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Journal of Visualized Experiments : JoVE Aug 2023Neutrophil extracellular traps (NETs) are released by neutrophils as a response to bacterial infection or traumatic tissue damage but also play a role in autoimmune...
Neutrophil extracellular traps (NETs) are released by neutrophils as a response to bacterial infection or traumatic tissue damage but also play a role in autoimmune diseases and sterile inflammation. They are web-like structures composed of double-stranded DNA filaments, histones, and antimicrobial proteins. Once released, NETs can trap and kill extracellular pathogens in blood and tissue. Furthermore, NETs participate in homeostatic regulation by stimulating platelet adhesion and coagulation. However, the dysregulated production of NETs has also been associated with various diseases, including sepsis or autoimmune disorders, which makes them a promising target for therapeutic intervention. Apart from electron microscopy, visualizing NETs using immunofluorescence imaging is currently one of the only known methods to demonstrate NET interactions in tissue. Therefore, various staining methods to visualize NETs have been utilized. In the literature, different staining protocols are described, and we identified four key components showing high variability between protocols: (1) the types of antibodies used, (2) the usage of autofluorescence-reducing agents, (3) antigen retrieval methods, and (4) permeabilization. Therefore, in vitro immunofluorescence staining protocols were systemically adapted and improved in this work to make them applicable for different species (mouse, human) and tissues (skin, intestine, lung, liver, heart, spinal disc). After fixation and paraffin-embedding, 3 µm thick sections were mounted onto slides. These samples were stained with primary antibodies for myeloperoxidase (MPO), citrullinated histone H3 (H3cit), and neutrophil elastase (NE) according to a modified staining protocol. The slides were stained with secondary antibodies and examined using a widefield fluorescence microscope. The results were analyzed according to an evaluation sheet, and differences were recorded semi-quantitatively. Here, we present an optimized NET staining protocol suitable for different tissues. We used a novel primary antibody to stain for H3cit and reduced non-specific staining with an autofluorescence-reducing agent. Furthermore, we demonstrated that NET staining requires a constant high temperature and careful handling of samples.
Topics: Humans; Animals; Mice; Extracellular Traps; Histones; Diagnostic Imaging; Fluorescent Antibody Technique; Neutrophils; Antibodies; Coloring Agents; Autoimmune Diseases
PubMed: 37677039
DOI: 10.3791/65272 -
PLoS Computational Biology Oct 2022Language interfaces with many other cognitive domains. This paper explores how interactions at these interfaces can be studied with deep learning methods, focusing on...
Language interfaces with many other cognitive domains. This paper explores how interactions at these interfaces can be studied with deep learning methods, focusing on the relation between language emergence and visual perception. To model the emergence of language, a sender and a receiver agent are trained on a reference game. The agents are implemented as deep neural networks, with dedicated vision and language modules. Motivated by the mutual influence between language and perception in cognition, we apply systematic manipulations to the agents' (i) visual representations, to analyze the effects on emergent communication, and (ii) communication protocols, to analyze the effects on visual representations. Our analyses show that perceptual biases shape semantic categorization and communicative content. Conversely, if the communication protocol partitions object space along certain attributes, agents learn to represent visual information about these attributes more accurately, and the representations of communication partners align. Finally, an evolutionary analysis suggests that visual representations may be shaped in part to facilitate the communication of environmentally relevant distinctions. Aside from accounting for co-adaptation effects between language and perception, our results point out ways to modulate and improve visual representation learning and emergent communication in artificial agents.
Topics: Language; Communication; Semantics; Cognition; Visual Perception
PubMed: 36315590
DOI: 10.1371/journal.pcbi.1010658 -
Medicine Nov 2018This study is to investigate the computed tomography (CT) image quality of the low- tube-voltage protocol with low contrast agent dose.CT portography was performed in... (Randomized Controlled Trial)
Randomized Controlled Trial
Computed tomography portography of patients with cirrhosis with normal body mass index: Comparison between low-tube-voltage CT with low contrast agent dose and conventional CT.
