Inter-organelle cross-talk supports acetyl-coenzyme A homeostasis and lipogenesis under metabolic stress.

Proliferating cells rely on acetyl-CoA to support membrane biogenesis and acetylation. Several organelle-specific pathways are available for provision of acetyl-CoA as nutrient availability fluctuates, so understanding how cells maintain acetyl-CoA homeostasis under such stresses is critically important. To this end, we applied 13C isotope tracing cell lines deficient in these mitochondrial [ATP-citrate lyase (ACLY)]-, cytosolic [acetyl-CoA synthetase (ACSS2)]-, and peroxisomal [peroxisomal biogenesis factor 5 (PEX5)]-dependent pathways. ACLY knockout in multiple cell lines reduced fatty acid synthesis and increased reliance on extracellular lipids or acetate. Knockout of both ACLY and ACSS2 (DKO) severely stunted but did not entirely block proliferation, suggesting that alternate pathways can support acetyl-CoA homeostasis. Metabolic tracing and PEX5 knockout studies link peroxisomal oxidation of exogenous lipids as a major source of acetyl-CoA for lipogenesis and histone acetylation in cells lacking ACLY, highlighting a role for inter-organelle cross-talk in supporting cell survival in response to nutrient fluctuations.

Plasma volume expansion reveals hidden metabolic acidosis in patients with diabetic ketoacidosis.

BACKGROUND: Hyperchloremic metabolic acidosis that develops during the treatment of diabetic ketoacidosis is usually attributed to the chloride content of resuscitation fluids. We explored an alternative explanation, namely that fluid-induced plasma volume expansion alters the absolute differences in the concentrations of sodium and chloride (the Na-Cl gap) enough to affect the acid-base balance. We analyzed data from a prospective single-center cohort study of 14 patients treated for diabetic ketoacidosis. All patients received 1 L of 0.9% saline over 30 min on two consecutive days. Blood gases were sampled before and after the infusions. RESULTS: The initial plasma volume was estimated to be 25 ± 13% (mean ± SD) below normal on admission to the intensive care unit. At that time, most patients had an increased actual Na-Cl gap, which counteracts acidosis. However, the correction of the plasma volume deficit revealed that these patients would have had a decreased Na-Cl gap upon admission if they had been normovolemic at that time; the estimated "virtual Na-Cl gap" of 29 ± 5 mmol/L was significantly lower than the uncorrected value, which was 39 ± 5 mmol/L (P < 0.001). On Day 2, most patients had a decreased actual Na-Cl gap (33 ± 5 mmol/L), approaching the corrected value on Day 1. CONCLUSIONS: The hyperchloremic acidosis commonly seen in diabetic ketoacidosis may not be primarily caused by the chloride content of resuscitation fluids but, rather, by the restoration of plasma volume, which reveals the hidden metabolic acidosis caused by a decreased Na-Cl gap. Trial registration Clinical Trials Identifier NCT02172092, registered June 24, 2014, https://www. CLINICALTRIALS: gov/NCT02172092.

Does it matter in what family constellations adolescents live? Reconsidering the relationship between family structure and delinquent behaviour.

OBJECTIVES: This study examines the associations between ten family structure types and delinquency, including four groups of symmetrical and asymmetrical living arrangements. We also adjust for attachment to parents and parental monitoring. METHODS: Data are drawn from four cross-sectional surveys conducted between 2016 and 2019 in southern Sweden. The sample consists of 3,838 adolescents, aged 14-15. Negative binomial models were used to calculate the associations between family structure and delinquency. RESULTS: The results show that those living in single-father, single-mother, father-stepmother, mother-stepfather families report significantly more delinquency than adolescents living with both their parents. Adolescents living in "symmetrical" family arrangements, i.e. both parents are single or have a new partner, reported lower levels of delinquency, whereas adolescents living in "asymmetrical" family arrangements, i.e. where either the mother or the father, but not both, have a new partner, reported higher levels of delinquency. Most of the associations between family structure and delinquency decline when adjusted for attachment to parents and parental monitoring. DISCUSSION: This study shows that it is important to move on to the use of more detailed categorisations of family structure in relation to delinquency. We need to increase our knowledge about the group of adolescents that moves between parents and especially about the different constellations of asymmetrical and symmetrical living arrangements.

