Myostatin gene knockdown through lentiviral-mediated delivery of shRNA for in vitro production of transgenic bovine embryos

Myostatin is described as a negative regulator of the skeletal muscle growth. Genetic engineering, in order to produce animals with double the muscle mass and that can transmit the characteristic to future progeny, may be useful. In this context, the present study aimed to analyse the feasibility of lentiviral-mediated delivery of short hairpin RNA (shRNA) targeting of myostatin into in vitro produced transgenic bovine embryos. Lentiviral vectors were used to deliver a transgene that expressed green fluorescent protein (GFP) and an shRNA that targeted myostatin. Vector efficiency was verified through in vitro murine myoblast (C2C12) cell morphology after inductive differentiation and by means of real-time PCR. The lentiviral vector was microinjected into the perivitellinic space of in vitro matured oocytes. Non-microinjected oocytes were used as the control. After injection, oocytes were fertilized and cultured in vitro. Blastocysts were evaluated by epifluorescence microscopy. Results demonstrated that the vector was able to inhibit myostatin mRNA in C2C12 cells, as the transducted group had a less amount of myostatin mRNA after 72 h of differentiation (p < 0.05) and had less myotube formation than the non-transduced group (p < 0.05). There was no difference in cleavage and blastocyst rates between the microinjected and control groups. After hatching, 3.07% of the embryos exhibited GFP expression, indicating that they expressed shRNA targeting myostatin. In conclusion, we demonstrate that a lentiviral vector effectively performed shRNA myostatin gene knockdown and gene delivery into in vitro produced bovine embryos. Thus, this technique can be considered a novel option for the production of transgenic embryos and double muscle mass animals.

Dressing liposomal particles with chitosan and poly(vinylic alcohol) for oral vaccine delivery

Liposomes have been used as adjuvants since 1974. One major limitation for the use of liposomes in oral vaccines is the lipid structure instability caused by enzyme activities. Our aim was to combine liposomes that could encapsulate antigens (i.e., Dtxd, diphtheria toxoid) with chitosan, which protects the particles and promotes mucoadhesibility. We employed physical techniques to understand the process by which liposomes (SPC: Cho, 3: 1) can be sandwiched with chitosan (Chi) and stabilized by PVA (poly-vinylic alcohol), which are biodegradable, biocompatible polymers. Round, smooth-surfaced particles of REVs-Chi (reversed-phase vesicles sandwiched by Chi) stabilized by PVA were obtained. The REVs encapsulation efficiencies (Dtxd was used as the antigen) were directly dependent on the Chi and PVA present in the formulation. Chi adsorption on the REVs surface was accompanied by an increase of zeta-potential. In contrast, PVA adsorption on the REVs-Chi surface was accompanied by a decrease of zeta-potential. The presence of Dtxd increased the Chi surface-adsorption efficiency. The PVA affinity by mucine was 2,000 times higher than that observed with Chi alone and did not depend on the molecule being in solution or adsorbed on the liposomal surface. The liberation of encapsulated Dtxd was retarded by encapsulation within REVs-Chi-PVA. These results lead us to conclude that these new, stabilized particles were able to be adsorbed by intestinal surfaces, resisted degradation, and controlled antigen release. Therefore, REVs-Chi-PVA particles can be used as an oral delivery adjuvant.

Delivery care in Quang Ninh province, Northern Vietnam : resources and access to safe care.

Every mother and child has the right to survive childbirth which requires skilled birth attendants together with referral and available emergency obstetric care (EmOC). The objective of the study was to describe delivery care routines at different levels in the health care system in Quang Ninh province, Northern Vietnam. The design was cross sectional using a structured questionnaire. Two districts in Quang Ninh province with 40 Community Health Centres (CHC), three district hospitals and one region hospital was included in the study, in total 138 (CHC n=105 and hospitals n=33) health care providers participated. In our study 20% (CHC) of the health care providers assisting deliveries at CHC were midwives and health care provider’s in our study further report to have assisted at less then 10 deliveries/year (81% of respondents at CHC). Findings show that the health care provider’s routines and care for women during labour and delivery vary and that there is a need for re-training and that women in labour should be cared for by health care providers with adequate training like midwifery. In our study CHC had poor resources to provide basic or comprehensive EmOC. Our findings indicate that there is a need for re-training in delivery care among health care providers and since the number of deliveries at CHC is few they should be handled by someone who is a skilled birth attendant. Our findings also show a variation in care routines during labour and delivery among health care providers at CHC and hospital levels and this also show the need for re-training and support from proper authorities in order to improve maternal and newborn health.

