Cytoplasmic DNA: sources, sensing, and role in aging and disease.

Endogenous cytoplasmic DNA (cytoDNA) species are emerging as key mediators of inflammation in diverse physiological and pathological contexts. Although the role of endogenous cytoDNA in innate immune activation is well established, the cytoDNA species themselves are often poorly characterized and difficult to distinguish, and their mechanisms of formation, scope of function and contribution to disease are incompletely understood. Here, we summarize current knowledge in this rapidly progressing field with emphases on similarities and differences between distinct cytoDNAs, their underlying molecular mechanisms of formation and function, interactions between cytoDNA pathways, and therapeutic opportunities in the treatment of age-associated diseases.

Histone chaperone HIRA, promyelocytic leukemia protein, and p62/SQSTM1 coordinate to regulate inflammation during cell senescence.

Cellular senescence, a stress-induced stable proliferation arrest associated with an inflammatory senescence-associated secretory phenotype (SASP), is a cause of aging. In senescent cells, cytoplasmic chromatin fragments (CCFs) activate SASP via the anti-viral cGAS/STING pathway. Promyelocytic leukemia (PML) protein organizes PML nuclear bodies (NBs), which are also involved in senescence and anti-viral immunity. The HIRA histone H3.3 chaperone localizes to PML NBs in senescent cells. Here, we show that HIRA and PML are essential for SASP expression, tightly linked to HIRA's localization to PML NBs. Inactivation of HIRA does not directly block expression of nuclear factor κB (NF-κB) target genes. Instead, an H3.3-independent HIRA function activates SASP through a CCF-cGAS-STING-TBK1-NF-κB pathway. HIRA physically interacts with p62/SQSTM1, an autophagy regulator and negative SASP regulator. HIRA and p62 co-localize in PML NBs, linked to their antagonistic regulation of SASP, with PML NBs controlling their spatial configuration. These results outline a role for HIRA and PML in the regulation of SASP.

Senescent glia link mitochondrial dysfunction and lipid accumulation.

Senescence is a cellular state linked to ageing and age-onset disease across many mammalian species1,2. Acutely, senescent cells promote wound healing3,4 and prevent tumour formation5; but they are also pro-inflammatory, thus chronically exacerbate tissue decline. Whereas senescent cells are active targets for anti-ageing therapy6-11, why these cells form in vivo, how they affect tissue ageing and the effect of their elimination remain unclear12,13. Here we identify naturally occurring senescent glia in ageing Drosophila brains and decipher their origin and influence. Using Activator protein 1 (AP1) activity to screen for senescence14,15, we determine that senescent glia can appear in response to neuronal mitochondrial dysfunction. In turn, senescent glia promote lipid accumulation in non-senescent glia; similar effects are seen in senescent human fibroblasts in culture. Targeting AP1 activity in senescent glia mitigates senescence biomarkers, extends fly lifespan and health span, and prevents lipid accumulation. However, these benefits come at the cost of increased oxidative damage in the brain, and neuronal mitochondrial function remains poor. Altogether, our results map the trajectory of naturally occurring senescent glia in vivo and indicate that these cells link key ageing phenomena: mitochondrial dysfunction and lipid accumulation.

Heme oxygenase-1 drives metaflammation and insulin resistance in mouse and man.

Obesity and diabetes affect more than half a billion individuals worldwide. Interestingly, the two conditions do not always coincide and the molecular determinants of "healthy" versus "unhealthy" obesity remain ill-defined. Chronic metabolic inflammation (metaflammation) is believed to be pivotal. Here, we tested a hypothesized anti-inflammatory role for heme oxygenase-1 (HO-1) in the development of metabolic disease. Surprisingly, in matched biopsies from "healthy" versus insulin-resistant obese subjects we find HO-1 to be among the strongest positive predictors of metabolic disease in humans. We find that hepatocyte and macrophage conditional HO-1 deletion in mice evokes resistance to diet-induced insulin resistance and inflammation, dramatically reducing secondary disease such as steatosis and liver toxicity. Intriguingly, cellular assays show that HO-1 defines prestimulation thresholds for inflammatory skewing and NF-κB amplification in macrophages and for insulin signaling in hepatocytes. These findings identify HO-1 inhibition as a potential therapeutic strategy for metabolic disease.

