United States v. Salvucci: The Problematic Absence of Automatic Standing

The United States Supreme Court recently abolished the automatic standing rule in United States v. Salvucci. The author analyzes the difficulties created for the criminal defendant charged with a possessory crime. In particular, this note focuses on the inequitable position the defendant is placed in when his suppression hearing testimony is used as a tool to impeach subsequent testimony offered at trial. The author continues by pointing out that the prosecutorial self-contradiction, sought to be abolished in Salvucci, remains a part of our present judicial system. In conclusion, the author offers several considerations that will necessarily be an integral part of any future decision regarding the impeachment use of suppression hearing testimony

Acute exercise enhances the expansion of cytotoxic T-cells specific to leukemia and melanoma antigens: implications for adoptive transfer immunotherapy?

INTRODUCTION: The ex vivo expansion of tumor-associated-antigen (TAA)-specific cytotoxic T-cells from healthy donors for adoptive transfer in cancer patients has been used successfully to prevent relapse after hematopoietic stem cell transplantation (HSCT). However, this therapy is limited by the difficulty in priming and expanding sufficient numbers of functional TAA-specific T-cells, as T-cells recognizing TAA are usually low in frequency and avidity in healthy donors. Furthermore, monocyte-derived dendritic cells (Mo-DC) are used for TAA-presentation, but their manufacture is limited by low blood monocyte numbers. Therefore, large and impractical numbers of blood cells are required to successfully expand TAA-specific T-cells. Acute exercise is well-known to transiently activate and increase the numbers of T-cells and monocytes in peripheral blood. We therefore hypothesized that the immune-enhancing effects of exercise could be harnessed to enhance the ex vivo expansion of TAA-specific T-cells for adoptive transfer immunotherapy. AIMS: To examine the effects of acute exercise on (1) the number and function of TAA-specific T-cells expanded ex vivo, and 2) the generation and function of mo-DC. METHODS: 12 healthy adults (mean ± SD: Age 27±2.6yrs) completed an acute bout of stair-running exercise (time: 104±17sec). Mo-DC generated from pre and post exercise blood samples were pulsed with the melanoma-associated-antigens MAGE-A4 and PRAME, the common tumor-antigen survivin, and the leukemia-associated-antigen WT-1. Autologous DC were used to expand TAA-specific T-cells obtained before and after exercise over 14-days. T-cells were enumerated and phenotyped by flow cytometry and function was assessed by ELISPOT and antigen-specific cytotoxicity. RESULTS: A greater number of mo-DC were generated from post-exercise blood samples (pre: 2.0±1.0 X106cells, post: 5.2±2.6 X106cells). This was due to the 1.7 fold increase in blood monocytes post-exercise, as the number of mo-DC generated per input CD14+cell did not differ (pre: 0.40±0.25, post: 0.59±0.36). Total T-cell expansion was increased post-exercise (fold-increase: pre: 2.48±0.75, post: 2.90±0.74). ELISPOT revealed that the majority of donors had enrichment in TAA-specific T-cells post-exercise, as T-cell lines expanded from post-exercise samples exhibited an increased interferon-gamma response to TAA compared to T-cell lines expanded from pre-exercise samples. Exercise had no effect on T-cell phenotype or antigen-specific cytotoxicity in the expanded cells. CONCLUSION: These data indicate that a single bout of exercise enhances mo-DC generation and the expansion of TAA-specific T-cells ex vivo. Exercise may therefore serve as an adjuvant to enhance the expansion of TAA-specific T-cells in healthy donors and improve the efficacy of adoptive transfer therapy in cancer patients

Evaluation of treatments for claw horn lesions in dairy cows in a randomized controlled trial

Lameness is one of the most significant endemic disease problems facing the dairy industry. Claw horn lesions (principally sole hemorrhage, sole ulcer, and white line disease) are some of the most prevalent conditions. Despite the fact that thousands of animals are treated for these conditions every year, experimental evidence is limited on the most effective treatment protocols. A randomized, positively controlled clinical trial was conducted to test the recovery of newly lame cows with claw horn lesions. Animals on 5 farms were locomotion scored every 2 wk. Cows were eligible for recruitment if they had 2 nonlame scores followed by a lame score and had a claw horn lesion on a single claw of a single foot. Following a therapeutic trim, enrolled cows were randomly allocated to 1 of 4 treatments: treatment 1—no further treatment (positive control; TRM), treatment 2—trim plus a block on the sound claw (TB), treatment 3—trim plus a 3-d course of the nonsteroidal anti-inflammatory drug (NSAID) ketoprofen (TN), treatment 4—trim plus a block plus ketoprofen (TBN). The primary outcome measure was locomotion score 35 d after treatment, by an observer blind to treatment group. Descriptive statistics suggested that treatment groups were balanced at the time of enrollment, that is, randomization was successful. Based on a sound locomotion score (score 0) 35 d after treatment, the number of cures was 11 of 45 (24.4%) for TRM, 14 of 39 (35.9%) for TB, 12 of 42 (28.6%) for TN, and 23 of 41 (56.1%) for TBN. The difference between TBN and TRM was significant. To test for confounding imbalances between treatment groups, logistic regression models were built with 2 outcomes, either sound (score 0) or nonlame (score 0 or 1) 35 d after treatment. Compared with TRM, animals that received TBN were significantly more likely to cure to a sound outcome. Farm, treatment season, lesion diagnosis, limb affected, treatment operator, and stage of lactation were included in the final models. Our work suggests that lameness cure is maximized with NSAID treatment in addition to the common practices of therapeutic trimming and elevation of the diseased claw using a block when cows are newly and predominantly mildly lame

