CSNK2B modulates IRF1 binding to functional DNA elements and promotes basal and agonist-induced antiviral signaling.

Interferon regulatory factor 1 (IRF1) is a critical component of cell-intrinsic innate immunity that regulates both constitutive and induced antiviral defenses. Due to its short half-life, IRF1 function is generally considered to be regulated by its synthesis. However, how IRF1 activity is controlled post-translationally has remained poorly characterized. Here, we employed a proteomics approach to identify proteins interacting with IRF1, and found that CSNK2B, a regulatory subunit of casein kinase 2, interacts directly with IRF1 and constitutively modulates its transcriptional activity. Genome-wide CUT&RUN analysis of IRF1 binding loci revealed that CSNK2B acts generally to enhance the binding of IRF1 to chromatin, thereby enhancing transcription of key antiviral genes, such as PLAAT4 (also known as RARRES3/RIG1/TIG3). On the other hand, depleting CSNK2B triggered abnormal accumulation of IRF1 at AFAP1 loci, thereby down-regulating transcription of AFAP1, revealing contrary effects of CSNK2B on IRF1 binding at different loci. AFAP1 encodes an actin crosslinking factor that mediates Src activation. Importantly, CSNK2B was also found to mediate phosphorylation-dependent activation of AFAP1-Src signaling and exert suppressive effects against flaviviruses, including dengue virus. These findings reveal a previously unappreciated mode of IRF1 regulation and identify important effector genes mediating multiple cellular functions governed by CSNK2B and IRF1.

Deletion of fatty acid transport protein 2 (FATP2) in the mouse liver changes the metabolic landscape by increasing the expression of PPARα-regulated genes.

Fatty acid transport protein 2 (FATP2) is highly expressed in the liver, small intestine, and kidney, where it functions in both the transport of exogenous long-chain fatty acids and the activation of very-long-chain fatty acids. Here, using a murine model, we investigated the phenotypic impacts of deleting FATP2, followed by a transcriptomic analysis using unbiased RNA-Seq to identify concomitant changes in the liver transcriptome. WT and FATP2-null (Fatp2-/-) mice (5 weeks) were maintained on a standard chow diet for 6 weeks. The Fatp2-/- mice had reduced weight gain, lowered serum triglyceride, and increased serum cholesterol levels and attenuated dietary fatty acid absorption. Transcriptomic analysis of the liver revealed 258 differentially expressed genes in male Fatp2-/- mice and a total of 91 in female Fatp2-/- mice. These genes mapped to the following gene ontology categories: fatty acid degradation, peroxisome biogenesis, fatty acid synthesis, and retinol and arachidonic acid metabolism. Targeted RT-quantitative PCR verified the altered expression of selected genes. Of note, most of the genes with increased expression were known to be regulated by peroxisome proliferator-activated receptor α (PPARα), suggesting that FATP2 activity is linked to a PPARα-specific proximal ligand. Targeted metabolomic experiments in the Fatp2-/- liver revealed increases of total C16:0, C16:1, and C18:1 fatty acids; increases in lipoxin A4 and prostaglandin J2; and a decrease in 20-hydroxyeicosatetraenoic acid. We conclude that the expression of FATP2 in the liver broadly affects the metabolic landscape through PPARα, indicating that FATP2 provides an important role in liver lipid metabolism through its transport or activation activities.

Background mortality drivers of European tree species: climate change matters.

Increases in tree mortality rates have been highlighted in different biomes over the past decades. However, disentangling the effects of climate change on the temporal increase in tree mortality from those of management and forest dynamics remains a challenge. Using a modelling approach taking tree and stand characteristics into account, we sought to evaluate the impact of climate change on background mortality for the most common European tree species. We focused on background mortality, which is the mortality observed in a stand in the absence of abrupt disturbances, to avoid confusion with mortality events unrelated to long-term changes in temperature and rainfall. We studied 372 974 trees including 7312 dead trees from forest inventory data surveyed across France between 2009 and 2015. Factors related to competition, stand characteristics, management intensity, and site conditions were the expected preponderant drivers of mortality. Taking these main drivers into account, we detected a climate change signal on 45% of the 43 studied species, explaining an average 6% of the total modelled mortality. For 18 out of the 19 species sensitive to climate change, we evidenced greater mortality with increasing temperature or decreasing rainfall. By quantifying the mortality excess linked to the current climate change for European temperate forest tree species, we provide new insights into forest vulnerability that will prove useful for adapting forest management to future conditions.