Mitochondria-to-nucleus retrograde signaling drives formation of cytoplasmic chromatin and inflammation in senescence.

Cellular senescence is a potent tumor suppressor mechanism but also contributes to aging and aging-related diseases. Senescence is characterized by a stable cell cycle arrest and a complex proinflammatory secretome, termed the senescence-associated secretory phenotype (SASP). We recently discovered that cytoplasmic chromatin fragments (CCFs), extruded from the nucleus of senescent cells, trigger the SASP through activation of the innate immunity cytosolic DNA sensing cGAS-STING pathway. However, the upstream signaling events that instigate CCF formation remain unknown. Here, we show that dysfunctional mitochondria, linked to down-regulation of nuclear-encoded mitochondrial oxidative phosphorylation genes, trigger a ROS-JNK retrograde signaling pathway that drives CCF formation and hence the SASP. JNK links to 53BP1, a nuclear protein that negatively regulates DNA double-strand break (DSB) end resection and CCF formation. Importantly, we show that low-dose HDAC inhibitors restore expression of most nuclear-encoded mitochondrial oxidative phosphorylation genes, improve mitochondrial function, and suppress CCFs and the SASP in senescent cells. In mouse models, HDAC inhibitors also suppress oxidative stress, CCF, inflammation, and tissue damage caused by senescence-inducing irradiation and/or acetaminophen-induced mitochondria dysfunction. Overall, our findings outline an extended mitochondria-to-nucleus retrograde signaling pathway that initiates formation of CCF during senescence and is a potential target for drug-based interventions to inhibit the proaging SASP.

Best practice approach for redo-surgeries after sleeve gastrectomy, an expert's modified Delphi consensus.

BACKGROUND: Sleeve gastrectomy (SG) is the most common metabolic and bariatric surgical (MBS) procedure worldwide. Despite the desired effect of SG on weight loss and remission of obesity-associated medical problems, there are some concerns regarding the need to do revisional/conversional surgeries after SG. This study aims to make an algorithmic clinical approach based on an expert-modified Delphi consensus regarding redo-surgeries after SG, to give bariatric and metabolic surgeons a guideline that might help for the best clinical decision. METHODS: Forty-six recognized bariatric and metabolic surgeons from 25 different countries participated in this Delphi consensus study in two rounds to develop a consensus on redo-surgeries after SG. An agreement/disagreement ≥ 70.0% on statements was considered to indicate a consensus. RESULTS: Consensus was reached for 62 of 72 statements and experts did not achieve consensus on 10 statements after two rounds of online voting. Most of the experts believed that multi-disciplinary team evaluation should be done in all redo-procedures after SG and there should be at least 12 months of medical and supportive management before performing redo-surgeries after SG for insufficient weight loss, weight regain, and gastroesophageal reflux disease (GERD). Also, experts agreed that in case of symptomatic GERD in the presence of adequate weight loss, medical treatment for at least 1 to 2 years is an acceptable option and agreed that Roux-en Y gastric bypass is an appropriate option in this situation. There was disagreement consensus on efficacy of omentopexy in rotation and efficacy of fundoplication in the presence of a dilated fundus and GERD. CONCLUSION: Redo-surgeries after SG is still an important issue among bariatric and metabolic surgeons. The proper time and procedure selection for redo-surgery need careful considerations. Although multi-disciplinary team evaluation plays a key role to evaluate best options in these situations, an algorithmic clinical approach based on the expert's consensus as a guideline can help for the best clinical decision-making.

Monitoring partially marked populations using camera and telemetry data.