Morphology, adipocyte size, and fatty acid analysis of dairy cattle digital cushions, and the effect of body condition score and age

The digital cushion is an essential part of maintaining a healthy foot, working to dissipate foot strike and body weight forces and lameness from claw horn disruption lesions. Despite the importance of the digital cushion, little is known about the basic anatomy, adipocyte morphology, and fatty acid composition in relation to age, limb position, and body condition score. In total, 60 claws (from 17 cows) were selected and collected from a herd, ensuring that body condition score data and computed micro-tomography were known for each animal. Digital cushion tissue underwent histological staining combined with stereology, systematic random sampling, and cell morphology analysis, in addition to lipid extraction followed by fatty acid analysis. The results describe digital cushion architecture and adipocyte sizes. Adipocyte size was similar across all 4 claws (distal left lateral and medial and distal right lateral and medial) and across the ages (aged 2–7 yr); however, animals with body condition score of 3.00 or more at slaughter had a significantly increased cell size in comparison to those with a score of less than 2.50. Of 37 fatty acid methyl esters identified, 5 differed between either the body condition score or different age groups. C10:0 capric acid, C14:0 myristic acid, C15:0 pentadecanoic acid, and C20:0 arachidic acid percentages were all lesser in lower body condition score cows, whereas C22:1n-9 erucic acid measurements were lesser in younger cows. Saturated fatty acid, monounsaturated fatty acid, and polyunsaturated fatty acid percentages were not altered in the different claws, ages, or body condition score groups. Triglyceride quantities did not differ for claw position or age but had decreased quantities in lower body condition score animals. Digital cushion anatomy, cellular morphology, and fatty acid composition have been described in general and also in animals with differing ages, body condition scores, and in the differing claws. Understanding fat deposition, mobilization, and composition are essential in not only understanding the roles that the digital cushion plays but also in preventing disorders and maintaining cattle health and welfare

Cytotoxic T Lymphocyte Therapy for Epstein-Barr Virus+ Hodgkin's Disease

Epstein Barr virus (EBV)+ Hodgkin's disease (HD) expresses clearly identified tumor antigens derived from the virus and could, in principle, be a target for adoptive immunotherapy with viral antigen–specific T cells. However, like most tumor-associated antigens in immunocompetent hosts, these potential targets are only weakly immunogenic, consisting primarily of the latent membrane protein (LMP)1 and LMP2 antigens. Moreover, Hodgkin tumors possess a range of tumor evasion strategies. Therefore, the likely value of immunotherapy with EBV-specific cytotoxic effector cells has been questioned. We have now used a combination of gene marking, tetramer, and functional analyses to track the fate and assess the activity of EBV cytotoxic T lymphocyte (CTL) lines administered to 14 patients treated for relapsed EBV+ HD. Gene marking studies showed that infused effector cells could further expand by several logs in vivo, contribute to the memory pool (persisting up to 12 mo), and traffic to tumor sites. Tetramer and functional analyses showed that T cells reactive with the tumor-associated antigen LMP2 were present in the infused lines, expanded in peripheral blood after infusion, and also entered tumor. Viral load decreased, demonstrating the biologic activity of the infused CTLs. Clinically, EBV CTLs were well tolerated, could control type B symptoms (fever, night sweats, and weight loss), and had antitumor activity. After CTL infusion, five patients were in complete remission at up to 40 mo, two of whom had clearly measurable tumor at the time of treatment. One additional patient had a partial response, and five had stable disease. The performance and fate of these human tumor antigen–specific T cells in vivo suggests that they might be of value for the treatment of EBV+ Hodgkin lymphoma

CAR-T cell. the long and winding road to solid tumors

Adoptive cell therapy of solid tumors with reprogrammed T cells can be considered the "next generation" of cancer hallmarks. CAR-T cells fail to be as effective as in liquid tumors for the inability to reach and survive in the microenvironment surrounding the neoplastic foci. The intricate net of cross-interactions occurring between tumor components, stromal and immune cells leads to an ineffective anergic status favoring the evasion from the host's defenses. Our goal is hereby to trace the road imposed by solid tumors to CAR-T cells, highlighting pitfalls and strategies to be developed and refined to possibly overcome these hurdles

Statistical HOmogeneous Cluster SpectroscopY (SHOCSY): an optimized statistical approach for clustering of ¹H NMR spectral data to reduce interference and enhance robust biomarkers selection.

We propose a novel statistical approach to improve the reliability of (1)H NMR spectral analysis in complex metabolic studies. The Statistical HOmogeneous Cluster SpectroscopY (SHOCSY) algorithm aims to reduce the variation within biological classes by selecting subsets of homogeneous (1)H NMR spectra that contain specific spectroscopic metabolic signatures related to each biological class in a study. In SHOCSY, we used a clustering method to categorize the whole data set into a number of clusters of samples with each cluster showing a similar spectral feature and hence biochemical composition, and we then used an enrichment test to identify the associations between the clusters and the biological classes in the data set. We evaluated the performance of the SHOCSY algorithm using a simulated (1)H NMR data set to emulate renal tubule toxicity and further exemplified this method with a (1)H NMR spectroscopic study of hydrazine-induced liver toxicity study in rats. The SHOCSY algorithm improved the predictive ability of the orthogonal partial least-squares discriminatory analysis (OPLS-DA) model through the use of "truly" representative samples in each biological class (i.e., homogeneous subsets). This method ensures that the analyses are no longer confounded by idiosyncratic responders and thus improves the reliability of biomarker extraction. SHOCSY is a useful tool for removing irrelevant variation that interfere with the interpretation and predictive ability of models and has widespread applicability to other spectroscopic data, as well as other "omics" type of data