Biallelic variants in SLC38A3 encoding a glutamine transporter cause epileptic encephalopathy

The solute carrier (SLC) superfamily encompasses >400 transmembrane transporters involved in the exchange of amino acids, nutrients, ions, metals, neurotransmitters and metabolites across biological membranes. SLCs are highly expressed in the mammalian brain; defects in nearly 100 unique SLC-encoding genes (OMIM: https://www.omim.org) are associated with rare Mendelian disorders including developmental and epileptic encephalopathy (DEE) and severe neurodevelopmental disorders (NDDs). Exome sequencing and family-based rare variant analyses on a cohort with NDD identified two siblings with DEE and a shared deleterious homozygous splicing variant in SLC38A3. The gene encodes SNAT3, a sodium-coupled neutral amino acid transporter and a principal transporter of the amino acids asparagine, histidine, and glutamine, the latter being the precursor for the neurotransmitters GABA and glutamate. Additional subjects with a similar DEE phenotype and biallelic predicted-damaging SLC38A3 variants were ascertained through GeneMatcher and collaborations with research and clinical molecular diagnostic laboratories. Untargeted metabolomic analysis was performed to identify novel metabolic biomarkers. Ten individuals from seven unrelated families from six different countries with deleterious biallelic variants in SLC38A3 were identified. Global developmental delay, intellectual disability, hypotonia, and absent speech were common features while microcephaly, epilepsy, and visual impairment were present in the majority. Epilepsy was drug-resistant in half. Metabolomic analysis revealed perturbations of glutamate, histidine, and nitrogen metabolism in plasma, urine, and cerebrospinal fluid of selected subjects, potentially representing biomarkers of disease. Our data support the contention that SLC38A3 is a novel disease gene for DEE and illuminate the likely pathophysiology of the disease as perturbations in glutamine homeostasis

AMFR dysfunction causes autosomal recessive spastic paraplegia in human that is amenable to statin treatment in a preclinical model

Hereditary spastic paraplegias (HSP) are rare, inherited neurodegenerative or neurodevelopmental disorders that mainly present with lower limb spasticity and muscle weakness due to motor neuron dysfunction. Whole genome sequencing identified bi-allelic truncating variants in AMFR, encoding a RING-H2 finger E3 ubiquitin ligase anchored at the membrane of the endoplasmic reticulum (ER), in two previously genetically unexplained HSP-affected siblings. Subsequently, international collaboration recognized additional HSP-affected individuals with similar bi-allelic truncating AMFR variants, resulting in a cohort of 20 individuals from 8 unrelated, consanguineous families. Variants segregated with a phenotype of mainly pure but also complex HSP consisting of global developmental delay, mild intellectual disability, motor dysfunction, and progressive spasticity. Patient-derived fibroblasts, neural stem cells (NSCs), and in vivo zebrafish modeling were used to investigate pathomechanisms, including initial preclinical therapy assessment. The absence of AMFR disturbs lipid homeostasis, causing lipid droplet accumulation in NSCs and patient-derived fibroblasts which is rescued upon AMFR re-expression. Electron microscopy indicates ER morphology alterations in the absence of AMFR. Similar findings are seen in amfra-/- zebrafish larvae, in addition to altered touch-evoked escape response and defects in motor neuron branching, phenocopying the HSP observed in patients. Interestingly, administration of FDA-approved statins improves touch-evoked escape response and motor neuron branching defects in amfra-/- zebrafish larvae, suggesting potential therapeutic implications. Our genetic and functional studies identify bi-allelic truncating variants in AMFR as a cause of a novel autosomal recessive HSP by altering lipid metabolism, which may potentially be therapeutically modulated using precision medicine with statins

Effectiveness of school food environment policies on children's dietary behaviors: A systematic review and meta-analysis.