Long-term monitoring is an important component of effective wildlife conservation. However, many methods for estimating density are too costly or difficult to implement over large spatial and temporal extents. Recently developed spatial mark-resight (SMR) models are increasingly being applied as a cost-effective method to estimate density when data include detections of both marked and unmarked individuals. We developed a generalized SMR model that can accommodate long-term camera data and auxiliary telemetry data for improved spatiotemporal inference in monitoring efforts. The model can be applied in two stages, with detection parameters estimated in the first stage using telemetry data and camera detections of instrumented individuals. Density is estimated in the second stage using camera data, with all individuals treated as unmarked. Serial correlation in detection and density parameters is accounted for using time-series models. The two-stage approach reduces computational demands and facilitates the application to large data sets from long-term monitoring initiatives. We applied the model to 3 years (2015-2017) of white-tailed deer (Odocoileus virginianus) data collected in three study areas of the Big Cypress Basin, Florida, USA. In total, 59 females marked with ear tags and fitted with GPS-telemetry collars were detected along with unmarked females on 180 remote cameras. Most of the temporal variation in density was driven by seasonal fluctuations, but one study area exhibited a slight population decline during the monitoring period. Modern technologies such as camera traps provide novel possibilities for long-term monitoring, but the resulting massive data sets, which are subject to unique sources of observation error, have posed analytical challenges. The two-stage spatial mark-resight framework provides a solution with lower computational demands than joint SMR models, allowing for easier implementation in practice. In addition, after detection parameters have been estimated, the model may be used to estimate density even if no synchronous auxiliary information on marked individuals is available, which is often the case in long-term monitoring.

The first modified Delphi consensus statement on sleeve gastrectomy.

INTRODUCTION: Sleeve gastrectomy (SG) is the commonest bariatric procedure worldwide. Yet there is significant variation in practice concerning its various aspects. This paper report results from the first modified Delphi consensus-building exercise on SG. METHODS: We established a committee of 54 globally recognized opinion makers in this field. The committee agreed to vote on several statements concerning SG. An agreement or disagreement amongst ≥ 70.0% experts was construed as a consensus. RESULTS: The committee achieved a consensus of agreement (n = 71) or disagreement (n = 7) for 78 out of 97 proposed statements after two rounds of voting. The committee agreed with 96.3% consensus that the characterization of SG as a purely restrictive procedure was inaccurate and there was 88.7% consensus that SG was not a suitable standalone, primary, surgical weight loss option for patients with Barrett's esophagus (BE) without dysplasia. There was an overwhelming consensus of 92.5% that the sleeve should be fashioned over an orogastric tube of 36-40 Fr and a 90.7% consensus that surgeons should stay at least 1 cm away from the angle of His. Remarkably, the committee agreed with 81.1% consensus that SG patients should undergo a screening endoscopy every 5 years after surgery to screen for BE. CONCLUSION: A multinational team of experts achieved consensus on several aspects of SG. The findings of this exercise should help improve the outcomes of SG, the commonest bariatric procedure worldwide, and guide future research on this topic.

Physiologic and blood gas effects of xylazine-ketamine versus xylazine-tiletamine-zolazepam immobilization of white-tailed deer before and after oxygen supplementation: a preliminary study.