This study is to investigate the computed tomography (CT) image quality of the low- tube-voltage protocol with low contrast agent dose.CT portography was performed in 118 cirrhosis patients with body mass index (BMI) less than 25 kg/m under 2 protocols: Protocol A, tube voltage of 90 kVp/395 mAs and contrast agent dosage of 1.2 mL/kg, and, Protocol B, tube voltage of 120 kVp/200 mAs and contrast agent dosage of 1.5 mL/kg.The number of patients in each protocol was 59. The CT value noise, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) in portal veins was comparatively analyzed between the 2 protocols. The subjective image quality was further assessed on 5-point scales. Radiation dose was also recorded and statistical analysis was performed.The CT value, CNR, and SNR of the images were higher at 90 kVp than those at 120 kVp (P < .05). There was no significant difference in image noise between the 2 protocols (P > .05). The CT dose index volume, dose-length product, and effective dose at 90 kVp were 18.2%, 16.0%, and 16.0% less than that at 120 kVp, respectively. There was no difference in image quality score between the 2 protocols (P > .05). The average amount of contrast agent was decreased by 17.8% when the 90 kVp protocol was used.CT portography at 90 kVp combined with low-dosage contrast agent leads to a significant reduction in radiation dose and improved SNR and CNR, without deterioration of image quality.
Topics: Adult; Aged; Body Mass Index; Contrast Media; Dose-Response Relationship, Drug; Female; Humans; Iohexol; Liver Cirrhosis; Male; Middle Aged; Portography; Radiation Dosage; Radiographic Image Interpretation, Computer-Assisted; Tomography, X-Ray Computed
PubMed: 30508890
DOI: 10.1097/MD.0000000000013141 -
Journal of Nuclear Medicine Technology Mar 2007Is there an ideal myocardial perfusion imaging protocol? In order to answer this question and choose a protocol for clinical use, one must understand the characteristics... (Review)
Review
Is there an ideal myocardial perfusion imaging protocol? In order to answer this question and choose a protocol for clinical use, one must understand the characteristics of the available radiopharmaceuticals, the protocol variables, and the advantages and disadvantages of each. After reading this article, the technologist should be able to list the ideal characteristics of a myocardial perfusion imaging agent, describe and compare the characteristics of myocardial perfusion imaging agents, discuss the relationship between coronary blood flow and myocardial uptake of various tracers, describe imaging protocols, and discuss the advantages and disadvantages of each protocol.
Topics: Coronary Artery Disease; Coronary Circulation; Coronary Vessels; Humans; Image Enhancement; Radiopharmaceuticals; Tomography, Emission-Computed; Ventricular Dysfunction, Left
PubMed: 17337651
DOI: No ID Found -
Nephrology, Dialysis, Transplantation :... Sep 2014Histopathological lesions in renal biopsy (RB) at presentation of ANCA-associated vasculitis (AAV) have been described in depth but repeat protocolized renal biopsies...
BACKGROUND
Histopathological lesions in renal biopsy (RB) at presentation of ANCA-associated vasculitis (AAV) have been described in depth but repeat protocolized renal biopsies are seldomly performed in AAV. In this study, we present a group of AAV patients with repeat protocolized biopsies, and we evaluate their clinical significance.
METHODS
A total of 17 consecutive patients diagnosed between 1991 and 1995 with AAV and renal involvement confirmed by biopsy at presentation in a single center underwent a protocol planned rebiopsy in remission after a median of 13 months (range 11-28) from diagnosis. Biopsies were assessed by two independent pathologists, blinded to patient data. Clinical data were collected retrospectively.
RESULTS
Patients were followed-up for a median of 189 months from diagnosis. Renal relapse was observed in eight patients (47.1%), seven patients died, three patients reached end-stage renal failure. There was a significant decrease in the percentage of acute lesions (cellular crescents, fibrinoid necrosis, P < 0.001) and a significant increase in chronic lesions (glomerulosclerosis, interstitial fibrosis, P ≤ 0.01) in the repeat RB compared with the first RB. This resulted in a class change over the biopsies within most patients. The percentage of normal glomeruli in the first biopsy positively correlated with estimated GFR at the end of follow-up (rs = 0.509, P = 0.05).
