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Introduction The COVID-19 pandemic has significantly affected everyday life in most countries of the world. Researches conducted in 2020 showed that COVID-19 was a traumatic experience for 18-60% of respondents in the general population, depending on the country where the research is conducted. Researches for later periods are rare, although we can expect significant changes. We found only one study from 2023 on this topic, although not on the general population. Objectives That is why we were interested in what the situation is like after 3 years of the pandemic, when we have been living without non-pharmacological anti-pandemic measures for almost a year. Methods The research was conducted at the beginning of 2023. 48 respondents who were not treated psychiatrically or are medical workers were surveyed, because it was shown that these groups were exposed to a greater risk of impaired mental health during the COVID-19 pandemic. To assess the level of traumatic experience, i.e. the risk of developing PTSD as a consequence of the COVID-19 pandemic, we used the Impact of Event Scale With Modifications for COVID-19 (IES-COVID19). A score on that scale of 27 to 34 indicates a clinically significant level of trauma, i.e. there is a 75% chance of developing PTSD. A result of 35 and above suggests that it is necessary to seek professional help. Results Our research included 19 (39.6%) men and 29 (60.4%) women. The average age of the respondents is 60.4 years. 29 (60.4%) respondents know that they have recovered from COVID-19. 2 (4.2%) subjects were treated in the hospital due to COVID-19. 8 (16.7%) respondents have a traumatic experience of the COVID-19 pandemic. 5 (10.4%) respondents are in the category of clinically significant level of trauma, while 3 (6.3%) respondents are in the category that should seek professional help. The group traumatized by COVID-19 does not have significantly more respondents who recovered from COVID-19 (p=0.510) nor does it differ in terms of gender representation (p=0.984). Conclusions At the beginning of the COVID-19 pandemic, there were discussions about whether it can even be classified as a traumatic experience and whether we can talk about PTSD as a consequence of the pandemic. With this time lag, it seems that in part of the population we are finding PTSD symptoms that are a consequence of the pandemic, but to a lesser extent than research at the beginning of the pandemic suggested. Certainly, additional research is needed on this topic. Also, it is necessary to examine risk factors for possible prevention, as well as therapeutic possibilities. Disclosure of Interest None Declared

Introduction Online group therapy has become more popular in the past few years. But as a result of the COVID-19-caused pandemic, it developed suddenly. Due to the conventional face-to-face format no longer being possible and the need for psychotherapists to conduct psychotherapy online, the pandemic has had significant effects on group psychotherapy and the interactions between group therapy members. While therapists are becoming accustomed to the modern form of psychotherapy, its efficacy is being questioned due to technical issues, the problem of the therapeutic alliance, the environment, the ability to read nonverbal signals, breaking group norms, etc. Since the pandemic did not abate, as a part of specialist education training groups were also held online. Objectives The pandemic changed the basic settings of our Group-Analytic Training Group, forcing us to switch to online sessions. This study aimed to find personal experiences that varied throughout online and face-to-face meetings. Methods Seven out of the twelve participants accepted to take part in the group therapy/training after it was recommended by the group leader that they write a paper. After 30 sessions, the group turned from face-to-face to online group therapy, and the members were asked how they felt about the difference between the two types of therapy. A questionnaire was produced by the group’s leader and a number of other participants, who then forwarded it through email to every group member. Results Everyone who participated thought that because one can more quickly pick up on non-verbal signs in a face-to-face scenario, it was simpler to notice feedback from the other group members. Most participant comments focused on the leader’s role. The majority of members claimed that taking part in the experiential group had benefited both their personal and professional lives.However they thought the in-person setting was better since it was more interesting and complex. Conclusions Since there were no other options during the epidemic, group therapy has moved to virtual environments, although there are still a lot of problems to this method. The formation of group cohesion becomes difficult by the absence of group members’ physical presence and by the inability to completely understand nonverbal communication. Disclosure of Interest None Declared

Introduction For many people, January is the most depressing month of the year. “Blue Monday” encompasses the generally accepted belief that Monday is the hardest day of the week compared to Friday and Saturday, which are the most anticipated days of the week. The connection between the color blue and Monday is in the emotional stage, which is presented as emotional anger. The third Monday in January is currently known as the most depressing day of the year. Speaker Cliff Arnall was the first to declare that day in 2014. The theory says that this is the time of the year when respiratory diseases are common, the day is shorter, the weather conditions are worse, and the time when people are burdened with guilt about whether they will achieve their New Year’s resolutions. Objectives The aim of this work was to investigate that on third Monday in January there were more suicide attempts and that there were more depressive disorders in emergency psychiatric admissions. Methods In the research, we included participants who were examined at the Emergency Psychiatric Admission of the Clinical Hospital Center in Split, in the period from 2019. until 2023. Inclusion criteria were respondents of both sexes, examined in the outpatient clinic on Mondays in January for five years. Results There were 198 of them in total. The primary outcome of the research is to determine the occurrence (incidence) of psychological deterioration in patients diagnosed with the anxiety-depressive spectrum. The secondary research outcomes are of a descriptive nature, patient follow-up, examination outcome, and psychiatric heredity. Conclusions For now, there are no strong scientific foundations that justify the formula of “the most depressing day” of the year, some scientists believe that it is a marketing trick to achieve higher tourist revenues. However, the post-holiday period can have an impact on individuals. Disclosure of Interest None Declared

Introduction The COVID-19 pandemic has had an enormous impact on both physical and mental health of people around the world. Objectives The aim of the study was to evaluate the number and characteristics of people seeking emergency psychiatric help during combined psychosocial stressful events in March 2020. Methods Data for 3927 patients seeking emergency psychiatric help were collected and analyzed for the months preceding, during and after lockdown due to COVID-19 pandemic and concomitant earthquake that took place on 22nd March 2020 in Zagreb, and compared with the same months of 2019. Results A significant decrease in both the number of visits and admissions to the hospital was found for the month of lockdown. There was a significant decrease in the number of out-patients visits and day hospital admissions. Compared with other months, more women and younger patients sought help. There was a significant rise in the number of patients presenting with suicidal thoughts, as well as a larger percentage of involuntary admissions. Conclusions Overall less people sought psychiatric help in the face of an unpredictable acute threat, which was interpreted in the light of prioritizing fear of infection over mental health issues. Alternatively, it is possible that people threatened with immediate danger mobilize short- term compensatory psychological resources which help deal with or put off mental illness. This research was conducted as part of the project of the Croatian Science Foundation CORONA-04-2086 Life in the time of COVID-19-social implications on the security and well-being of vulnerable groups in the European context. Disclosure No significant relationships.

Background: Breast conservation therapy is a widely accepted approach in treating breast cancer, yet the average re-excision rates are approximately 25% despite surgical advancements. The Food and Drug Administration (FDA)-approved MarginProbe ® device uses radiofrequency spectroscopy for intraoperative margin assessment, potentially reducing re-excision rates. This study evaluated the effectiveness of MarginProbe ® in reducing re-excisions compared with standard of care (SOC)., Methods: A prospective cohort with MarginProbe ® usage during partial mastectomies from June 2019 to July 2023 (153 patients) was compared with a retrospective control group without the device from January 2015 to May 2019 (300 patients). Both groups underwent partial mastectomies performed by two breast surgeons. Positive margins were defined as tumor on ink for invasive cancers and within 2 mm for ductal carcinoma in situ., Results: When control was used for patient demographics and tumor characteristics, the findings showed that MarginProbe ® significantly decreased the probability of re-excision by 58% (p < 0.001), although it led to a higher shave volume, with an average of 9.8 cc additional tissue removed compared with SOC (p < 0.001). Human epidermal growth factor 2 (HER2) positivity was significantly associated with increased odds of re-excision (p = 0.036). MarginProbe ® demonstrated a sensitivity of 70.1% and a specificity of 47.5%., Conclusions: MarginProbe ® is an effective adjunct for intraoperative margin assessment to decrease re-excision rates. However, patient selection is paramount. Given its significant increase in shave volume, women with small breasts may be at higher risk for poor cosmesis. Surgeons should exercise clinical judgement when determining the suitability of MarginProbe ® use for patients undergoing breast conservation. Further research is necessary to refine MarginProbe ® 's specificity and to optimize its clinical application., (© 2024. Society of Surgical Oncology.)