OBJECTIVE: To compare oxygenation and ventilation in white-tailed deer (Odocoileus virginianus) anesthetized with two treatments with and without oxygen supplementation. STUDY DESIGN: Randomized, blinded, crossover study. ANIMALS: A total of eight healthy adult white-tailed deer weighing 49-62 kg. METHODS: Each deer was anesthetized twice intramuscularly: 1) treatment XK, xylazine (2 mg kg-1) and ketamine (6 mg kg-1) and 2) treatment XTZ, xylazine (2 mg kg-1) and tiletamine-zolazepam (4 mg kg-1). With the deer in sternal position, arterial and venous blood was collected before and at 30 minutes during administration of oxygen at 1 L minute-1 through a face mask. PaO2 and heart rate (HR) were compared using two-way repeated measures anova. pH, PaCO2 and lactate concentration were analyzed using mixed-effects linear models, p < 0.05. RESULTS: When breathing air, PaO2 was < 80 mmHg (10.7 kPa) in six and seven deer with XK and XTZ, respectively, and of these, PaO2 was < 60 mmHg (8.0 kPa) in three and five deer, respectively. With oxygen supplementation, PaO2 increased to 128 ± 4 and 140 ± 5 mmHg (17.1 ± 0.5 and 18.7 ± 0.7 kPa), mean ± standard error, with XK and XTZ, respectively (p < 0.001). PaO2 was not significantly different between treatments at either time point. HR decreased during oxygen supplementation in both treatments (p < 0.001). Lactate was significantly lower (p = 0.047) with XTZ than with XK (2.2 ± 0.6 versus 3.5 ± 0.6 mmol L-1) and decreased (p < 0.001) with oxygen supplementation (4.1 ± 0.6 versus 1.6 ± 0.6 mmol L-1). PaCO2 increased in XTZ during oxygen breathing. CONCLUSIONS AND CLINICAL RELEVANCE: Treatments XK and XTZ resulted in hypoxemia, which responded to oxygen supplementation. Both treatments are suitable for immobilization of white-tailed deer under the study circumstances.

Mercury Concentrations in Feathers of Adult and Nestling Osprey (Pandion haliaetus) from Coastal and Freshwater Environments of Florida.

We determined mercury (Hg) concentrations in feathers of osprey (Pandion haliaetus), both nestlings (n = 95) and adults (n = 110), across peninsular Florida and the Florida Keys during February-August 2014. Feathers plucked from nestlings, aged 3-7 weeks, contained Hg concentrations that ranged from 0.338 to 45.79 mg/kg and averaged 6.92 ± 7.58 mg/kg (mean ± 1SD). Feathers shed from adults contained significantly higher concentrations ranging from 0.375 to 93.65 mg/kg, with an average of 17.8 ± 16.1 mg/kg. These levels were in the upper range of previously reported feather Hg concentrations of osprey and clearly show that Florida continues to have Hg hotspots that are elevated compared with many other regions. While these concentrations did not exceed levels previously reported in osprey from heavily Hg contaminated areas that showed no evidence of reproductive impairments, we cannot rule out potential individual-level effects to highly exposed nestlings after fledging. Mercury concentrations in nestlings were highest in coastal habitats of Collier and Monroe counties, where ongoing declines in osprey populations also have been documented.

Isolation and genotypic characterization of Trueperella (Arcanobacterium) pyogenes recovered from active cranial abscess infections of male white-tailed deer (Odocoileus virginianus).

Trueperella (Arcanobacterium) pyogenes is a causative agent of suppurative infections in domestic and wild animals. In some populations of captive or free-ranging white-tailed deer (Odocoileus virginianus), cranial abscess disease is an important source of mortality in adult males. Although the pathogenesis of this disease is poorly understood, T. pyogenes has been isolated from active infections with other opportunistic bacteria. In this study, bacteria associated with cranial abscess infections were identified, the prevalence of T. pyogenes associated with these infections was determined, and the presence of known virulence determinants in T. pyogenes isolates was ascertained. Using routine biochemical techniques seven bacterial species were identified from 65 samples taken from active cranial abscess infections of 65 male white-tailed deer. Trueperellapyogenes was recovered from 46 samples; in 32 samples it was the only bacterium species detected. Staphylococcus aureus was detected in 26 samples. From these samples, the presence of known and putative virulence genes of T. pyogenes--plo, nanH, nanP, cbpA, fimA, fimC, fimE, and fimG--was examined by conventional polymerase chain reaction. All T. pyogenes isolates were positive for the pyolysin genes plo, nanP, and fimA. Furthermore, nanH, fimA, fimC, and fimE were detected in over 70% of isolates. Of the isolates tested, 48% had genotypes containing all virulence genes except cbpA. The suggestive virulence potential of all isolates, coupled with the large number of pure cultures obtained, implies that T. pyogenes is a causative agent of cranial abscess disease.