Does gender matter? The association between different digital media activities and adolescent well-being.

BACKGROUND: Previous research on the relationship between social media use and well-being in adolescents has yielded inconsistent results. We addressed this issue by examining the association between various digital media activities, including a new and differentiated measure of social media use, and well-being (internalizing symptoms) in adolescent boys and girls. METHOD: The sample was drawn from the four cross-sectional surveys from the Öckerö project (2016-2019) in eight municipalities in southern Sweden, consisting of 3957 adolescents in year 7 of compulsory education, aged 12-13. We measured the following digital media activities: playing games and three different activities of social media use (chatting, online sociability, and self-presentation). Our outcome measure was internalizing symptoms. Hypotheses were tested with linear regression analysis. RESULTS: Social media use and playing games were positively associated with internalizing symptoms. The effect of social media use was conditional on gender, indicating that social media use was only associated with internalizing symptoms for girls. Of the social media activities, only chatting and self-presentation (posting information about themselves) were positively associated with internalizing symptoms. Self-presentation was associated with internalizing symptoms only for girls. CONCLUSION: Our study shows the importance of research going beyond studying the time spent on social media to examine how different kinds of social media activities are associated with well-being. Consistent with research in psychology, our results suggest that young girls posting information about themselves (i.e. self-presentation) might be especially vulnerable to display internalizing symptoms.

Trends in alcohol intoxication among native and immigrant youth in Sweden, 1999-2017: A comparison across family structure and parental employment status.

BACKGROUND: Developing a better understanding of drinking patterns across immigrant generations and how these change over time is important for the development of effective alcohol polices. This study investigates the direction and rate of change in youth alcohol intoxication over time, based on immigrant status, and by family structure and parental employment status. METHOD: The study is based on eight nationally representative school surveys conducted by the Swedish National Council for Crime Prevention between 1999 and 2017, with a combined sample of 50,657 adolescents. Group by time interactions were examined to compare rates of change of alcohol intoxication over time across immigrant generations. RESULTS: The results show a decreasing trend in alcohol intoxication among both first and second generation immigrant youth, and also among immigrant youth across different family structures and parental employment statuses. The results also show that the decrease in alcohol intoxication over time is greater for youths born abroad and for youths with two immigrant parents than for native Swedes, and that the decrease over time is greater for youths from intact families than for native Swedish youths from non-intact families and youths with one immigrant parent. CONCLUSION: Native and first- and second-generation immigrant youth may differ substantially from one another in many ways, and may therefore manifest different patterns of drinking behaviours. From a policy and prevention perspective, the data in this study imply that native youths and youths with one immigrant parent should be a central target group for alcohol prevention policy in Sweden.

A global analysis of the impact of COVID-19 stay-at-home restrictions on crime.

The stay-at-home restrictions to control the spread of COVID-19 led to unparalleled sudden change in daily life, but it is unclear how they affected urban crime globally. We collected data on daily counts of crime in 27 cities across 23 countries in the Americas, Europe, the Middle East and Asia. We conducted interrupted time series analyses to assess the impact of stay-at-home restrictions on different types of crime in each city. Our findings show that the stay-at-home policies were associated with a considerable drop in urban crime, but with substantial variation across cities and types of crime. Meta-regression results showed that more stringent restrictions over movement in public space were predictive of larger declines in crime.

A community intervention to reduce alcohol consumption and drunkenness among adolescents in Sweden: a quasi-experiment.

BACKGROUND: Several studies have examined the effect of community interventions on youth alcohol consumption, and the results have often been mixed. The aim of this study is to evaluate the effectiveness of a community intervention known as the Öckerö Method on adolescent alcohol consumption and perceived parental attitudes towards adolescent drinking. METHOD: The study is based on a quasi-experimental design, using matched controls. Self-report studies were conducted among adolescents in grades 7-9 of compulsory education in four control and four intervention communities in the south of Sweden in 2016-2018. Baseline measures were collected in autumn 2016 before the intervention was implemented in the intervention communities. Outcomes were the adolescents' alcohol consumption, past-year drunkenness, past-month drunkenness and perceived parental attitudes towards alcohol. RESULTS: Estimating Difference-in-Difference models using Linear Probability Models, we found no empirical evidence that the intervention has any effect on adolescents' drinking habits, or on their perceptions of their parents' attitudes towards adolescent drinking. CONCLUSION: This is the first evaluation of this method, and we found no evidence that the intervention had any effect on the level of either young people's alcohol consumption or their past-year or past-month drunkenness, nor on their parents' perceived attitudes toward adolescent drinking. A further improvement would be to employ a follow-up period that is longer than the three-year period employed in this study. TRIAL REGISTRATION: ISRCTN registry: Study ID: 51635778 , 31th March 2021 (Retrospectively registered).

