tbuckley/cabot-search huggingface dataset:
Mitochondria Are a Subset of Extracellular Vesicles Released by Activated Monocytes and Induce Type I IFN and TNF Responses in Endothelial Cells
Authors: Florian Puhm, Taras Afonyushkin, Ulrike Resch, Georg Obermayer, Manfred Rohde, Thomas Penz, Michael Schuster, Gabriel Wagner, Andre F. Rendeiro, Imene Melki, Christoph Kaun, Johann Wojta, Christoph Bock, Bernd Jilma, Nigel Mackman, Eric Boilard, Christoph J. Binder
Journal: Circulation Research (2019)

Rationale: Extracellular vesicles, including microvesicles, are increasingly recognized as important mediators in cardiovascular disease. The cargo and surface proteins they carry are considered to define their biological activity, including their inflammatory properties. Monocyte to endothelial cell signaling is a prerequisite for the propagation of inflammatory responses. However, the contribution of microvesicles in this process is poorly understood. Objective: To elucidate the mechanisms by which microvesicles derived from activated monocytic cells exert inflammatory effects on endothelial cells. Methods and Results: LPS (lipopolysaccharide)-stimulated monocytic cells release free mitochondria and microvesicles with mitochondrial content as demonstrated by flow cytometry, quantitative polymerase chain reaction, Western Blot, and transmission electron microscopy. Using RNAseq analysis and quantitative reverse transcription-polymerase chain reaction, we demonstrated that both mitochondria directly isolated from and microvesicles released by LPS-activated monocytic cells, as well as circulating microvesicles isolated from volunteers receiving low-dose LPS-injections, induce type I IFN (interferon), and TNF (tumor necrosis factor) responses in endothelial cells. Depletion of free mitochondria significantly reduced the ability of these microvesicles to induce type I IFN and TNF-dependent genes. We identified mitochondria-associated TNFα and RNA from stressed mitochondria as major inducers of these responses. Finally, we demonstrated that the proinflammatory potential of microvesicles and directly isolated mitochondria were drastically reduced when they were derived from monocytic cells with nonrespiring mitochondria or monocytic cells cultured in the presence of pyruvate or the mitochondrial reactive oxygen species scavenger MitoTEMPO. Conclusions: Mitochondria and mitochondria embedded in microvesicles constitute a major subset of extracellular vesicles released by activated monocytes, and their proinflammatory activity on endothelial cells is determined by the activation status of their parental cells. Thus, mitochondria may represent critical intercellular mediators in cardiovascular disease and other inflammatory settings associated with type I IFN and TNF signaling.
Abstract P2-05-08: An assessment of the potential role of intracellular Ca2+ store regulators in breast cancer cells
Authors: AH Bong, SJ Roberts-Thomson, MJ Milevskiy, GR Monteith
Journal: Cancer Research (2019)

