bluuebunny/crossref_metadata_2025 huggingface dataset:
Autoantibodies directed against apolipoprotein A-1 as a potential contributor to non-alcoholic fatty liver disease
Authors: Pagano S , Alfaiate D , Juillard C , Frias M , Magenta A , Martino F , Sciaqua A , Succurro E , Burger F , Brandt K J , Andreozzi F , Mach F , Perticone M , Vuilleumier N
Published: October 2021

Background

Non-Alcoholic Fatty Liver Disease (NAFLD) represents an increasing cause of liver disease worldwide. Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in these patients. Although NAFLD pathophysiology is not fully understood alterations in fat metabolism seem to play a role. Autoantibodies against apolipoprotein A-1 (anti-apoA-1 IgG) are a novel cardiovascular risk factor to which have been recently attributed a metabolic role in addition to a well-established macrophage-mediated inflammatory effect and have a function as a disruptor of the cholesterol pathway.

Purpose

This study aims at evaluating a possible role of anti-apoA-1 IgG in NAFLD.

Methods

Serum from 137 NAFLD patients were tested for anti-apoA-1 IgG prevalence. In vitro, SREBP1, SREBP2 expressions were assessed in the human hepatic cell line HepaRG by western blot analysis and bodipy staining was used to evaluate the lipid droplet content. Mescoscale technology platform was used to measure TNF-α, IL-6 and IL-8 cytokines/chemokines in HepaRG supernatants. Oil Red O staining was used to detect lipid accumulation in liver sections from ApoE−/− mice.

Results

Elevated anti-apoA-1 IgG seropositivity was found in patients with NAFLD (46%). In vitro, anti-apoA-1 IgG and not control IgG induced lipid accumulation in hepatic cells (5.9 vs 2.5, P=0.0008) and this lipid overload was associated with a high SREBP1 but not SREBP2 expression. Furthermore, anti-apoA-1 IgG and not control antibodies caused a significant large increase of the proinflammatory cytokines IL-6 (680 vs. 163 pg/mL, P=0.03) and TNF-α (391 vs 266 pg/mL, P=0.04) as well as of the chemokine IL-8 (174.1 vs. 72.6 ng/mL, P=0.03) detected in the hepatic cell supernatants. In vivo, anti-apoA-1 IgG and not control IgG also induced higher lipid accumulation in the livers of ApoE−/− mice (1.23 vs 0.53, P=0.03).

Conclusion

Anti-apoA-1 IgG are frequent in NAFLD, cause a strong inflammatory response and promote lipid accumulation through SREBP1 activation in human hepatic cells. We hypothesize that anti-apoA1 IgG may be a potential contributor in the development of NAFLD.

Funding Acknowledgement

Type of funding sources: Public hospital(s). Main funding source(s): Geneva University Hospital
Mid-term endothelial dysfunction post COVID-19
Authors: Lampsas S , Oikonomou E , Siasos G , Souvaliotis N , Goliopoulou A , Mistakidi C V , Theofilis P , Vogiatzi G , Kalogeras K , Katsianos E , Tousoulis D , Vavuranakis M
Published: October 2021

Introduction

Cardiovascular complications of Coronavirus disease (COVID-19), resulting from the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), have been documented. Endothelium-induced “cytokine storm” in critically ill COVID-19 patients is one of the leading causes of morbidity and mortality. Vascular endothelial damage caused by COVID-19 emphasizes the crucial role of endothelium in COVID-19 clinical impact.

Purpose

To examine the mid-term (1-month) impact of COVID-19 in endothelial function.

Methods

In this case control study, 20 consecutive patients who were hospitalized for COVID-19 either on Intensive Care Unit (ICU) or non-ICU were examined one month following hospital discharge. In the control group we recruited 12 consecutive subjects from the outpatient cardiology clinic. Demographic and clinical data were collected, and endothelial function was evaluated by brachial artery flow-mediated dilation (FMD).

Results

There was no difference in age between COVID-19 patients and control subjects (66±12 years vs. 71±5 years, p<0.17), in male sex (63% vs. 54%, p=0.66) in history of diabetes mellitus (27% vs. 36%, p=0.64), hypertension (36% vs. 54%, p=0.39), cardiovascular disease (27% vs.18%, p=0.61). From the COVID-19 subjects 65% were overweight or obese. During their hospitalization [3 ICU (15%)/17 non-ICU (85%), mean days: 17±6.7], 4 (20%) of COVID-19 patients developed ARDS, while single cases of stress-induced cardiomyopathy, pulmonary embolism, and acute coronary syndrome were detected. One month post discharge D-dimers (0.71±0.55 μg/ml) levels were above upper reference limit. Importantly, FMD one month after hospital discharge date, was significantly impaired in the COVID-19 group (3.59±1.63% vs. 9.31±2.98%, p<0.001) compared to control group.

Conclusion

Post COVID-19 subjects one month post discharge have significant impaired endothelial function compared to control subjects. These findings highlight the significant interaction of COVID-19 with arterial endothelium and merit further research to conclude on the exact impact of vascular endothelium in physical history of SARS-CoV-2 infection.

Funding Acknowledgement

Type of funding sources: None.
A multivariate analysis identifies genetic loci associated with atherosclerotic plaque composition and cardiovascular disease trajectory
Authors: Cui K , Mekke J , Haitjema S , Pasterkamp G , Asselbergs F W , Mokry M , Van Der Laan S W
Published: October 2021

Background

From cross-sectional studies we have learned that composition of atherosclerotic plaques differs between individuals, and this contributes to the inter-individual differences in susceptibility to incident coronary and cerebral events. In pathological studies the extent and type of atherosclerosis is commonly assessed based on histological plaque characteristics that are linked to plaque rupture and erosion. A better understanding of the biology underlying variability in plaque composition will provide insights into the progression of cardiovascular diseases.