Enhancing radiotherapy response via intratumoral injection of the TLR9 agonist CpG to stimulate CD8 T cells in an autochthonous mouse model of sarcoma.

Radiation therapy is frequently used to treat cancers including soft tissue sarcomas. Prior studies established that the toll-like receptor 9 (TLR9) agonist cytosine-phosphate-guanine oligodeoxynucleotide (CpG) enhances the response to radiation therapy (RT) in transplanted tumors, but the mechanism(s) remain unclear. Here, we used CRISPR/Cas9 and the chemical carcinogen 3-methylcholanthrene (MCA) to generate autochthonous soft tissue sarcomas with high tumor mutation burden. Treatment with a single fraction of 20 Gy RT and two doses of CpG significantly enhanced tumor response, which was abrogated by genetic or immunodepletion of CD8+ T cells. To characterize the immune response to RT + CpG, we performed bulk RNA-seq, single-cell RNA-seq, and mass cytometry. Sarcomas treated with 20 Gy and CpG demonstrated increased CD8 T cells expressing markers associated with activation and proliferation, such as Granzyme B, Ki-67, and interferon-γ. CpG + RT also upregulated antigen presentation pathways on myeloid cells. Furthermore, in sarcomas treated with CpG + RT, TCR clonality analysis suggests an increase in clonal T-cell dominance. Collectively, these findings demonstrate that RT + CpG significantly delays tumor growth in a CD8 T cell-dependent manner. These results provide a strong rationale for clinical trials evaluating CpG or other TLR9 agonists with RT in patients with soft tissue sarcoma.

Shining a LIGHT on myeloid cell targeted immunotherapy.

Despite over a decade of clinical trials combining inhibition of emerging checkpoints with a PD-1/L1 inhibitor backbone, meaningful survival benefits have not been shown in PD-1/L1 inhibitor resistant or refractory solid tumours, particularly tumours dominated by a myelosuppressive microenvironment. Achieving durable anti-tumour immunity will therefore likely require combination of adaptive and innate immune stimulation, myeloid repolarisation, enhanced APC activation and antigen processing/presentation, lifting of the CD47/SIRPα (Cluster of Differentiation 47/signal regulatory protein alpha) 'do not eat me' signal, provision of an apoptotic 'pro-eat me' or 'find me' signal, and blockade of immune checkpoints. The importance of effectively targeting mLILRB2 and SIRPAyeloid cells to achieve improved response rates has recently been emphasised, given myeloid cells are abundant in the tumour microenvironment of most solid tumours. TNFSF14, or LIGHT, is a tumour necrosis superfamily ligand with a broad range of adaptive and innate immune activities, including (1) myeloid cell activation through Lymphotoxin Beta Receptor (LTßR), (2) T/NK (T cell and natural killer cell) induced anti-tumour immune activity through Herpes virus entry mediator (HVEM), (3) potentiation of proinflammatory cytokine/chemokine secretion through LTßR on tumour stromal cells, (4) direct induction of tumour cell apoptosis in vitro, and (5) the reorganisation of lymphatic tissue architecture, including within the tumour microenvironment (TME), by promoting high endothelial venule (HEV) formation and induction of tertiary lymphoid structures. LTBR (Lymphotoxin beta receptor) and HVEM rank highly amongst a range of costimulatory receptors in solid tumours, which raises interest in considering how LIGHT-mediated costimulation may be distinct from a growing list of immunotherapy targets which have failed to provide survival benefit as monotherapy or in combination with PD-1 inhibitors, particularly in the checkpoint acquired resistant setting.