Electrolyte-based calculation of fluid shifts after infusing 0.9% saline in severe hyperglycemia.

BACKGROUND: Early treatment of severe hyperglycemia involves large shifts of body fluids that entail a risk of hemodynamic instability. We studied the feasibility of applying a new electrolyte equation that estimates the degree of volume depletion and the distribution of infused 0.9% saline in this setting. METHODS: The new equation was applied to plasma and urinary concentrations of sodium and chloride measured before and 30 min after a 30-min infusion of 1 L of 0.9% saline on two consecutive days in 14 patients with severe hyperglycemia (mean age 50 years). The extracellular fluid (ECF) volume was also estimated based on the volume dilution kinetics of chloride. RESULTS: On day 1, the baseline ECF volume amounted to 11.5 L. The saline infusion expanded the ECF space by 160 mL and the intracellular fluid space by 375 mL. On day 2, the ECF volume was 15.5 L, and twice as much of the infused fluid remained in the ECF space. The chloride dilution kinetics yielded baseline ECF volumes of 11.6 and 15.2 L on day 1 and day 2, respectively. No net uptake of glucose to the cells occurred during the two 1-h measurement periods despite insulin administration in the intervening time period. CONCLUSIONS: The electrolyte equation was feasible to apply in a group of hyperglycemic patients. The ECF space was 3 L smaller than expected on admission but normal on the second day. Almost half of the infused fluid was distributed intracellularly.

Locally invasive, castrate-resistant prostate cancer in a Pten/Trp53 double knockout mouse model of prostate cancer monitored with non-invasive bioluminescent imaging.

Here we have improved an existing mouse model of prostate cancer based on prostate-specific deletion of Pten and Trp53 by incorporating a Cre-activatable luciferase reporter. By coupling the deletion of those genes to the activation of a luciferase reporter, we were able to monitor tumor burden non-invasively over time. We show that, consistent with previous reports, deletion of both Pten and Trp53 on a C57BL/6 background accelerates tumor growth and results in both the loss of androgen receptor expression and castrate resistant tumors as compared with loss of Pten alone. Loss of Trp53 results in the development of sarcomatoid histology and the expression of markers of epithelial-to-mesenchymal transition Zeb1 and vimentin, with kinetics and penetrance dependent on whether one or both alleles of Trp53 were deleted. Homozygous deletion of Trp53 and Pten resulted in uniformly lethal disease by 25 weeks. While we were able to detect locally invasive disease in the peritoneal cavity in aggressive tumors from the double knockout mice, we were unable to detect lymphatic or hematogenous metastatic disease in lymph nodes or at distant sites.

Internet use and adolescent drinking: Does it matter what young people do online?

BACKGROUND: In this study we examine whether the association between internet use and drinking could be different for different types of internet activities among adolescents. We also adjust for a number of theoretically relevant factors such as peer influence, unstructured activities, impulsivity and parental monitoring. METHOD: The data are drawn from four cross-sectional surveys from the years 2016-2019 in eight municipalities in southern Sweden. The sample consist of 3733 adolescents in year 9 of compulsory education, aged 14-15. RESULTS: The results show that there is an association between internet activities and drinking and that there are differences depending on what young people do online. Self-presentation and online sociality are both positively associated with drinking, whereas news consumption and playing games are negatively associated with drinking. The results also show that the association between the different internet activities and drinking becomes weaker when adjusting for the control variables. CONCLUSION: This study suggests that more research is needed to examine the correlations between different forms of internet activities and drinking among adolescents in more detail.