Objectives

We investigated the genetics of the plaque through multivariate and integrative genome-wide analyses (GWAS) of individual plaque characteristics.

Methods

We included carotid endarterectomy patients from the Athero-Express Biobank Study (n=2,124) with high-density imputed data and extensive histochemical plaque phenotyping available. We used slideToolKit to quantify the number of endothelial cells, macrophages and smooth muscle cells (SMCs), and manually assessed the number of intraplaque vessels, the amount of collagen and calcification, the atheroma size, and the presence of plaque hemorrhage. We ran GWAS on all traits correcting for age, sex, array used, and genetic ancestry.

Results

We identified 3 loci that significantly associate with CD68+ macrophages and ACTA2+ SMCs, p<5x10–8. Statistical finemapping revealed 9 variants in the 95% credible set and functional annotation linked these to genes associated with malignant neoplasms, circulating cholesterol, and transmembrane proteins, suggesting an effect on cellular proliferation and cholesterol metabolism.

Conclusions

We provide evidence for 3 loci that modulate plaque composition through macrophages and smooth muscle cell plaque proliferation and cell-cell interactions.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): European Research Area Network on Cardiovascular Diseases (ERA-CVD, druggable-MI-genes),China Scholarship Council
Protective effects of dapagliflozin on vascular remodeling in the carotid artery following balloon injury – potential role of angiotensin and purinergic signaling
Authors: Matsushita K , Sato C , Bruckert C , Gong D , Hmadeh S , Fakih W , Remila L , Auger C , Jesel L , Ohlmann P , Kauffenstein G , Schini-Kerth V , Morel O
Published: October 2021

Introduction

Sodium-glucose co-transporter 2 (SGLT2) inhibitors have been shown to reduce the risk of cardiovascular events independently of glycemic control. The possibility that SGLT2 inhibitors improve endothelial regeneration and vascular restenosis is unknown.

Purpose

To examine whether dapagliflozin, a selective SGLT2 inhibitor, could prevent neointima thickening induced by balloon injury and, if so, to determine the underlying mechanisms. The effect of dapagliflozin was compared to that of losartan, an angiotensin type 1 receptor (AT1R) antagonist.

Methods

Saline, dapagliflozin (1.5 mg/kg/day), or losartan (30 mg/kg/day) were administered orally for 5 weeks to male Wistar rats. Balloon injury of the left carotid artery was performed 1 week after starting the treatment and sacrificed 4 weeks later. Vascular reactivity was assessed on left (injured) and right (healthy) carotid artery rings. The extent of neointima was assessed by histomorphometric analysis, changes of target factors by immunofluorescence, RT-qPCR and histochemistry.

Results

Dapagliflozin and losartan treatments reduced neointima thickening by 32% and 27%, respectively. Blunted contractile responses to phenylephrine and relaxations to acetylcholine and down-regulation of eNOS were observed in the injured artery. These effects were not modified by the dapagliflozin or the losartan treatments. RT-qPCR investigations indicated an increased in gene expression of inflammatory (IL-1beta, ITGAM, VCAM-1), oxidative (p47phox, p22phox) and fibrotic (TGF-beta1) markers and a decreased of eNOS in the injured carotid. However, these changes were not affected by the pharmacological treatments. By contrast, significant increased levels of AT1R angiotensin receptor and NTPDase1 (CD39) ectonucleotidase were observed in the restenotic carotid artery of the dapagliflozin group. Histochemical analysis evidenced important NTPDase1 activity in the neointima.

Conclusions

Dapagliflozin effectively reduced neointimal thickening. As the contribution of AT1R and P2Y2 ATP receptor in smooth muscle cell proliferation and neointima formation has been reported in the literature, the present data suggest that dapagliflozin prevents restenosis through interfering with angiotensin and/or extracellular nucleotides signaling. SGLT2 transporter represent potential new target for limiting vascular restenosis.

Funding Acknowledgement

Type of funding sources: Private company. Main funding source(s): This work was supported by AstraZeneca
PI 3-kinase isoform PI3Kalpha controls smooth muscle cell functionality and protects against aortic aneurysm formation
Authors: Vantler M , Schorscher M , Berghausen E , Zierden M , Wissmueller M , Mollenhauer M , Mehrkens D , Adam M , Baldus S , Rosenkranz S
Published: October 2021

Background

Class I PI 3-kinase isoform PI3Kα is a lipid kinase and signals downstream of receptor tyrosine kinases. Smooth muscle cells (SMCs) lacking PI3Kα are characterized by impaired proliferation, migration and survival. Mice, harbouring a smooth muscle specific PI3Kα deficiency (SM-PI3Kα−/−), display reduced vascular wall thickness and impaired vascular remodeling in response to vessel injury. We hypothesize that SM-PI3Kα−/− mice are prone to aortic aneurysm (AA) formation due to impaired SMC functions. Herein, we investigated, how PI3Kα-dependent signaling in SMCs affect aortic aneurysm (AA) formation, aortic wall structure, and expression of extracellular matrix (ECM) components.

