Pavlovské vrchy Hills represent a distinctive elevation near the Czech-Austrian border where the active, dormant and relict landslides cover 12% of the area. Here we focused on the chronology of ...landsliding in this area using geological, archaeological and historical evidence. The earliest records of landsliding were determined in locations underlying the dated archaeological settlements. The Upper Paleolithic settlement complex dated between 37-24 ka cal BP, was originally deposited over these landslides. It was consequently destroyed in certain places by additional landslides preceding the last (Upper Pleniglacial) loess deposition (22 ka cal BP). These landslides took place before and after the Upper Paleolithic occupation of this area. This Pleistocene landslide event ranks among the oldest (albeit indirectly) dated landslide within the Czech part of the Western Carpathian Flysch Belt. The chronology of later, historical, landsliding was determined using written records (chronicles, official reports, archival evidence, etc.). Continuous records of landsliding were available as of the middle of the seventeenth century. The major concentration of landslides occurred at the beginning of the twentieth century (1910-1915). The 1663 landslide is currently the oldest landslide, in the Czech part of the Western Carpathian Flysch Belt, which was dated on the basis of documentary data.
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A low-cost Digital Signal Processor (DSP) unit for advanced Scanning Probe Microscopy measurements is presented. It is based on Red Pitaya board and custom built electronic boards ...with additional high bit depth AD and DA converters. By providing all the necessary information (position and time) with each data point collected it can be used for any scan path, using either existing libraries for scan path generation or creating adaptive scan paths using Lua scripting interface. The DSP is also capable of performing statistical calculations, that can be used for decision making during scan or for the scan path optimisation on the DSP level.
Backround. Capsule colonoscopy might present an alternative to colonoscopy for colorectal neoplasia screening. Aim. To assess the accuracy of second-generation capsule colonoscopy (CCE2) for ...colorectal neoplasia detection compared with conventional colonoscopy (CC). Methods. From 2011–2015, we performed a multicenter, prospective, cross-over study evaluating the use of CCE2 as a possible colorectal cancer (CRC) screening test based on the assessment of the method’s characteristics (accuracy) and safety and patient acceptance of the routine. Enrolled participants fulfilled the CRC screening population criteria if they were asymptomatic, were older than 50, and had no personal or familial history of colorectal neoplasia. The primary outcome was accuracy for the detection of polyps≥6 mm. Secondary outcomes were accuracy for all polyps, polyps≥10 mm, adenomas≥10 mm, and cancers, the quality of bowel cleansing, safety, and CCE2 acceptability by the screening population. Results. A total of 236 individuals were examined; 11 patients (5%) were excluded. Therefore, 225 subjects (95%) were considered in the intention-to-screen (ITS) group. A total of 201 patients (89%) completed both examinations successfully (per protocol group). In the ITS group, polyps were diagnosed during CC in 114 subjects (51%); polyps≥6 mm, polyps≥10 mm, and adenomas≥10 mm were diagnosed in 34 (15%), 16 (7%), and 11 (5%) patients, respectively. The sensitivity of CCE2 for polyps≥6 mm, polyps≥10 mm, and adenomas≥10 mm was 79% (95% confidence interval (CI): 62–91%), 88% (95% CI: 62–98%), and 100% (95% CI: 72–100%), respectively. Conclusion. Second-generation capsule colonoscopy is a safe, noninvasive, and sensitive method for colorectal neoplasia detection although CC remains the preferred method for considerable proportion of subjects. CCE2 may therefore be accepted as the primary screening test for colorectal cancer screening.
Cellular immunity against SARS-CoV-2 is an important component of the immune response to the virus. At present, two such tests based on interferon-gamma release (interferon-γ release assays, IGRAs) ...are available-Quan-T-Cell SARS-CoV-2 by EUROIMMUN and T-SPOT.COVID by Oxford Immunotec. In this paper, we compared the results of these two tests in 90 subjects employed at the Public Health Institute Ostrava who had previously undergone COVID-19 infection or were vaccinated against that disease. To the best of our knowledge, this is the first head-to-head comparison of these two tests evaluating T-cell-mediated immunity against SARS-CoV-2. In addition, we also evaluated humoral immunity in the same individuals using the in-house virus neutralization test and IgG ELISA assay. The evaluation yielded similar results for both IGRAs, with Quan-T-Cell appearing to be insignificantly (
= 0.08) more sensitive (all 90 individuals were at least borderline positive) than T-SPOT.COVID (negative results found in five patients). The overall qualitative (presence/absence of immune response) agreement of both tests with virus neutralization test and anti-S IgG was also excellent (close or equal to 100% in all subgroups, with the exception of unvaccinated Omicron convalescents, a large proportion of whom, i.e., four out of six subjects, were IgG negative while at least borderline positive for T-cell-mediated immunity measured by Quan-T). This implies that the evaluation of T-cell-mediated immunity is a more sensitive indicator of immune response than the evaluation of IgG seropositivity. This is true at least for unvaccinated patients with a history of being infected only by the Omicron variant, but also likely for other groups of patients.