Prenatal multivitamins, including folic acid, are commonly consumed in excess, whereas choline, an essential nutrient and an important source of labile methyl groups, is underconsumed. Here, we characterized profiles of one-carbon metabolism and related pathways and patterns of DNA methylation in offspring exposed to excess or imbalanced micronutrients prenatally. Pregnant Wistar rats were fed either recommended 1× vitamins (RV), high 10× vitamins (HV), high 10× folic acid with recommended choline (HFolRC), or high 10× folic acid with no choline (HFolNC). Offspring were weaned to a high-fat diet for 12 weeks. Circulating metabolites were analyzed with a focus on the hypothalamus, an area known to be under epigenetic regulation. HV, HFolRC, and HFolNC males had higher body weight (BW) and lower plasma choline and methionine consistent with lower hypothalamic S-adenosylmethionine (SAM):S-adenosylhomocysteine (SAH) and global DNA methylation compared with RV. HV and HFolNC females had higher BW and lower plasma 5-methyltetrahydrofolate and methionine consistent with lower hypothalamic global DNA methylation compared with RV. Plasma dimethylglycine (DMG) and methionine were higher as with hypothalamic SAM:SAH and global DNA methylation in HFolRC females without changes in BW compared with RV. Plasma trimethylamine and trimethylamine-N-oxide were higher in males but lower in females from HFolRC compared with RV. Network modeling revealed a link between the folate-dependent pathway and SAH, with most connections through DMG. Final BW was negatively correlated with choline, DMG, and global DNA methylation. In conclusion, prenatal intake of excess or imbalanced micronutrients induces distinct metabolic and epigenetic perturbations in offspring that reflect long-term nutritional programming of health., (© 2024 The Author(s). The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

Herein, we present the case of accidental intravenous injection of gasoline in a 62-year-old male who was admitted to a dialysis center for his regular hemodialysis. Due to previous contact with another SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) positive patient, the hemodialysis was conducted in an isolated room. At the end of the procedure, the nurse, wearing all necessary personal protective equipment (PPE), in the intent to clean the dialysis catheter, applied medical gasoline, instead of 0.9% sodium chloride, intravenously to the patient. Soon afterwards, the patient's clinical condition deteriorated, and cardiopulmonary resuscitation was started. Despite the immediate reaction of the medical staff, after two successful cardiopulmonary reanimation and necessary intensive care measures, the patient suffered respiratory, metabolic, and lactic acidosis, hypotension, and tachyarrhythmia and ultimately died 7 h after the incident. The autopsy was conducted under the order of the district attorney. Main autopsy findings were marked congestion; right pleural and pericardial effusion; brain and lung edema; enlarged heart with left ventricle thickening and mild perivascular fibrosis; nephrosclerosis; tubular thyroidization; and interstitial fibrosis with inflammation. Gasoline presence was indisputably proven by conducted toxicology analysis in lung, bile, and brain samples. Traces of gasoline could be noted in the patient's blood sample in comparison to the blood that did not contain gasoline, but it was not possible to confidently claim that gasoline was present in the blood. Based on relevant findings, we concluded that the death of the patient was violent and that the cause of death was acute intoxication by gasoline., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: Disparities in insurance and socioeconomic status (SES) may impact surgical management and subsequent postoperative outcomes for patients with craniosynostosis. This systematic review summarizes the evidence on possible differences in surgical care, including procedure type, age at surgery, and differences in surgical outcomes such as complications, length of hospital stay, and child development based on SES., Design: The databases Scopus, PubMed, and CINAHL were searched between May and July 2022. Following PICO criteria, studies included focused on patients diagnosed with craniosynostosis; corrective surgery for craniosynostosis; comparison of insurance, income, or zip code; and surgical management of postoperative outcomes., Results: The initial search yielded 724 articles. After three stages of screening, 13 studies were included. Assessed outcomes included: type of procedure (6 articles), age at time of surgery (3 articles), post-operative complications (3 articles), referral delay (2 articles), length of stay (2 articles), hospital costs (2 articles), and child development (1 article). Of the studies with significant results, insurance type was the main SES variable of comparison. While some findings were mixed, these studies indicated that patients with public medical insurance were more likely to experience a delay in referral, undergo an open rather than minimally-invasive procedure, and have more complications, longer hospitalization, and higher medical charges., Conclusions: This study demonstrated that SES may be associated with several differences in the management of patients with craniosynostosis. Further investigation into the impact of SES on the management of patients with craniosynostosis is warranted to identify possible interventions that may improve overall care., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Simultaneous free flap breast reconstruction combined with contralateral mastopexy or breast reduction can increase patient satisfaction and minimize the need for a second procedure. Surgeon concerns of increases in operative time, postoperative complications, and final breast symmetry may decrease the likelihood of these procedures being done concurrently. This study analyzed postoperative outcomes of simultaneous contralateral mastopexy or breast reduction with free flap breast reconstruction., Methods: By using the American College of Surgeons National Surgical Quality Improvement Program database (2010-2020), we analyzed 2 patient cohorts undergoing (A) free flap breast reconstruction only and (B) free flap breast reconstruction combined with contralateral mastopexy or breast reduction. The preoperative variables assessed included demographic data, comorbidities, and perioperative data. Using a neighbor matching algorithm, we performed a 1:1 propensity score matching of 602 free flap breast reconstruction patients and 621 with concurrent contralateral operation patients. Bivariate analysis for postoperative surgical and medical complications was performed for outcomes in the propensity-matched cohort., Results: We identified 11,308 cases who underwent microsurgical free flap breast reconstruction from the American College of Surgeons National Surgical Quality Improvement Program database from the beginning of 2010 to the end of 2020. A total of 621 patients underwent a free flap breast reconstruction combined with contralateral mastopexy or breast reduction. After propensity score matching, there were no significant differences in patient characteristics, perioperative variables or postoperative medical complications between the 2 cohorts., Conclusions: Simultaneous free flap breast reconstruction combined with contralateral mastopexy or breast reduction can be performed safely and effectively without an increase in postoperative complication rates. This can improve surgeon competence in offering this combination of procedures as an option to breast cancer survivors, leading to better patient outcomes in terms of symmetrical and aesthetically pleasing results, reduced costs, and elimination of the need for a second operation., Competing Interests: Conflicts of interest and sources of funding: none declared., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

This study aimed to investigate the serum metabolomic profile of obese and lean cats as well as obese cats before and after energy restriction for weight loss. Thirty cats, 16 obese (body condition score 8 to 9/9) and 14 lean (body condition score 4 to 5/9), were fed a veterinary weight loss food during a 4-week period of weight maintenance (L-MAINT and O-MAINT). The 16 obese cats were then energy restricted by a 60% energy intake reduction with the same food for a 10-week period (O-RESTRICT). Fasted serum metabolites were measured using nuclear magnetic resonance and direct infusion mass spectrometry after the maintenance period for L-MAINT and O-MAINT cats and after the energy restriction period for O-RESTRICT and compared between groups using a two-sided t-test. Obese cats lost 672 g ± 303 g over the 10-week restriction period, representing a weight loss rate of 0.94 ± 0.28% per week. Glycine, l-alanine, l-histidine, l-glutamine, 2-hydroxybutyrate, isobutryric acid, citric acid, creatine, and methanol were greater in O-RESTRICT compared to O-MAINT. There was a greater concentration of long-chain acylcarnitines in O-RESTRICT compared to both O-MAINT and L-MAINT, and greater total amino acids compared to O-MAINT. Glycerol and 3-hydroxybutyric acid were greater in O-MAINT compared to L-MAINT, as were several lysophosphatidylcholines. Thus, energy restriction resulted in increased dispensable amino acids in feline serum which could indicate alterations in amino acid partitioning. An increase in lipolysis was not evident, though greater circulating acylcarnitines were observed, suggesting that fatty acid oxidation rates may have been greater under calorie restriction. More research is needed to elucidate energy metabolism and substrate utilization, specifically fatty acid oxidation and methyl status, during energy restriction in strict carnivorous cats to optimize weight loss., Competing Interests: The authors declare no conflicts of interest. CG held the Nestle Purina Professorship in Companion Animal Nutrition at the Ontario Veterinary College at the time of manuscript drafting, is the owner of Grant Veterinary Nutrition Services and consults with Simmons Pet Food. MT is currently employed by Royal Canin. The study was conducted before this employment during MT’s Doctor Veterinary Sciences Degree at the Ontario Veterinary College. HG declares that they have participated in paid internships and engagements with pet food companies within Canada. AKS is the Champion Petfoods Chair in Canine and Feline Nutrition, Physiology and Metabolism, consults for Champion Petfood, was previously employed by P&G and Mars Pet Care, serves on the Scientific Advisory Board for Trouw Nutrition, and has received honoraria and research funding from various commodity groups, pet food manufacturers, and ingredient suppliers. AV is the Royal Canin Veterinary Diets Endowed Chair in Canine and Feline Clinical Nutrition and declares that they serve on the Health and Nutrition Advisory Board for Vetdiet. AV has also received honoraria and research funding from various pet food manufacturers and ingredient suppliers. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Grant et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Dermatofibrosarcoma protuberans (DFSP) is a rare malignant fibroblastic tumor. DFSP has an insidious onset, slow growth, and heterogeneous presentation that can create a delay in diagnosis and increase morbidity. In this case report, we present a child with DFSP that presented as a large, slow-growing mass over the dorsum of the left foot. She underwent successful surgical excision with no functional sequelae., Competing Interests: The authors have no financial interest to declare in relation to the content of this article., (Copyright © 2024 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of The American Society of Plastic Surgeons.)