Methods and results

AA formation in SM-PI3Kα−/− mice and wild-type littermates was examined by means of the “porcine pancreatic elastase” (PPE) AA model. PPE was infused into the infrarenal aorta to induce AA formation. Ultrasound examination revealed a significantly increased aortic diameter in SM-PI3Kα−/− mice (1.22±0.12 mm) compared to wild-type animals (0.96±0.02 mm, p=0.014). These data indicate a protective function of SM-PI3Kα in AA formation. In addition, the media thickness in the abdominal aorta was significantly reduced in SM-PI3Kα−/− mice (29.0±3.1 vs. 42.5±4.1 μm). Ultrastructural analysis of aortic wall morphology in SM-PI3Kα−/−mice using transmission electron microscopy (TEM) showed a deranged tunica media with detached SMCs and increased apoptotic cell death. Consequently, SM-PI3Kα deficiency significantly diminished responsiveness of aortic rings to vasodilator acetylcholine and NO-donor nitroglycerin, further indicating impaired aortic wall structure. Western blots demonstrated a reduced elastin and fibrillin expression in SMCs from SM-PI3Kα−/− mice. Furthermore, immunofluorescence stainings of PI3Kα−/− and wild-type SMCs, cultured for seven days under 10% fetal calf serum containing DMEM medium, showed significantly disturbed structures of elastin-, fibrillin-1- and collagen-1-fibers. These data indicate that PI3Kα signaling contributes to elastic fiber homeostasis thus affecting SMC phenotypic modulation. Immunoblots demonstrated that PDGF and insulin induced phosphorylation and inactivation of key regulators of SMC differentiation and dedifferentiation including FoxO1, FoxO3a, Foxo4, and GSK3b, respectively, were reduced or even abrogated in PI3Kα−/− SMCs.

Conclusion

These data show that deficiency of PI3Kα in SMCs promotes the formation and progression of AA. Causative is a deranged aortic structure of SM-PI3Kα−/− aortae which can likely be attributed to an impaired production of elastic fiber components by PI3Kα−/− SMCs.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft
The m6A methyltransferase METTL3 promotes hypoxic pulmonary arterial hypertension via targeting PTEN
Authors: Yuhan Q I N , Chengchun T A N G
Published: October 2021

Background

N6-methyladenosine (m6A) is the most prevalent internal RNA modification in mammal mRNAs. Accumulating evidence has indicated the crucial role of m6A modification in cardiovascular diseases including cardiac hypertrophy, heart failure, ischemic heart disease, vascular calcification, restenosis, and aortic aneurysm. However, the role of m6A methylation in the occurrence and development of hypoxic pulmonary hypertension (HPH) remains largely unknown.

Purpose

The present study aims to explore the role of key transferase METTL3, in the development of HPH.

Methods

Hypoxic rat models and pulmonary artery smooth muscle cells (PASMCs) and were used to research the METTL3-mediated m6A in HPH in vivo and in vitro. CCK-8, EdU, PCNA, transwell and TUNEL assay were performed to evaluate the proliferation, migration and apoptosis rates of PASMCs. m6A RNA Methylation Quantification Kit and m6A-qPCR were utilized to measure the total m6A level and m6A-PTEN mRNA expression. RNA immunoprecipitation and RNA pull down were used to detect the interaction between METTL3 and PTEN mRNA. The half-life of mRNA was detected through actinomycin D assay.

Results

Both METTL3 mRNA and protein were found abnormally upregulated in pulmonary arteries of HPH rats and hypoxia induced PASMCs. Furthermore, downregulation of METTL3 attenuated PASMCs proliferation and migration exposed to hypoxia. In addition, m6A binding protein YTHDF2 was found significantly increased in HPH group in vivo and in vitro. Mechanistically, YTHDF2 recognized METTL3 mediated m6A-PTEN mRNA and promoted the degradation of PTEN. Decreased PTEN led to over-proliferation of PASMCs through activation of PI3K/Akt signaling pathway.

Conclusion

METTL3/YTHDF2/PTEN axis exerts a significant role in hypoxia induced PASMCs proliferation, providing a novel therapeutic target for HPH.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): National Natural Science Foundation of China Figure 1
Parathyroid hormone affects right heart hemodynamics and exacerbates pulmonary hypertension
Authors: Joki Y J , Konishi H K , Takasu K T , Minamino T M
Published: October 2021

Background

Pulmonary hypertension (PH) is characterized by increased pulmonary artery pressure and develops right heart failure. Parathyroid hormone (PTH) is secreted from parathyroid gland and regulates a calcium homeostasis. Recent studies have suggested that PTH also acts on the cardiovascular system and affects cardiovascular prognosis. We hypothesized that PTH would play a role in the pathogenesis of PH.

Purpose

This study aimed to investigate the effect of PTH on pulmonary hemodynamics.

Method

We measured serum PTH levels in patients who were suspected of PH and underwent right heart catheter examination. We used two types of PH animal models, hypoxia (Hx)-induced PH mouse model and Sugen/hypoxia (SuHx)-induced PH rat model. Hx mice were administered PTH daily for 3 weeks. SuHx rats underwent parathyroidectomy, after which they received SuHx treatment for 10weeks. We measured physical data and right ventricular systolic pressure (RVSP) in these models. We cultured pulmonary artery smooth muscle cell (PASMC) treated with PTH to analyze cell signaling, proliferation and migration.

Result

We enrolled 20 participants. PTH concentration was significantly correlated with mean pulmonary artery pressure (r=0.58, p=0.006) as well as with pulmonary vascular resistance (r=0.61, p=0.04). Receiver operating characteristic curve displayed a cut-off PTH level of 48.0pg/ml that offered optimal differentiation between patients with and without PH (100% sensitivity, 73% specificity). PTH treatment exacerbated right ventricular hypertrophy and increased RVSP (33.6mmHg vs. 48.2mmHg) in Hx mice compared with non-treated Hx mice (Figure 1). Conversely, parathyroidectomy significantly attenuated right ventricular hypertrophy and reduced RVSP (54.2mmHg vs. 29.3mmHg) in SuHx rats compared with sham-operated SuHx rats. PTH promoted migration and proliferation through ERK signaling in PASMC.

Conclusion

Our clinical and experimental data demonstrated a critical role of PTH in the development of PH and suggested that PTH would be a novel therapeutic target for PH treatment.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Grant-in-Aid for Young Scientists Figure 1. PTH treatment exacerbated RVSP
Senescent endothelial cells exacerbate pulmonary hypertension through notch-mediated juxtacrine interaction with pulmonary artery smooth muscle cells
Authors: Ramadhiani R , Ikeda K , Miyagawa K , Ryanto G R T , Tamada N , Suzuki Y , Hirata K , Emoto N
Published: October 2021

Background

Despite recently developed clinical therapies, vascular remodelling in pulmonary arterial hypertension (PAH) progressively worsen. Hemodynamic unloading has been proposed to normalize the remodelled pulmonary vascular structures in the lungs. Recently, it has been reported that cellular senescence was associated with the irreversibility of pulmonary vascular structures after hemodynamic unloading.

