A Simple and Effective Flow Cytometry-Based Method for Identification and Quantification of Tissue Infiltrated Leukocyte Subpopulations in a Mouse Model of Peripheral Arterial Disease

Arteriogenesis, the growth of a natural bypass from pre-existing arteriolar collaterals, is an endogenous mechanism to compensate for the loss of an artery. Mechanistically, this process relies on a locally and temporally restricted perivascular infiltration of leukocyte subpopulations, which mediate arteriogenesis by supplying growth factors and cytokines. Currently, the state-of-the-art method to identify and quantify these leukocyte subpopulations in mouse models is immunohistology. However, this is a time consuming procedure. Here, we aimed to develop an optimized protocol to identify and quantify leukocyte subpopulations by means of flow cytometry in adductor muscles containing growing collateral arteries. For that purpose, adductor muscles of murine hindlimbs were isolated at day one and three after induction of arteriogenesis, enzymatically digested, and infiltrated leukocyte subpopulations were identified and quantified by flow cytometry, as exemplary shown for neutrophils and macrophages (defined as CD45+/CD11b+/Ly6G+ and CD45+/CD11b+/F4/80+ cells, respectively). In summary, we show that flow cytometry is a suitable method to identify and quantify leukocyte subpopulations in muscle tissue, and provide a detailed protocol. Flow cytometry constitutes a timesaving tool compared to histology, which might be used in addition for precise localization of leukocytes in tissue samples.

Quality-of-Service monitoring of hybrid industrial communication networks

Today many hybrid (wired & wireless) industrial communication networks with a huge variety of heterogeneous technologies and protocols are present in the manufacturing and automation domain. The increasing requirements regarding e. g., latency, reliability, or determinism create the need for a holistic network management concept in order to assure a network-wide Quality-of-Service (QoS) resource provisioning and the assurance of the admissioned resources. Consequently, a monitoring of the whole network is required to feed the network management system with the needed information about the underlying network processes. Various technical approaches using different methods of extracting the information from network traffic are available for the purpose of QoS parameter observance and measurement at the moment. Therefore, this paper provides a state of the art research about network management and QoS provisioning respectively QoS assurance concepts. In addition, the passive network monitoring approach using the flow export technique based on the Internet Protocol Flow Information Export (IPFIX) is investigated for a utilisation in the nowadays industry domain based on a conceptual case study with a wireless protocol. As a conclusion, an evaluation is performed in order to clarify the limits and the overall usability of IPFIX for the monitoring of industrial networks in order to support future network management systems.

METHODS AND PROTOCOL FLOW CELLS: TECHNIQUE USED FOR STUDYING MICROBIAL BIOFILMS

Biofilms are sessile communities of microorganisms growing on material surfaces and embedded in self-accumulated extracellular polymers. A comprehensive analysis of physical, chemical and biological factors including hydrodynamic and nutrient conditions that regulate their formation is required to adequately gain insight to this complex multicellular microbial life style. Reproducible experimental models that consider all the conditions under which they grow and develop also remain a required tool for studying the biofilms. As a result of its ability to create hydrodynamic and nutrient conditions coupled with continuous and non-destructive ability to grow biofilms, flow cell technology has become one of the most recently patronised models used to study microbial biofilms. This article focuses on recent advancements, principles and practical application of flow cell technology to study microbial biofilms.

Women in situations of violence: limits of assistance

This paper analyzes the limitations of the care of professionals of family health teams provided to women in situations of violence in the integrated care network in a city in the northwest region of Rio Grande do Sul state, Brazil. The data is part of a survey conducted with six teams participating in family health care in the course of eight meetings, in which the group identified the limits and potential of the practice. The causes of limits, solutions and improvement actions were discussed in a movement of reflection, action and learning. The empirical material was recorded, transcribed and subjected to Bardin's thematic content analysis. The results show the lack of coordination between the sectors of the network; lack of protocol, flow and organization of reference and counter-reference; inefficiency of police, judiciary and security sectors; and the lack of professionals to work on violence against women and in the perspective of the network as restrictive conditions for efficient practice. The need is highlighted for greater support of municipal managers to the professionals for networking and encouragement through the qualification and ongoing education courses that address issues such as gender equity, guarantee of rights and a care network.