Aerosol transmission of chronic wasting disease in white-tailed deer.

While the facile transmission of chronic wasting disease (CWD) remains incompletely elucidated, studies in rodents suggest that exposure of the respiratory mucosa may be an efficient pathway. The present study was designed to address this question in the native cervid host. Here, we demonstrate aerosol transmission of CWD to deer with a prion dose >20-fold lower than that used in previous oral inoculations. Inhalation of prions may facilitate transmission of CWD and, perhaps, other prion infections.

Exploring sleepiness and stress among London bus drivers: An on-road observational study.

Bus driver sleepiness is commonplace but often goes unreported within the industry. Whilst past research has begun to shed a light on the prevalence, potential causes, and consequences of bus driver sleepiness, this is often done using self-report methods. This is the first study to investigate sleepiness amongst city bus drivers on-road using a live bus route with drivers' regular schedules. A total of 16 participants completed two drives of their regular bus route once during an early morning shift and once during a daytime shift whilst physiological and self-report measures of sleep and stress were taken. Prior to these drives, drivers recorded their sleep in a diary and wore an actigraph to obtain objective sleep measures. Results showed that most drivers did not obtain sufficient sleep prior to early morning shifts, and often did not obtain as much sleep as they would need in order to feel rested before work. Sleepiness and stress were observed in both shifts. During early morning shifts sleepiness was likely a result of working during circadian lows and not obtaining enough sleep prior to the shift. In contrast, sleepiness during the daytime shift was likely a result of completing a highly demanding task in complex traffic which not only contributed to fatigue, but also led to increased levels of stress. As well as demonstrating the prevalence of sleepiness amongst bus drivers, these findings show that the causes of sleepiness can be multifaceted and often come about due to a combination of work and personal factors. In addition, the experience of sleepiness is not the same for all drivers, with individual differences in the experience of sleepiness playing a large role. These differences highlight the need for individualised interventions which should be considered by policymakers alongside the combination of causal factors within a larger systems approach.

Measuring the benefit of a defensive trait: Vigilance and survival probability.

Defensive traits are hypothesized to benefit prey by reducing predation risk from a focal predator but come at a cost to the fitness of the prey. Variation in the expression of defensive traits is seen among individuals within the same population, and in the same individual in response to changes in the environment (i.e., phenotypically plastic responses). It is the relative magnitude of the cost and benefit of the defensive trait that underlies the defensive trait expression and its consequences to the community. However, whereas the cost has received much attention in ecological research, the benefit is seldom examined. Even in a defensive trait as extensively studied as vigilance, there are few studies of the purported benefit of the behavior, namely that vigilance enhances survival. We examined whether prey vigilance increased survival and quantified that benefit in a natural system, with white-tailed deer (Odocoileus virginianus) experiencing unmanipulated levels of predation risk from Florida panther (Puma concolor coryi). Deer that spent more time vigilant (as measured by head position using camera trap data) had a higher probability of survival. Indeed, an individual deer that was vigilant 75% of the time was more than three times as likely to be killed by panthers over the course of a year than a deer that was vigilant 95% of the time. Our results therefore show that within-population variation in the expression of a defensive trait has profound consequences for the benefit it confers. Our results provide empirical evidence supporting a long-held but seldom-tested hypothesis, that vigilance is a behavior that reduces the probability of predation and quantifies the benefit of this defensive trait. Our work furthers an understanding of the net effects of a trait on prey fitness and predator-prey interactions, within-population variation in traits, and predation risk effects.

Female Deer Movements Relative to Firearms Hunting in Northern Georgia, USA.