Domestic violence reporting during the COVID-19 pandemic: evidence from Latin America.

This article examines changes in the frequency and characteristics of domestic violence reports following the onset of the COVID-19 pandemic and the imposition of mobility restrictions in six Latin American countries. We find significantly different patterns between reports of psychological and physical violence, non-cohabitant and cohabitant violence, and across alternative reporting channels (domestic violence hotlines, emergency lines, and police reports). Calls to domestic violence hotlines soared, suggesting that this channel was best suited to respond to victims' needs during the pandemic. In turn, calls to emergency lines and police complaints declined (especially in the first weeks of the pandemic), consistent with an increase in the perceived (relative) cost of using these channels. The results reveal how the pandemic altered domestic violence victims' demand for institutional help and highlight the relevance of domestic violence hotlines as an accessible and valuable service.

Real-life impact of uncontrolled severe asthma on mortality and healthcare use in adolescents and adults: findings from the retrospective, observational RESONANCE study in France.

OBJECTIVE: To characterise uncontrolled severe asthma and compare the disease burden with the general and asthmatic populations. DESIGN: Retrospective observational study using a national sample of a French healthcare database (Echantillon Généraliste des Bénéficiaires (EGB)). SETTING: The EGB, an anonymised permanent sample of health insurance databases, representing 1/97th of the French population. PARTICIPANTS: Patients (≥12 years) were selected in year 2014 and followed 2 years. A cohort of patients with uncontrolled severe asthma was defined using an algorithm based on peer-reviewed literature and Global Initiative for Asthma recommendations. Index date was the occurrence of the first marker of uncontrolled asthma. This cohort was matched with two control cohorts, general population and asthmatic controls, on baseline characteristics. MAIN OUTCOMES MEASURES: Mortality, healthcare use and associated costs were studied in the 2 years of follow-up. RESULTS: Among 467 716 individuals in the EGB, 16 588 patients with asthma were identified, including 739 (4.5%) with uncontrolled severe disease. The survival probability at 2 years for patients with uncontrolled severe asthma (92.0%) was lower than in the general population cohort (96.6%; relative risk of death: 2.35; 95% CI: 1.70 to 3.29; p<0.0001) and tended to be lower than in the control asthmatic cohort (94.3%; p=0.07). Emergency department visits and hospitalisations were higher in patients with uncontrolled severe asthma than in the general population (64.7% vs 34.9%; p<0.0001) and asthmatic controls (64.7% vs 55.2%; p=0.0002). Other components of healthcare use (medical and paramedical visits, medications) were increased in patients with uncontrolled severe asthma compared with control populations. These increases translated into higher costs (p<0.0001 for both comparisons). CONCLUSIONS: This study demonstrates the huge burden of uncontrolled severe asthma in terms of mortality, morbidity and healthcare resource consumption compared with other patients with asthma and with the general population and emphasises the importance of appropriate management in this high-risk population.

The PDAC Extracellular Matrix: A Review of the ECM Protein Composition, Tumor Cell Interaction, and Therapeutic Strategies.

Pancreatic ductal adenocarcinoma (PDAC) is notorious for a dense fibrotic stroma that is interlaced with a collagen-based extracellular matrix (ECM) that plays an important role in tumor biology. Traditionally thought to only provide a physical barrier from host responses and systemic chemotherapy, new studies have demonstrated that the ECM maintains biomechanical and biochemical properties of the tumor microenvironment (TME) and restrains tumor growth. Recent studies have shown that the ECM augments tumor stiffness, interstitial fluid pressure, cell-to-cell junctions, and microvascularity using a mix of biomechanical and biochemical signals to influence tumor fate for better or worse. In addition, PDAC tumors have been shown to use ECM-derived peptide fragments as a nutrient source in nutrient-poor conditions. While collagens are the most abundant proteins found in the ECM, several studies have identified growth factors, integrins, glycoproteins, and proteoglycans in the ECM. This review focuses on the dichotomous nature of the PDAC ECM, the types of collagens and other proteins found in the ECM, and therapeutic strategies targeting the PDAC ECM.