Choline is beneficial for energy metabolism and growth in various species. Choline may work similarly in kittens at risk of obesity. Direct infusion MS (Di-MS) and NMR spectroscopy were used to investigate the metabolomic signatures of kittens supplemented with or without additional dietary choline for 12 weeks. Fifteen intact male kittens consumed a base diet (3310 mg choline/kg DM) to their daily metabolisable energy requirement (DER) over an 11-week acclimation. Kittens were gonadectomised and assigned, based on body weight, to the base diet (CONTROL, n 7) or the base diet with 300 mg/kgBW 0·75 additional choline as choline chloride (CHOLINE, n 8) and offered three times their individual energy requirement divided into three meals. At weeks -1 and 12, fasted blood was sampled and serum analysed for 130 metabolites via Di-MS and fifty-one metabolites via NMR spectroscopy. Changes in fasted metabolites were assessed using a repeated-measures GLIMMIX procedure with time and group as fixed effects, and time as a repeated measure. Metabolites of one-carbon metabolism and lipids increased, and medium-chain acyl carnitines decreased from week -1 to 12 for CHOLINE ( P < 0·05), but not CONTROL ( P > 0·05). Increases in amino acid, biogenic amine and organic compound concentrations were observed in both groups ( P < 0·05). The results suggest impacts of dietary choline at greater intakes than currently recommended on one-carbon metabolism and fatty acid oxidation, and these may promote healthy growth in post-gonadectomy kittens.

Introduction: Due to the involvement in one-carbon metabolism and lipid mobilization, choline and L-carnitine supplementation have been recommended to minimize hepatic lipid accumulation and support fat oxidation, respectively. This study investigated the lipotropic benefits of choline or L-carnitine supplementation in lean and obese cats maintaining body weight (BW)., Methods: Lean [ n = 9; body condition score (BCS): 4-5/9] and obese ( n = 9; BCS: 8-9/9) adult male neutered colony cats were used in a replicated 3 x 3 complete Latin square design. Treatments included choline (378 mg/kg BW 0.67 ), L-carnitine (200 mg/kg BW) and control (no supplement). Treatments were supplemented to the food for 6 weeks each, with a 2-week washout between treatments. Cats were fed once daily to maintenance energy requirements, and BW and BCS were assessed weekly. Fasted blood collection, indirect calorimetry, and dual-energy X-ray absorptiometry occurred at the end of each treatment period. Serum was analyzed for cholesterol (CHOL), high-density lipoprotein CHOL (HDL-C), triglycerides (TAG), non-esterified fatty acids (NEFA), glucose, creatinine (CREAT), urea, alkaline phosphatase (ALP) and alanine aminotransferase (ALT). Very low-density lipoprotein CHOL (VLDL) and low-density lipoprotein CHOL (LDL-C) were calculated. Data were analyzed using proc GLIMMIX, with group and period as random effects, and treatment, body condition, and their interaction as fixed effects, followed by a Tukey's post-hoc test when significance occurred., Results: Cats supplemented choline had lower food intake ( P = 0.025). Treatment did not change BW, BCS and body composition ( P > 0.05). Obese cats had greater ALP, TAG, and VLDL, and lower HDL-C compared to lean cats ( P < 0.05). Choline resulted in greater CHOL, HDL-C, LDL-C and ALT ( P < 0.05). L-carnitine resulted in lower CREAT ( P = 0.010). Following the post-hoc test, differences between treatment means were not present for ALP ( P = 0.042). No differences were found for glucose, urea or NEFA ( P > 0.05). Obese cats had a lower fed respiratory quotient (RQ), regardless of treatment ( P = 0.045). Treatment did not affect fed or fasted RQ and energy expenditure ( P > 0.05)., Discussion: Choline appeared to increase circulating lipid and lipoprotein concentrations regardless of body condition, likely through enhanced lipid mobilization and hepatic elimination. Neither dietary choline or L-carnitine altered body composition or energy metabolism in the lean or obese cats, as compared to control., Competing Interests: AR declares that they have participated in paid engagements with and received scholarships from pet food companies within Canada. At the time of the study, SV-S held the position of Registered Veterinary Technician in Clinical Nutrition funded by Nestlé Purina Proplan Veterinary Diets, they were previously employed by Hill's Pet Nutrition, and received honoraria from pet food manufacturers. AS is the Champion Petfoods Chair in Canine and Feline Nutrition, Physiology and Metabolism, consults for Champion Petfood, was previously employed by P&G and Mars Pet Care, serves on the Scientific Advisory Board for Trouw Nutrition, and has received honoraria and research funding from various commodity groups, pet food manufacturers, and ingredient suppliers. AV is the Royal Canin Veterinary Diets Endowed Chair in Canine and Feline Clinical Nutrition, serves on the Health and Nutrition Advisory Board for Vetdiet and has received honoraria and research funding from various pet food manufacturers and ingredient suppliers. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Rankovic, Verton-Shaw, Shoveller, Bakovic, Kirby and Verbrugghe.)

The maintenance of phospholipid homeostasis is increasingly being implicated in metabolic health. Phosphatidylethanolamine (PE) is the most abundant phospholipid on the inner leaflet of cellular membranes, and we have previously shown that mice with a heterozygous ablation of the PE synthesizing enzyme, Pcyt2 ( Pcyt2 +/- ), develop obesity, insulin resistance, and NASH. Skeletal muscle is a major determinant of systemic energy metabolism, making it a key player in metabolic disease development. Both the total PE levels and the ratio of PE to other membrane lipids in skeletal muscle are implicated in insulin resistance; however, the underlying mechanisms and the role of Pcyt2 regulation in this association remain unclear. Here, we show how reduced phospholipid synthesis due to Pcyt2 deficiency causes Pcyt2 +/- skeletal muscle dysfunction and metabolic abnormalities. Pcyt2 +/- skeletal muscle exhibits damage and degeneration, with skeletal muscle cell vacuolization, disordered sarcomeres, mitochondria ultrastructure irregularities and paucity, inflammation, and fibrosis. There is intramuscular adipose tissue accumulation, and major disturbances in lipid metabolism with impaired FA mobilization and oxidation, elevated lipogenesis, and long-chain fatty acyl-CoA, diacylglycerol, and triacylglycerol accumulation. Pcyt2 +/- skeletal muscle exhibits perturbed glucose metabolism with elevated glycogen content, impaired insulin signaling, and reduced glucose uptake. Together, this study lends insight into the critical role of PE homeostasis in skeletal muscle metabolism and health with broad implications on metabolic disease development., Competing Interests: The authors declare no competing interests., (© The Author(s) 2023. Published by Oxford University Press on behalf of American Physiological Society.)

