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Objective: Lipoprotein assembly and secretion in the small intestine are critical for dietary fat absorption. Surfeit locus protein 4 (SURF4) serves as a cargo receptor, facilitating the cellular transport of multiple proteins and mediating hepatic lipid secretion in vivo. However, its involvement in intestinal lipid secretion is not fully understood. In this study, we investigated the role of SURF4 in intestinal lipid absorption. Methods: We generated intestine-specific Surf4 knockout mice and characterized the phenotypes. Additionally, we investigated the underlying mechanisms of SURF4 in intestinal lipid secretion using proteomics and cellular models. Results: We unveiled that SURF4 is indispensable for apolipoprotein transport and lipoprotein secretion. Intestine-specific Surf4 knockout mice exhibited ectopic lipid deposition in the small intestine and hypolipidemia. Deletion of SURF4 impeded the transport of apolipoprotein A1 (ApoA1), proline-rich acidic protein 1 (PRAP1), and apolipoprotein B48 (ApoB48) and hindered the assembly and secretion of chylomicrons and high-density lipoproteins. Conclusions: SURF4 emerges as a pivotal regulator of intestinal lipid absorption via mediating the secretion of ApoA1, PRAP1 and ApoB48.

The brain's primary immune cells, microglia, are a leading causal cell type in Alzheimer's disease (AD). Yet, the mechanisms by which microglia can drive neurodegeneration remain unresolved. Here, we discover that a conserved stress signaling pathway, the integrated stress response (ISR), characterizes a microglia subset with neurodegenerative outcomes. Autonomous activation of ISR in microglia is sufficient to induce early features of the ultrastructurally distinct "dark microglia" linked to pathological synapse loss. In AD models, microglial ISR activation exacerbates neurodegenerative pathologies and synapse loss while its inhibition ameliorates them. Mechanistically, we present evidence that ISR activation promotes the secretion of toxic lipids by microglia, impairing neuron homeostasis and survival in vitro. Accordingly, pharmacological inhibition of ISR or lipid synthesis mitigates synapse loss in AD models. Our results demonstrate that microglial ISR activation represents a neurodegenerative phenotype, which may be sustained, at least in part, by the secretion of toxic lipids., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Lipoprotein assembly and secretion in the small intestine are critical for dietary fat absorption. Surfeit locus protein 4 (SURF4) serves as a cargo receptor, facilitating the cellular transport of multiple proteins and mediating hepatic lipid secretion in vivo. However, its involvement in intestinal lipid secretion is not fully understood. In this study, we investigated the role of SURF4 in intestinal lipid absorption. We generated intestine-specific Surf 4 knockout mice and characterized the phenotypes. Additionally, we investigated the underlying mechanisms of SURF4 in intestinal lipid secretion using proteomics and cellular models. We unveiled that SURF4 is indispensable for apolipoprotein transport and lipoprotein secretion. Intestine-specific Surf 4 knockout mice exhibited ectopic lipid deposition in the small intestine and hypolipidemia. Deletion of SURF4 impeded the transport of apolipoprotein A1 (ApoA1), proline-rich acidic protein 1 (PRAP1), and apolipoprotein B48 (ApoB48) and hindered the assembly and secretion of chylomicrons and high-density lipoproteins. SURF4 emerges as a pivotal regulator of intestinal lipid absorption via mediating the secretion of ApoA1, PRAP1 and ApoB48. [Display omitted] • Intestinal Surf4 knockout mice recapitulate the phenotypes of chylomicron retention disease. • SURF4 regulates intestinal lipid secretion by mediating the transport of apolipoproteins PRAP1, ApoA1, and ApoB48. • Female Surf4 heterozygous mice are resistant to high-fat diet-induced hypercholesterolemia. [ABSTRACT FROM AUTHOR]

The mechanism of lipotropic action provided by betaine in early diet-induced liver steatosis was investigated. Male rats received a high-fat liquid diet (HF) for 3 wk. HF intake resulted in a significant accumulation of triacyglycerol and cholesterol in the liver. Phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) was decreased, while protein expression of liver-X-receptor α (LXRα) and nuclear sterol regulatory element binding protein-1c (nSREBP-1c) was enhanced. Betaine supplement at 1% in HF inhibited hepatic triacyglycerol accumulation, but elevated serum cholesterol and triacyglycerol levels. Phosphorylation of AMPK and ACC was normalized, and induction of LXRα and nSREBP-1c was prevented. HF intake down-regulated betaine–homocysteine methyltransferase and increased homocysteine levels, both of which were blocked by betaine. Betaine supplementation elevated hepatic apolipoprotein B and serum phosphatidylcholine levels. The results suggest that both inhibition of lipid synthesis and enhancement of its secretion are involved in the anti-steatotic activity of betaine.

