Linking Innate and Adaptive Immunity: Human Vγ9Vδ2 T Cells Enhance CD40 Expression and HMGB-1 Secretion

γδ T cells play an important role in regulating the immune response to stress stimuli; however, the mean by which these innate lymphocytes fulfill this function remains poorly defined. The main subset of human peripheral blood γδ T cells responds to nonpeptidic antigens, such as isopentylpyrophosphate (IPP), a metabolite in the mevalonate pathway for both eukaryote and prokaryote cells. IPP-primed γδ T cells significantly augment the inflammatory response mediated by monocytes and αβ T cells to TSST-1, the staphylococcal superantigen that is the major causative agent of toxic shock syndrome. Here we show that the small pool of activated peripheral γδ T cells induces an early upregulation of CD40 on monocytes and the local release of High Mobility Group Box-1 (HMGB-1), the molecule designated as the late mediator of systemic inflammation. This finding provides a new basis for how γδ T cells may serve as influential modulators of both endogenous and exogenous stress stimuli

Staphylococcal Toxic Shock Syndrome Toxin-1 Induces the Translocation and Secretion of High Mobility Group-1 Protein from Both Activated T Cells and Monocytes

High mobility group box-1 (HMGB-1) is a DNA-binding protein secreted by activated monocytes and has been identified as a key late mediator of endotoxic shock. We investigated the regulation of HMGB-1 in human peripheral blood mononuclear cells (PBMCs) following stimulation with the staphylococcal superantigen, toxic shock syndrome toxin-1 (TSST-1), and found that TSST-1, like LPS, induced the secretion of HMGB-1 from human PBMC. However, unlike monocyte-driven sepsis caused by endotoxin, translocation and secretion of HMGB-1 mediated by TSST-1 was dependent on the presence of both activated T cells and monocytes. Furthermore, we show that nuclear HMGB-1 is released from TSST-1 stimulated T cells. This finding presents a basis for investigating the potential of targeting HMGB-1 for the treatment of toxic shock syndrome, and provides further insight on the role of HMGB-1 in the cross-talk between activated monocytes and T cells

T helper cell polarisation as a measure of the maturation of the immune response.

BACKGROUND: T helper cell polarisation is important under chronic immune stimulatory conditions and drives the type of the evolving immune response. Mice treated with superantigens in vivo display strong effects on Th subset differentiation. The aim of the study was to detect the intrinsic capacity of T cells to polarise under various ex vivo conditions. METHODS: Purified CD4+ T cells obtained from super-antigen-treated mice were cultured under Th polarising conditions in vitro. By combining intracellular cytokine staining and subsequent flow cytometric analysis with quantitative cytokine measurements in culture supernatants by enzyme-linked immunosorbent assay (ELISA), the differential Th polarising capacity of the treatment can be detected in a qualitative and quantitative manner. RESULTS AND CONCLUSIONS: BALB/c mice were shown to be biased to develop strong Th2 polarised immune responses using Th0 stimulation of purified CD4+ T cells from phosphate-buffered saline-treated mice. Nevertheless, our analysis methodology convincingly showed that even in these mice, Toxic Shock Syndrome Toxin-1 treatment in vivo resulted in a significantly stronger Th1 polarising effect than control treatment. Our results indicate that populations of Th cells can be assessed individually for their differential Th1 or Th2 maturation capacity in vivo by analysing robust in vitro polarisation cultures combined with intracellular cytokine staining and ELISA

Ocular Penetration of N-Formimidoyl Thienamycin (MK-787) and Potentiation by Dipeptidase Inhibitor (MK-791)

N-formimidoyl thienamycin (MK-787) is a new /?-lactam with potent activity against both aerobic and anaerobic gram-positive and gram-negative bacteria. Its spectrum and activity suggest it may be useful in treatment of complicated intraocular infections. Its ocular penetration was studied in New Zealand white rabbits immediately before and after the third dose of 40 mg/kg administered intravenously at q6h intervals. Plasma, aqueous humor, and vitreous humor were obtained by direct aspiration, and antibiotic levels were asayed using an agar well diffusion method. MK-787 penetrated uninflamed intraocular fluids, including vitreous humor, although vitreous concentrations achieved (0.1-0.2 Mg/ml) were significantly lower than the mean peak plasma (15 Mg/ml) and aqueous concentrations (7 Mg/ml). Nevertheless, the intraocular levels attained approached or exceeded the MIC 90 and markedly enhanced spectrum of activity against both gram-positive and gram-negative bacteria

Domestic Violence Service Providers’ Needs and Perceptions of Technology: A Qualitative Study