Muscle degeneration is the most prevalent cause for frailty and dependency in inherited diseases and ageing. Elucidation of pathophysiological mechanisms, as well as effective treatments for muscle diseases, represents an important goal in improving human health. Here, we show that the lipid synthesis enzyme phosphatidylethanolamine cytidyltransferase (PCYT2/ECT) is critical to muscle health. Human deficiency in PCYT2 causes a severe disease with failure to thrive and progressive weakness. pcyt2-mutant zebrafish and muscle-specific Pcyt2-knockout mice recapitulate the participant phenotypes, with failure to thrive, progressive muscle weakness and accelerated ageing. Mechanistically, muscle Pcyt2 deficiency affects cellular bioenergetics and membrane lipid bilayer structure and stability. PCYT2 activity declines in ageing muscles of mice and humans, and adeno-associated virus-based delivery of PCYT2 ameliorates muscle weakness in Pcyt2-knockout and old mice, offering a therapy for individuals with a rare disease and muscle ageing. Thus, PCYT2 plays a fundamental and conserved role in vertebrate muscle health, linking PCYT2 and PCYT2-synthesized lipids to severe muscle dystrophy and ageing., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

In mammalian cells, mitochondrial respiration produces reactive oxygen species (ROS) such as superoxide (O 2 - ), which is then converted by the SOD1 enzyme into hydrogen peroxide (H 2 O 2 ), the predominant form of cytosolic ROS. ROS at high levels can be toxic, but below this threshold are important for physiological processes acting as a second messenger similar to Ca 2+ . Mitochondrial Ca 2+ influx from the ER increases ATP and ROS production, while ATP and ROS can regulate Ca 2+ homeostasis, leading to an intricate interplay between Ca 2+ , ROS, and ATP synthesis. The Unfolded Protein Response (UPR) proteins ATF6α and XBP1 contribute to protection from oxidative stress through upregulation of Sod1 and Catalase genes. Here, UPR-associated protein CREB3 is shown to play a role in balancing Ca 2+ , ROS, and ATP homeostasis. Creb3-deficient mouse embryonic fibroblast cells (MEF -/- ) were susceptible to H 2 O 2 -induced oxidative stress while having a functioning antioxidant gene expression response compared to MEF +/+ . MEF -/- cells also contained elevated basal cytosolic ROS levels, which was attributed to drastically increased basal mitochondrial respiration and spare respiratory capacity relative to MEF +/+ . MEF -/- cells also showed an increase in endoplasmic reticulum Ca 2+ release and mitochondrial Ca 2+ levels hinting at a potential cause for MEF -/- cell mitochondrial dysfunction. These results suggest that CREB3 is essential for maintaining proper Ca 2+ , ATP, and ROS homeostasis in mammalian cells., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. and Mitochondria Research Society. All rights reserved. All rights reserved.)

Choline participates in methyl group metabolism and has been recognized for its roles in lipid metabolism, hepatic health and muscle function in various species. Data regarding the impacts of choline on feline metabolic pathways are scarce. The present study investigated how choline intake affects the metabolomic profile of overweight cats fed at maintenance energy. Overweight (n = 14; body condition score:6-8/9) male adult cats were supplemented with five doses of choline in a 5x5 Latin Square design. Cats received a daily dose of choline on extruded food (3620 mg choline/kg diet) for three weeks at maintenance energy requirements (130 kcal/kgBW0.4). Doses were based on body weight (BW) and the daily recommended allowance (RA) for choline for adult cats (63 mg/kg BW0.67). Treatment groups included: Control (no additional choline, 1.2 x NRC RA, 77 mg/kg BW0.67), 2 x NRC RA (126 mg/kg BW0.67), 4 x NRC RA (252 mg/kg BW0.67), 6 x RA (378 mg/kg BW0.67), and 8 x NRC RA (504 mg/kg BW0.67). Serum was collected after an overnight fast at the end of each treatment period and analyzed for metabolomic parameters through nuclear magnetic resonance (NMR) spectroscopy and direct infusion mass spectrometry (DI-MS). Data were analyzed using GLIMMIX, with group and period as random effects, and dose as the fixed effect. Choline up to 8 x NRC RA was well-tolerated. Choline at 6 and 8 x NRC RA resulted in greater concentrations of amino acids and one-carbon metabolites (P < 0.05) betaine, dimethylglycine and methionine. Choline at 6 x NRC RA also resulted in greater phosphatidylcholine and sphingomyelin concentrations (P < 0.05). Supplemental dietary choline may be beneficial for maintaining hepatic health in overweight cats, as it may increase hepatic fat mobilization and methyl donor status. Choline may also improve lean muscle mass in cats. More research is needed to quantify how choline impacts body composition., Competing Interests: The authors declare no conflicts of interest. AR and HG declare that they have participated in paid internships and engagements with pet food companies within Canada. CG holds the Nestle Purina Professorship in Companion Animal Nutrition at the Ontario Veterinary College, is the owner of Grant Veterinary Nutrition Services and consults with Simmons Pet Food. AKS is the Champion Petfoods Chair in Canine and Feline Nutrition, Physiology and Metabolism, consults for Champion Petfood, was previously employed by P&G and Mars Pet Care, serves on the Scientific Advisory Board for Trouw Nutrition, and has received honoraria and research funding from various commodity groups, pet food manufacturers, and ingredient suppliers. AV is the Royal Canin Veterinary Diets Endowed Chair in Canine and Feline Clinical Nutrition and declares that they serve on the Health and Nutrition Advisory Board for Vetdiet. AV has also received honoraria and research funding from various pet food manufacturers and ingredient suppliers. At the time of the study, the Ontario Veterinary College received funding from Nestlé Purina Proplan Veterinary Diets to support a Registered Veterinary Technician in Clinical Nutrition, who helped perform the described animal trial. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Rankovic et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Phosphatidylethanolamine is the major inner-membrane lipid in the plasma and mitochondrial membranes. It is synthesized in the endoplasmic reticulum from ethanolamine and diacylglycerol (DAG) by the CDP-ethanolamine pathway and from phosphatidylserine by decarboxylation in the mitochondria. Recently, multiple genetic disorders that impact these pathways have been identified, including hereditary spastic paraplegia 81 and 82, Liberfarb syndrome, and a new type of childhood-onset neurodegeneration-CONATOC. Individuals with these diseases suffer from multisystem disorders mainly affecting neuronal function. This indicates the importance of maintaining proper phospholipid homeostasis when major biosynthetic pathways are impaired. This study summarizes the current knowledge of phosphatidylethanolamine metabolism in order to identify areas of future research that might lead to the development of treatment options., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 St Germain, Iraji and Bakovic.)

Background: The endoplasmic reticulum senses alterations to cellular homeostasis that activates the unfolded protein response (UPR). UPR proteins are known to aid in regulating glucose and lipid metabolism. CREB3 is a UPR-associated transcription factor whose potential role in regulating energy metabolism remains unclear., Methods: Eight-week-old wild-type (WT) and Creb3 +/- mice were placed on control and high-fat diets (HFD) for 8 weeks, and metabolic phenotypes characterized by weekly weighing, indirect calorimetry, body composition scans, glucose tolerance tests, plasma analysis, tissue lipid quantifications and gene/protein expression analysis., Results: HFD weight gain in Creb3 +/- males was reduced by 34% (p < 0.0001) and females by 39.5% (p = 0.014) from their WT counterparts. No differences were found in HFD food intake or total fecal lipids between genotypes. Creb3 +/- mice had increased energy expenditure and respiratory exchange ratios (p = 0.002) relative to WT. Creb3 +/- mice had significant reductions in absolute fat and lean tissue, while Creb3 +/- females had significant reductions in body fat% and increased lean% composition (p < 0.0001) compared to WT females. Creb3 +/- mice were protected from HFD-induced basal hyperglycemia (males p < 0.0001; females p = 0.0181). Creb3 +/- males resisted HFD-induced hepatic lipid accumulation (p = 0.025) and glucose intolerance compared to WT (p < 0.0001) while Creb3 +/- females were protected from lipid accumulation in skeletal muscle (p = 0.001). Despite the metabolic differences of Creb3 +/- mice on HFD, lipid plasma profiles did not significantly differ from WT. Fasted Creb3 +/- mice additionally revealed upregulation of hepatic energy expenditure and gluconeogenic genes such as Pgc-1a and Gr (glucocorticoid receptor) (p < 0.05), respectively., Conclusions: Reduced expression of CREB3 increased energy expenditure and the respiratory exchange ratio, and protected mice from HFD-induced weight gain, basal hyperglycemia, and sex-specific tissue lipid accumulation. We postulate that CREB3 is a novel key regulator of diet-induced obesity and energy metabolism that warrants further investigation as a potential therapeutic target in metabolic disorders., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Choline is an essential nutrient linked to hepatic lipid metabolism in many animal species, including cats. The current study investigated the serum lipid profiles, serum liver enzymes, respiratory quotients, and energy expenditures of overweight cats fed maintenance diets, in response to graded doses of supplemental dietary choline. Overweight (body condition score [BCS]: ≥6/9) adult male neutered cats (n = 14) were supplemented with five choline chloride doses for 3-wk periods, in a 5 × 5 Latin square design. Doses were based on individual body weight (BW) and the daily recommended allowance (RA) for choline (63 mg/kg BW0.67) according to the National Research Council. Doses were control (no additional choline: 1.2 × RA, 77 mg/kg BW0.67), 2 × RA (126 mg/kg BW0.67), 4 × RA (252 mg/kg BW0.67), 6 × RA (378 mg/kg BW0.67), and 8 × RA (504 mg/kg BW0.67). Choline was top-dressed over the commercial extruded cat food (3,620 mg choline/kg diet), fed once a day at maintenance energy requirements (130 kcal/kgBW0.4). Body weight and BCS were assessed weekly. Fasted blood samples were taken and indirect calorimetry was performed at the end of each 3-wk period. Serum was analyzed for cholesterol, high-density lipoprotein cholesterol (HDL-C), triglycerides, non-esterified fatty acids, glucose, creatinine, blood urea nitrogen (BUN), alkaline phosphatase (ALP), and alanine aminotransferase. Very low-density lipoprotein cholesterol (VLDL) and low-density lipoprotein cholesterol were calculated. Data were analyzed via SAS using proc GLIMMIX, with group and period as the random effects, and treatment as the fixed effect. Statistical significance was considered at P < 0.05. Body weight and BCS did not change (P > 0.05). Serum cholesterol, HDL-C, triglycerides, and VLDL increased with 6 × RA (P < 0.05). Serum ALP decreased with 8 × RA (P = 0.004). Choline at 4 × and 6 × RA decreased serum BUN (P = 0.006). Fed or fasted respiratory quotient and energy expenditure did not differ among dietary choline doses (P > 0.05). These results suggest that dietary choline supplementation at 6 × RA may increase hepatic fat mobilization through increased lipoprotein transport and beneficially support hepatic health in overweight cats. Future studies that combine these results with existing knowledge of feline weight loss and hepatic lipidosis are warranted., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Phospholipids represent a crucial component for the structure of cell membranes. Phosphatidylcholine and phosphatidylethanolamine are two phospholipids that comprise the majority of cell membranes. De novo biosynthesis of phosphatidylcholine and phosphatidylethanolamine occurs via the Kennedy pathway, and perturbations in the regulation of this pathway are linked to a variety of human diseases, including cancer. Altered phosphatidylcholine and phosphatidylethanolamine membrane content, phospholipid metabolite levels, and fatty acid profiles are frequently identified as hallmarks of cancer development and progression. This review summarizes the research on how phospholipid metabolism changes over oncogenic transformation, and how phospholipid profiling can differentiate between human cancer and healthy tissues, with a focus on colorectal cancer, breast cancer, and non-small cell lung cancer. The potential for phospholipids to serve as biomarkers for diagnostics, or as anticancer therapy targets, is also discussed.

