Understanding response to rituximab treatment in rheumatoid arthritis through immune fingerprinting of T and B cells

Despite the great research advances in dissecting the mechanisms underlying rheumatoid arthritis onset and development, the exact pathophysiology of this disease remains unsolved. In the past years, the introduction of biologicals, and in particular of therapeutic monoclonal antibodies (mAb), has constituted a major breakthrough in the clinical management of the disease. Although these new drugs have proven effective, in some patients, remission or low disease activity is only partially or temporally achieved. Understanding the mechanism behind this incomplete response might lead to improved therapies and in ultimately to improved quality of life for patients. This thesis describes our efforts to elucidate the mechanisms behind response to B-cell depletion therapy using rituximab in rheumatoid arthritis. We evaluated how both direct and indirect effects can influence clinical response to the treatment. To achieve this we applied AIRR sequencing or Adaptive Immune Receptor Repertoire sequencing, which allows to identify and quantify all T- and B-cell receptors within an individual. This methodology allowed us to fingerprint and monitor ongoing immune responses in cohorts of rheumatoid arthritis patients undergoing rituximab treatment

Understanding response to rituximab treatment in rheumatoid arthritis through immune fingerprinting of T and B cells

Despite the great research advances in dissecting the mechanisms underlying rheumatoid arthritis onset and development, the exact pathophysiology of this disease remains unsolved. In the past years, the introduction of biologicals, and in particular of therapeutic monoclonal antibodies (mAb), has constituted a major breakthrough in the clinical management of the disease. Although these new drugs have proven effective, in some patients, remission or low disease activity is only partially or temporally achieved. Understanding the mechanism behind this incomplete response might lead to improved therapies and in ultimately to improved quality of life for patients. This thesis describes our efforts to elucidate the mechanisms behind response to B-cell depletion therapy using rituximab in rheumatoid arthritis. We evaluated how both direct and indirect effects can influence clinical response to the treatment. To achieve this we applied AIRR sequencing or Adaptive Immune Receptor Repertoire sequencing, which allows to identify and quantify all T- and B-cell receptors within an individual. This methodology allowed us to fingerprint and monitor ongoing immune responses in cohorts of rheumatoid arthritis patients undergoing rituximab treatment

Effects of polysaccharides from Botryotinia fuckeliana (Botrytis cinerea) on in vitro culture of table and wine grapes (Vitis vinifera)

Shoots of several table and wine grape cultivars were cultured in vitro on a medium supplemented with polysaccharides obtained from a culture filtrate of Botryotinia fuckeliana through differential ethanolic precipitations. The general effects of polysaccharides resulted in leaf yellowness and in a reduction of fresh and dry weight. Differential response of assayed cultivars to polysaccharides seemed to be not related to their bunch susceptibility to grey mould under field conditions

A ready-to-use single- and Duplex-TaqMan-qPCR assay to detect and quantify the biocontrol agents Trichoderma asperellum and Trichoderma gamsii

Trichoderma asperellum strain icc012 and Trichoderma gamsii strain icc080, the microbial active ingredients of Remedier™ (ISAGRO, Novara, Italy), are biocontrol agents (BCAs) employable for crop protection against a wide range of fungal pathogens, including soil-borne pathogens and fungi involved in grapevine trunk disease. In this study, single and duplex real-time quantitative PCR (qPCR) methods to detect and quantify T. asperellum and T. gamsii were developed. Primers/probe sets were designed on the T. asperellum and T. gamsii rpb2 genes and tested for specificity on a panel of microorganisms commonly associated with grape wood and soil. No differences were observed comparing single- and duplex-qPCR assays on different BCAs, 1 pg of target DNA was detected approximately at Cq= 34. R2-values and the efficiency were always equal to 0.99 and > 80%, respectively. The detection limit of the duplex-qPCR assay on artificially inoculated samples was 2 × 103and 4 × 104conidia g-1of grape wood tissue and soil, respectively. The methods will be useful to better schedule BCA application in the field and in grapevine nurseries, as well as for investigating the dynamic of BCA populations