Prevalence and correlates of disability in Latin America and the Caribbean: Evidence from 8 national censuses.

We estimate disability prevalence rates and gaps in social conditions in eight Latin America and the Caribbean (LAC) countries and project current and future disability prevalence rates in the region. Using data from representative samples of the population in eight countries, we find that reported disability prevalence varies widely across countries, ranging between 4.5 percent in Trinidad and Tobago (2011) to 24.9 percent in Brazil (2010). Differences in surveying approaches and demographic structures likely explain a part of this variation. We find marked sociodemographic gradients for disability. We also report significant disability gaps: people living with disabilities have lower educational attendance and completion rates and lower employment rates. We use age and sex-specific disability rates from our sample of countries and information on the current and future demographic structures in LAC countries to project disability prevalence for the whole region. We project that the total number of people with disabilities in this region will increase by approximately 60 million between 2020 and 2050. Our projections suggest that countries need to systematically plan and implement inclusion policies to adequately address the growing population of people with disabilities in the years to come.

Crime under lockdown: The impact of COVID-19 on citizen security in the city of Buenos Aires.

RESEARCH SUMMARY: This paper studies the impact of the COVID-19 pandemic and the subsequent lockdown on criminal activity in the City of Buenos Aires, Argentina. Following quarantine restrictions, we find a large, significant, robust, and immediate decline in property crime reported to official agencies, police arrests, and crime reported in victimization surveys. We observe no significant change in homicides, and a significant increase in arrests for "resistance to authorities". The decrease in criminal activity was greater in business and transportation areas, but still large in commercial and residential areas (including informal settlements). After the sharp and immediate fall, crime recovered but, by the end of 2020, it had not reached its initial levels. The arrest data additionally shows a reduction in the distance from the detainee's address to the crime location, and a fall in the number of detainees from outside the City of Buenos Aires. Crime became more local as mobility was restricted. POLICY IMPLICATIONS: We find no evidence that the reduction in the number of detainees from outside the City of Buenos Aires led to a displacement of crimeto suburban areas. This result aligns with the hypothesis that focalized place-based interventions have the potential to reduce overall crime rates. Moreover, the increase in arrests for "resistance to authorities" at the checkpoints set up during the lockdown shows that the enforcement of mobility restrictions can cause frictions between citizens and police, negatively affecting police's legitimacy. We also find that the increased government presence for the provision of health and social services in informal settlements during the pandemic led, as a positive externality, to an additional decrease in crime.

Secretion and proteolysis of heterologous proteins fused to the Escherichia coli maltose binding protein in Pichia pastoris.

The Escherichia coli maltose binding protein (MBP) has been utilized as a translational fusion partner to improve the expression of foreign proteins made in E. coli. When located N-terminal to its cargo protein, MBP increases the solubility of intracellular proteins and improves the export of secreted proteins in bacterial systems. We initially explored whether MBP would have the same effect in the methylotrophic yeast Pichia pastoris, a popular eukaryotic host for heterologous protein expression. When MBP was fused as an N-terminal partner to several C-terminal cargo proteins expressed in this yeast, proteolysis occurred between the two peptides, and MBP reached the extracellular region unattached to its cargo. However, in two of three instances, the cargo protein reached the extracellular region as well, and its initial attachment to MBP enhanced its secretion from the cell. Extensive mutagenesis of the spacer region between MBP and its C-terminal cargo protein could not inhibit the cleavage although it did cause changes in the protease target sites in the fusion proteins, as determined by mass spectrometry. Taken together, these results suggested that an uncharacterized P. pastoris protease attacked at different locations in the region C-terminal of the MBP domain, including the spacer and cargo regions, but the MBP domain could still act to enhance the secretion of certain cargo proteins.