Abstract Background: Calcium (Ca2+) regulates many crucial cellular processes including cell survival, proliferation and death. Endoplasmic reticulum (ER) Ca2+ store levels are critical in the death-inducing effects of some anti-cancer agents. Modulators of ER Ca2+ signalling, such as neuronal calcium sensor-1 (NCS-1) may therefore represent new therapeutic opportunities to enhance the effect of some anti-cancer agents. NCS-1 is associated with poorer survival in breast cancer patients. However, the expression of NCS-1 in specific breast cancer molecular subtypes and its potential role in intracellular Ca2+ signalling in breast cancer cells has not been fully explored. Aim: To assess expression of NCS-1 in breast cancer molecular subtypes and assess the effect of silencing NCS-1 on intracellular Ca2+ homeostasis and on sensitivity to doxorubicin treatment in MDA-MB-231 breast cancer cells. Methods: NCS-1 levels were assessed in breast cancer molecular subtypes based on PAM50 subtyping using the TCGA public breast cancer database. Intracellular Ca2+ changes as a consequence of siRNA-mediated silencing of NCS-1 were evaluated using a Fluorescence Imaging Plate Reader (FLIPR) in MDA-MB-231 cells expressing the genetically-encoded Ca2+ indicator GCaMP6m. The effect of NCS-1 silencing on MDA-MB-231 cells treated with doxorubicin (0.03 and 1 μM, 24 h) was evaluated by cell nuclear enumeration (Hoechst 33342 staining) and the percentage of dead cells (propidium iodide staining). Images were acquired using an automated epifluorescence microscope (ImageXpress Micro). Results: Levels of NCS-1 were higher in the basal molecular subtype compared to other molecular subtypes. NCS-1 silencing promoted cell death induced by 1 μM doxorubicin. NCS-1 silencing had no major effect on cytosolic free Ca2+ levels as a result of either IP3-mediated Ca2+ store release with the purinergic receptor activator ATP or the protease activated receptor activator trypsin. However, NCS-1 silencing suppressed constitutive Ca2+ influx in MDA-MB-231 breast cancer cells. The expression of NCS-1 was positively correlated with the Ca2+ entry channel Orai1 in breast cancers on the TCGA database. Orai1 is associated with increased migration and invasiveness in some breast cancers. Discussion: These studies suggest that elevation in NCS-1 is a feature of breast cancers of the basal molecular subtype and that NCS-1 is a regulator of the cytotoxic effects of doxorubicin. Further studies are required to determine if this effect is related to changes in constitutive Ca2+ influx that may be mediated via Orai1. Citation Format: Bong AH, Roberts-Thomson SJ, Milevskiy MJ, Monteith GR. An assessment of the potential role of intracellular Ca2+ store regulators in breast cancer cells [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-05-08.
The plausibility of a chaotic brain theory
Authors: Ichiro Tsuda
Journal: Behavioral and Brain Sciences (2001)

We consider the significance of high-dimensional transitory dynamics in the brain and mind. In particular, we highlight the roles of high-dimensional chaotic dynamical systems as an “adequate language” (Gelfand 1989), which should possess both explanatory and predictive power of description. We discuss the methods of description of dynamic behavior of the brain. These methods have been adopted to capture the averaged or deterministic complexity, and further to allow for discussion of a new approach to capture the complexity of the deviation from such an averaged complexity and also the complexity of interactive modes. We also give arguments in defense of our models for dynamic memory with chaotic itinerancy and Cantor coding. In addition, we discuss the reality that a model of the brain and mind should reflect.
Abstract A04: Structural and functional caracterization of BARD1/CDK13 interaction
Authors: Vanessa C. Fernandes, Thales C. Nepomuceno, Renato S. Carvalho, Guilherme Suarez-Kurtz, Alvaro N. Monteiro, Marcelo A. Carvalho
Journal: Clinical Cancer Research (2018)

Abstract DNA damage response (DDR) is crucial for cell survivor through genomic integrity maintenance. Dysfunctions in proteins implicated in DDR prompt cell to diseases, such as cancer. BRCA1 is a central protein involved in DDR and, once damaged, plays a pivotal role in cancer development. BRCA1 structure comprises two major domains, a RING finger domain and two tandem BRCT domains (tBRCT). This structure is shared by BRCA1 major interaction partner: BARD1. The tBRCT domain is commonly found in DDR-associated proteins. To study interactions in DDR, our group charted the tBRCT interactome for 7 different proteins, using three different strategies: tandem affinity purification followed by mass spectrometry (TAP-MS), yeast two hybrid (Y2H) screening, and literature curation. CDK13 was identified as a putative BARD1 tBRCT interactor by both TAP-MS and Y2H strategies. CDK13 is described as involved in several processes, such as chromatin remodeling, transcription regulation, and splicing coordination; however, its biological role remains unclear. To confirm BARD1/CDK13 interaction, whole cell extracts obtained from MCF-7 cells were used in coimmunoprecipitation assays using anti-CDK13 and anti-BARD1. Constructs encoding the HA-tagged regions 1-706aa (N-terminal), 706-982aa (kinase domain), and 1006-1452aa (C-terminal) of CDK13 were ectopically coexpressed with Flag-tagged full-length BARD1 or GFP-tagged BARD1 tBRCT in HEK293Tcells; whole cellular extracts were used in coimmunoprecipitation assays. BARD1 and CDK13 interaction was confirmed both in ectopic and constitutive expression approaches. CDK13 silencing was performed using lentiviral constructs encoding five different short hairpin RNAs (shRNA); MCF-7 cells were transduced and submitted to puromycin selection; extracts were used to evaluate CDK13 silencing by immunoblotting. Our data suggest that BARD1 tBRCT and the CDK13 N-terminal region, as well as the kinase domain, are critical for the formation of BARD1/CDK13 complex. MCF-7 cells lacking CDK13 expression will be used to investigate the kinase role during DDR. Citation Format: Vanessa C. Fernandes, Thales C. Nepomuceno, Renato S. Carvalho, Guilherme Suarez-Kurtz, Alvaro N. Monteiro, Marcelo A. Carvalho. Structural and functional caracterization of BARD1/CDK13 interaction [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A04.
COPD: spirometry does not occur in 60% of admissions, audit shows
Authors: Jacqui Wise
Journal: BMJ (2019)