PP272—Migraine and parthenolide inhibition of transient receptor potential ankyrin 1

2013 e103 emerged as a major complication of bortezomib therapy, which usually appears in the first courses of therapy with a number of sensory and painful symptoms, including reduced threshold to mechanical and cold stimuli. No satisfactory explanation or effective treatment exists for bortezomib-evoked CIPN. Patients (or Materials) and Methods: In this study, we evaluated whether TRPA1 acted as a critical mediator of CIPN by bortezomib or oxaliplatin in a mouse model system. Results: Our data demonstrated that CIPN hypersensitivity phenotype that was stably established by bortezomib could be transiently reverted by systemic or local treatment with the TRPA1 antagonist HC-030031. A similar effect was produced by the oxidative stress scavenger α -lipoic acid. Notably, the CIPN phenotype was abolished completely in mice that were genetically deficient in TRPA1, highlighting its essential role. Administration of bortezomib or oxaliplatin, which also elicits TRPA1-dependent hypersensitivity, produced a rapid, transient increase in plasma of carboxy-methyllysine, a byproduct of oxidative stress. Short-term systemic treatment with either HC-030031 or α -lipoic acid could completely prevent hypersensitivity if administered before the cytotoxic drug. Conclusion: Our findings highlight a key role for early activation/ sensitization of TRPA1 by oxidative stress by-products in producing CIPN. Furthermore, they suggest prevention strategies for CIPN in patients through the use of early, short-term treatments with TRPA1 antagonists. Disclosure of Interest: None declared

Absinthin, an agonist of the bitter taste receptor hTAS2R46, uncovers an ER-to-mitochondria Ca2–shuttling event

Type 2 taste receptors (TAS2R) are G protein-coupled receptors first described in the gustatory system, but have also been shown to have extra-oral localizations, including airway smooth muscle (ASM) cells, in which TAS2R have been reported to induce relaxation. TAS2R46 is an unexplored subtype that responds to its highly specific agonist absinthin. Here, we first demonstrate that, unlike other bitter-taste receptor agonists, absinthin alone (1 μM) in ASM cells does not induce Ca2+ signals, but reduces histamine-induced cytosolic Ca2+ increases. To investigate this mechanism, we introduced into ASM cells aequorin-based Ca2+ probes targeted to the cytosol, sub-plasma membrane domain, or the mitochondrial matrix. We show that absinthin reduces cytosolic histamine-induced Ca2+-rises and simultaneously increases Ca2+-influx into mitochondria. We found that this effect is inhibited by the potent human TAS2R46 (hTAS2R46) antagonist 3β-hydroxydihydrocostunolide and is no longer evident in hTAS2R46-silenced ASM cells, indicating that it is hTAS2R46-dependent. Furthermore, these changes were sensitive to the mitochondrial uncoupler carbonyl cyanide p-(trifluoromethoxy)phenyl-hydrazone (FCCP); the mitochondrial calcium uniporter inhibitor KB-R7943 (carbamimidothioic-acid); the cytoskeletal disrupter latrunculin; and an inhibitor of the exchange protein directly activated by cAMP (EPAC), ESI-09. Similarly, the β2 agonist salbutamol also could induce Ca2+ shuttling from cytoplasm to mitochondria, suggesting that this new mechanism might be generalizable. Moreover, forskolin and an EPAC activator mimicked this effect in HeLa cells. Our findings support the hypothesis that plasma membrane receptors can positively regulate mitochondrial Ca2+ uptake, adding a further facet to the ability of cells to encode complex Ca2+ signals

Patient-Reported Outcomes in the Translational Breast Cancer Research Consortium

Members of the Translational Breast Cancer Research Consortium conducted an expert-driven literature review to identify a list of domains and to evaluate potential measures of these domains for inclusion in a list of preferred measures. Measures were included if they were easily available, free of charge, and had acceptable psychometrics based on published peer-reviewed analyses. A total of 22 domains and 52 measures were identified during the selection process. Taken together, these measures form a reliable and validated list of measurement tools that are easily available and used in multiple cancer trials to assess patient-reported outcomes in relevant patients