NIS-Seq enables cell-type-agnostic optical perturbation screening.

Optical pooled screening offers a broader-scale alternative to enrichment-based perturbation screening, using fluorescence microscopy to correlate phenotypes and perturbations across single cells. Previous methods work well in large, transcriptionally active cell lines, because they rely on cytosolic detection of endogenously expressed barcoded transcripts; however, they are limited by reliable cell segmentation, cytosol size, transcriptional activity and cell density. Nuclear In-Situ Sequencing (NIS-Seq) expands this technology by creating bright sequencing signals directly from nuclear genomic DNA to screen nucleated cells at high density and high library complexity.

Monitoring of Inflammasome Activation of Macrophages and Microglia In Vitro, Part 2: Assessing Inflammasome Activation.

Inflammasomes are macromolecular complexes that assemble upon the detection of cytoplasmic pathogen-associated or danger-associated signals and induce a necrotic type of cell death termed pyroptosis, facilitating pro-inflammatory cytokine release. Inflammasomes play a critical role in innate immunity and inflammatory response; however, they have also been associated with multiple diseases, including autoinflammatory and neurodegenerative conditions. In the following chapter, we describe methods to detect inflammasome activation and its downstream effects, including detection of ASC oligomerization, detection of activated caspase-1 and cleaved IL-1β, as well as read-outs for inflammasome-mediated cell death.

Monitoring of Inflammasome Activation of Macrophages and Microglia In Vitro, Part 1: Cell Preparation and Inflammasome Stimulation.

Inflammasomes are intracellular, multiprotein supercomplexes that mediate a post-translational inflammatory response to both pathogen and endogenous danger signals. They consist of a sensor, the adapter ASC, and the protease caspase 1 and, following their activation, lead to cl1β, as well as lytic cell death. Due to this potent inflammatory capacity, understanding inflammasome biology is important in many pathological conditions.

Mitochondrial damage activates the NLRP10 inflammasome.

Upon detecting pathogens or cell stress, several NOD-like receptors (NLRs) form inflammasome complexes with the adapter ASC and caspase-1, inducing gasdermin D (GSDMD)-dependent cell death and maturation and release of IL-1β and IL-18. The triggers and activation mechanisms of several inflammasome-forming sensors are not well understood. Here we show that mitochondrial damage activates the NLRP10 inflammasome, leading to ASC speck formation and caspase-1-dependent cytokine release.

How location and cellular signaling combine to activate the NLRP3 inflammasome.

NOD-, LRR-, and pyrin domain-containing 3 (NLRP3) is a cytosolic innate immune sensor of cellular stress signals, triggered by infection and sterile inflammation. Upon detection of an activating stimulus, NLRP3 transitions from an inactive homo-oligomeric multimer into an active multimeric inflammasome, which promotes the helical oligomeric assembly of the adaptor molecule ASC. ASC oligomers provide a platform for caspase-1 activation, leading to the proteolytic cleavage and activation of proinflammatory cytokines in the IL-1 family and gasdermin D, which can induce a lytic form of cell death.

Development of Fluorescent and Biotin Probes Targeting NLRP3.

Extracellular signals drive the nucleation of the NLRP3 inflammasome which leads to the release of cytokines and causes inflammatory events. Hence, the inflammasome has gained enormous momentum in biomedical basic research. The detailed mechanisms of inflammasome generation and regulation remain to be elucidated.

Partial Endograft Removal Preserves the Aortic Walls During Delayed Open Conversions of Endovascular Aortic Repair.

Background: Open conversion of endovascular aortic repair (EVAR) is the first-choice treatment in case of endograft failure or high-flow endoleak. However, the traditional technique based on the total removal of the endograft can produce injuries of the aortic walls, with severe consequences on the anastomoses quality. Our aim is to show the advantages of the partial endograft removal on the aortic integrity by reporting a case series including 25 delayed open conversion performed with this technique.

The Bolton Treo Endograft: A Single-Center Preliminary Experience.

Background: Endovascular repair is currently the most frequently used treatment of abdominal aortic aneurysms, but its feasibility and success highly depend on the characteristics and the correct choice of the endograft to be used. Bolton Treo is one of the most popular endografts of newer generation which have been launched in the past few years, and this study aims to analyze the preliminary outcomes (8 months in average) of a single-center experience with this device.

Methods: Thirty-seven consecutive abdominal aortic aneurysms, treated with Treo between June 2016 and December 2017, were followed up every 3, 6, and 12 months, and any kind of endograft-related complications was recorded.

Near infrared spectroscopy monitoring during carotid endarterectomy: which threshold value is critical?

Objectives: Retrospectively to verify which decreasing percentage in regional oxygen saturation (rSO(2)) identified patients with good collateralisation during carotid artery cross clamp.

Materials And Methods: During 594 endarterectomies under general anaesthesia the decreasing percentage from preclamp value to value detected in the first 2 min after clamping the CCA and/or ICA was calculated in real time. No temporary shunt was placed in any case.

Efficacy of external cephalic version performed at term.

Objective: To evaluate the efficacy and complications of external cephalic version in term (37 or more weeks) gestation.

Study Design: A case series from February 1990 until December 1994 studied 160 patients with term singleton breech presentation. External cephalic version (ECV) with prophylactic tocolysis was the method used.

Prevalence of fetal left ventricular hyperechogenic foci in a low risk population.

Objective: To ascertain the prevalence and clinical significance of small hyperechogenic foci detected in the fetal left ventricle in routine ultrasound screening of pregnant women attending our hospital.

Population And Methods: From April 1994 to April 1995, 1135 consecutive pregnant women examined at the obstetric ultrasound unit of our hospital were studied prospectively. A postnatal cardiologic examination was performed when signs of cardiopathy were observed at the prenatal ultrasound investigation or the neonatal examination.

A case of congenital hypoplasia of the right external iliac artery.

A 18 years male had congenital hypoplasia of the right external iliac artery and thrombosis of its narrowest part. Congenital megacolon, anal atresia and hyposomical nanism were also present. Surgical intervention was indicated because he was developing arterial insufficiency and muscular hypotrophy of the right leg.

Antiphospholipid antibodies and recurrent abortion.

We examined the association between anticardiolipin antibodies, lupus anticoagulant, and the risk of recurrent spontaneous abortion in a case-control study conducted in a network of general and teaching hospitals in northern Italy. Subjects consisted of 220 women with two or more unexplained consecutive spontaneous abortions and 193 controls admitted for acute conditions other than immunologic, infective, gynecologic, or cardiovascular. Lupus anticoagulant was detected in 16 of 220 cases (7%, 95% confidence interval 4-11%) but in none of the 193 controls (Fisher exact test, P less than .