Kinetics of crystalloid fluid in hyperglycemia; an open-label exploratory clinical trial.

BACKGROUND: Infusion with 0.9% saline is a mainstay in the treatment of severe hyperglycemia, but the kinetics of the saline volume in this setting has not been studied. METHODS: An intravenous infusion of 1 L of 0.9% saline over 30 minutes was given on 31 occasions to 17 patients with hyperglycemia due to poorly controlled diabetes (mean age 51 years). A two-volume kinetic model was fitted to serial data on the hemodilution and urinary excretion, using mixed-effects modeling software. RESULTS: Plasma glucose was 36 ± 9 mmol/L on arrival to the hospital. The central volume of distribution (the plasma) was only 2.38 L (mean; 95% confidence interval 1.73-3.04) on the day of admission. Uptake into a remote compartment, believed to be the cells, amounted to 300 mL of the first liter of saline, although only small amounts of insulin were given. Plasma glucose, plasma bicarbonate, urine glucose, and plasma creatinine served as covariates in the kinetic model and mathematically affected the urinary excretion. For example, elimination of the infused fluid tripled from an increase in plasma glucose from 5 to 35 mmol/L and doubled from a reduction in plasma bicarbonate from 24 to 5 mmol/L. CONCLUSIONS: The excretion of 0.9% saline was increased depending on the degree of hyperglycemia. The kinetics was characterized by glucose-accelerated diuresis, and an intracellular uptake that occurred at two thirds the urine flow rate. These data could help to determine appropriate volumes and rates of infusion of crystalloids in hyperglycemia.

The CREB coactivator CRTC2 promotes oncogenesis in LKB1-mutant non-small cell lung cancer.

The LKB1 tumor suppressor is often mutationally inactivated in non-small cell lung cancer (NSCLC). LKB1 phosphorylates and activates members of the AMPK family of Ser/Thr kinases. Within this family, the salt-inducible kinases (SIKs) modulate gene expression in part via the inhibitory phosphorylation of the CRTCs, coactivators for CREB (cAMP response element-binding protein). The loss of LKB1 causes SIK inactivation and the induction of the CRTCs, leading to the up-regulation of CREB target genes. We identified CRTC2 as a critical factor in LKB1-deficient NSCLC. CRTC2 is unphosphorylated and therefore constitutively activated in LKB1-mutant NSCLC, where it promotes tumor growth, in part via the induction of the inhibitor of DNA binding 1 (ID1), a bona fide CREB target gene. As ID1 expression is up-regulated and confers poor prognosis in LKB1-deficient NSCLC, our results suggest that small molecules that inhibit CRTC2 and ID1 activity may provide therapeutic benefit to individuals with NSCLC.

The AMPK-Related Kinases SIK1 and SIK3 Mediate Key Tumor-Suppressive Effects of LKB1 in NSCLC.

Mutations in the LKB1 (also known as STK11) tumor suppressor are the third most frequent genetic alteration in non-small cell lung cancer (NSCLC). LKB1 encodes a serine/threonine kinase that directly phosphorylates and activates 14 AMPK family kinases ("AMPKRs"). The function of many of the AMPKRs remains obscure, and which are most critical to the tumor-suppressive function of LKB1 remains unknown. Here, we combine CRISPR and genetic analysis of the AMPKR family in NSCLC cell lines and mouse models, revealing a surprising critical role for the SIK subfamily. Conditional genetic loss of Sik1 revealed increased tumor growth in mouse models of Kras-dependent lung cancer, which was further enhanced by loss of the related kinase Sik3. As most known substrates of the SIKs control transcription, gene-expression analysis was performed, revealing upregulation of AP1 and IL6 signaling in common between LKB1- and SIK1/3-deficient tumors. The SIK substrate CRTC2 was required for this effect, as well as for proliferation benefits from SIK loss. SIGNIFICANCE: The tumor suppressor LKB1/STK11 encodes a serine/threonine kinase frequently inactivated in NSCLC. LKB1 activates 14 downstream kinases in the AMPK family controlling growth and metabolism, although which kinases are critical for LKB1 tumor-suppressor function has remained an enigma. Here we unexpectedly found that two understudied kinases, SIK1 and SIK3, are critical targets in lung cancer.This article is highlighted in the In This Issue feature, p. 1469.

