Tasnim Mohaissen

This study aimed to identify reported causes of neonatal death across the European Economic Area (EEA) member states and Switzerland as of 1 January 2010 (hereafter referred to as the European Region) and, where possible, examine the cause-specific distribution of neonatal deaths over time. We conducted a two-pronged analysis: (1) database analysis using WHO public datasets (2000–2021) for 28 countries in the European Region and (2) a systematic review and meta-analysis of pertinent records from PubMed, Embase, Web of Science, Scopus, Cochrane, and Google Scholar, which were comprehensively reviewed against inclusion criteria up to 10 August 2024, following PRISMA guidelines. The average neonatal mortality rate (NMRs) between 2000 and 2021 in WHO data was 2.63 per 1000 live births, with a significant decline (−0.074 per year) across the region. Prematurity (41.2%) and congenital anomalies (28.9%) were the most common registered causes of neonatal death; however, WHO data lacked etiological detail to reliably analyze cause-specific trends. The systematic review identified 41 eligible studies, of which 15 were included in meta-analyses. Pooled estimates showed that congenital anomalies and prematurity accounted for 30% (95% CI 17–46) and 31% (95% CI 18–49) of deaths, respectively. Among extremely preterm neonates, infections, respiratory, and cardiovascular disorders were the most common reported causes of death.      Conclusion: NMRs across the European Region are declining, with prematurity and congenital anomalies being leading causes of neonatal death. Current reporting frameworks lack granularity, and a wider adoption of standardized classification systems is critical to improving surveillance and data comparability, especially for preterm infants. What is Known: • Neonatal mortality rates have declined across Europe over recent decades. • There are considerable regional disparities in both causes and rates of neonatal death. What is New: • Prematurity and congenital anomalies each account for ~30% of neonatal deaths. • Greater use of standardized classifications of neonatal death causes is urgently needed.

A bstract: Purpose: Despite considerable progress in reducing neonatal mortality, significant work remains. The regional differences in both the rates and causes of neonatal mortality, necessitates addressing this issue within a regional context. This study aimed to identify reported causes of neonatal death in the European Region and examine the cause-specific distribution of neonatal deaths over time. Methods: We conducted a two-pronged analysis: (1) database analysis using WHO public datasets (2000–2021) for 28 countries in the European Region and (2) a systematic review and meta-analysis of pertinent records from various databases which were comprehensively reviewed against inclusion criteria up to 10 ^th August 2024, following PRISMA guidelines. Results: The average neonatal mortality rate between 2000–2021 in WHO data was 2.63 per 1,000 live births, with a significant decline (-0.074 per year) across the European Region. Prematurity (41.2%) and congenital anomalies (28.9%) were the most common registered causes of neonatal death; no detailed etiological breakdown was available. The systematic review identified 41 eligible studies; 14 could be included in meta- analyses. Pooled estimates showed that congenital anomalies and prematurity each accounted for 30% of deaths. Among extremely preterm neonates, infections, respiratory and cardiovascular disorders were the most common reported causes of death. Conclusions: NMRs across the European Region is declining (rapidly) with prematurity and congenital anomalies being leading causes of neonatal death. Current mortality reporting frameworks inadequately capture the diagnostic complexity of neonatal death causes. Adoption of detailed, standardized classification systems is critical to improving surveillance and data comparability, especially for preterm infants. W hat is K nown: Neonatal mortality rates have declined across Europe over recent decades. There are considerable regional disparities in both causes and rates of neonatal death. W hat is N ew: Prematurity and congenital anomalies each account for ∼30% of neonatal deaths. In extremely preterm infants, infections and cardiorespiratory disorders predominate. Greater use of standardised classifications of neonatal death causes is urgently needed.