Primary human lung organoid-derived air-liquid interface (ALI) cultures serve as a physiologically relevant model to study human airway epithelium in vitro. Here, we present a protocol for ...establishing these cultures from cryopreserved human lung tissue. We describe steps for lung tissue cryostorage, tissue dissociation, lung epithelial organoid generation, and ALI culture differentiation. We also include quality control steps and technical readouts for monitoring virus response. This protocol demonstrates severe acute respiratory syndrome coronavirus 2 infection in these cultures as an example of their utility. For complete details on the use and execution of this protocol, please refer to Diana Cadena Castaneda et al. (2023).
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We describe a pipeline for optimized and streamlined multiplexed immunofluorescence-guided laser capture microdissection allowing the harvest of individual cells based on their phenotype and tissue ...localization for transcriptomic analysis with next-generation RNA sequencing. Here, we analyze transcriptomes of CD3+ T cells, CD14+ monocytes/macrophages, and melanoma cells in non-dissociated metastatic melanoma tissue. While this protocol is described for melanoma tissues, we successfully applied it to human tonsil, skin, and breast cancer tissues as well as mouse lung tissues.
For complete details on the use and execution of this protocol, please refer to Martinek et al. (2022).
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•Assessing the topology and transcriptome of individual cells in human melanoma tissue•Optimized immunofluorescence staining protocol for RNA preservation•Multiplexed immunofluorescence-based harvest of individual cells from tissues•RNA library preparation for next-generation sequencing
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
We describe a pipeline for optimized and streamlined multiplexed immunofluorescence-guided laser capture microdissection allowing the harvest of individual cells based on their phenotype and tissue localization for transcriptomic analysis with next-generation RNA sequencing. Here, we analyze transcriptomes of CD3+ T cells, CD14+ monocytes/macrophages, and melanoma cells in non-dissociated metastatic melanoma tissue. While this protocol is described for melanoma tissues, we successfully applied it to human tonsil, skin, and breast cancer tissues as well as mouse lung tissues.
BackgroundInteractions between immune and tumor cells are critical to determining cancer progression and response. In addition, preclinical prediction of immune-related drug efficacy is limited by ...interspecies differences between human and mouse, as well as inter-person germline and somatic variation. To address these gaps, we developed an autologous system that models the tumor microenvironment (TME) from individual patients with solid tumors.MethodWith patient-derived bone marrow hematopoietic stem and progenitor cells (HSPCs), we engrafted a patient’s hematopoietic system in MISTRG6 mice, followed by transfer of patient-derived xenograft (PDX) tissue, providing a fully genetically matched model to recapitulate the individual’s TME. We used this system to prospectively study tumor-immune interactions in patients with solid tumor.ResultsAutologous PDX mice generated innate and adaptive immune populations; these cells populated the TME; and tumors from autologously engrafted mice grew larger than tumors from non-engrafted littermate controls. Single-cell transcriptomics revealed a prominent vascular endothelial growth factor A (VEGFA) signature in TME myeloid cells, and inhibition of human VEGF-A abrogated enhanced growth.ConclusionsHumanization of the interleukin 6 locus in MISTRG6 mice enhances HSPC engraftment, making it feasible to model tumor-immune interactions in an autologous manner from a bedside bone marrow aspirate. The TME from these autologous tumors display hallmarks of the human TME including innate and adaptive immune activation and provide a platform for preclinical drug testing.