Perceived risk associated with hunters can cause white-tailed deer (Odocoileus virginianus) to shift their activity away from key foraging areas or alter normal movements, which are important considerations in managing hunting and its effects on a population. We studied the effects of seven firearms hunts on the movements of 20 female deer in two Wildlife Management Areas within the Chattahoochee National Forest of northern Georgia, USA, during the 2018-2019 and 2019-2020 hunting seasons. Deer populations and the number of hunters in our study area have declined significantly since the 1980s. In response, hunting regulations for the 2019-2020 hunting season eliminated opportunities for harvesting female deer. To evaluate the indirect effects of antlered deer hunting on non-target female deer, we calculated 90% utilization distributions (UDs), 50% UDs, and step lengths for pre-hunt, hunt, and post-hunt periods using the dynamic Brownian bridge movement model. Data included 30 min GPS locations for 44 deer-hunt combinations. Pre-hunt 50% UDs (x- = 7.0 ha, SE = 0.4 ha) were slightly greater than both hunt (x- = 6.0 ha, SE = 0.3 ha) and post-hunt (x- = 6.0 ha, SE = 0.2 ha) 50% UDs (F = 3.84, p = 0.03). We did not detect differences in step length, nor did we detect differences in size or composition of 90% UDs, among the periods. Overall, our results suggest that the low level of hunting pressure in our study area and lack of exposure to hunters led to no biologically significant changes in female deer movements. To the extent of the findings presented in this paper, adjustments to the management of hunting in our study area do not appear to be necessary to minimize hunting-related disturbances for female deer. However, managers should continue to consider female deer behavior when evaluating future changes to hunting regulations.

Histone chaperone HIRA, Promyelocytic Leukemia (PML) protein and p62/SQSTM1 coordinate to regulate inflammation during cell senescence.

Cellular senescence, a stress-induced stable proliferation arrest associated with an inflammatory Senescence-Associated Secretory Phenotype (SASP), is a cause of aging. In senescent cells, Cytoplasmic Chromatin Fragments (CCFs) activate SASP via the anti-viral cGAS/STING pathway. PML protein organizes PML nuclear bodies (NBs), also involved in senescence and anti-viral immunity. The HIRA histone H3.3 chaperone localizes to PML NBs in senescent cells. Here, we show that HIRA and PML are essential for SASP expression, tightly linked to HIRA's localization to PML NBs. Inactivation of HIRA does not directly block expression of NF-κB target genes. Instead, an H3.3-independent HIRA function activates SASP through a CCF-cGAS-STING-TBK1-NF-κB pathway. HIRA physically interacts with p62/SQSTM1, an autophagy regulator and negative SASP regulator. HIRA and p62 co-localize in PML NBs, linked to their antagonistic regulation of SASP, with PML NBs controlling their spatial configuration. These results outline a role for HIRA and PML in regulation of SASP.

Revision/Conversion Surgeries After One Anastomosis Gastric Bypass-An Experts' Modified Delphi Consensus.

PURPOSE: There is a lack of evidence for treatment of some conditions including complication management, suboptimal initial weight loss, recurrent weight gain, or worsening of a significant obesity complication after one anastomosis gastric bypass (OAGB). This study was designed to respond to the existing lack of agreement and to provide a valuable resource for clinicians by employing an expert-modified Delphi consensus method. METHODS: Forty-eight recognized bariatric surgeons from 28 countries participated in the modified Delphi consensus to vote on 64 statements in two rounds. An agreement/disagreement among ≥ 70.0% of the experts was regarded to indicate a consensus. RESULTS: A consensus was achieved for 46 statements. For recurrent weight gain or worsening of a significant obesity complication after OAGB, more than 85% of experts reached a consensus that elongation of the biliopancreatic limb (BPL) is an acceptable option and the total bowel length measurement is mandatory during BPL elongation to preserve at least 300-400 cm of common channel limb length to avoid nutritional deficiencies. Also, more than 85% of experts reached a consensus on conversion to Roux-en-Y gastric bypass (RYGB) with or without pouch downsizing as an acceptable option for the treatment of persistent bile reflux after OAGB and recommend detecting and repairing any size of hiatal hernia during conversion to RYGB. CONCLUSION: While the experts reached a consensus on several aspects regarding revision/conversion surgeries after OAGB, there are still lingering areas of disagreement. This highlights the importance of conducting further studies in the future to address these unresolved issues.