Little is known, how life-long hyperlipidaemia affects vascular ageing, before atherosclerosis. Here, we characterise effects of mild, life-long hyperlipidaemia on age-dependent endothelial dysfunction (ED) in humanised dyslipidaemia model of E3L.CETP mice. Vascular function was characterised using magnetic resonance imaging in vivo and wire myograph ex vivo. Plasma endothelial biomarkers and non-targeted proteomics in plasma and aorta were analysed. Early atherosclerosis lesions were occasionally present only in 40-week-old or older E3L.CETP mice. However, age-dependent ED developed earlier, in 14-week-old male and 22-week-old female E3L.CETP mice as compared with 40-week-old female and male C57BL/6J mice. Acetylcholine-induced vasodilation in 8-week-old E3L.CETP, especially female mice, was blocked by catalase and attributed to H₂O₂. In 8-week-old female E3L.CETP mice, changes in plasma proteome in response to hyperlipidaemia were modest, while in male mice a number of differentially expressed proteins were identified that were involved in oxidative stress response, inflammation and regulation of metabolic pathways. In contrast, in older E3L.CETP and C57BL/6J mice, either plasma or aortic proteome displayed similar pattern of vascular ageing, dominating over hyperlipidaemia-induced changes. Interestingly, in 48-week-old male but not female E3L.CETP mice, vascular mitochondrial functional response was impaired. Early resilience of hyperlipidaemia-induced detrimental effects in young female E3L.CETP mice on a functional level was associated with a switch in vasodilation mechanism, blunted systemic proteomic response in plasma and slower ED development as compared to male E3L.CETP mice. The results indicate that profile of early vascular response to risk factors in young age may determine level of ED in older age before atherosclerosis development.

Phylloquinon (PK) and menaquinones (MK) are both naturally occurring compounds belonging to vitamin K group. Present study aimed to comprehensively analyze the influence of PK in several models of vascular dysfunction to determine whether PK has vasoprotective properties, similar to those previously described for MK. Effects of PK and MK on endothelial dysfunction were studied in ApoE/LDLR^−/− mice in vivo, in the isolated aorta incubated with TNF, and in vascular cells as regard inflammation and cell senescence (including replicative and stress-induced models of senescence). Moreover, the vascular conversion of exogenous vitamins to endogenous MK-4 was analyzed. PK, as well as MK, given for 8 weeks in diet (10 mg/kg) resulted in comparable improvement in endothelial function in the ApoE/LDLR^−/− mice. Similarly, PK and MK prevented TNF-induced impairment of endothelium-dependent vasorelaxation in the isolated aorta. In in vitro studies in endothelial and vascular smooth muscle cells, we identified that both PK and MK displayed anti-senescence effects via decreasing DNA damage while in endothelial cells anti-inflammatory activity was ascribed to the modulation of NFκB activation. The activity of PK and MK was comparable in terms of their effect on senescence and inflammation. Presence of endogenous synthesis of MK-4 from PK in aorta and endothelial and smooth muscle cells suggests a possible involvement of MK in vascular effects of PK. In conclusion, PK and MK display comparable vasoprotective effects, which may be ascribed, at least in part, to the inhibition of cell senescence and inflammation. The vasoprotective effect of PK in the vessel wall can be related to the direct effects of PK, as well as to the action of MK formed from PK in the vascular wall.

Adipose triglyceride lipase (ATGL) is involved in lipolysis and displays a detrimental pathophysiological role in cardio-metabolic diseases. However, the organo-protective effects of ATGL-induced lipolysis were also suggested. The aim of this work was to characterize the function of lipid droplets (LDs) and ATGL-induced lipolysis in the regulation of endothelial function. ATGL-dependent LDs hydrolysis and cytosolic phospholipase A₂ (cPLA₂)-derived eicosanoids production were studied in the aorta, endothelial and smooth muscle cells exposed to exogenous oleic acid (OA) or arachidonic acid (AA). Functional effects of ATGL-dependent lipolysis and subsequent activation of cPLA₂/PGI₂ pathway were also studied in vivo in relation to postprandial endothelial dysfunction.The formation of LDs was invariably associated with elevated production of endogenous AA-derived prostacyclin (PGI₂). In the presence of the inhibitor of ATGL or the inhibitor of cytosolic phospholipase A₂, the production of eicosanoids was reduced, with a concomitant increase in the number of LDs. OA administration impaired endothelial barrier integrity in vitro that was further impaired if OA was given together with ATGL inhibitor. Importantly, in vivo, olive oil induced postprandial endothelial dysfunction that was significantly deteriorated by ATGL inhibition, cPLA₂ inhibition or by prostacyclin (IP) receptor blockade.In summary, vascular LDs formation induced by exogenous AA or OA was associated with ATGL- and cPLA₂-dependent PGI₂ production from endogenous AA. The inhibition of ATGL resulted in an impairment of endothelial barrier function in vitro. The inhibition of ATGL-cPLA₂-PGI₂ dependent pathway resulted in the deterioration of endothelial function upon exposure to olive oil in vivo. In conclusion, vascular ATGL-cPLA₂-PGI₂ dependent pathway activated by lipid overload and linked to LDs formation in endothelium and smooth muscle cells has a vasoprotective role by counterbalancing detrimental effects of lipid overload on endothelial function.

