transcription is down-regulated and vimentin is up-regulated across kidney diseases, (iv) describe previously unrecognized feedback mechanisms within PTEC; Smad-3 is down-regulated in many kidney diseases suggesting a possible negative feedback loop for TGF-β in the disease state, whilst tight junction protein-1 is up-regulated in many kidney diseases, suggesting feedback interactions with vimentin expression. These data demonstrate that the combined techniques of laser capture microdissection and multiplex-tandem PCR have the power to study molecular signaling within single cell populations derived from clinically sourced tissue.

Up the junction? Exploiting knowledge-based development through supply chain and SME cluster interactions

Maximisation of Knowledge-Based Development (KBD) benefits requires effective dissemination and utilisation mechanisms to accompany the initial knowledge creation process. This work highlights the potential for interactions between Supply Chains (SCs) and Small and Medium sized Enterprise Clusters (SMECs), (including via ‘junction’ firms which are members of both networks), to facilitate such effective dissemination and utilisation of knowledge. In both these network types there are firms that readily utilise their relationships and ties for ongoing business success through innovation. The following chapter highlights the potential for such beneficial interactions between SCs and SMECs in key elements of KBD, particularly knowledge management, innovation and technology transfer. Because there has been little focus on the interactions between SCs and SMECs, particularly when firms simultaneously belong to both, this chapter examines the conduits through which information and knowledge can be transferred and utilised. It shows that each network type has its own distinct advantages in the types of information searched for and transferred amongst network member firms. Comparing and contrasting these advantages shows opportunities for both networks to leverage the knowledge sharing strengths of each other, through these ‘junctions’ to address their own weaknesses, allowing implications to be drawn concerning new ways of utilising relationships for mutual network gains.

Coordinated road-junction traffic control by dynamic programming

This paper presents a novel approach to road-traffic control for interconnected junctions. With a local fuzzy-logic controller (FLC) installed at each junction, a dynamic-programming (DP) technique is proposed to derive the green time for each phase in a traffic-light cycle. Coordination parameters from the adjacent junctions are also taken into consideration so that organized control is extended beyond a single junction. Instead of pursuing the absolute optimization of traffic delay, this study examines a practical approach to enable the simple implementation of coordination among junctions, while attempting to reduce delays, if possible. The simulation results show that the delay per vehicle can be substantially reduced, particularly when the traffic demand reaches the junction capacity. The implementation of this controller does not require complicated or demanding hardware, and such simplicity makes it a useful tool for offline studies or realtime control purposes.

Railway junction traffic control by heuristic methods

Traffic conflicts at railway junctions are very conmon, particularly on congested rail lines. While safe passage through the junction is well maintained by the signalling and interlocking systems, minimising the delays imposed on the trains by assigning the right-of-way sequence sensibly is a bonus to the quality of service. A deterministic method has been adopted to resolve the conflict, with the objective of minimising the total weighted delay. However, the computational demand remains significant. The applications of different heuristic methods to tackle this problem are reviewed and explored, elaborating their feasibility in various aspects and comparing their relative merits for further studies. As most heuristic methods do not guarantee a global optimum, this study focuses on the trade-off between computation time and optimality of the resolution.

Fuzzy logic traffic control at a road junction with time-varying flow rates

Fuzzy logic has been applied to control traffic at road junctions. A simple controller with one fixed rule-set is inadequate to minimise delays when traffic flow rate is time-varying and likely to span a wide range. To achieve better control, fuzzy rules adapted to the current traffic conditions are used.

Railway junction conflict resolution by genetic algorithm

In general, simple and traditional methods are applied to resolve traffic conflicts at railway junctions. They are, however, either inefficient or computationally demanding. A simple genetic algorithm is presented to enable a search for a near optimal resolution to be carried out while meeting the constraints on generation evolution and minimising the search time.