Purpose

This study aims to elucidate the role of senescent endothelial cells (ECs) in the pathogenesis of PAH.

Methods

We generated EC-specific progeroid mice in which ECs undergo premature senescence by overexpressing the dominant-negative form of telomere repeat-binding factor 2 under the control of the VE-cadherin promoter. Following three weeks of hypoxia exposure, the PH phenotypes were assessed by RVSP, lung histology, and RT-qPCR. The interaction of human pulmonary artery ECs (hPAECs) and human pulmonary artery smooth muscle cells (hPASMCs) was indirectly and directly explored through the co-culture system. Gamma-secretase inhibitor (DAPT) was administrated to inhibit Notch signalling both in the in-vitro and in-vivo study.

Results

EC-specific progeroid mice showed exacerbated pulmonary hypertension after chronic hypoxia exposure, accompanied by the enhanced medial SMCs proliferation in the distal pulmonary arteries. Contact-mediated interaction with senescent hPAECs increased proliferation and migration capacities in hPASMCs, while no such effects were detected in the absence of ECs-SMCs contact. Consistently, senescent ECs highly expressed Notch ligands, thus activated Notch signalling in hPASMCs, leading to increased Notch target genes in hPASMCs. Pharmacological inhibition of Notch signalling attenuated the enhanced SMCs proliferation and migration induced by senescent hPAECs, as well as the worsened PH phenotypes in EC-specific progeroid mice.

Conclusions

Our data established a crucial role of senescent ECs in the PAH pathogenesis through the dysregulated SMC functions via juxtacrine signaling. Senescent ECs are attracting targets for further pathological-targeted therapy to cure PAH completely.

Funding Acknowledgement

Type of funding sources: None.
Efficacy and safety of direct oral anticoagulants in morbidly obese patients with non-valvular atrial fibrillation: a systematic review and meta-analysis
Authors: Mhanna M , Beran A , Al-Abdouh A , Srour O , Abdulsattar W , Srour O , Altujjar M , Alom M , Abumoawad A , Assaly R
Published: October 2021

Introduction

Atrial fibrillation (AF) is the most common arrhythmia, with an estimated prevalence between 1–4%. On the other hand, obesity continued to be a prevalent health issue worldwide. Direct oral anticoagulants (DOACs) have been increasingly preferred over warfarin; however, The International Society of Thrombosis and Hemostasis (ISTH) recommended avoiding the use of DOACs in patients with a BMI >40 or weight >120 kg because of limited clinical data in these patients. In this meta-analysis, we aimed to evaluate the efficacy and safety of DOACs in morbidly obese patients with non-valvular AF.

Method

We performed a comprehensive literature search using multiple databases from database inception through January 2021, for all the studies that evaluated the efficacy and safety of DOACs in morbidly obese patients with non-valvular AF. The primary outcome of interest was stroke or systemic embolism (SSE) rate. The secondary outcome was major bleeding (MB). All meta-analyses were conducted using a random-effect model.

Results

A total of 10 studies including 89,494 morbidly obese patients (BMI >40 or weight >120 kg) with non-valvular AF on oral anticoagulation therapy (45427 on DOACs vs. 44067 on warfarin) were included in the final analysis. One included study was a randomized controlled trial (RCT), another study was a post hoc analysis of an RCT and the rest were retrospective cohort studies. The mean follow-up period was 1.8 years (range 8 months to 3.1 years). The SSE rate was significantly lower in DOACs group compared to warfarin group (odds ratio (OR): 0.71; 95% confidence interval (CI): 0.62, 0.81; p<0.0001; I2=0%). MB rate was also significantly lower in DOACs group compared to the warfarin group (OR 0.60, 95% CI 0.46–0.78, P<0.0001, I2=86%). Subgroup analysis in the rivaroxaban and apixaban AF cohort showed a statistically significant difference in SSE and MB event rates favoring both over warfarin therapy. Dabigatran showed non-inferiority to warfarin in SSE rate but superiority in the safety outcome.

Conclusions

Our meta-analysis demonstrated that DOACs are effective and safe when compared to warfarin in morbidly obese patients. However, more large scale randomized clinical trials are needed to further evaluate the efficacy and safety of DOACs compared to warfarin in this cohort of patients.

Funding Acknowledgement

Type of funding sources: None. Stroke and systemic embolism eventsMajor bleeding events
URAT1-selective inhibition ameliorates insulin resistance by attenuating diet-induced hepatic steatosis and BAT whitening in mice
Authors: Tanaka Y , Nagoshi T , Yoshii A , Oi Y , Takahashi H , Kimura H , Kashiwagi Y , Tanaka T D , Yoshimura M
Published: October 2021

Background

Accumulating evidence suggests that high uric acid is strongly associated with obesity and metabolic syndrome and drives the development of non-alcoholic fatty liver disease (NAFLD) and insulin resistance. Although urate transporter-1 (URAT1), which is primarily expressed in the kidney, plays a critical role in the development of hyperuricemia, its pathophysiological implication in NAFLD and insulin resistance remains unclear.

Objectives

We hypothesizes that URAT1 plays an important role in obesity-induced metabolic disorders, and URAT1-selective inhibitor treatment ameliorates systemic insulin resistance, NAFLD and adipose tissue dysfunction using diet-induced obese mice.

Methods

Mice fed a high-fat diet (HFD) for 16 to 18 weeks or a normal-fat diet (NFD) were treated with or without a novel oral URAT1-selective inhibitor (dotinurad [50 mg/kg/day]) for another 4 weeks.