Vascular ageing is associated with increased arterial stiffness and cardiovascular mortality that might be linked to altered vascular energy metabolism. The aim of this study was to establish a Seahorse XFe96 Analyzer-based methodology for the reliable, functional assessment of mitochondrial respiration and glycolysis in single murine aortic rings and to validate this functional assay by characterising alterations in vascular energy metabolism in aged mice. Healthy young and old C57BL/6 mice were used for the analyses. An optimised setup consisting of the Seahorse XFe96 Analyzer and Seahorse Spheroid Microplates was applied for the mitochondrial stress test and the glycolysis stress test on the isolated murine aortic rings, supplemented with analysis of NAD content in the aorta. To confirm the age-dependent stiffness of the vasculature, pulse wave velocity was measured in vivo. In addition, the activity of vascular nitric oxide synthase and vascular wall morphology were analysed ex vivo. The vascular ageing phenotype in old mice was confirmed by increased aortic stiffness, vascular wall remodelling, and nitric oxide synthase activity impairment. The rings of the aorta taken from old mice showed changes in vascular energy metabolism, including impaired spare respiratory capacity, maximal respiration, glycolysis, and glycolytic capacity, as well as a fall in the NAD pool. In conclusion, optimised Seahorse XFe96-based analysis to study energy metabolism in single aortic rings of murine aorta revealed a robust impairment of functional vascular respiratory and glycolytic capacity in old mice linked to NAD deficiency that coincided with age-related aortic wall remodelling and stiffness.

Aim: Chronic heart failure (CHF) is often linked to liver malfunction and systemic endothelial dysfunction. However, whether cardio‐hepatic interactions in heart failure involve dysfunction of liver sinusoidal endothelial cells (LSECs) is not known. Here we characterize LSECs phenotype in early and end stages of chronic heart failure in a murine model.Methods: Right ventricle (RV) function, features of congestive hepatopathy, and the phenotype of primary LSECs were characterized in Tgαq*44 mice, with cardiomyocyte‐specific overexpression of the Gαq protein, at the age of 4‐ and 12‐month representative for early and end‐stage phases of CHF, respectively.Results: 4‐ and 12‐month‐old Tgαq*44 mice displayed progressive impairment of RV function and alterations in hepatic blood flow velocity resulting in hepatic congestion with elevated GGT and bilirubin plasma levels and decreased albumin concentration without gross liver pathology. LSECs isolated from 4‐ and 12‐month‐old Tgαq*44 mice displayed significant loss of fenestrae with impaired functional response to cytochalasin B, significant changes in proteome related to cytoskeleton remodeling, and altered vasoprotective function. However, LSECs barrier function and bioenergetics were largely preserved. In 4‐ and 12‐month‐old Tgαq*44 mice, LSECs defenestration was associated with prolonged postprandial hypertriglyceridemia and in 12‐month‐old Tgαq*44 mice with proteomic changes of hepatocytes indicative of altered lipid metabolism.Conclusion: Tgαq*44 mice displayed right‐sided HF and altered hepatic blood flow leading to LSECs dysfunction involving defenestration, shift in eicosanoid profile, and proteomic changes. LSECs dysfunction appears as an early and persistent event in CHF, preceding congestive hepatopathy and contributing to alterations in lipoprotein transport and CHF pathophysiology.