Gonadectomy is a major risk factor for feline obesity. The lipotropic effects of choline have demonstrated benefits for growth and carcass composition in livestock. The consumption of supplemental choline on body weight (BW), body composition, lipid metabolism, energy expenditure (EE), and serum satiety hormones were evaluated in 15 gonadectomized male kittens. Kittens were offered a base diet formulated for growth (3310mg choline/kg dry matter [DM]) to daily energy requirements (DER) over an 11-week acclimation. Post-gonadectomy, kittens were assigned to a base diet (CONTROL, n = 7) or choline group (base diet with additional choline at 300mg/kg BW0.75 as a top dress) (CHOLINE, n = 8). For 12-weeks post-neuter, kittens were offered three times their DER over three meals to mimic ad libitum feeding. At week -1 and 12, body composition was assessed using dual energy x-ray absorptiometry (DXA), 24-hour indirect calorimetry was performed for EE and respiratory quotients (RQ), and fasted serum samples were analyzed for lipid compounds and satiety hormones. Daily food intake (FI) and weekly BW were measured. Data was analyzed as a repeated measures of variance (ANCOVA) using the GLIMMIX procedure with time and group as fixed effects. CHOLINE had lower mean daily FI and lower rates of BW accretion (P<0.05) in contrast to CONTROL. All absolute body composition data increased over time for both groups, with lower increases in total tissue mass (P = 0.031) and fat mass (P = 0.005) in CHOLINE. Serum satiety hormones and lipid compounds did not differ (P>0.05) between groups, but both groups experienced a decrease in low-density lipoproteins and increase in high-density lipoproteins (P<0.05). Primary substrate utilization showed lipid use when fasted and use of protein or mixed macronutrients in the fed state. Fed state EE decreased post-gonadectomy (P = 0.004), however, CHOLINE did not affect total EE or RQ. These results suggest that supplemental dietary choline reduces FI, BW, and fat mass and may help to reduce the propensity of weight gain and subsequent obesity in gonadectomized feline populations., Competing Interests: The authors declare no conflicts of interest. AV is the Royal Canin Veterinary Diets Endowed Chair in Canine and Feline Clinical Nutrition and declares that they serve on the Health and Nutrition Advisory Board for Vetdiet. AV has received honoraria and research funding from various pet food manufacturers and ingredient suppliers. HG declares that they have paid engagements with pet food companies within Canada. AR declares that they have participated in paid internships and engagements with pet food companies within Canada. CG holds the Nestlé Purina Professorship in Companion Animal Nutrition at the Ontario Veterinary College, is the owner of Grant Veterinary Nutrition Services and consults with Simmons Pet Food. The Ontario Veterinary College received funding from Nestlé Purina Proplan Veterinary Diets to support a Registered Veterinary Technician in Clinical Nutrition, who assisted with this study. AKS was previously employed by P&G and Mars Pet Care, serves on the Scientific Advisory Board for Trouw Nutrition, and has received honoraria and research funding from various commodity groups, pet food manufacturers, and ingredient suppliers. SKA is the owner of Sit, Stay Speak Nutrition LLC and provides nutrition consultation to industry partners. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: The high prevalence of uncontrolled hypertension (systolic blood pressure [SBP] ≥140 mmHg or diastolic blood pressure [DBP] ≥90 mmHg) in Black patients represents a significant racial health disparity in the United States., Aims: This study evaluated the efficacy of a telephone-based strategy for inviting high-risk patients with severe hypertension to weekly self-management education classes. Further, the study assessed how the outreach intervention correlated with relevant quality improvement outcomes, including improved blood pressure and primary care follow-up among our clinic population of Black men with severe hypertension., Methods: A cohort of 265 Black men aged ≥18 years with SBP ≥160 mmHg or DBP ≥100 mmHg at the most recent clinic visit were identified using Epic reports formatted for Federal Uniformed Data Set annual reporting. Telephone outreach was used to invite the cohort to attend weekly in-person classes facilitated by various healthcare professionals. Logistic regression was performed to determine the associations between being reached by phone with (1) class attendance and (2) follow-up appointment attendance., Results: Most of the Black men were single (57.4%, n = 152), 49.1% had history of alcohol or substance use (n = 130), and 35.8% (n = 95) was uninsured. The average age was 55.6 years (SD = 11.6). After controlling for sociodemographic factors, being reached by phone was significantly associated with an increased likelihood of patient attendance at follow-up appointments (OR = 1.91, p = .038) but not with class attendance (OR = 2.45, p = .155). Patients who attended a follow-up appointment experienced significant reductions in both SBP and DBP at 9 months., Linking Evidence to Action: Telephone outreach was labor-intensive but effective in keeping under-resourced patient populations engaged in primary care. Future work should aim to develop more efficient strategies for engaging high-risk patients in self-monitoring education to manage hypertension., (© 2022 Sigma Theta Tau International.)

Background: Uncontrolled hypertension disproportionately affects Black men. Patient engagement with health promotion and health behavior programs for hypertension control is low. We held evidence-based hypertension classes at a Federally Qualified Health Center., Aims: The aim of this project was to elicit patient preferences and expectations for community-based hypertension classes., Methods: Group hypertension classes were held at a Federally Qualified Health Center. The priority population was Black men with hypertension. Semi-structured interviews were conducted with the group class attendees. Two independent coders performed content analysis using field notes from the interviews., Results: Seven group class attendees were interviewed. Six interviewees were Black males, one was a Black female, and the mean age was 65.7 years. Content analysis of the interview field notes revealed five content areas: (1) Recommendation of Group Classes to a Friend, (2) Expectations for the Group Classes, (3) Reasons for Continued Attendance, (4) Lifestyle Changes and Adopting the Group Class Lessons, and (5) Feedback and Recommendations to Improve Group Classes. Findings suggest that creating a cultural context in which open, transparent discussion of blood pressure occur can be achieved. Thus, regarding acceptability, classes fulfilled and often exceeded the attendees' expectations and needs., Linking Evidence to Action: Our findings suggest that community-based group classes are a viable intervention to contribute toward addressing the disparity of hypertension control among Black men. When designing peer-group interventions, taking patient preferences and expectations into account increases the effectiveness of these interventions. The qualitative narrative provided in this paper contributes to the development of similar community-based group classes for the management of severe hypertension., (© 2022 Sigma Theta Tau International.)