Current recommendations for procedure selection in class I and II obesity developed by an expert modified Delphi consensus.

Metabolic and bariatric surgery (MBS) is widely considered the most effective option for treating obesity, a chronic, relapsing, and progressive disease. Recently, the American Society of Metabolic and Bariatric Surgery (ASMBS) and the International Federation for the Surgery of Obesity and Metabolic Disorders (IFSO) issued new guidelines on the indications for MBS, which have superseded the previous 1991 National Institutes of Health guidelines. The aim of this study is to establish the first set of consensus guidelines for selecting procedures in Class I and II obesity, using an Expert Modified Delphi Method. In this study, 78 experienced bariatric surgeons from 32 countries participated in a two-round Modified Delphi consensus voting process. The threshold for consensus was set at an agreement or disagreement of ≥ 70.0% among the experts. The experts reached a consensus on 54 statements. The committee of experts reached a consensus that MBS is a cost-effective treatment option for Class II obesity and for patients with Class I obesity who have not achieved significant weight loss through non-surgical methods. MBS was also considered suitable for patients with Type 2 diabetes mellitus (T2DM) and a body mass index (BMI) of 30 kg/m2 or higher. The committee identified intra-gastric balloon (IGB) as a treatment option for patients with class I obesity and endoscopic sleeve gastroplasty (ESG) as an option for patients with class I and II obesity, as well as for patients with T2DM and a BMI of ≥ 30 kg/m2. Sleeve gastrectomy (1) and Roux-en-Y gastric bypass (RYGB) were also recognized as viable treatment options for these patient groups. The committee also agreed that one anastomosis gastric bypass (OAGB) is a suitable option for patients with Class II obesity and T2DM, regardless of the presence or severity of obesity-related medical problems. The recommendations for selecting procedures in Class I and II obesity, developed through an Expert Modified Delphi Consensus, suggest that the use of standard primary bariatric endoscopic (IGB, ESG) and surgical procedures (SG, RYGB, OAGB) are acceptable in these patient groups, as consensus was reached regarding these procedures. However, randomized controlled trials are still needed in Class I and II Obesity to identify the best treatment approach for these patients in the future.

Potential role of upstream stimulatory factor 1 gene variant in familial combined hyperlipidemia and related disorders.

OBJECTIVE: Genetic studies implicated upstream stimulatory factor 1 (USF1) in familial combined hyperlipidemia because the rs2073658 minor allele was associated with reduced risk of familial combined hyperlipidemia and related disorders. The molecular mechanisms whereby rs2073658 influences trait expression have remained elusive. METHODS AND RESULTS: Plasma lipids, rs2073658 genotypes (N=372), and hepatic transcript levels (N=96) of USF1 and genes involved in hepatic lipoprotein production were determined in obese subjects. The rs2073658 minor allele was associated with reduced plasma triglycerides (TGs) (P<0.001), hepatic USF1 (P<0.01), and microsomal TG transfer protein transcript levels (P<0.05). Functional studies in human hepatocellular carcinoma cells showed that rs2073658 is located in a forkhead box A2 (FOXA2) binding site and that major allele constructs displayed higher transcriptional activity than minor allele constructs. Knockdown of FOXA2 reduced the activity of major, but not minor allele constructs. Furthermore, an interaction between hepatic FOXA2 transcript levels and rs2073658 minor allele carrier status on hepatic USF1 transcript levels was observed in vivo (P<0.05). USF1 activated the transcription of FOXA2 and FOXA2 strongly activated the transcription of microsomal TG transfer protein. CONCLUSIONS: A feed-forward loop comprising activation of USF1 transcription by FOXA2 and activation of FOXA2 transcription by USF1, driving microsomal TG transfer protein expression, is modulated by rs2073658. Hence, rs2073658 likely influences hepatic TG secretion.