Results

Dotinurad administration significantly ameliorated HFD-induced obesity and insulin resistance. We found that URAT1 was also expressed in the liver and brown adipose tissue (BAT) other than kidney. HFD markedly induced NAFLD, which was characterized by severe hepatic steatosis, as well as the elevation of serum ALT activity and tissue inflammatory cytokine genes (Ccl2 and TNFα), all of which were attenuated by dotinurad. Likewise, HFD significantly increased URAT1 expression in BAT, resulting in the lipid accumulation (whitening of BAT) and increased production of tissue reactive oxygen species, which were reduced by dotinurad via UCP1 activation.

Conclusions

A novel URAT1-selective inhibitor, dotinurad, ameliorates insulin resistance by attenuating hepatic steatosis and promoting rebrowning of lipid-rich BAT in HFD-induced obese mice. URAT1 serves as a key regulator of the pathophysiology of metabolic syndrome, and may be a new therapeutic target for insulin-resistant individuals, particularly those with concomitant NAFLD.

Funding Acknowledgement

Type of funding sources: None.
GPNMB protects against obesity-related metabolic disorders by attenuating macrophage-derived inflammation in white adipose tissue through CD44 receptor-dependent mechanism
Authors: Prabata A , Ikeda K , Hirata K , Emoto N
Published: October 2021

Introduction

Obesity is a worldwide health issue frequently linked to cardiovascular disease. Obesity is closely associated with chronic low-grade inflammation in adipose tissue caused by macrophage infiltration and activation, which leads to metabolic disorders. We discovered glycoprotein non-metastatic melanoma protein B (GPNMB) as a gene that is robustly expressed in white adipose tissue of obese mice.

Purpose

We aim to elucidate the role of GPNMB in obesity and its related metabolic disorders.

Methods

We generated GPNMB-knockout (KO) mice and assessed body weight, insulin sensitivity, and inflammation-related phenotypes after 12 weeks of high fat-diet (HFD) feeding. For in vitro experiment, we isolated peritoneal macrophages from wild-type (WT) and GPNMB-KO mice to explore their inflammatory phenotypes.

Results

After HFD feeding, obesity-related metabolic disorders were markedly worsened in GPNMB-KO mice, assessed by insulin and glucose tolerance test, although the weight gain and adiposity were similar between WT and GPNMB-KO mice. Expression of inflammatory cytokines was significantly increased in WAT of GPNMB-KO mice, in addition to a remarkable increase in crown-like structures. Peritoneal macrophages isolated from GPNMB-KO mice exhibited higher inflammatory cytokine levels, and significantly impaired the adipocytes insulin sensitivity in an endocrine manner. Recombinant soluble GPNMB attenuated the inflammatory capabilities in peritoneal macrophages isolated from GPNMB-KO mice. We identified that GPNMB inhibits the NF-κB signaling through binding to the CD44 receptor, and therefore negatively regulates inflammatory responses in macrophages.

Conclusion

Our study demonstrates GPNMB as an adipokine that protects against obesity-related metabolic disorders by reducing macrophage-derived inflammation, which in turn improves adipocytes functions and consequently preserves systemic metabolic health. Therefore, GPNMB is an attractive therapeutic target for the treatment of obesity-related metabolic disorders.

Funding Acknowledgement

Type of funding sources: None.
Functional and transcriptional differences in monocytes from children with obesity compared to children of healthy weight
Authors: Bekkering S , Saner C , Novakovic B , McCallum Z , Netea M G , Riksen N P , Sabin M A , Saffery R , Burgner D P
Published: October 2021

Background and aim

Cardiometabolic risk accrues across the entire life course and childhood is a key epoch for effective prevention. Obesity in childhood is the most prevalent modifiable risk factor for later cardiovascular disease (CVD). Inflammatory biomarkers and innate immune capacity are increased in adults with obesity, but childhood data are scarce. We aimed to investigate (i) innate immune cell activation in children with and without obesity; and (ii) whether weight loss impacts the innate immune inflammatory phenotype.

Methods

The innate immune phenotype of Peripheral Blood Mononuclear Cells (PBMCs) from 31 children with obesity (BMI z-score>2.5) and 22 children of healthy weight (−1.5≤BMIz≤1.5, sex, age and pubertal stage matched) was characterized by high dimensional flow cytometry, ex vivo stimulation assays with subsequent 27-plex cytokine measurements, and transcriptome analysis using RNA sequencing (Figure 1). Children with obesity participated to the Royal Children's Hospital Weight Management Service (median 5 years) and at follow-up, PBMCs were obtained again as well as anthropometric data and subclinical cardiovascular phenotypes.

Results

Flow cytometric analysis showed marked differences in cell composition between children with obesity and children of healthy weight. Specifically, children with obesity have significant changes in monocyte subsets and an increased expression of monocyte activation markers. Upon stimulation, monocytes of children with obesity show an increased cytokine production capacity. Finally, transcriptomic analysis shows significant differences between monocytes from obese children and healthy controls. Effects of weight loss on these immune parameters and correlations with preclinical CVD phenotypes are currently being analysed.

Conclusions

Monocytes from children with obesity have a pro-inflammatory phenotype compared to children of normal weight. Heightened inflammation may contribute to increased CVD risk later in life and may offer opportunities for early intervention.