A spirometry test is essential to confirm a diagnosis of chronic obstructive pulmonary disorder (COPD), but a new audit shows that it was not recorded in 60% of patients admitted to hospital with an acute exacerbation of the condition.1

A large clinical audit of COPD, run by the Royal College of Physicians, included data from almost 75 000 admissions to 179 hospitals in England and Wales. It found better care of patients with COPD than the previous audit in 2018 but still noted significant areas for improvement.

On the positive side, 75% of patients are …
Large local reactions to Hymenoptera stings: Outcome of re‐stings in real life
Authors: Maria Beatrice Bilò, Matteo Martini, Valerio Pravettoni, Donatella Bignardi, Patrizia Bonadonna, Gabriele Cortellini, Magdalena Kosinska, Donatella Macchia, Marina Mauro, Elisa Meucci, Marita Nittner‐Marszalska, Vincenzo Patella, Roberta Pio, Oliviero Quercia, Federico Reccardini, Erminia Ridolo, Michael Rudenko, Maurizio Severino
Journal: Allergy (2019)

Large local reaction to Hymenoptera stings is usually defined as a swelling >10 cm which lasts longer than 24 hours, sometimes associated with erythema, pruritus and blisters. Currently, the risk of subsequent systemic reactions after re-stings is considered low (2%-15%). Therefore, a diagnostic workup in case of large local reaction is often judged unnecessary, as well as adrenaline auto-injector and venom immunotherapy prescription. The aim of this study was to prospectively evaluate the outcome of re-stings in a real-world setting, in patients with a history of one previous large local reaction.We consecutively enrolled patients who experienced their first large local reaction (as per EAACI definition), treated with antihistamine and steroids. They were followed for field re-stings and assessed for risk of subsequent systemic reactions.We enrolled 662 patients. Out of the 225 re-stung subjects, 24% did not experience reactions, 52% reported a second large local reaction and 24% had systemic reactions. The risk of subsequent systemic reactions was higher in case of skin test reactivity to Apis mellifera or Vespula species (OR 2.1 and 3.8, respectively), in particular if positive at 0.001 µg/mL concentration (OR 13.4 and 16.5, respectively).Systemic reactions, after a previous large local reaction, occur more frequently than that reported by literature. After analysing the predictive role of large local reactions for systemic reactions, we demonstrated that an accurate diagnostic workup may be considered, particularly skin tests. Further studies in different countries are needed to confirm these results and large local reaction management.
Abstract 5568: Elevated interleukin-12 is a plasma marker of poor prognosis in stage III melanoma patients
Authors: Yun S Chun, Yuling Wang, Amy H Vo, Huey Liu, Daniel Y Wang, Jeffrey E Gershenwald, Janice N Cormier, Merrick I Ross, Dana M McClain, Christopher W Schacherer, John D Reveille, Jeffrey E Lee
Journal: Cancer Research (2008)

