A novel role for granzymes in anti-tumor immunity

The cytotoxic properties of granzymes are well established, though recent publications suggest additional roles for granzymes in immunity. We demonstrated that granzymes can act as regulators of cross-presentation by dendritic cells by inducing critical “eat-me” signals on the dying tumor cell, resulting in efficient phagocytosis of cell-associated tumor antigen

Eliciting third person perspectives in social work case discussions:A device for reflective supervision?

Reflective supervision is widely recommended as an effective way to support social workers to think about their practice and to make better decisions. Although previous research has proposed methods of pursuing reflective supervision, little is known about how supervisors attempt this in actual supervision meetings. One proposed method for supervisors is to elicit third person perspectives so that social workers can consider a situation from different points of view. In this article, we examine this method by analysing audio recordings of 12 supervision meetings from one local authority Children and Families Social Work team. Using Conversation Analysis (CA), we explore supervisors’ attempts to elicit other people’s perspectives, focusing on how such requests were formed and how the social worker responded. We found 35 instances of supervisors attempting to elicit third person perspectives and identified four different ways that supervisors designed these enquiries. Supervisors oriented to two concerns, based on whether they enquired about a perspective that was currently established or projected into the future, and whether the information was verifiable through speech or actions, or imagined based on the other person’s thoughts or understanding. We draw on CA work on epistemics and stance to show how these different approaches have implications for what the social worker is expected to know and how both speakers orient to the accountability of the social worker. We conclude by considering the epistemic friction between the design of these enquiries, the relevance of professional accountability and the possibilities for pursuing reflective supervision in practice

Neurocognitive Correlates of Treatment Response in Children with Tourette\u27s Disorder

This paper examined neurocognitive functioning and its relationship to behavior treatment response among youth with Tourette\u27s Disorder (TD) in a large randomized controlled trial. Participants diagnosed with TD completed a brief neurocognitive battery assessing inhibitory functions, working memory, and habit learning pre- and post-treatment with behavior therapy (CBIT, Comprehensive Behavioral Intervention for Tics) or psychoeducation plus supportive therapy (PST). At baseline, youth with tics and Attention Deficit Hyperactivity Disorder (ADHD) exhibited some evidence of impaired working memory and simple motor inhibition relative to youth with tics without ADHD. Additionally, a small negative association was found between antipsychotic medications and youth\u27s performance speed. Across treatment groups, greater baseline working memory and aspects of inhibitory functioning were associated with a positive treatment response; no between-group differences in neurocognitive functioning at post-treatment were identified. Within the behavior therapy group, pre-treatment neurocognitive status did not predict outcome, nor was behavior therapy associated significant change in neurocognitive functioning post-treatment. Findings suggest that co-occurring ADHD is associated with some impairments in neurocognitive functioning in youth with Tourette\u27s Disorder. While neurocognitive predictors of behavior therapy were not found, participants who received behavior therapy exhibited significantly reduced tic severity without diminished cognitive functioning

An Empirical Examination of Symptom Substitution Associated with Behavior Therapy for Tourette\u27s Disorder

Over the past six decades, behavior therapy has been a major contributor to the development of evidence-based psychotherapy treatments. However, a long-standing concern with behavior therapy among many nonbehavioral clinicians has been the potential risk for symptom substitution. Few studies have been conducted to evaluate symptom substitution in response to behavioral treatments, largely due to measurement and definitional challenges associated with treated psychiatric symptoms. Given the overt motor and vocal tics associated with Tourette’s disorder, it presents an excellent opportunity to empirically evaluate the potential risk for symptom substitution associated with behavior therapy. The present study examined the possible presence of symptom substitution using four methods: (a) the onset of new tic symptoms, (b) the occurrence of adverse events, (c) change in tic medications, and (d) worsening of co-occurring psychiatric symptoms. Two hundred twenty-eight participants with Tourette’s disorder or persistent motor or vocal tic disorders were randomly assigned to receive behavioral therapy or supportive therapy for tics. Both therapies consisted of eight sessions over 10 weeks. Results indicated that participants treated with behavior therapy were not more likely to have an onset of new tic symptoms, experience adverse events, increase tic medications, or have an exacerbation in co-occurring psychiatric symptoms relative to participants treated with supportive therapy. Further analysis suggested that the emergence of new tics was attributed with the normal waxing and waning nature of Tourette’s disorder. Findings provide empirical support to counter the long-standing concern of symptom substitution in response to behavior therapy for individuals with Tourette\u27s disorder