CONCLUSIONS
This is the first study on protocolized repeat biopsies in AAV, giving insight into disease activity under immunosuppressive treatment. Apparently, many AAV patients have grumbling disease with ongoing activity, eventually leading to an increased amount of chronic lesions.
Topics: Adult; Aged; Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis; Biopsy; Clinical Protocols; Female; Humans; Immunosuppressive Agents; Kidney; Kidney Diseases; Kidney Glomerulus; Male; Middle Aged; Retrospective Studies; Severity of Illness Index; Young Adult
PubMed: 24578468
DOI: 10.1093/ndt/gfu042 -
Journal of the Mechanical Behavior of... Sep 2021To simulate removing luting agent remnants from crowns fixed onto implant-abutment analogs using a standardized machine-driven protocol including a scaler and air...
INTRODUCTION
To simulate removing luting agent remnants from crowns fixed onto implant-abutment analogs using a standardized machine-driven protocol including a scaler and air polishing or sonic.
MATERIAL AND METHODS
A motor-driven device was constructed that controlled the rotational speed of the specimens, machining distance, contact pressure, and working time. A standardized layer of cement (Provicol, VOCO; Cuxhaven, G; Ketac Cem, 3MEspe; Seefeld, G; or Rely X Unicem, 3MEspe, Seefeld, G) was placed onto the finishing line of the crowns luted onto titanium-abutment analogs. The cement layer was scaled with a fresh titanium scaler maneuvered by the motor-driven device and treated with air polishing or sonic. Protocol 1: Scaling only for 20s, 40s, or 60s; n=20; protocol 2: 40s of scaling plus 20s of air polishing; protocol 3: 20s of scaling plus 40s of air polishing; protocol 4: 20s of scaling plus 40s of sonic; protocol 5: 40s of scaling plus 20s of sonic; protocols 2-5: n=10. Cement remnants were counted digitally as "percentage of remnants".
STATISTICS
mean, standard deviation, Bonferroni post hoc tests; α=0.05.
RESULTS
Ketac Cem was easily removed by scaling only and Provicol by scaling and air polishing, but the self-adhesive resin composite cement Rely X Unicem was not removable with the device. Only remnants of Provicol could be significantly reduced by further treatment after scaling (p<0.001).
CONCLUSION
The presented motor-driven device enables reproducible investigations of various cleaning protocols and is thus useful to create an overview of cleaning protocols needed for the different types of cement.
Topics: Composite Resins; Crowns; Dental Cements; Glass Ionomer Cements; Materials Testing; Resin Cements; Titanium
PubMed: 34090119
DOI: 10.1016/j.jmbbm.2021.104584 -
Cancer Chemotherapy and Pharmacology Mar 2001A case of haemolytic anaemia in a patient under treatment with UFT for metastatic colon cancer is reported. Haemolytic anaemia has previously been associated with many...
A case of haemolytic anaemia in a patient under treatment with UFT for metastatic colon cancer is reported. Haemolytic anaemia has previously been associated with many other chemotherapeutic agents, but not with UFT, an oral anticancer agent combining tegafur (Ftorafur, a prodrug of 5-fluorouracil) and uracil.