Abstract Background: The heterodimeric cytokine interleukin-12 (IL-12) can both induce and suppress cellular immunity; the IL-12 p40 subunit is also a component of IL-23, which can promote tumor growth. IL-12 plasma levels have been reported to rise with age. Since survival in melanoma patients also depends on age, we investigated the relationship between IL-12, age and survival in a large cohort of melanoma patients. Methods: Melanoma patients underwent blood draw for determination of combined IL-12-p40 levels by ELISA. Clinical data was obtained from a prospective database. AJCC stage of disease, age and overall survival (OS) were determined from the date of blood draw. Results: Plasma IL-12 levels were determined in 658 patients (445 stages I-II, 150 stage III, 63 stage IV). IL-12 levels rose with age, with mean levels of 75, 84, 96 and 112 pg/ml in ages &amp;lt;40, 40-59, 60-79 and ≥80 yr, respectively (p=0.012). After median follow-up of 13 mo, age was confirmed as a predictor of shorter OS on univariate analysis (p=0.009). However, in a multivariate model, while both advanced stage (p&amp;lt;0.0001) and elevated IL-12 (p=0.006) were independent predictors of shorter OS, age (p=0.339) was not. Stage-specific analysis demonstrated that the association between IL-12 and OS was limited to patients with regional metastasis (stage III, table). The risk of death among stage III patients with elevated IL-12 (&amp;gt; 150 pg/ml) was &amp;gt;4-fold greater than that of patients with normal IL-12. Furthermore, the association of elevated IL-12 with OS in stage III patients was independent of the extent of regional disease. Conclusions: An elevated plasma level of IL-12 is an independent predictor of poor prognosis in melanoma patients with regional metastasis. The association between IL-12 and advanced age could help explain the link between age and poorer prognosis in melanoma. Elevated IL-12 may be a marker of an immunosuppressive, tumor-promoting response in melanoma patients. Prognostic factorUnivariate PMultivariate PHazard ratio (95% CI)Gender (M v. F)0.461--Age (yr)0.0300.349-Extent of disease (stage III A v. B v. C)0.0310.031Elevated IL-12 (&amp;gt; 150 pg/ml)0.0010.0044.763 (1.660-13.669)
Online question banks
Authors: Guy Rughani
Journal: BMJ (2013)

Attempting mock exam questions is often touted as the best way to revise. The idea is that they help you to identify areas that need work, and provide you with a more memorable experience than wading through textbooks. Although multiple choice question books remain popular, an array of online resources exists. The resources cater mainly for finals students, and they provide more flexibility, more questions, and more features than their paper counterparts. With exams coming up, we’ve looked at four of the main subscription resources available online: PasTest, OnExamination, Examdoctor, and Passmedicine.

The advantage of online tools over question books is that you can easily track your progress over time, compare modules, and see how you fared alongside fellow subscribers. You can also design tests, choose to sit only new questions, or questions that you’ve previously answered …
Abstract 2807: Coordinated upregulation of peroxiredoxins in breast cancer
Authors: Jillian Muhlbauer, Shelley A. Phelan
Journal: Cancer Research (2016)

Abstract Peroxiredoxins (Prdxs) are a group of thiol-specific antioxidant proteins that are involved in many cellular processes, including growth, proliferation, differentiation, and the stress response. The family includes six proteins in humans, which show differences in substrate specificity, tissue expression, and cellar localization and distribution. Cancer cells commonly exhibit aberrantly regulated expression of antioxidants, which can support cancer cell survival in the face of elevated levels of reactive oxygen species. Breast cancers exhibit significantly elevated Prdx levels of Prdx. We previously reported overexpression of many Prdx proteins in the MCF-7 breast cancer line, and further demonstrated their importance in cell survival and doxorubicin-resistance in this line. To further investigate the misregulation of these proteins in human breast cancer, we compared Prdx levels between breast tumor and adjacent normal breast tissue from 20 breast cancer patients. We showed that most patients exhibited elevated levels of several Prdx proteins in the tumor tissue. We have gone on to analyze the promoters of these genes in an effort to identify potential mechanisms of coordinate regulation, as well as downstream targets that may mediate their role in cell survival. Together, we feel that this family of proteins may be useful biomarkers, and also targets, in breast cancer biology. Citation Format: Jillian Muhlbauer, Shelley A. Phelan. Coordinated upregulation of peroxiredoxins in breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2807.
Abstract A29: Gram-negative bacterial infection enhances the potential of gastric adenocarcinoma peritoneal metastasis via TNFR1 dependent manner
Authors: Malak Alzahrani, Betty Giannias, France Bourdeau, Rafael Kayano, Lorenzo Ferri
Journal: Cancer Research (2016)