Production and Delivery (Optimization of production system and reliability)

This thesis is done to solve two issues for Sayid Paper Mill Ltd Pakistan. Section one deals with a practical problem arise in SPM that is cutting a given set of raw paper rolls of known length and width, and a set of product paper rolls of known length (equal to the length of raw paper rolls) and width, practical cutting constraints on a single cutting machine, according to demand orders for all customers. To solve this problem requires to determine an optimal cutting schedule to maximize the overall cutting process profitability while satisfying all demands and cutting constraints. The aim of this part of thesis is to develop a mathematical model which solves this problem.Second section deals with a problem of delivering final product from warehouse to different destinations by finding shortest paths. It is an operational routing problem to decide the daily routes for sending trucks to different destination to deliver their final product. This industrial problem is difficult and includes aspect such as delivery to a single destination and multiple destinations with limited resources. The aim of this part of thesis is to develop a process which helps finding shortest path.

Amningsduration vid sectio och vaginalförlossning : Breastfeeding duration at cesarean section and vaginal delivery. Apopulationbased register study.

Bakgrund: Amning är av stor betydelse för det nyfödda barnet och mamma, både ur närings- och trygghetsaspekter. Tidigare studier om sambandet mellan förlossningssätt och amning visar motstridiga resultat; vissa studier indikerar att sectio påverkar amningen negativt medan andra studier inte visar på något samband.Syftet: Att undersöka huruvida det finns demografiska och hälsorelaterade skillnader hos mammor gällande förlossningssätt samt om förlossningssätt påverkar amningsdurationen upp till två månaders ålder.Metod: Studien har en populationsbaserad kohortdesign och omfattar n= 35250 mamma-barn par i Örebro- och Uppsala län, där barnet fötts åren 1993-2001. Statistiska analyser har genomförts med Chi-square test, binär logistisk regressionsanalys och multivariat logistik regressionsanalys.Resultat: Visade att mammor som förlösts med sectio ammade i lägre utsträckning vid två månaders ålder i jämförelse med mammor som förlösts vaginalt. Många riskfaktorer identifierades för en ökad risk att förlösas med sectio: län, hälsofaktorer hos mamma och barn, paritet, mammans ålder, rökning samt socioekonomiska faktorer. Resultatet kan hjälpa barnmorskan i hennes profession, genom att kunskap erhålls om dessa samband. Barnmorskan kan därmed identifiera riskfaktorer, arbeta förebyggande och underlätta initieringen av amning.Konklusion: Barnmorskor kan med hjälp av denna studie öka medvetenheten hos vårdpersonal angående sectioförlösta mammors behov av extra stöd vid initiering av amning.

Maternal health care professionals perspective on the provision and use of antenatal and delivery care : a qualitative descriptive study in rural Vietnam

Background. High quality maternal health care is an important tool to reduce maternal and neonatal mortality. Services offered should be evidence based and adapted to the local setting. This qualitative descriptive study explored the perspectives and experiences of midwives, assistant physicians and medical doctors on the content and quality of maternal health care in rural Vietnam. Method. The study was performed in a rural district in northern Vietnam. Four focus group discussions with health care professionals at primary health care level were conducted. The data was analysed using qualitative manifest and latent content analysis. Result. Two main themes emerged: "Contextual conditions for maternal health care" and "Balancing between possibilities and constraints". Contextual conditions influenced both pregnant women's use of maternal health care and health care professionals' performance. The study participants stated that women's uses of maternal health care were influenced by economical constraints and cultural norms that impeded their autonomy in relation to childbearing. Structural constraints within the health care system included inadequate financing of the primary health care, resulting in lack of human resources, professional re-training and adequate equipment. Conclusion. Contextual conditions strongly influenced the performance and interaction between pregnant women and health care professionals within antenatal care and delivery care in a rural district of Vietnam. Although Vietnam is performing comparatively well in terms of low maternal and child mortality figures, this study revealed midwives' and other health care professionals' perceived difficulties in their daily work. It seemed maternal health care was under-resourced in terms of staff, equipment and continuing education activities. The cultural setting in Vietnam constituting a strong patriarchal society and prevailing Confucian norms limits women's autonomy and reduce their possibility to make independent decisions about their own reproductive health. This issue should be further addressed by policy-makers. Strategies to reduce inequities in maternal health care for pregnant women are needed. The quality of client-provider interaction and management of pregnancy may be strengthened by education, human resources, re-training and provision of essential equipment.

Describing Web Service Architectures through Design-by-Contract

Architectural description languages (ADLs) are used to specify a high-level, compositional view of a software application, specifying how a system is to be composed from coarse-grain components. ADLs usually come equipped with a formal dynamic semantics, facilitating specification and analysis of distributed and event-based systems. In this paper, we describe the TrustME, an ADL framework that provides both a process and a structural view of web service-based systems. We use Petri-net descriptions to give a dynamic view of business workflow for web service collaboration. We adapt the approach of Schmidt to define a form of Meyer's design-by-contract for configuring workflow architectures. This serves as a configuration-level means of constructing safer, more robust systems.