Topics: Aged; Aged, 80 and over; Anemia, Hemolytic; Antineoplastic Combined Chemotherapy Protocols; Colonic Neoplasms; Humans; Male; Tegafur; Uracil
PubMed: 11320674
DOI: 10.1007/s002800000195 -
Seminars in Oncology Apr 2001The methotrexate/vinblastine/doxorubicin/cisplatin (MVAC) regimen has been considered standard therapy for advanced bladder cancer. However, the toxicity of this regimen... (Review)
Review
The methotrexate/vinblastine/doxorubicin/cisplatin (MVAC) regimen has been considered standard therapy for advanced bladder cancer. However, the toxicity of this regimen motivated the development of safer and/or more effective therapy. Gemcitabine (Gemzar; Eli Lilly and Company, Indianapolis, IN) is a promising new agent with activity in advanced disease. Early phase studies yielded response rates of 23% to 29% with single-agent treatment and 41% to 57% with the combination of gemcitabine/cisplatin (GC). In a randomized phase III trial involving 405 randomized patients with locally advanced or metastatic bladder cancer, data comparing gemcitabine at 1,000 mg/m(2) on days 1, 8, and 15 plus cisplatin at 70 mg/m(2) on day 2 every 28 days with standard MVAC demonstrated that the two regimens were associated with comparable response rates, time to disease progression, and overall survival. The GC regimen was associated with superior safety and tolerability, including reduced frequencies of grade 3/4 mucositis (1% v 22%), neutropenic fever (2% v 14%), and neutropenic sepsis (1% v 12%). These findings suggested that on the basis of superior risk to benefit ratio, the GC regimen should be favored as standard treatment in advanced bladder cancer. Other promising combinations include gemcitabine/cisplatin/paclitaxel, and a phase III trial comparing this triple-agent combination with the GC regimen in advanced bladder cancer patients has been initiated. Semin Oncol 28 (suppl 7):11-14.
Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Cisplatin; Clinical Trials as Topic; Deoxycytidine; Doxorubicin; Humans; Methotrexate; Urinary Bladder Neoplasms; Vinblastine; Gemcitabine
PubMed: 11372046
DOI: 10.1053/sonc.2001.24369 -
Current Opinion in Critical Care Apr 2019In this article, the specific and general indications for sedatives in the neurocritical care unit are discussed, together with an overview on current insights in... (Review)
Review
PURPOSE OF REVIEW
In this article, the specific and general indications for sedatives in the neurocritical care unit are discussed, together with an overview on current insights in sedative protocols for these patients. In addition, physiological effects of sedative agents on the central nervous system are reviewed.
RECENT FINDINGS
In the general ICU population, a large body of evidence supports light protocolized sedation over indiscriminate deep sedation. Unfortunately, in patients with severe acute brain injury, the evidence from randomized controlled trials is scarce to nonexistent, and practice is supported by expert opinion, physiological studies and observational or small interventional trials. The different sedatives each have different beneficial effects and side-effects.
SUMMARY
Extrapolating the findings from studies in the general ICU population suggests to reserve deep continuous sedation in the neuro-ICU for specific indications. Although an improved understanding of cerebral physiological changes in patients with brain injury may be helpful to guide individualized sedation, we still lack the evidence base to make broad recommendations for specific patient groups.
Topics: Anesthesia; Conscious Sedation; Critical Care; Humans; Hypnotics and Sedatives; Intensive Care Units; Nervous System Diseases; Respiration, Artificial
PubMed: 30672819
DOI: 10.1097/MCC.0000000000000592 -
Dermatology Online Journal Jul 2021Periungual pyogenic granulomas are benign vascular tumors that present as painful, round, spontaneously bleeding lesions composed of rapidly proliferating capillaries...
Periungual pyogenic granulomas are benign vascular tumors that present as painful, round, spontaneously bleeding lesions composed of rapidly proliferating capillaries and excess tissue. The vast majority of pyogenic granulomas are caused by physical trauma or infectious agents and they may resolve spontaneously. Herein, we highlight a very rare case of periungual pyogenic granulomas induced by the regularly prescribed oral retinoid acitretin during treatment for congenital palmoplantar keratoderma. This unique case showed that it is feasible to continue acitretin therapy in the presence of pyogenic granuloma development if proper dose reduction and topical therapies are utilized. The patient's lesions resolved within two weeks of this protocol's initiation and the pyogenic granulomas did not recur over the course of a six-month follow-up observation period. In addition, we performed a systematic review of the literature using PubMed databases for the clinical features and treatments in other reported acitretin-induced pyogenic granuloma cases; we compiled a comprehensive list of other prescription drugs known to cause pyogenic granulomas up-to-date.
Topics: Acitretin; Administration, Oral; Adult; Anti-Bacterial Agents; Clobetasol; Glucocorticoids; Granuloma, Pyogenic; Humans; Keratoderma, Palmoplantar; Keratolytic Agents; Male; Mupirocin; Nail Diseases
PubMed: 34391333
DOI: 10.5070/D327754369