Funding Acknowledgement

Type of funding sources: Other. Main funding source(s): Dutch Scientific Organisation (NWO) - Rubicon grant to S.B. Dutch Heart Foundation - CVON IN CONTROL II to N.P.R. and D.B. Figure 1. Schematical overview of study
Wastewater surveillance of SARS-CoV-2 corroborates heightened community infection during the initial peak of COVID-19 in Bexar County, Texas
Authors: Al-Duroobi Haya , Moghadam Sina V , Phan Duc C , Jafarzadeh Arash , Matta Akanksha , Kapoor Vikram
Published: October 2021

The purpose of this study was to conduct a preliminary assessment of the levels of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in wastewater at the Salitrillo Wastewater Treatment Plant in Texas during the initial peak of coronavirus disease 2019 (COVID-19) outbreak. Raw wastewater influent (24 h composite, time-based 1 L samples, n = 13) was collected weekly during June–August 2020. We measured SARS-CoV-2 RNA in wastewater by reverse transcription droplet digital PCR using the same N1 and N2 primer sets as employed in COVID-19 clinical testing. Virus RNA copies for positive samples (77%) ranged from 1.4 × 102 to 4.1 × 104 copies per liter of wastewater, and exhibited both increasing and decreasing trends, which corresponded well with the COVID-19 weekly infection rate (N1: ρ = 0.558, P = 0.048; N2: ρ = 0.487, P = 0.092). A sharp increase in virus RNA concentrations was observed during July sampling dates, consistent with the highest number of COVID-19 cases reported. This could be attributed to an increase in the spread of COVID-19 infection due to the Fourth of July holiday week gatherings (outdoor gatherings were limited to 100 people during that time). Our data show that wastewater surveillance is an effective tool to determine trends in infectious disease prevalence, and provide complementary information to clinical testing.
De novowhole-genome assembly ofChrysanthemum makinoi, a key wild chrysanthemum
Authors: van Lieshout Natascha , van Kaauwen Martijn , Kodde Linda , Arens Paul , Smulders Marinus J M , Visser Richard G F , Finkers Richard
Published: October 2021

Chrysanthemum is among the top 10 cut, potted, and perennial garden flowers in the world. Despite this, to date, only the genomes of two wild diploid chrysanthemums have been sequenced and assembled. Here, we present the most complete and contiguous chrysanthemum de novo assembly published so far, as well as a corresponding ab initio annotation. The cultivated hexaploid varieties are thought to originate from a hybrid of wild chrysanthemums, among which the diploid Chrysanthemum makinoi has been mentioned. Using a combination of Oxford Nanopore long reads, Pacific Biosciences long reads, Illumina short reads, Dovetail sequences, and a genetic map, we assembled 3.1 Gb of its sequence into nine pseudochromosomes, with an N50 of 330 Mb and a BUSCO complete score of 92.1%. Our ab initio annotation pipeline predicted 95,074 genes and marked 80.0% of the genome as repetitive. This genome assembly of C. makinoi provides an important step forward in understanding the chrysanthemum genome, evolution, and history.
Gender differences in informal payments for healthcare: evidence from 34 African countries
Authors: Rajan Selina , Santoso Cornelia , Abba-Aji Mohammed , Stuckler David , McKee Martin , Hutchinson Eleanor , Onwujekwe Obinna , Balabanova Dina
Published: October 2021

Informal payments are widespread in many healthcare systems and can impede access to healthcare and thwart progress to achieving universal health coverage, a major element of the health-related Sustainable Development Goals. Gender may be an important driver in determining who pays informally for care, but few studies have examined this, particularly in low- and middle-income countries. Our study aimed to examine gender disparities in paying informally for healthcare in Africa. We used Afrobarometer Round 7 survey data collected between September 2016 and August 2018 from 34 African countries. The final sample was composed of 44 715 adults. We used multiple logistic regression to evaluate associations between gender and paying informally to obtain healthcare. Our results show that 12% of women and 14% of men reported paying informally for healthcare. Men were more likely to pay informally for healthcare than women in African countries [odds ratio 1.22 (95% confidence interval 1.13–1.31)], irrespective of age, residential location, educational attainment, employment status, occupation and indicators of poverty. To make meaningful progress towards improving universal healthcare coverage in African countries, we must improve our understanding of the gendered aspects of informal payments in healthcare, which can act as both a barrier to accessing care and a determinant of poor health.
Successful use of bio plugs for delayed bronchial closure after pneumonectomy in experimental settings
Authors: Moriyama Masaaki , Matsumoto Keitaro , Taniguchi Daisuke , Machino Ryusuke , Tsuchiya Tomoshi , Nakayama Koichi , Nagayasu Takeshi
Published: October 2021

OBJECTIVES

Cell therapies, such as stem cell suspension injection, are used to treat bronchopleural fistula. Although it is safe and effective, injected cells cannot remain within the bronchioles of the fistula due to cell leakage into the thoracic cavity. Here, we inserted a ‘bio plug’ into the fistula, produced using cells and a bio-3D printer, to examine the effectiveness of bio plugs for the closure of bronchopleural fistulas, the optimal cell source and the closure mechanism.

METHODS

Bio plugs were made with mesenchymal stem (stromal) cells derived from bone marrow (MSCBM), fibroblasts and rat lung micro-vessel endothelial cells using a bio-3D printer with different cell mixing ratios. Six groups, according to the presence or absence and the type of bio plugs, were compared. The plugs were inserted into the bronchi of F344 rats. The obstruction ratio and histological and immunohistochemical findings were evaluated.

RESULTS

MSCBM+ rat lung micro-vessel endothelial cell group exhibited a higher obstruction ratio among all groups excluding the MSCBM group (P = 0.039). This group had fibrosis and CD31-positive cells and fewer CD68-positive cells than MSCBM and MSCBM+ fibroblast groups.