There is growing recognition of the need to better understand the intersections between the work of domestic violence service providers and technology. Professionals who work with clients impacted by domestic violence are increasingly using technologies across different aspects of their work, including communicating with other professionals and seeking information and resources via the Internet. The current study used qualitative data from two sources—individual interviews and two focus groups—to learn about domestic violence service providers’ needs and perceptions related to technology use. The results provide insights about technologies used currently, expected benefits of future technological advances, barriers to using technology, and participants’ self-rated levels of comfort with technology

A community considers a Family Justice Center: perspectives of stakeholders during the early phases of development

Purpose– Family Justice Centers, or “one-stop shops” that enable domestic violence victims to access a range of services at one location, are becoming increasingly common. However, there is a limited body of research examining the outcomes and planning processes of these Centers. The early phases of planning Centers are critical to their initial and ongoing success. The purpose of this paper is to discuss these issues.Design/methodology/approach– In total, 15 stakeholders in a community in the early phases of planning a Center were interviewed.Findings– Content analysis procedures were used to identify themes related to participants’ ideas about what the Family Justice Center should look like (e.g. services to include and perceived benefits and challenges for the Center), the steps required for planning it (e.g. identifying the purpose of the Center, getting key people involved, and building collaborations), and desired technologies.Originality/value– This paper is the first known research effort to examine the early phases of development in constructing a Family Justice Center

Non-intrusive assessment of photosystem II and photosystem I in whole coral tissues

© 2017 Szabó, Larkum, Suggett, Vass, Sass, Osmond, Zavafer, Ralph and Chow. Reef building corals (phylum Cnidaria) harbor endosymbiotic dinoflagellate algae (genus Symbiodinium) that generate photosynthetic products to fuel their host's metabolism. Non-invasive techniques such as chlorophyll (Chl) fluorescence analyses of Photosystem II (PSII) have been widely used to estimate the photosynthetic performance of Symbiodinium in hospite. However, since the spatial origin of PSII chlorophyll fluorescence in coral tissues is uncertain, such signals give limited information on depth-integrated photosynthetic performance of the whole tissue. In contrast, detection of absorbance changes in the near infrared (NIR) region integrates signals from deeper tissue layers due to weak absorption and multiple scattering of NIR light. While extensively utilized in higher plants, NIR bio-optical techniques are seldom applied to corals. We have developed a non-intrusive measurement method to examine photochemistry of intact corals, based on redox kinetics of the primary electron donor in Photosystem I (P700) and chlorophyll fluorescence kinetics (Fast-Repetition Rate fluorometry, FRRf). Since the redox state of P700 depends on the operation of both PSI and PSII, important information can be obtained on the PSII-PSI intersystem electron transfer kinetics. Under moderate, sub-lethal heat stress treatments (33◦ C for~20 min), the coral Pavona decussata exhibited down-regulation of PSII electron transfer kinetics, indicated by slower rates of electron transport from QA to plastoquinone (PQ) pool, and smaller relative size of oxidized PQ with concomitant decrease of a specifically-defined P700 kinetics area, which represents the active pool of PSII. The maximum quantum efficiency of PSII (Fv /Fm ) and functional absorption cross-section of PSII (σPSII ) remained unchanged. Based on the coordinated response of P700 parameters and PSII-PSI electron transport properties, we propose that simple P700 kinetics parameters as employed here serve as indicators of the integrity of PSII-PSI electron transfer dynamics in corals

A 21-Year-Old Pregnant Woman with Hypertension and Proteinuria

Ronald Ma and colleagues describe the differential diagnosis, investigation, and management of a 21-year-old pregnant woman presenting with hypertension and proteinuria at 20 weeks of gestation

JWalk: a tool for lazy, systematic testing of java classes by design introspection and user interaction

Popular software testing tools, such as JUnit, allow frequent retesting of modified code; yet the manually created test scripts are often seriously incomplete. A unit-testing tool called JWalk has therefore been developed to address the need for systematic unit testing within the context of agile methods. The tool operates directly on the compiled code for Java classes and uses a new lazy method for inducing the changing design of a class on the fly. This is achieved partly through introspection, using Java’s reflection capability, and partly through interaction with the user, constructing and saving test oracles on the fly. Predictive rules reduce the number of oracle values that must be confirmed by the tester. Without human intervention, JWalk performs bounded exhaustive exploration of the class’s method protocols and may be directed to explore the space of algebraic constructions, or the intended design state-space of the tested class. With some human interaction, JWalk performs up to the equivalent of fully automated state-based testing, from a specification that was acquired incrementally