BACKGROUND: School food environment policies may be a critical tool to promote healthy diets in children, yet their effectiveness remains unclear. OBJECTIVE: To systematically review and quantify the impact of school food environment policies on dietary habits, adiposity, and metabolic risk in children. METHODS: We systematically searched online databases for randomized or quasi-experimental interventions assessing effects of school food environment policies on children's dietary habits, adiposity, or metabolic risk factors. Data were extracted independently and in duplicate, and pooled using inverse-variance random-effects meta-analysis. Habitual (within+outside school) dietary intakes were the primary outcome. Heterogeneity was explored using meta-regression and subgroup analysis. Funnel plots, Begg's and Egger's test evaluated potential publication bias. RESULTS: From 6,636 abstracts, 91 interventions (55 in US/Canada, 36 in Europe/New Zealand) were included, on direct provision of healthful foods/beverages (N = 39 studies), competitive food/beverage standards (N = 29), and school meal standards (N = 39) (some interventions assessed multiple policies). Direct provision policies, which largely targeted fruits and vegetables, increased consumption of fruits by 0.27 servings/d (n = 15 estimates (95%CI: 0.17, 0.36)) and combined fruits and vegetables by 0.28 servings/d (n = 16 (0.17, 0.40)); with a slight impact on vegetables (n = 11; 0.04 (0.01, 0.08)), and no effects on total calories (n = 6; -56 kcal/d (-174, 62)). In interventions targeting water, habitual intake was unchanged (n = 3; 0.33 glasses/d (-0.27, 0.93)). Competitive food/beverage standards reduced sugar-sweetened beverage intake by 0.18 servings/d (n = 3 (-0.31, -0.05)); and unhealthy snacks by 0.17 servings/d (n = 2 (-0.22, -0.13)), without effects on total calories (n = 5; -79 kcal/d (-179, 21)). School meal standards (mainly lunch) increased fruit intake (n = 2; 0.76 servings/d (0.37, 1.16)) and reduced total fat (-1.49%energy; n = 6 (-2.42, -0.57)), saturated fat (n = 4; -0.93%energy (-1.15, -0.70)) and sodium (n = 4; -170 mg/d (-242, -98)); but not total calories (n = 8; -38 kcal/d (-137, 62)). In 17 studies evaluating adiposity, significant decreases were generally not identified; few studies assessed metabolic factors (blood lipids/glucose/pressure), with mixed findings. Significant sources of heterogeneity or publication bias were not identified. CONCLUSIONS: Specific school food environment policies can improve targeted dietary behaviors; effects on adiposity and metabolic risk require further investigation. These findings inform ongoing policy discussions and debates on best practices to improve childhood dietary habits and health

Effect of carotenoids, extracted from dry tomato waste, on the stability and characteristics of various vegetable oils

In this study, various vegetable oils were enriched with carotenoids originating from dry tomato waste using ultrasound-assisted extraction, microwave-assisted extraction and maceration. Analyses of total carotenoids, color and oxidative stability (determined by free radical scavenger activity, peroxide value, Rancimat method and DSC analysis) were carried out. Commercial oils without enrichment were used for comparison. The results showed that the carotenoid contents of the oils increased significantly by increasing incorporation of dry tomato waste. Also, maceration proved to extract significantly higher amounts of these compounds. In some oils the extraction of dry tomato waste improved their oxidative and thermal stability while in others it caused an increase in the peroxide value and a decrease in induction time. Finally, the color parameters of the oils were significantly influenced. Enriched oils could be a potential source of bioactive compounds and might have significant antioxidant activity when ingested as part of a dietary regime

Anticancer and biochemical effects of Viscum album L. protein extracts on HeLa cells

European mistletoe (Viscum album) is a medicinal plant with significant anticancer properties. In vitro callus production provides an essential alternative for biomass production and medicinal compounds propagation. In this study, we investigated (a) the biotechnological production of somaclonal proteins from mistletoe callus propagation and (b) the in vitro cytotoxic properties of these mistletoe protein extracts against cervical cancer. Mistletoe proteins (MPEs) were extracted from natural resources (plant leaves and stems) and in vitro propagated variant calluses. Potential cytotoxic effects of the extracted proteins on HeLa cells were assessed by the MTT proliferation assay, whereas oxidative damage and induction of apoptosis were measured fluorometrically by the reactive oxygen species (ROS) sensitive carboxy-H2-DCFDA method and caspase-3 activation assays, respectively. Electrophoretic results indicated differences in the protein bands between callus and donor plant tissues. Leaf and stem protein extracts (PEs) demonstrated a strong impact on HeLa cells viability by promoting oxidative damage and inducing apoptosis in contrast to callus PEs. This study shows that MPEs application in HeLa cells causes oxidative alterations and apoptosis through caspase-3 activation and increased intracellular ROS levels. These findings may be useful for uncovering novel bioactive products of pharmaceutical interest as well as for gaining an understanding of the biochemical pathways of mistletoe proteins in cancer treatment. © 2019, Springer Nature B.V

