Waltonfagan3450

We found significantly higher concentrations in railway-associated samples of dandelion and grain for both metals (particularly lead, iron, and chromium), and the sum of 16 PAHs. Several metals and PAHs in railway-associated samples exceeded regulatory standards for soil or animal feed. Mycotoxins were detectable in grain samples, but occurred well below permissible standards. Metal concentrations in bear hair were not predicted by railway use, but higher metal concentrations occurred in male bears and two individuals that used ski hills during fall. As mitigation to reduce wildlife exposure to contaminants, particularly in protected areas, we encourage removal of railway grain deposits, regular maintenance of railway infrastructure, such as lubricating stations, and investigation of contaminants associated with other human infrastructures, such as ski hills.BACKGROUND Second-generation cryoballoon ablation is safe and effective in patients with persistent atrial fibrillation (AF). The aim of this study is to report the real long-term AF burden and freedom from AF post-cryoablation using continuous monitoring, and to assess whether intraoperative confirmation of pulmonary vein isolation using electrical mapping is necessary. METHODS A total of 33 patients (mean age 75.7 ± 5.6 years, 16 men) with persistent AF who underwent second-generation cryoablation without electrical mapping were reviewed. All patients had a cardiac implantable device and were followed up for a mean of 755 ± 170 days. RESULTS AF burden significantly decreased from 67.51% ± 34.90% to 18.28% ± 26.65% at 1-year follow-up, and this reduction was maintained at final follow-up (18.26% ± 23.70%, p  less then  0.001). Continuous monitoring revealed a freedom from AF rate of 33% and 24% at 1-year and full follow-up, respectively. Patients who remained in persistent AF at final follow-up had a trend towards higher pre-ablation AF burden (81.6% ± 29.7% vs 57.3% ± 36.4%, p = 0.08). CONCLUSION Second-generation cryoablation without confirming pulmonary vein isolation using electrical mapping is effective leading to significant reductions in AF burden based on continuous beat-to-beat monitoring at 1-year and long-term follow-up.Natural killer (NK) cells are cytotoxic lymphocytes of our immune system with the ability to identify and kill certain virally infected and tumor-transformed cells. During the past 15 years, it has become increasingly clear that NK cells are involved in tumor immune surveillance and that they can be utilized to treat cancer patients. GSK461364 manufacturer However, their ability to induce durable responses in settings of adoptive cell therapy needs to be further improved. One possible approach is to genetically engineer NK cells to augment their cytotoxicity per se, but also their ability to persist in vivo and home to the tumor-bearing tissue. In recent years, investigators have explored the potential of viral transduction and mRNA electroporation to modify NK cells. Although these methods have generated promising data, they are associated with certain limitations. With the increasing advances in the CRISPR/Cas9 technology, investigators have now turned their attention toward using this technology with NK cells as an alternative method. In this book chapter, we introduce NK cells and provide an historical overview of techniques to genetically engineer lymphocytes. Further, we elucidate protocols for inducing double-strand breaks in NK cells via CRISPR/Cas9 together with readouts to address its efficacy and functional outcome. We also discuss the pros and cons of the described readouts. The overall aim of this book chapter is to help introduce the CRISPR/Cas9 technology to the broader audience of NK cell researchers.Innate lymphoid cells (ILCs) are emerging as important effectors of innate immunity and play a critical role in maintaining intestinal immune homeostasis. They are tissue-residing immune cells that can be subdivided based on master transcription factor and cytokine expression, bearing striking resemblance to their CD4+ T helper (Th) cell counterparts. ILCs are increasingly recognized as potential mediators of inflammatory bowel disease (IBD) providing a need to explore their functional and phenotypic differences in health vs. disease. In this chapter we outline protocols for the characterization of human ILCs and intracellular cytokine expression using flow cytometry. We include protocols for isolating human peripheral blood and colonic lamina propria mononuclear cells essential for evaluating human IBD specimens.Self-organizing mini-intestines cultured ex vivo from intestinal biopsy/resected samples, termed intestinal organoids or enteroids, present a unique opportunity for mechanistic investigation of health and disease of the intestinal epithelium. These patient-derived epithelial cultures are nontransformed, retain the genetic background of the patient, maintain regional specificity, differentiate into all major cell types of the intestinal epithelium, and are physiologically active. The biological relevance of human intestinal enteroids also circumvents the need for animal models for studies on the human gastrointestinal epithelium. Coculture with human endogenous microbes allows for exciting new studies on microbial-host interactions.While the popularity of organoids/enteroids for human research has risen drastically over the past decade, existing work and published methods are primarily limited to adult tissue. Here, we describe a concise and effective method for the establishment neonatal enteroids (including preterm and term) from surgically resected tissue or biopsy material. While the protocol works on adult tissue/biopsies, it has been specifically adopted and optimised for neonatal tissue. We detail the procedure at each stage ranging from human tissue collection and extraction of stem cells from the tissue, to passaging and general maintenance of organoid/enteroid lines, and how to freeze and revive lines as needed.The highly parallel nature of sequencing by synthesis (SBS) allows millions of amplicons to be sequenced simultaneously, which has led to enormous interest in the investigation of bacterial communities (often referred to as the microbiota). In this protocol, we describe a method for the 'universal' amplification of the v4 region of the bacterial 16S rRNA gene from genomic DNA and prepare these amplicons so that they can be sequenced using the MiSeq system (Illumina). The protocol provides instruction on sequencing of 188 genomic DNA samples plus PCR positive and negative controls, which can be applied to any sample type where bacterial DNA may be of interest.