Trust-By-Contract: Modelling, Analysing and Predicting Behaviour in Software Architectures

Architecture description languages (ADLs) are used to specify high-level, compositional views of a software application. ADL research focuses on software composed of prefabricated parts, so-called software components. ADLs usually come equipped with rigorous state-transition style semantics, facilitating verification and analysis of specifications. Consequently, ADLs are well suited to configuring distributed and event-based systems. However, additional expressive power is required for the description of enterprise software architectures – in particular, those built upon newer middleware, such as implementations of Java’s EJB specification, or Microsoft’s COM+/.NET. The enterprise requires distributed software solutions that are scalable, business-oriented and mission-critical. We can make progress toward attaining these qualities at various stages of the software development process. In particular, progress at the architectural level can be leveraged through use of an ADL that incorporates trust and dependability analysis. Also, current industry approaches to enterprise development do not address several important architectural design issues. The TrustME ADL is designed to meet these requirements, through combining approaches to software architecture specification with rigorous design-by-contract ideas. In this paper, we focus on several aspects of TrustME that facilitate specification and analysis of middleware-based architectures for trusted enterprise computing systems.

Modelling and Administration of Contract-Based Systems

Mirroring the paper versions exchanged between businesses today, electronic contracts offer the possibility of dynamic, automatic creation and enforcement of restrictions and compulsions on agent behaviour that are designed to ensure business objectives are met. However, where there are many contracts within a particular application, it can be difficult to determine whether the system can reliably fulfil them all; computer-parsable electronic contracts may allow such verification to be automated. In this paper, we describe a conceptual framework and architecture specification in which normative business contracts can be electronically represented, verified, established, renewed, etc. In particular, we aim to allow systems containing multiple contracts to be checked for conflicts and violations of business objectives. We illustrate the framework and architecture with an aerospace example.

Towards a Monitoring Framework for Agent-Based Contract Systems

The behaviours of autonomous agents may deviate from those deemed to be for the good of the societal systems of which they are a part. Norms have therefore been proposed as a means to regulate agent behaviours in open and dynamic systems, and may be encoded in electronic contracts in order to specify the obliged, permitted and prohibited behaviours of agents that are signatories to such contracts. Enactment and management of electronic contracts thus enables the use of regulatory mechanisms to ensure that agent behaviours comply with the encoded norms. To facilitate such mechanisms requires monitoring in order to detect and explain violation of norms. In this paper we propose a framework for monitoring that is to be implemented and integrated into a suite of contract enactment and management tools. The framework adopts a non-intrusive approach to monitoring, whereby the states of a contract with respect to its contained norms can be inferred on the basis of messages exchanged. Specifically, the framework deploys agents that observe messages sent between contract signatories, where these messages correspond to agent behaviours and therefore indicate whether norms are, or are in danger of, being violated.

A contract-based system for aerospace aftercare

The logistics of the aerospace aftermarket raises a number of very interesting challenges from the perspective of electronic contracting. This is a highly dynamic domain, where contracts are established between airlines and engine manufacturers, as well as between engine manufacturers all the way down the supply lines, providing a particularly illustrative showcase for the technologies developed in the CONTRACT project. In this paper, we describe such a domain, as well as our modelling of it as a multiagent simulator where the CONTRACT framework has been used to monitor for compliance with norms.

Monitoring and Explanation of Contract Execution: A Case Study in the Aerospace Domain

In the domain of aerospace aftermarkets, which often has long supply chains that feed into the maintenance of aircraft, contracts are used to establish agreements between aircraft operators and maintenance suppliers. However, violations at the bottom of the supply chain (part suppliers) can easily cascade to the top (aircraft operators), making it difficult to determine the source of the violation, and seek to address it. In this context, we have developed a global monitoring architecture that ensures the detection of norm violations and generates explanations for the origin of violations. In this paper, we describe the implementation and deployment of a global monitor in the aerospace domain of [8] and show how it generates explanations for violations within the maintenance supply chain. We show how these explanations can be used not only to detect violations at runtime, but also to uncover potential problems in contracts before their deployment, thus improving them.

Case studies for contract-based systems

Of the ways in which agent behaviour can be regulated in a multiagent system, electronic contracting – based on explicit representation of different parties' responsibilities, and the agreement of all parties to them – has significant potential for modern industrial applications. Based on this assumption, the CONTRACT project aims to develop and apply electronic contracting and contract-based monitoring and verification techniques in real world applications. This paper presents results from the initial phase of the project, which focused on requirements solicitation and analysis. Specifically, we survey four use cases from diverse industrial applications, examine how they can benefit from an agent-based electronic contracting infrastructure and outline the technical requirements that would be placed on such an infrastructure. We present the designed CONTRACT architecture and describe how it may fulfil these requirements. In addition to motivating our work on the contract-based infrastructure, the paper aims to provide a much needed community resource in terms of use case themselves and to provide a clear commercial context for the development of work on contract-based system.