The mechanisms of NASH development in the context of age and genetics are not fully elucidated. This study investigates the age-dependent liver defects during NASH development in mice with heterozygous deletion of Pcyt2 (Pcyt2 +/- ), the rate limiting enzyme in phosphatidylethanolamine (PE) synthesis. Further, the therapeutic potential of Pcyt2 substrate, phosphoethanolamine (PEtn), is examined. Pcyt2 +/- were investigated at 2 and 6-8 months (mo) of age and in addition, 6-mo old Pcyt2 +/- with developed NASH were supplemented with PEtn for 8 weeks and glucose and fatty acid metabolism, insulin signaling, and inflammation were examined. Heterozygous ablation of Pcyt2 causes changes in liver metabolic regulators from young age, prior to the development of liver disease which does not occur until adulthood. Only older Pcyt2 +/- experiences perturbed glucose and fatty acid metabolism. Older Pcyt2 +/- liver develops NASH characterized by increased glucose production, accumulation of TAG and glycogen, and increased inflammation. Supplementation with PEtn reverses Pcyt2 +/- steatosis, inflammation, and other aspects of NASH, showing that was directly caused by Pcyt2 deficiency. Pcyt2 deficiency is a novel mechanism of metabolic dysregulation due to reduced membrane ethanolamine phospholipid synthesis, and the metabolite PEtn offers therapeutic potential for NASH reversion., (© 2022. The Author(s).)

Grain-based ingredients are replaced in part by pulse ingredients in grain-free pet foods. Pulse ingredients are lower in methionine and cysteine, amino acid (AA) precursors to taurine synthesis in dogs. Although recent work has investigated plasma and whole blood taurine concentrations when feeding grain-free diets, supplementation of a grain-free diet with various nutrients involved in the biosynthesis of taurine has not been evaluated. This study aimed to investigate the effects of supplementing a complete grain-free dry dog food with either methionine (MET), taurine (TAU), or methyl donors (choline) and methyl receivers (creatine and carnitine; CCC) on postprandial AA concentrations. Eight healthy Beagle dogs were fed one of the three treatments or the control grain-free diet (CON) for 7 d in a 4 × 4 Latin square design. On day 7, cephalic catheters were placed and one fasted sample (0 min) and a series of nine post-meal blood samples were collected at 15, 30, 60, 90, 120, 180, 240, 300, and 360 min. Data were analyzed as repeated measures using the PROC GLIMMIX function in SAS (Version 9.4). Dogs fed MET had greater plasma and whole blood methionine concentrations from 30 to 360 min after a meal (P < 0.0001) and greater plasma homocysteine concentrations from 60 to 360 min after a meal (P < 0.0001) compared with dogs fed CON, TAU, and CCC. Dogs fed TAU had greater plasma taurine concentrations over time compared with dogs fed CON (P = 0.02) but were not different than dogs fed MET and CCC (P > 0.05). In addition, most AAs remained significantly elevated at 6 h post-meal compared with fasted samples across all treatments. Supplementation of creatine, carnitine, and choline in grain-free diets may play a role in sparing the methionine requirement without increasing homocysteine concentrations. Supplementing these nutrients could also aid in the treatment of disease that causes metabolic or oxidative stress, including cardiac disease in dogs, but future research is required., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Obesity is a health concern for domestic cats. Obesity and severe energy restriction predispose cats to feline hepatic lipidosis. As choline is linked to lipid metabolism, we hypothesized that dietary choline supplementation would assist in reducing hepatic fat through increased lipoprotein transport and fatty acid oxidation. Twelve obese cats (body condition score [BCS] ≥ 8/9) were split into two groups. Cats were fed a control ( n = 6; 4587 mg choline/kg dry matter [DM]) or a high choline diet ( n = 6; 18,957 mg choline/kg DM) for 5 weeks, for adult maintenance. On days 0 and 35, fasted blood was collected, and the body composition was assessed. Serum lipoprotein and biochemistry profiles, plasma amino acids and plasma acylcarnitines were analyzed. The body weight, BCS and body composition were unaffected ( p > 0.05). Choline increased the serum cholesterol, triacylglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol and plasma methionine ( p < 0.05) and decreased the serum blood urea nitrogen and alkaline phosphatase ( p < 0.05). Choline also reduced the plasma acylcarnitine to free carnitine ratio ( p = 0.006). Choline may assist in eliminating hepatic fat through increased fat mobilization and enhanced methionine recycling.

Structural racism has contributed to persistent racial disparities in hypertension control, with Black men suffering the highest prevalence of uncontrolled hypertension. Lincoln Community Health Center, our urban Federally Qualified Health Center (FQHC), aimed to use hypertension self-management classes to improve hypertension control among our clinic patients, particularly Black men. Patients attending classes learned about hypertension, were given blood pressure cuffs to use at home, and had the opportunity to speak to physicians in a group setting. We used a nonexperimental quality improvement intervention design to identify baseline differences between participants who attended multiple classes and those who attended only 1 class. Participants who attended multiple classes, most of whom were Black men, achieved an average blood pressure reduction of 19.1/14.8 mm Hg. Although the classes were effective, current policies around health insurance reimbursement and federal quality reporting standards hamper the ability of health care providers to implement such patient education initiatives.

Due to the impact of persistent structural racism, Black men have low rates of hypertension treatment and control despite having high rates of hypertension-related mortality. Peer-based education can improve blood pressure monitoring and lower blood pressure in Black men with hypertension. To address this disparity, we implemented weekly community-based group classes for severe hypertension at a Federally Qualified Health Center. After 9 months, 28 classes were held and 96 individuals were served. Fifty-six percent of the person-hours of attendance have been by Black men. Seven individuals were interviewed about their perspectives and preferences for the classes. They reported that the peer-based model of the group classes was advantageous, and they would recommend the classes to a friend. Successes of implementation included (a) ability to facilitate medication adjustments for participants with blood pressure readings that were over target goal, (b) capacity to give participants take-home blood pressure monitors, (c) and the community-centered approach. Challenges included variability in attendance by participants and retention of individuals with uncontrolled hypertension in primary care. These findings have implications for other Federally Qualified Health Centers seeking to design similar health promotion programs and policy makers evaluating the funding and reimbursement models for community-based disease management programs., (© 2021 Wiley Periodicals, Inc.)

The membrane phospholipids phosphatidylcholine and phosphatidylethanolamine (PE) are synthesized de novo by the CDP-choline and CDP-ethanolamine (Kennedy) pathway, in which the extracellular substrates choline and ethanolamine are transported into the cell, phosphorylated, and coupled with diacylglycerol to form the final phospholipid product. Although multiple transport systems have been established for choline, ethanolamine transport is poorly characterized and there is no single protein assigned a transport function for ethanolamine. The solute carriers 44A (SLC44A) known as choline transporter-like proteins-1 and -2 (CTL1 and CTL2) are choline transporter at the plasma membrane and mitochondria. We report a novel function of CTL1 and CTL2 in ethanolamine transport. Using the lack or the gain of gene function in combination with specific antibodies and transport inhibitors we established two distinct ethanolamine transport systems of a high affinity, mediated by CTL1, and of a low affinity, mediated by CTL2. Both transporters are Na + -independent ethanolamine/H + antiporters. Primary human fibroblasts with separate frameshift mutations in the CTL1 gene (M1= SLC44A1 ΔAsp517 and M2= SLC44A1 ΔSer126 ) are devoid of CTL1 ethanolamine transport but maintain unaffected CTL2 transport. The lack of CTL1 in M2 cells reduced the ethanolamine transport, the flux through the CDP-ethanolamine Kennedy pathway, and PE synthesis. In contrast, overexpression of CTL1 in M2 cells improved ethanolamine transport and PE synthesis. These data firmly establish that CTL1 and CTL2 are the first identified ethanolamine transporters in whole cells and mitochondria, with intrinsic roles in de novo PE synthesis by the Kennedy pathway and intracellular redistribution of ethanolamine., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: This study aimed to determine if obese cats undergoing energy restriction for weight loss would meet the National Research Council's (NRC) indispensable amino acid and vitamin recommendations when fed a purpose-formulated diet. Thirty cats were placed into one of two groups; obese (BCS 8 to 9/9; n = 16) and lean (BCS 4 to 5/9; n = 14) and included in a non-randomized retrospective observational study. Cats were fed a veterinary weight loss food during a 4-week period of weight maintenance. Obese cats (O-MAINT) refers to obese cats during this period, L-MAINT to lean cats. After this initial 4-week period, the lean cats finished the study at this time and the 16 obese cats continued and were energy restricted for a 10-week period (O-RESTRICT). Analysis for dietary concentrations of indispensable amino acid and vitamin contents were performed. Daily food intakes were used to determine minimum, maximum and average daily intakes of individual nutrients for all three groups and compared against NRC 2006 minimum requirements (MR), adequate intakes (AI) and recommended allowances (RA) for adult cats., Results: Over 10 weeks, O-RESTRICT cats lost 672 g ± 303 g, representing a weight loss rate of 0.94 ± 0.28% per week. Daily intake of the majority of indispensable amino acids and vitamins was greater than the NRC 2006 recommended allowance (RA per kg ideal body weight ^0.67), except for arginine, choline, crude protein, phenylalanine plus tyrosine and threonine. All O-RESTRICT cats had minimum, average, and maximum arginine intakes less than the NRC AI. Minimum daily intake of choline was below NRC RA for all O-RESTRICT cats and below NRC MR for two. All, except one, O-RESTRICT cats had a maximum and average choline intake below RA., Conclusions: All cats remained clinically healthy and showed no clinical signs of deficiency. Dietary choline and arginine requirements of obese cats as well as health risks associated with low dietary intake during energy restriction warrant further investigation.