Evaluation of the Quality of Evidence of the Association of Foods and Nutrients with Cardiovascular Disease and Diabetes: A Systematic Review

Importance: Poor diet is a leading global factor associated with cardiometabolic disease (CMD). Understanding the quality of evidence of the associations between specific dietary factors and CMD, including effect size (relative risk [RR]) and uncertainty, is essential to guide policy and consumer actions to achieve healthy diet and public health goals. Objective: To assess the quality of evidence of the associations between specific dietary factors and CMD as well as the quantitative evidence for RRs and the uncertainty of these risk estimates. Evidence Review: PubMed and the reference lists of eligible articles were searched between May 1, 2015, and February 26, 2021, for systematic reviews with meta-analyses of randomized clinical trials and prospective cohort studies that analyzed the consumption of 1 or more of the dietary factors of interest; reported dose-response meta-analyses; included healthy adults; and assessed 1 or more of the outcomes of interest. Study characteristics and RR estimates were extracted in duplicate. For identified associations, quality of evidence was assessed using the Bradford-Hill criteria for causation. Findings: A total of 2058 potentially relevant reports were identified, from which 285 full-text articles were assessed for eligibility. The final selection of articles included 28 meta-analyses representing 62 associations between diet and CMD. Among these associations, 10 foods, 3 beverages, and 12 nutrients had at least probable evidence of associations with coronary heart disease, stroke, and/or diabetes. Most RRs ranged from 0.87 to 0.96 per daily serving change for protective associations and from 1.06 to 1.15 per daily serving change for harmful associations. Most identified associations were protective (n = 38) and a smaller number were harmful (n = 24), with a higher risk associated with higher intake. Conclusions and Relevance: This systematic review summarized the current quality of evidence of the associations of specific dietary factors with coronary heart disease, stroke, and diabetes. These findings may inform dietary guidance, the assessment of disease burden in specific populations, policy setting, and future research. © 2022 American Medical Association. All rights reserved

Nutritional Characterization of Leaves and Herbal Tea of Moringa oleifera Cultivated in Greece

Moringa oleifera tree was cultivated for the first time in the mild temperate climate of Greece. Moringa oleifera leaf protein and amino acid content, total phenols and flavonols, lipids, fatty acids, ash and minerals, dietary fiber and carbohydrates, carotenoids, and vitamins C and E were determined. Leaves were high in protein content (26.3%), including all the essential amino acids. They also contained 4512.2 mg GAE.100 g–1 DM of total phenols and were rich in flavonols (myricetin, quercetin, and kaempherol at 649.8, 77.2, and 37.2 mg.100 g–1 DM, respectively). Their lipid content was 5.6% (saturated fatty acids 28.33%, mono-unsaturated fatty acids 8.54%, and polyunsaturated fatty acids 61.27%). Leaves were rich in Ca, K, Mg, and P (1712.6, 1002.9, 460.4, and 194.2mg.100 g–1 DM, respectively), while other minerals (Mn, Na, Fe, Zn, Cu, and Se) were found in lower concentrations. The dietary fiber content of dried leaves was 34.1%, while the nonstructural carbohydrate content was 22.0%. They also had high carotenoid content (lutein, zeaxanthin, and β-carotene at 10.03, 1.52, and 2.02 mg.100 g–1 DM, respectively). The vitamins C and E concentrations were 203.1 and 104 mg.100 g–1 DM, respectively. The herbal tea was high in total phenols (736.9 mg GAE.L–1) and flavonols (myricetin, quercetin, and kaempherol at 203.7, 17.4, and 10.6 mg.L–1, respectively), and had antioxidant activity (protection factor: 4.2). © 2017 Taylor & Francis Group, LLC