Temporal and spatial dynamics of CO2 air-sea flux in the Gulf of Maine

Ocean surface layer carbon dioxide (CO2) data collected in the Gulf of Maine from 2004 to 2008 are presented. Monthly shipboard observations are combined with additional higher‐resolution CO2 observations to characterize CO2 fugacity ( fCO2) and CO2 flux over hourly to interannual time scales. Observed fCO2 andCO2 flux dynamics are dominated by a seasonal cycle, with a large spring influx of CO2 and a fall‐to‐winter efflux back to the atmosphere. The temporal results at inner, middle, and outer shelf locations are highly correlated, and observed spatial variability is generally small relative to the monthly to seasonal temporal changes. The averaged annual flux is in near balance and is a net source of carbon to the atmosphere over 5 years, with a value of +0.38 mol m−2 yr−1. However, moderate interannual variation is also observed, where years 2005 and 2007 represent cases of regional source (+0.71) and sink (−0.11) anomalies. We use moored daily CO2 measurements to quantify aliasing due to temporal undersampling, an important error budget term that is typically unresolved. The uncertainty of our derived annual flux measurement is ±0.26 mol m−2 yr−1 and is dominated by this aliasing term. Comparison of results to the neighboring Middle and South Atlantic Bight coastal shelf systems indicates that the Gulf of Maine exhibits a similar annual cycle and range of oceanic fCO2 magnitude but differs in the seasonal phase. It also differs by enhanced fCO2 controls by factors other than temperature‐driven solubility, including biological drawdown, fall‐to‐winter vertical mixing, and river runoff

Morphological Effects of Lonidamine on Two Human-Tumor Cell Culture Lines

Lonidamine, 1-(2-4-dichlorobenzyl)-1-H-indazol-3-carboxylic acid. is an anticancer drug that has its primary action on cellular metabolism rather than cell division. Since lonidamine is not effective in all tumor cells, we have tested it in two human-tumor cell culture lines: MOLT-4, a T-leukemia and U-87 MG. a glioma. Lonidamine exposure of MOLT-4 cells at 50 μg/mL and pH 6.7 disrupted the mitochondria within 1 h of treatment. The mitochondria were swollen and the cristae were disrupted. When the treated cells were re-incubated in fresh medium at pH 7.4 the mitochondria rapidly returned to their normal morphology. The U-87 MG glioma cells did not show ultrastructural disruption after 1-h treatment with lonidamine at concentrations up to 200 μg/mL at pH 6.7 In the concentration range of 25 μg/mL to 200 μg/mL. lonidamine did not produce any cell killing in MOLT-4 after a 1-h exposure at pH 7.4, although the drug had some limited effectiveness at pH 6.7. Compared to sham-treated controls, long exposures to 100 μg/mL of lonidamine at pH 6.7 reduced survival in MOLT-4 to 92% and 53% after 6-h and 24-h exposures, respectively. Survival of U-87 MG glioma cells was also strongly pH dependent, a 2-h exposure to 50 μg/mL lonidamine at pH 7.4 did not cause cell death; however, survival dropped to 84% of the control at pH 6.65

Expression of Zm13, a pollen specific maize protein, in Escherichia coli reveals IgE-binding capacity and allergenic potential