Abstract Adenocarcinoma of the proximal stomach is the fastest rising malignancy in North America, and is associated with a high rate of recurrence to the peritoneum. As part of cancer treatment, the majority of patients undergo at least one invasive surgical procedure. Recent clinical data has linked postoperative infection complications with adverse oncologic outcomes; however, the underlying mechanisms are unclear. Emerging evidence suggests that the release of TNFa, a key inflammatory cytokine during infection facilitates cancer progression. The role and mechanisms the gram-negative bacterial infections play in facilitating the metastatic potential of gastric cancer to the peritoneum is entirely unknown. We hypothesized that incubation of gastric cancer cells and MC with heat-inactivated E. Coli or LPS can augment gastric cancer cell adhesion and invasion via TNFR1 signaling and increase the potential of peritoneal metastasis. Incubation of human gastric cancer cells or/and MC with heat inactivated E. coli, LPS or TNFa significantly increased in vitro adhesion 3-4 folds to MC and enhanced in vitro invasion. These enhanced cell adhesion and invasion phenotypes following incubation with LPS or E. coli were attenuated at three levels: inhibition of TLR4 (Eritoran), inhibition of TNFR1 (anti-TNFR1/isotype control antibodies) or p38 MAPK inhibitor (BIRB0796). TNFa treatment also increases CD54, CD44 and CD29 expression on cancer cells and MC. To further validate in vitro results, a novel ex-vivo murine peritoneal metastasis model is developed. We report that ex vivo gastric cancer cells adhesion to murine peritoneum is augmented by overnight LPS, E. Coli and TNFa treatments and this effect was abrogated when TNFR1-/- mouse peritoneum is used or in the presence of TNFR1 monoclonal or isotype control antibodies. These findings implicate TNFa in the process of gastric cancer metastasis to peritoneum in the context of systemic infection and identify TNFa as potential therapeutic targets. Citation Format: Malak Alzahrani, Betty Giannias, France Bourdeau, Rafael Kayano, Lorenzo Ferri. Gram-negative bacterial infection enhances the potential of gastric adenocarcinoma peritoneal metastasis via TNFR1 dependent manner. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A29.
Getting real about evidence in surgery
Authors: Peter McCulloch
Journal: BMJ (2013)

When evidence based medicine (EBM) began in the 1990s, great emphasis was placed on the “right” methodology for studying different questions. For comparing treatments, the arguments for randomised controlled trials (RCTs) were irrefutable. Randomisation was the only reliable way of minimising bias in treatment allocation.

The way this was interpreted, however, led to difficulties. As RCTs were the only “good” design, non-RCTs were regarded as having little value. Subjects like surgery, where RCTs were rare and often small, were criticised,1 and surgeons were portrayed as lacking scientific …
You’ve been bleeped: the drop after the op
Authors: Flavia Serrelli, Andrew Clarke, Sunjay Parmar, Saima Ehsan, Rakesh Patel
Journal: BMJ (2015)

#### Scenario

It’s 8 pm and you are the night foundation year 1 (FY1) doctor on call for the surgical wards for the first time. In the handover room you meet the rest of the on-call team: the surgical core trainee year 1 who will deal with the new admissions, and the surgical registrar, who will divide her time between reviewing new patients on the surgical assessment unit and dealing with emergency cases in theatre.

After picking up your bleep you make your way to the wards to start working through the job list that was handed over by the day team. Things are going well, and you are about to stop for a break at 1 30 am when the bleep goes off again.

#### Bleep

“Hello, my name is Martha, staff nurse on the colorectal unit. I’m calling about Miss Lowe, who is 28 years old and has low blood pressure. She had a subtotal colectomy with an ileostomy for her ulcerative colitis two days ago. The operation went smoothly and she has been otherwise well, but on checking her observations I’ve noticed her blood pressure is low, about 84/62 mm Hg. Could you come and see her please?”

### How would you react to this bleep on the phone?

Several pathologies can cause hypotension, and therefore low blood pressure may indicate a problem that requires immediate attention. On the phone you will need to ask questions that differentiate between these various causes, specifically inquiring about signs or symptoms that help you formulate an initial impression about the most likely diagnosis. You should prioritise this referral alongside other requests for help and your time.

Seeing patients in the middle of the night is challenging. Patients might be irritated by being woken and not be cooperative, especially if they do not have ill effects from the low blood pressure. A patient objecting to being …