Genetic Analysis Reveals AMPK Is Required to Support Tumor Growth in Murine Kras-Dependent Lung Cancer Models.

AMPK, a conserved sensor of low cellular energy, can either repress or promote tumor growth depending on the context. However, no studies have examined AMPK function in autochthonous genetic mouse models of epithelial cancer. Here, we examine the role of AMPK in murine KrasG12D-mediated non-small-cell lung cancer (NSCLC), a cancer type in humans that harbors frequent inactivating mutations in the LKB1 tumor suppressor-the predominant upstream activating kinase of AMPK and 12 related kinases. Unlike LKB1 deletion, AMPK deletion in KrasG12D lung tumors did not accelerate lung tumor growth. Moreover, deletion of AMPK in KrasG12D p53f/f tumors reduced lung tumor burden. We identified a critical role for AMPK in regulating lysosomal gene expression through the Tfe3 transcription factor, which was required to support NSCLC growth. Thus, AMPK supports the growth of KrasG12D-dependent lung cancer through the induction of lysosomes, highlighting an unrecognized liability of NSCLC.

An examination of the interaction between morality and self-control in offending: A study of differences between girls and boys.

BACKGROUND: There is a well-documented gender difference in offending, with evidence that boys, on average, are more involved in crime than girls. Opinions differ, however, on whether the causes of crime apply to girls and boys similarly. AIMS: Our aim is to explore crime propensity in boys and girls. Our research questions were (1) are there differences between boys and girls in moral values and self-control; (2) are these attributes similarly correlated with offending among girls and boys; and (3) is any interaction effect between morality and self-control identical for girls and boys. METHODS: Data were drawn from the Malmö Individual and Neighbourhood Development Study, which includes 481 girls and boys aged 16-17. An 8-item self-control scale was derived from Grasmick's self-control instrument; we created a 16-item morality scale. Analysis of variance was used to test for differences in scale scores. RESULTS: There were significant gender differences in moral values but not self-control. Moral values and self-control were significantly correlated with offending among both girls and boys. In the multiple regression analysis, the three-way interaction term used to test the interaction between gender, self-control and moral values was non-significant, indicating that the magnitude of the self-control-moral value interaction is not affected by gender. CONCLUSIONS: Our findings indicate that effects of morality and self-control are general and apply to girls and boys similarly, so more research is needed to explain gender differences in crime prevalence. © 2018 The Authors Criminal Behaviour and Mental Health Published by John Wiley & Sons Ltd.

Chemical Proteomics Identifies Druggable Vulnerabilities in a Genetically Defined Cancer.

The transcription factor NRF2 is a master regulator of the cellular antioxidant response, and it is often genetically activated in non-small-cell lung cancers (NSCLCs) by, for instance, mutations in the negative regulator KEAP1. While direct pharmacological inhibition of NRF2 has proven challenging, its aberrant activation rewires biochemical networks in cancer cells that may create special vulnerabilities. Here, we use chemical proteomics to map druggable proteins that are selectively expressed in KEAP1-mutant NSCLC cells. Principal among these is NR0B1, an atypical orphan nuclear receptor that we show engages in a multimeric protein complex to regulate the transcriptional output of KEAP1-mutant NSCLC cells. We further identify small molecules that covalently target a conserved cysteine within the NR0B1 protein interaction domain, and we demonstrate that these compounds disrupt NR0B1 complexes and impair the anchorage-independent growth of KEAP1-mutant cancer cells. Our findings designate NR0B1 as a druggable transcriptional regulator that supports NRF2-dependent lung cancers.

LKB1 promotes metabolic flexibility in response to energy stress.