By replacing lost or dysfunctional myocardium, tissue regeneration is a promising approach to treat heart failure. However, the challenge of detecting bona fide heart regeneration limits the validation of potential regenerative factors. One method to detect new cardiomyocytes is multicolor lineage tracing with clonal analysis. Clonal analysis experiments can be difficult to undertake, because labeling conditions that are too sparse lack sensitivity for rare events such as cardiomyocyte proliferation, and diffuse labeling limits the ability to resolve clones. Presented here is a protocol to undertake clonal analysis of the neonatal mouse heart by using statistical modeling of nearest neighbor distributions to resolve cardiomyocyte clones. This approach enables resolution of clones over a range of labeling conditions and provides a robust analytical approach for quantifying cardiomyocyte proliferation and regeneration. This protocol can be adapted to other tissues and can be broadly used to study tissue regeneration.

Unlike polyphenols, which are widely available in the diet, polyacetylenes are available only from the Apiaceae family vegetables, including carrot, parsnip, fennel, celery, and many herbs (parsley, lovage, etc). The aim of this study was to investigate the hypothesis that polyacetylene falcarinol (FA) reduces intestinal inflammation and examine its similarity of effect to isothiocyanate R-sulforaphane during the late phase of acute inflammation. To this end, 3-month-old male CB57BL/6 mice were fed twice daily for 1 week with 5 mg/kg of FA, sulforaphane, or vehicle before receiving an intraperitoneal injection of 5 mg/kg endotoxin (lipopolysaccharide [LPS]) to induce modest acute inflammation. The expression of intestinal and hepatic heme oxygenase-1 at the mRNA and protein levels, circulating cytokines, as well as intestinal and mesenteric n-6 and n-3 fatty acid lipid mediators was compared 24 hours after LPS administration to examine its effects on the late phase of inflammation. Intestinal nuclear factor (erythroid-derived 2)-like 2 target enzyme heme oxygenase-1 was upregulated 8.42-fold at the mRNA level and 10.7-fold at the protein level by FA-supplemented diet. However, the FA-supplemented diet produced a unique type-2 plasma cytokine skew after LPS treatment. Plasma cytokines interleukin (IL)-4, IL-13, IL-9, and IL-10 were upregulated, reflecting the cytokine profile of reduced type 1 inflammation. A detailed lipidomic analysis of n-6 and n-3 fatty acid pro- and anti-inflammatory pathways in the mesentery and intestinal mucosa showed that FA diet was more similar to the control groups than to other LPS treated groups. In this study, we demonstrated that FA-supplemented diet produced a unique immunomodulatory effect not observed with sulforaphane in late phases of inflammation. These results support the hypothesis that FA may have role as a dietary immunosuppressant in patients with inflammatory gastrointestinal as well as other inflammatory disorders that may be alleviated by increasing consumption of carrot or other FA-containing food sources., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Cerebral choline metabolism is crucial for normal brain function, and its homoeostasis depends on carrier-mediated transport. Here, we report on four individuals from three families with neurodegenerative disease and homozygous frameshift mutations (Asp517Metfs*19, Ser126Metfs*8, and Lys90Metfs*18) in the SLC44A1 gene encoding choline transporter-like protein 1. Clinical features included progressive ataxia, tremor, cognitive decline, dysphagia, optic atrophy, dysarthria, as well as urinary and bowel incontinence. Brain MRI demonstrated cerebellar atrophy and leukoencephalopathy. Moreover, low signal intensity in globus pallidus with hyperintensive streaking and low signal intensity in substantia nigra were seen in two individuals. The Asp517Metfs*19 and Ser126Metfs*8 fibroblasts were structurally and functionally indistinguishable. The most prominent ultrastructural changes of the mutant fibroblasts were reduced presence of free ribosomes, the appearance of elongated endoplasmic reticulum and strikingly increased number of mitochondria and small vesicles. When chronically treated with choline, those characteristics disappeared and mutant ultrastructure resembled healthy control cells. Functional analysis revealed diminished choline transport yet the membrane phosphatidylcholine content remained unchanged. As part of the mechanism to preserve choline and phosphatidylcholine, choline transporter deficiency was implicated in impaired membrane homeostasis of other phospholipids. Choline treatments could restore the membrane lipids, repair cellular organelles and protect mutant cells from acute iron overload. In conclusion, we describe a novel childhood-onset neurometabolic disease caused by choline transporter deficiency with autosomal recessive inheritance., (© The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: The aim of this study was to investigate the effects of variety (Nantes and Imperator), soil fertility status (high and low) and agronomic treatments on yield and quality traits of carrot composition and sensory factors. The treatments compared synthetic nitrogen at conventionally recommended amounts with compost-sourced nitrogen (high and low rates) and a range of amendments (compost, compost tea, micronutrients and foliar treatments). Additionally, we intended to identify factors affecting polyacetylene accumulation in carrots, owing to the growing interest in their health effects and paucity of agronomic information on their bioaccumulation in carrots., Results: Imperator accumulated more minerals, produced more phytochemicals and had higher antioxidant capacity than Nantes, which had more carotenoids. However, consumers preferred the flavor of Nantes over Imperator. High-fertility soil produced carrots of superior nutritional quality than did low-fertility soil, regardless of year-of-application amendments, the effects of which were often variety-dependent. High soil biological activity was able to overcome low fertility status and stimulate greater yield. Carrot phosphorus was correlated with falcarindiol biosynthesis. Chlorogenic acid and falcarindiol were correlated with antioxidant capacity, but not falcarinol or total phenolic compounds., Conclusion: Carrots were not strongly affected by agronomic treatments in terms of yield or phytochemical status; however, soil biological activity overcame a soil nutrient deficit in terms of yield. Phenolic compounds and antioxidant status were generally higher in the high-fertility site, whereas polyacetylenes were variety-dependent. Chlorogenic acid and falcarindiol were associated with antioxidant capacity, but not total phenolic compounds and carotenoids. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Phosphatidylcholine (PC) and phosphatidylethanolamine (PE), which make up the bulk of mammalian cell membrane phospholipids, are recognized for their importance in metabolic health. Perturbations in the ratio of PC:PE can affect membrane integrity and function, which thus have serious health consequences. Imbalance in the hepatic PC and PE membrane content can be linked to metabolic disturbances such as ER stress, fatty liver and insulin resistance. Given that impaired insulin sensitivity underlies the pathology of many metabolic disorders and skeletal muscle is a significant regulator of energy metabolism, it is likely that aberrant phospholipid metabolism in skeletal muscle affects whole-body insulin sensitivity. Sarco/endoplasmic reticulum Ca 2+ ATPase (SERCA) activity and mitochondrial function respond to alterations in PC:PE ratio and are associated with glucose homeostasis. Moreover, PC and PE content within the mitochondrial membrane influence mitochondrial respiration and biogenesis and thus, metabolic function. As skeletal muscle phospholipids respond to stimuli such as diet and exercise, understanding the implications of imbalances in PC:PE ratio is of great importance in the face of the rising epidemic of obesity related diseases. This review will summarize the current state of knowledge signifying the links between skeletal muscle PC:PE ratio and insulin sensitivity with respects to PC and PE metabolism, SERCA activity, mitochondrial function and exercise.

Impact Statement: This review will provide a summary of recent advances in choline transport research and highlight important novel areas of focus in the field.