Simulating phenological shifts in French temperate forests under two climatic change scenarios and four driving global circulation models.

After modeling the large-scale climate response patterns of leaf unfolding, leaf coloring and growing season length of evergreen and deciduous French temperate trees, we predicted the effects of eight future climate scenarios on phenological events. We used the ground observations from 103 temperate forests (10 species and 3,708 trees) from the French Renecofor Network and for the period 1997-2006. We applied RandomForest algorithms to predict phenological events from climatic and ecological variables. With the resulting models, we drew maps of phenological events throughout France under present climate and under two climatic change scenarios (A2, B2) and four global circulation models (HadCM3, CGCM2, CSIRO2 and PCM). We compared current observations and predicted values for the periods 2041-2070 and 2071-2100. On average, spring development of oaks precedes that of beech, which precedes that of conifers. Annual cycles in budburst and leaf coloring are highly correlated with January, March-April and October-November weather conditions through temperature, global solar radiation or potential evapotranspiration depending on species. At the end of the twenty-first century, each model predicts earlier budburst (mean: 7 days) and later leaf coloring (mean: 13 days) leading to an average increase in the growing season of about 20 days (for oaks and beech stands). The A2-HadCM3 hypothesis leads to an increase of up to 30 days in many areas. As a consequence of higher predicted warming during autumn than during winter or spring, shifts in leaf coloring dates appear greater than trends in leaf unfolding. At a regional scale, highly differing climatic response patterns were observed.

Prevalence of Gout in the Adult Population of France.

OBJECTIVE: To estimate adult gout prevalence in France. METHODS: We used a previously established phone questionnaire that allowed for classifying patients as gouty or nongouty by 2 logistic regression models and 1 classification and regression tree (CART) model, the sensitivity and specificity of which were all more than 80%. The full questionnaire was administered by phone to subjects who acknowledged present or past nontraumatic acute pain in a peripheral joint, the others being classified as nongouty. A random sample of adults residing in France was derived from the national telephone directory (home and mobile) by the quota method and further redressed to match the French population. The target size for the interview survey conducted in March and June 2013 was 10,000 participants. RESULTS: We interviewed 10,026 participants. All 3 models (2 logistic regression models and a CART model) converged to an estimated gout prevalence of 0.9%. This prevalence was lower than that estimated by self-reporting only (3.7% [95% confidence interval 3.3-4.1]). The prevalence was higher for men than women and increased with age but did not differ by area of France. CONCLUSION: Gout prevalence in the adult population of France in 2013 was estimated at 0.9%. Studies using self-reporting only might overestimate the prevalence.

Radioisotope cisternography: a potentially useful tool for headache diagnosis in patients with an IDD pump.

BACKGROUND: Spontaneous intracranial hypotension (SIH) and related headache after intrathecal pump implantation is a possibility. Knowing the exact location of the cerebrospinal fluid (CSF) leak is mandatory for managing with interventional therapies. OBJECTIVES: Radioisotope cisternography (RICG) is a common procedure in nuclear medicine and may be used as an additional procedure in the differential diagnosis of headaches due to intracranial liquor hypotension syndrome caused by microleakage of CSF. STUDY DESIGN: We report an imaging case of complex chronic headache in a patient with an implanted drug delivery pump. The symptomatology of the patient was considered typical of spontaneous intracranial hypotension (SIH). Therefore, the initial suspicion was the loss of CSF due to the implanted pump catheter. The negativity of the routine headache test can exclude any leakage of CSF at the site of catheter insertion. SETTING: Anesthesia, Critical Care, and Multidisciplinary Pain Management Department, General University General Hospital, Valencia, Spain. RESULTS: Many patients are treated with implanted intrathecal infusion systems. In these patients, any neurological symptoms are readily attributed to the invasive neuroaxial system, especially severe cephalalgia. RICG was helpful in this particular case to make a diagnosis of exclusion of the diagnosis of SIH due to CSF leakage. CONCLUSION: RICG may be considered as an additional diagnostic technique in patients with chronic pain and implanted spinal infusion devices for whom standard detection procedures have been exhausted.