The Liver Kinase B1 (LKB1) tumor suppressor acts as a metabolic energy sensor to regulate AMP-activated protein kinase (AMPK) signaling and is commonly mutated in various cancers, including non-small cell lung cancer (NSCLC). Tumor cells deficient in LKB1 may be uniquely sensitized to metabolic stresses, which may offer a therapeutic window in oncology. To address this question we have explored how functional LKB1 impacts the metabolism of NSCLC cells using 13C metabolic flux analysis. Isogenic NSCLC cells expressing functional LKB1 exhibited higher flux through oxidative mitochondrial pathways compared to those deficient in LKB1. Re-expression of LKB1 also increased the capacity of cells to oxidize major mitochondrial substrates, including pyruvate, fatty acids, and glutamine. Furthermore, LKB1 expression promoted an adaptive response to energy stress induced by anchorage-independent growth. Finally, this diminished adaptability sensitized LKB1-deficient cells to combinatorial inhibition of mitochondrial complex I and glutaminase. Together, our data implicate LKB1 as a major regulator of adaptive metabolic reprogramming and suggest synergistic pharmacological strategies for mitigating LKB1-deficient NSCLC tumor growth.

Inhibition of acetyl-CoA carboxylase suppresses fatty acid synthesis and tumor growth of non-small-cell lung cancer in preclinical models.

Continuous de novo fatty acid synthesis is a common feature of cancer that is required to meet the biosynthetic demands of a growing tumor. This process is controlled by the rate-limiting enzyme acetyl-CoA carboxylase (ACC), an attractive but traditionally intractable drug target. Here we provide genetic and pharmacological evidence that in preclinical models ACC is required to maintain the de novo fatty acid synthesis needed for growth and viability of non-small-cell lung cancer (NSCLC) cells. We describe the ability of ND-646-an allosteric inhibitor of the ACC enzymes ACC1 and ACC2 that prevents ACC subunit dimerization-to suppress fatty acid synthesis in vitro and in vivo. Chronic ND-646 treatment of xenograft and genetically engineered mouse models of NSCLC inhibited tumor growth. When administered as a single agent or in combination with the standard-of-care drug carboplatin, ND-646 markedly suppressed lung tumor growth in the Kras;Trp53-/- (also known as KRAS p53) and Kras;Stk11-/- (also known as KRAS Lkb1) mouse models of NSCLC. These findings demonstrate that ACC mediates a metabolic liability of NSCLC and that ACC inhibition by ND-646 is detrimental to NSCLC growth, supporting further examination of the use of ACC inhibitors in oncology.

Lipid Synthesis Is a Metabolic Liability of Non-Small Cell Lung Cancer.

The renaissance in the study of cancer metabolism has refocused efforts to identify and target metabolic dependencies of tumors as an approach for cancer therapy. One of the unique metabolic requirements that cancer cells possess to sustain their biosynthetic growth demands is altered fatty acid metabolism, in particular the synthesis of de novo fatty acids that are required as cellular building blocks to support cell division. Enhanced fatty acid synthesis that is observed in many tumor types has been postulated to open a therapeutic window for cancer therapy and, correspondingly, efforts to pharmacologically inhibit key enzymes of fatty acid synthesis are being pursued. However, despite these efforts, whether inhibition of fatty acid synthesis stunts tumor growth in vivo has been poorly understood. In this review, we focus on the recent evidence that pharmacologic inhibition of acetyl-CoA carboxylase, the enzyme that regulates the rate-limiting step of de novo fatty acid synthesis, exposes a metabolic liability of non-small cell lung cancer and represses tumor growth in preclinical models.

An AMPK-independent signaling pathway downstream of the LKB1 tumor suppressor controls Snail1 and metastatic potential.

The serine/threonine kinase LKB1 is a tumor suppressor whose loss is associated with increased metastatic potential. In an effort to define biochemical signatures of metastasis associated with LKB1 loss, we discovered that the epithelial-to-mesenchymal transition transcription factor Snail1 was uniquely upregulated upon LKB1 deficiency across cell types. The ability of LKB1 to suppress Snail1 levels was independent of AMPK but required the related kinases MARK1 and MARK4. In a screen for substrates of these kinases involved in Snail regulation, we identified the scaffolding protein DIXDC1. Similar to loss of LKB1, DIXDC1 depletion results in upregulation of Snail1 in a FAK-dependent manner, leading to increased cell invasion. MARK1 phosphorylation of DIXDC1 is required for its localization to focal adhesions and ability to suppress metastasis in mice. DIXDC1 is frequently downregulated in human cancers, which correlates with poor survival. This study defines an AMPK-independent phosphorylation cascade essential for LKB1-dependent control of metastatic behavior.