Hierarchical fuzzy logic traffic control at a road junction

Traffic control at road junctions is one of the major concerns in most metropolitan cities. Controllers of various approaches are available and the required control action is the effective green-time assigned to each traffic stream within a traffic-light cycle. The application of fuzzy logic provides the controller with the capability to handle uncertain natures of the system, such as drivers’ behaviour and random arrivals of vehicles. When turning traffic is allowed at the junction, the number of phases in the traffic-light cycle increases. The additional input variables inevitably complicate the controller and hence slow down the decision-making process, which is critical in this real-time control problem. In this paper, a hierarchical fuzzy logic controller is proposed to tackle this traffic control problem at a 2-way road junction with turning traffic. The two levels of fuzzy logic controllers devise the minimum effective green-time and fine-tune it respectively at each phase of a traffic-light cycle. The complexity of the controller at each level is reduced with smaller rule-set. The performance of this hierarchical controller is examined by comparison with a fixed-time controller under various traffic conditions. Substantial delay reduction has been achieved as a result and the performance and limitation of the controller will be discussed.

Railway junction conflict resolution by local search method

This study investigates the application of local search methods on the railway junction traffic conflict-resolution problem, with the objective of attaining a quick and reasonable solution. A procedure based on local search relies on finding a better solution than the current one by a search in the neighbourhood of the current one. The structure of neighbourhood is therefore very important to an efficient local search procedure. In this paper, the formulation of the structure of the solution, which is the right-of-way sequence assignment, is first described. Two new neighbourhood definitions are then proposed and the performance of the corresponding local search procedures is evaluated by simulation. It has been shown that they provide similar results but they can be used to handle different traffic conditions and system requirements.

Hierarchical fuzzy logic traffic control at a road junction using genetic algorithms

Traffic control at a road junction by a complex fuzzy logic controller is investigated. The increase in the complexity of junction means more number of input variables must be taken into account, which will increase the number of fuzzy rules in the system. A hierarchical fuzzy logic controller is introduced to reduce the number of rules. Besides, the increase in the complexity of the controller makes formulation of the fuzzy rules difficult. A genetic algorithm based off-line leaning algorithm is employed to generate the fuzzy rules. The learning algorithm uses constant flow-rates as training sets. The system is tested by both constant and time-varying flow-rates. Simulation results show that the proposed controller produces lower average delay than a fixed-time controller does under various traffic conditions.

Substrate recognition and catalysis by the Holliday junction resolving enzyme Hje

Two archaeal Holliday junction resolving enzymes, Holliday junction cleavage (Hjc) and Holliday junction endonuclease (Hje), have been characterized. Both are members of a nuclease superfamily that includes the type II restriction enzymes, although their DNA cleaving activity is highly specific for four-way junction structure and not nucleic acid sequence. Despite 28% sequence identity, Hje and Hjc cleave junctions with distinct cutting patterns—they cut different strands of a four-way junction, at different distances from the junction centre. We report the high-resolution crystal structure of Hje from Sulfolobus solfataricus. The structure provides a basis to explain the differences in substrate specificity of Hje and Hjc, which result from changes in dimer organization, and suggests a viral origin for the Hje gene. Structural and biochemical data support the modelling of an Hje:DNA junction complex, highlighting a flexible loop that interacts intimately with the junction centre. A highly conserved serine residue on this loop is shown to be essential for the enzyme's activity, suggesting a novel variation of the nuclease active site. The loop may act as a conformational switch, ensuring that the active site is completed only on binding a four-way junction, thus explaining the exquisite specificity of these enzymes.

Managing older workers during a period of tight labour supply

This article reports on a recent survey of employer attitudes and policies towards older workers in Australia at a time of sustained economic growth and ongoing concerns about labour shortages. Findings from a survey of 590 employers with more than 50 employees in the State of Queensland point to an unusually strong orientation towards the recruitment of older workers among respondents, although the retraining of older workers is not prioritised by the majority. The issue of workforce ageing is viewed as being of medium-term importance by the majority of respondents, although for a substantial number the issue is of immediate concern. Both sector and organisation size are predictive of the application of a broad range of policies targeting older workers, with public-sector and larger organisations more likely to be active. Concerns about workforce ageing and labour supply are predictive of employer behaviours regarding older workers, suggesting that sustained policy making may be emerging in response to population ageing over and above more immediate concerns about labour shortages and that this broad thrust of organisational policy making may be immune to the point in the economic cycle. This study found no evidence that the flexible firm will not countenance an ageing workforce.