CONCLUSIONS

Bio plugs with mixed cells, including stem cells, contribute to bronchial closure in the current experimental setting. Endothelial cells effectively maintain the structure in this model. Although bronchial closure for bronchopleural fistula could not be described as clinical conditions were not reproduced, we collected essential data on bronchial closure; however, further experiments are warranted.
Combination of Clofazimine and Atovaquone as a Potent Therapeutic Regimen for the Radical Cure of Babesia microti Infection in Immunocompromised Hosts
Authors: Tuvshintulga Bumduuren , Sivakumar Thillaiampalam , Nugraha Arifin Budiman , Ahedor Believe , Batmagnai Enkhbaatar , Otgonsuren Davaajav , Liu MingMing , Xuan Xuenan , Igarashi Ikuo , Yokoyama Naoaki
Published: October 2021

Human babesiosis caused by Babesia microti can be fatal in immunocompromised patients, and the currently used drugs are often ineffective. A recent study found that clofazimine clears B. microti Munich strain in immunocompromised mice. In the present study, we investigated the efficacies of clofazimine and 2-drug combinations involving clofazimine, atovaquone, and azithromycin against B. microti Peabody mjr strain in immunocompromised mice. Treatment with clofazimine alone, clofazimine plus azithromycin, and atovaquone plus azithromycin was ineffective and failed to eliminate the parasites completely, while a 44-day treatment with clofazimine plus atovaquone was highly effective and resulted in a radical cure.
PSVI-12 The effects of road transportation with or without homeopathic remedy supplementation on growth performance, nutrient digestibility, microbiota, serum cortisol and superoxide dismutase in growing pigs
Authors: Deokho Koo , De Xin Dang , Thanapal Palanisamy , Ju Park Hyun , Je Min Ahn , Inho Kim
Published: October 2021

This study investigated the effects of road transportation with or without homeopathic remedy (Convermax) supplementation on growth performance, nutrient digestibility, fecal microbiota, and serum cortisol and superoxide dismutase (SOD) concentrations in growing pigs. A total of 180 49-day old growing pigs [(Yorkshire × Landrace) × Duroc] with an initial body weight of 13.17 ± 0.02 kg were randomly allotted to 2 groups based on the initial body weight, containing 18 replicates with 5 pigs (mixed sex) in each. The pigs were fed dietary supplementation of a homeopathic remedy (Convermax) (0 or 200 mg/kg of feed, as-fed) for 35 days. On day 21, forty-five pigs (70-day old; 25.25 ± 0.37 kg) were randomly selected from each group and assigned to either 2 h of road transportation or no road transportation, resulting in a 2×2 factorial design. Results showed that road transportation led to an increase in the fecal coliform bacteria counts (P = 0.023) and serum cortisol concentration (P = 0.039) and a decrease in the serum superoxide dismutase (SOD) concentration (P < 0.001). However, supplementing homeopathic remedy (Convermax) to the diet of growing pigs increased gain to feed ratio (P = 0.042), apparent nitrogen digestibility (P = 0.019), and serum SOD concentration (P = 0.007), whereas decreased serum cortisol concentration (P = 0.022). In brief, road transportation induced stress response for growing pigs and increased harmful bacteria counts in their intestines. Dietary supplementation of homeopathic remedy (Convermax) alleviated stress response, improved apparent nitrogen digestibility, and increased gain to feed ratio. However, no significant interactive effects between road transportation with or without dietary homeopathic remedy (Convermax) levels were observed on the detected parameters in growing pigs.
PSVII-3 Evaluation of dietary OmniGen AF supplementation and PG600 injection on reproductive characteristics of gilts
Authors: Chen Jun , Lee Jung Wook , Monegue Jim J , Niblett Richard , Williams Kim , Estienne Mark J , Chapman Jim , Humphrey Brooke , Lindemann Merlin D
Published: October 2021

A cooperative study evaluated effects of supplemental dietary OmniGen AF (OG; Phibro Animal Health Corporation) on reproductive characteristics of gilts. Crossbred gilts (n = 56; initial body weight = 152.3 ± 9.7 kg) were assigned to treatments in a 2 x 2 factorial design [diet (OG or control) and PG600 (Merck Animal Health, El Dora, KS) injection (Yes or No)]. Dietary treatments were: 1) corn-SBM basal diet (control) that met NRC (2012) requirement estimates, or 2) control plus 0.75% OG and were fed 35 to 40 d prior to breeding. A common diet formulation was used by both stations. Gilts were estrous-synchronized by 14-d feeding of Matrix (Merck Animal Health). PG600 injections occurred 24 hours after Matrix withdrawal. Gilts were weighed at breeding and preslaughter (at d 39 to 48 postbreeding) and were euthanized to evaluate reproductive characteristics. There were no OG x PG600 interactions on any response measures (P > 0.15). Mean responses did not differ (P > 0.20) when gilts were fed control or OG diets, respectively, for total uterus weight (6,065 vs. 5,883 g), uterine horn length (129.2 vs. 123.9 cm), ovary weight (9.66 vs. 9.94 g), total corpora lutea (CL) number (25.0 vs. 25.3), individual CL weight (0.41 vs. 0.43 g), total follicular fluid weight (3.17 vs. 3.03 g), individual placenta weight (86.3 vs. 86.4 g), total fetuses (14.2 vs. 13.0), and fetus crown-rump length (7.97 vs. 7.66 cm). Supplementation with OG reduced (P = 0.04) individual placenta fluid volume of gilts (150.6 vs. 115.4 mL). PG600 injection increased (P < 0.001) ovary weight (9.03 vs. 10.57 g), total CL (18.09 vs 32.27) and reduced CL weight (0.46 vs. 0.38 g). The results demonstrated classic PG600 responses on reproductive characteristics but no effects of 0.75% OG supplementation nor an interaction between the two factors.
PSIX-9 Effect of calcium nitrate fed to dairy cows on nitrate residues, fatty acids profile, and antioxidant capacity in milk
Authors: Almeida Kleves V , Osorio Jesus A C , Marchi Francilaine E , Silva Junior Ranulfo C , Sippert Micheli , Pinto Regina C C , Tuzzi Beatriz , Araujo Rafael C , Daniel João L P , Santos Geraldo T
Published: October 2021