Computer-tomographic verification of ultrasound-guided piriformis muscle injection: a feasibility study.

BACKGROUND: Approximately 6% to 8% of lumbar pain cases, whether associated with radicular pain or not, may be attributed to the presence of piriformis muscle syndrome. Available treatments, among others, include pharmacotherapy, physical therapy, and injections of different substances into the muscle. Various methods have been used to confirm correct needle placement during these procedures, including electromyography (EMG), fluoroscopy, computed tomography (CT), or magnetic resonance imaging (MRI). Ultrasonography (US) has now become a widely used technique and therefore may be an attractive alternative for needle guidance when injecting this muscle. OBJECTIVE: The objective of this study was to assess the reliability of US in piriformis injection of patients with piriformis syndrome. STUDY DESIGN: Feasibility study; 10 patients with piriformis muscle syndrome were injected with botulinum toxin A using a US-guided procedure. Then patients were administered 2 mL iodinated contrast and were then transferred to the CT scanner, where they underwent pelvic and hip imaging to assess intramuscular distribution of the iodinated contrast. SETTING: Multidisciplinary Pain Management Department in Spain. RESULTS: Of all 10 study patients (8 women, 2 men), 9 had intramuscular or intrafascial contrast distribution. Distribution did not go deeper than the piriformis muscle in any of the patients. The absence of contrast (intravascular injection) was not observed in any case. LIMITATIONS: The main limitation of our study is the use of ionizing radiation as confirmation technique. CONCLUSION: Ultrasound-guided puncture may be a reliable and simple procedure for injection of the piriformis muscle, as long as good education and training are provided to the operator. US has a number of advantages over traditional approaches, including accessibility and especially no ionizing radiation exposure for both health care providers and patients.

Role of lumbosacral retrograde neuromodulation in the treatment of painful disorders.

BACKGROUND: Neuromodulation is an effective and reversible treatment option for chronic intractable pain. Spinal cord stimulation (SCS) represents a field of application of neuromodulation and is known to be effective for several conditions including complex regional pain syndrome (CRPS), failed back surgery syndrome (FBSS), and chronic leg and back pain. SCS has some technical limitations that can be bypassed through retrograde neuromodulation. OBJECTIVE: To examine the safety and efficacy of retrograde neuromodulation in consecutive patients with neuropathic pain in the perineum or lower limb. STUDY DESIGN: Prospective chart review analyzing one year of retrograde stimulation in our department. METHODS: We present a series of 10 patients who underwent retrograde neuromodulation at the University General Hospital of Valencia (Spain). We analyzed the variables that can improve the outcome and help physicians choose retrograde neuromodulation. RESULTS: Seven of 10 patients had an effective treatment and 3 patients had an ineffective stimulation. In the group with the effective treatment the most represented type of pain was radiculopathy and perineal pain. LIMITATIONS: This is a prospective, single-center study with a relatively small number of patients and no control group. CONCLUSION: Retrograde neuromodulation seems to be effective in patients that present with a well localized pain with a clear dermatome distribution. We found retrograde neuromodulation to be effective in radiculopathy related to FBSS. We found it to be limited in the treatment of perineal pain probably due to technical limitations and anatomical reasons besides the lack of knowledge of the etiology of this pain model and the exact mechanisms of action of neuromodulation.

Gastric pneumatosis following nasogastric tube placement: a case report with literature review.

We report a case of gastric emphysema following placement of nasogastric (NG) tube. Gas in the wall of the stomach is a rare finding seen in various clinical situations. The reported cases fall into two different categories: "gastric emphysema" and "emphysematous gastritis". Differentiating these two entities is important, as the first one is usually a benign condition, but the second one carries a poor prognosis.