AbstractPlant proteins belong to the most frequent elicitors of type I allergic symptoms in industrialized countries. Several relevant plant allergens have been found to be either specifically expressed or highly upregulated in mature pollen. The cDNA coding for a pollen specific maize protein, Zm13, shows significant sequence homology with a number of pollen or anther specific proteins from monocot and dicot plants as well as with recently described allergens from olive and rye grass. To test whether the Zm13 protein might possess IgE-binding capacity, Zm13 was expressed in E coli. The coding region of Zm13 was PCR amplified from a genomic clone and expressed as as a glutathione-S-transferase fusion protein. The recombinant Zm13 fusion protein bound a Zm13 specific rabbit antiserum and reacted with serum IgE from grass pollen allergic patients indicating that Zm13 and homologous proteins represent a family of conserved plant allergens

Granzyme B (GraB) Autonomously Crosses the Cell Membrane and Perforin Initiates Apoptosis and GraB Nuclear Localization

Granzyme B (GraB) induces apoptosis in the presence of perforin. Perforin polymerizes in the cell membrane to form a nonspecific ion pore, but it is not known where GraB acts to initiate the events that ultimately lead to apoptosis. It has been hypothesized that GraB enters the target cell through a perforin channel and then initiates apoptosis by cleaving and activating members of the ICE/Ced-3 family of cell death proteases. To determine if GraB can enter the cell, we treated YAC-1 or HeLa cells with FITC-labeled GraB and measured intracellular fluorescence with a high sensitivity CCD camera and image analyzer. GraB was internalized and found diffusely dispersed in the cell cytoplasm within 10 min. Uptake was inhibited at low temperature (4°C) and by pretreatment with metabolic inhibitors, NaF and DNP, or cytochalasin B, a drug that both blocks microfilament formation, and FITC-GraB remained on the cell membrane localized in patches. With the simultaneous addition of perforin and FITC-GraB, no significant increase in cytoplasmic fluorescence was observed over that found in cells treated only with FITC-GraB. However, FITC-GraB was now detected in the nucleus of apoptotic cells labeling apoptotic bodies and localized areas within and along the nuclear membrane. The ability of GraB to enter cells in the absence of perforin was reexamined using anti-GraB antibody immunogold staining of ultrathin cryosections of cells incubated with GraB. Within 15 min, gold particles were detected both on the plasma membrane and in the cytoplasm of cells with some gold staining adjacent to the nuclear envelope but not in the nucleus. Cells internalizing GraB in the absence of perforin appeared morphologically normal by Hoechst staining and electron microscopy. GraB directly microinjected into the cytoplasm of B16 melanoma cells induced transient plasma membrane blebbing and nuclear coarsening but the cells did not become frankly apoptotic unless perforin was added. We conclude that GraB can enter cells autonomously but that perforin initiates the apoptotic process and the entry of GraB into the nucleus

Statistical approaches to access the effect of Lactobacillus sakei culture and ultrasound frequency on fatty acid profile of beef jerky

The objective of this study was to investigate the effect of ultrasonic frequencies and drying time on fatty acid profiles of beef jerky samples possessing different microbial compositions. Beef slices were cured using curing solutions formulated both with and without Lactobacillus sakei. Curing was carried out for 18 h at 4 °C prior to hot air drying at 60 °C for 4 h. Jerky samples from both treatment groups were then subjected to ultrasonic frequencies of 25 kHz, 33 kHz and 45 kHz for 30 min. Beef jerky samples were subsequently analysed for fatty acid profile using Gas Chromatography. In the present study, beef slices showed a high level of MUFAs, which accounted for 45.6–53.8%, followed by the SFAs (36.3–47.8%) and PUFAs (4.8–13.7%), respectively. Results demonstrated a significant effect of beef jerky processing on fatty acid profile. Various correlation analyses showed that changes in fatty acid profiles were significantly affected by individual and/or interactive effects of L. sakei, drying time and ultrasonic frequency