Six lactating cows (106.3 ± 14.8 DIM; 550.7 ± 21.8 kg BW) were enrolled in a replicated 3 × 3 Latin square to evaluate the supplementation of double salt of calcium ammonium nitrate decahydrate (CAN), composed by 16.5% N, 76.5% of nitrate (NO3 -), and 85.0% DM, on NO3 - and nitrite (NO2 -) residues, fatty acids (FA) proportion, and antioxidant capacity in milk. Each period lasted 21 d, with 14 d for adaptation and 7 d for sampling. Treatments were URE (11.9 g/DM of urea as a control), CAN15: 15 g of CAN (11.5 g of NO3 - on DM), and CAN30: 30 g of CAN (23 g of NO3 - on DM). Milk samples were collected on d 15–16 of each period, frozen at –20°C, and after analyzed for NO3 - and NO2 - residues, fatty acids, and antioxidant capacity. Data were analyzed using PROC MIXED of SAS. Supplemental CAN increased by 26% NO3 - residue in milk (P = 0.02), whereas did not affect (P < 0.05) NO2 - concentration (average of 0.042 mg/L). Total CLA and short-chain FA were not affected (P > 0.05) by CAN. Medium-chain FA decreased linearly (P < 0.05), while long-chain FA (P = 0.06) and monounsaturated FA (P = 0.08) tended to increase linearly. Saturated FA reduced (P < 0.05) linearly; however, the total of polyunsaturated FA was not affected (P > 0.05). There were no effects of CAN (P > 0.05) on n-3, n-6, and n-6/n-3 ratio. Milk reducing power and TBARS concentration were not affected (P > 0.05) by CAN, whereas conjugated dienes increased (P = 0.02) linearly (URE= 47.6 vs. CAN15= 52.7 vs. CAN30= 63.4 mmol/kg of fat). Supplementing CAN at 30 g/DM reduced total saturated FA and increased conjugated dienes, caused possibly by lower rumen biohydrogenation, which might increase lipid oxidation in milk.
PSX-B-7 Effect of supplementation of a robotic milker pellet on mixed diet intake, milk production, and blood metabolites of lactating cows
Authors: Uzee Nick , Rathert Abigail R , Salisbury Carlee M , Montgomery Dagan , Foote Andrew P
Published: October 2021

The objective of this experiment was to determine the effect of concentrate pellets, typically supplied by a voluntary robotic milking system (VMS), on milk production and composition, nutrient metabolism, and diet dry matter intake (DMI) of lactating dairy cows. Holstein cows (n = 24) were assigned to receive either no supplemental pellets (CON), 2 kg/d (LP), or 6 kg/d (HP) in addition to ad libitum diet access, with treatments balanced by lactation number and days in milk. The LP group was allotted 1 kg of pellets every 12 hours and the HP group was allotted 2 kg every 8 hours, to simulate the amount consumed while visiting a VMS unit two or three times daily. Pellet and diet consumption were measured using the Insentec feeding system. Milk production was measured daily, utilizing a milk meter in a conventional parlor. Milk samples were collected twice weekly for 35 days and analyzed for components. Blood samples, body weight, and body condition scores were obtained weekly after the morning milking. Data were analyzed using a mixed model (SAS 9.4) with fixed effects of treatment, day, and their interaction, with day as a repeated measure. No treatment × day interaction was present for diet DMI (P = 0.72); however, addition of pellets to the LP and HP treatment groups decreased diet DMI when compared to CON (P < 0.006). Plasma lactate tended to be greater for CON (P = 0.09) than LP and HP treatment groups with no treatment effect on other blood metabolites (P ≥ 0.30). There was no effect of treatment on milk yield, milk components, or body weight and condition (P ≥ 0.23). These data suggest that feeding a pellet similar to conditions in a robotic milking system could alter DMI without negatively impacting metabolism or milk production.
PSXIV-6 País grape marc (Vitis vinifera, País) inclusion in concentrate diets reduces ammonia nitrogen but not methane production in an in vitro batch culture
Authors: Suescun-Ospina Sandra , Vera Nelson , Astudillo Rita , Avila-Stagno Jorge
Published: October 2021

País Grape (Vitis vinifera L.) is an ancestral variety used in Chilean wine industry. It has a higher content of proanthocyanidins than commercial varieties such as Carmenère or Pinot Noir, resulting in País grape marc (PGM) with high contents of condensed tannins. As such, PGM inclusion in ruminant diets would have the potential to reduce enteric methane (CH4) emissions and decrease urinary N excretion. The objective of this study was to evaluate the effects of substitution of mixed hay (MH) with PGM in a high concentrate diet [65% dry matter (DM)] on in vitro dry matter disappearance (IVDMD), ruminal fermentation parameters, gas and CH4 production. Treatments were: T1 (Control) = 20% MH, 15% corn silage, 65% concentrate; T2 = 10% MH, 10% PGM, 15% corn silage, 65% concentrate; T3 = 20% PGM, 15% corn silage, 65% concentrate. The study was a randomized complete design with 3 treatment and 3 replicates, incubated for 24 h at 39º C. Data were compared by Tukey test and polynomial contrasts. There was a linear reduction in NH3-N (P = 0.001) as dietary PGM increased. Inclusion of PGM reduced NH3-N by 50% when added at 10% DM, and 71.7% at 20% DM. However, there also was 4% reduction in IVDMD (P ≤ 0.001) and gas production (P = 0.012) in the 20% GM diet. There were no treatment effects (P ≥ 0.05) on CH4 production or yield. Fermentation efficiency determined by the partition factor increased linearly (P = 0.013) as PGM inclusion increased, suggesting that it increases organic matter to be degraded. Based on this study it could be concluded that PGM is an alternative source of fiber for ruminants on concentrate diets, as it can result in improved rumen fermentation efficiency and a substantial reduction in ruminal ammonia nitrogen concentration (NH3-N).