Objective: The objective of our study was to determine the prevalence of alcohol and drug intoxication among fatally injured motorcyclists in a wide urban area of Zagreb, Croatia. Methods: We conducted a single-center observational retrospective study over a 10-year period (2007-2016) in 3 counties covering an area including 1.2 million residents. We reviewed the records on fatally injured motorcyclists, collecting information relating to sex, age, cause of death, time of death in relation to the time of the crash, and the circumstances of the crash (time of day, day of the week, season). Blood alcohol concentration (BAC) and toxicology analysis results were collected and analyzed. Results: We identified 163 deaths (95.7% males, 4.3% females). Overall, 64.2% of the victims were 20 to 39 years old. The majority (50.9%) of those fatally injured were responsible for causing the traffic crash; the rest were determined not to be responsible or the responsibility could not be determined. The most frequent causes of death were multiple injuries (55.8%) and isolated head trauma (23.3%). The rider's BAC was above the legal limit for driving (>0.50 g/kg) in 53.8% of cases, with a mean BAC of 1.91 g/kg. There was no difference in riding a motorcycle with a BAC above the legal limit between groups defined as younger (≤39 years of age) and older (≥40 years of age). The number of people with an illegal BAC was significantly higher during weekends than during the work week. The BAC of riders who were responsible for the crash was significantly higher than that of those who were not responsible or whose responsibility could not be determined. Use of illegal drugs or nontherapeutic use of legal drugs was not common and was detected in 10.4% of fatally injured riders. Conclusions: Alcohol intoxication has a major role in motorcycle crash-related mortality. A significant difference in BAC between fatally injured riders responsible for the accident and those who were not responsible implies that measures directed toward prevention of drinking and driving behavior could lower the number of fatal motorcycle crashes. Weekend measures, especially during spring and summer, could have particularly significant effects.

Studies over the past two decades have shown heart regeneration in zebrafish to be a dynamic process, choreographed by multiple cell types. In particular, recent work has identified revascularization of the wound to be a sentinel event during heart regeneration. The cardiac endothelium has emerged as a key orchestrator of heart regeneration, influencing cardiomyocyte hyperplasia and tissue morphogenesis. Here, we review how the coronary vasculature regenerates after injury, how signaling pathways link the cardiac endothelium to heart regeneration, and how understanding these signaling dynamics can lead to targeted therapies for heart regeneration.

Nuclear factor- (erythroid-derived 2) like 2 (Nrf2) is a transcription factor that regulates the expression of a battery of antioxidant, anti-inflammatory, and cytoprotective enzymes including heme oxygenase-1 (Hmox1, Ho-1) and NADPH:quinone oxidoreductase-1 (Nqo1). The isothiocyanate sulforaphane (SF) is widely understood to be the most effective natural activator of the Nrf2 pathway. Falcarinol (FA) is a lesser studied natural compound abundant in medicinal plants as well as dietary plants from the Apiaceae family such as carrot. We evaluated the protective effects of FA and SF (5 mg/kg twice per day in CB57BL/6 mice) pretreatment for one week against acute intestinal and systemic inflammation. The phytochemical pretreatment effectively reduced the magnitude of intestinal proinflammatory gene expression (IL-6, Tnf α /Tnf α r, Inf γ , STAT3, and IL-10/IL-10r) with FA showing more potency than SF. FA was also more effective in upregulating Ho-1 at mRNA and protein levels in both the mouse liver and the intestine. FA but not SF attenuated plasma chemokine eotaxin and white blood cell growth factor GM-CSF, which are involved in the recruitment and stabilization of first-responder immune cells. Phytochemicals generally did not attenuate plasma proinflammatory cytokines. Plasma and intestinal lipid peroxidation was also not significantly changed 4 h after LPS injection; however, FA did reduce basal lipid peroxidation in the mesentery. Both phytochemical pretreatments protected against LPS-induced reduction in intestinal barrier integrity, but FA additionally reduced inflammatory cell infiltration even below negative control.

We investigated the metabolic effects of betaine (Bet) supplementation on CTP:phosphoethanolamine cytidylyltransferase/Pcyt2 heterozygous mice (HET). HET received either no treatment or were allowed access to 1% Bet supplemented water for 8 weeks. As we previously showed with choline (Cho), Bet improved hypertriglyceridemia, and hepatic steatosis in HET. The protection from obesity associated with reduced hepatic steatosis and increased lipid breakdown in adipocytes was attributed to increased energy requirements for metabolism and elimination of supplemented Bet and Cho. 1 H-NMR-based profiling revealed metabolic changes caused by Bet and Cho supplementation. Cho increased the citric acid cycle intermediate succinic acid while reducing isoleucine, valine, threonine, and lysine. Bet increased α-ketoglutaric acid and did not stimulate catabolism of amino acids. Increased histidine and alanine are specific biomarkers for Bet treatment. Cho and Bet caused glycerol accumulation and reduced sarcosine, taurine, acetate, and β-hydroxybutyrate levels. These data provide new insights on how Cho and Bet supplementation can aid in treatment of obesity related disorders due to their positive effects on lipolysis, the citric acid cycle, and mitochondrial oxidative demethylation.

Aims: Circadian rhythms are important for healthy cardiovascular physiology and they are regulated by the molecular circadian mechanism. Previously, we showed that disruption of the circadian mechanism factor CLOCK in male ClockΔ19/Δ19 mice led to development of age-dependent cardiomyopathy. Here, we investigate the role of biological sex in protecting against heart disease in aging female ClockΔ19/Δ19 mice., Methods and Results: Female ClockΔ19/Δ19 mice are protected from the development of cardiomyopathy with age, as heart structure and function are similar to 18 months of age vs. female WT mice. We show that female ClockΔ19/Δ19 mice maintain normal glucose tolerance as compared with female WT. Tissue metabolic profiling revealed that aging female ClockΔ19/Δ19 mice maintain normal cardiac glucose uptake, whereas the male ClockΔ19/Δ19 mice have increased cardiac glucose uptake consistent with pathological remodelling. Shotgun lipidomics revealed differences in phospholipids that were sex and genotype specific, including cardiolipin CL76:11 that was increased and CL72:8 that was decreased in male ClockΔ19/Δ19 mice. Additionally, female ClockΔ19/Δ19 mice show increased activation of AKT signalling and preserved cytochrome c oxidase activity compared with male ClockΔ19/Δ19 mice, which can help to explain why they are protected from heart disease. To determine how this protection occurs in females even with the Clock mutation, we examined the effects of ovarian hormones. We show that ovarian hormones protect female ClockΔ19/Δ19 mice from heart disease as ovariectomized female ClockΔ19/Δ19 mice develop cardiac dilation, glucose intolerance and reduced cardiac cytochrome c oxidase; this phenotype is consistent with the age-dependent decline observed in male ClockΔ19/Δ19 mice., Conclusions: These data demonstrate that ovarian hormones protect female ClockΔ19/Δ19 mice from the development of age-dependent cardiomyopathy even though Clock function is disturbed. Understanding the interaction of biological sex and the circadian mechanism in cardiac growth, renewal and remodelling opens new doors for understanding and treating heart disease., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2017. For permissions please email: journals.permissions@oup.com.)

Changes in sensory and sympathetic innervation during diabetes mellitus (DM) can be a predictor of arrhythmias, silent myocardial ischemia, and chronic heart failure, but knowledge about these changes is still unsatisfactory. We analyzed whether prolonged DM induces changes in density of sensory and sympathetic nerve terminals of rat's heart and whether it contributes to cardiomyopathy during aging. DM was induced by i/p injecting 55 mg/kg streptozotocin to male Sprague-Dawley rats, while a control group received a citrate buffer. DM in the rats was validated by measuring blood glucose level. Animals were sacrificed after 2 weeks, 2 months, 6 months, and 12 months. Five areas of cardiac sections were analyzed. Antibodies raised against tyrosine hydroxylase (TH) and neurofilament 200 kDa (NF 200) were used to detect sympathetic and sensory fibers. TH immunoreactive fiber density increased in DM groups 2 weeks after induction, reaching a peek after 2 months, while in the later stages of DM (6 and 12 months), there was no significant difference compared to control. NF 200 immunoreactive fiber density increased 2 weeks after induction compared to control. There was no consistent pattern of change during the given period in both the DM or control groups. In the DM group, we found thickening of the left ventricle wall (P<.05) as the sign of cardiomyopathy. Our findings suggest that hyperglycemia as a hallmark of DM in early stages can lead to proliferation of sympathetic and sensory nerve terminals. This finding can contribute to a better understanding of the occurrence of arrhythmias and silent myocardial ischemia in DM., (Copyright © 2017 Elsevier Inc. All rights reserved.)