Lipophilic bisphosphonium salts are among the most promising antiprotozoal leads currently under investigation. As part of their preclinical evaluation we here report on their mode of action against ...African trypanosomes, the etiological agents of sleeping sickness. The bisphosphonium compounds CD38 and AHI-9 exhibited rapid inhibition of Trypanosoma brucei growth, apparently the result of cell cycle arrest that blocked the replication of mitochondrial DNA, contained in the kinetoplast, thereby preventing the initiation of S-phase. Incubation with either compound led to a rapid reduction in mitochondrial membrane potential, and ATP levels decreased by approximately 50% within 1 h. Between 4 and 8 h, cellular calcium levels increased, consistent with release from the depolarized mitochondria. Within the mitochondria, the Succinate Dehydrogenase complex (SDH) was investigated as a target for bisphosphonium salts, but while its subunit 1 (SDH1) was present at low levels in the bloodstream form trypanosomes, the assembled complex was hardly detectable. RNAi knockdown of the SDH1 subunit produced no growth phenotype, either in bloodstream or in the procyclic (insect) forms and we conclude that in trypanosomes SDH is not the target for bisphosphonium salts. Instead, the compounds inhibited ATP production in intact mitochondria, as well as the purified F1 ATPase, to a level that was similar to 1 mM azide. Co-incubation with azide and bisphosphonium compounds did not inhibit ATPase activity more than either product alone. The results show that, in T. brucei, bisphosphonium compounds do not principally act on succinate dehydrogenase but on the mitochondrial FoF1 ATPase.
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•Bisphosphonium salts display highly promising antiprotozoal activity.•It has been reported that, in Leishmania, they act on the mitochondrial SDH complex.•We show that in Trypanosoma brucei SDH is not essential and not the drug target.•Instead, we present strong evidence that the F1F0 ATPase is the target.
The book Geography of Barriers: Examples of Good Barrier-free Practice offers a collection of texts that are interested in making space, services, and information accessible. Thus, the collection is ...primarily about barriers, their nature, localization, and possible effects. Under barriers, most people imagine material barriers for wheelchair users in the form of steps or sidewalk curbs. Only a few would think that barriers can be represented by loosely placed rugs, unfit lighting, large glass surface, a doorbell without visual output, unwanted attention, frequent asking and offering help, lack of language knowledge, pity, etc. Thus, the book broadens awareness of what can be considered a barrier. The texts are divided into three parts – theoretical, methodic, and empirical – and their arrangement follows the logic of thematic focusing from the most general to the most concrete types of barriers. While the theoretical part places the policies of accessibility into the wider context of scientific disciplines and their development – such as disability studies, disability geography or service design, the methodic part discusses the introduction of accessibility policies in the city of Brno, describing how it tried to make its space, services, and information more accessible. Finally, the empirical part offers 18 examples of relatively good barrier-free implementations that have been recently realized in Brno. This division reflects an effort to target several groups of readers. The first part thus addresses mainly students interested in accessibility issues, the second, methodic part is intended for politicians and municipalities searching for inspiration to implement their own accessibility measures and policies. The last part stating concrete examples of good barrier-free implementations is aimed at general public, at people who like to think about a whole range of topics related to accessibility of space, services, and information.
Abstract
Background and study aims
Circular ESD (CESD) is a treatment option for patients with extensive early esophageal cancer. Its major drawback is the development of a stricture. Stenting may ...represent an attractive prevention strategy. We designed an experimental study to assess the effect of stents covered with acellular biomatrix (AB) and a drug-eluting stent.
Materials and methods
Thirty-five 35 pigs underwent CESD and were randomized into six groups: G1 (control), G2 (SEMS), G3 (SEMS + AB), G4 (SEMS + AB + steroid-eluting layer), G5 (biodegradable stent BD), G6 (BD + AB). SEMS were placed alongside the post-CESD defect, fixed and removed after 21 days. The main outcomes were stricture development, severity, and histopathology.
Results
Pigs with BD stents (G5, 6) experienced severe inflammation and hypergranulation without biodegradation, therefore, these groups were closed prematurely. Significant strictures developed in 29 of 30 pigs (96.7 %). The most severe stricture developed in G2 and G4 (narrowest diameter (mm) 8.5 ± 3, 3 (G2) and 8.6 ± 2.1 (G4) vs. 17 ± 7.3 (G1) and 13.5 ± 8.3 (G3);
P
< 0.01. Signs of re-epithelization were present in 67 % and 71 % in G1 and G2 and in 100 % in G3 and G4. The most robust re-epithelization layer was present in G4. The inflammation was the most severe in G1 (mean score 2.3) and least severe in G4 (0.4).
Conclusions
Stenting did not effectively prevent development of post-CESD esophageal stricture. SEMS with AB resulted in improved re-epithelization and decreased stricture severity. Steroid-eluting SEMS suppressed inflammation. BD stents seem inappropriate for this indication.
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