Main conclusion Based on 3D sections through cells of Botryococcus braunii, the structure of three domains of endoplasmic reticulum, and their spatial and functional relationships to other organelles are clarified. Oil production by photosynthetic microalgae has attracted attention since these oils can be converted into renewable, carbonneutral fuels. The green alga B. braunii accumulates large amounts of hydrocarbons, 30-50% of cell dry weight, in extracellular spaces rather than its cytoplasm. To advance the knowledge of hydrocarbon biosynthesis and transport pathways in this alga, we utilized transmission EM combined with rapid freezing and image reconstruction. We constructed detailed 3D maps distinguishing three ER domains: rdER with ribosomes on both sides, rsER with ribosomes on one side, and sER without ribosomes. The rsER and sER domains were especially prominent during the oil body formation and oil secretion stages. The ER contacted the chloroplasts, oil bodies, or plasma membrane via the rsER domains, oriented with the ribosomefree surface facing the organelles. We discuss the following transport pathway for hydrocarbons and their precursors in the cytoplasm: chloroplast → endoplasmic reticulum (ER) → oil bodies → ER → plasma membrane → secretion. This study represents the first 3D study of the three-domain classification (rdER, rsER and sER) of the ER network among eukaryotic cells. Finally, we propose the novel features of the ERs in plant cells that are distinct from the latest proposed model for the ERs in mammalian cells. [ABSTRACT FROM AUTHOR]

Lipid metabolism has important roles in animal growth, development, and reproduction. As a regulator of lipid metabolism, CIDEc promotes unilocular development of lipid droplets and stimulates intracellular lipid deposition, and has two isoforms, CIDEc-l and CIDEc-s. CIDEc-l has ten more N-terminal amino acids than CIDEc-s. However, the functions of two isoforms are largely unknown. In this study, the expression profiles of two isoforms in Bama pigs differed, with cidec-l dominant in the liver and small intestine, and cidec-s dominant in muscle and adipose tissue. Fasting and consuming a high-fat diet resulted in changes in the expression of the two isoforms that were closely related to changes in blood and muscle triglyceride (TG) concentrations. Comparison of gene expression and TG concentration suggested that CIDEc-l accelerated lipid secretion and that CIDEc-s promoted lipid deposition, implying that the two isoforms had different functions. Study In vitro confirmed that CIDEc-s stimulated lipid deposition in C2C12 muscle cells and CIDEc-l promoted lipid secretion in HepG2 liver cells. The results showed that two tissue-specific CIDEc isoforms had different roles in lipid deposition and secretion. They may be potential targets for regulation of fat content. [ABSTRACT FROM AUTHOR]

α-Linolenic acid (ALA) supplementation or exercise training can independently prevent hepatic lipid accumulation and reduced insulin signaling; however, this may occur through different mechanisms of action. In the current study, obese Zucker rats displayed decreased phospholipid (PL) content in association with hepatic lipid abundance, and therefore, we examined whether ALA and exercise training would prevent these abnormalities differently to reveal additive effects on the liver. To achieve this aim, obese Zucker rats were fed control diet alone or supplemented with ALA and were sedentary or exercise trained for 4 wk (C-Sed, ALA-Sed, C-Ex, and ALA-Ex). ALA-Sed rats had increased microsomal-triglyceride transfer protein (MTTP), a protein required for lipoprotein assembly/secretion, as well as modestly increased PL content in the absence of improvements in mitochondrial content, lipid accumulation, or insulin sensitivity. In contrast, C-Ex rats had increased mitochondrial content and insulin sensitivity; however, this corresponded with minimal improvements in PL content and hepatic lipid accumulation. Importantly, ALA-Ex rats demonstrated additive improvements in PL content and hepatic steatosis, which corresponded with increased mitochondrial content, MTTP and apolipoprotein B100 content, greater serum triacylglyceride, and insulin sensitivity. Overall, these data demonstrate additive effects of ALA and exercise training on hepatic lipid accumulation, as exercise training preferentially increased mitochondrial content, while ALA promoted an environment conducive for lipid secretion. These data highlight the potential for combination therapy to mitigate liver disease progression. [ABSTRACT FROM AUTHOR]

Shikonin derivatives are specialized lipophilic metabolites, secreted in abundant amounts from the root epidermal cells of Lithospermum erythrorhizon. Because they have anti-microbial activities, these compounds, which are derivatives of red naphthoquinone, are thought to serve as a chemical barrier for plant roots. The mechanism by which they are secreted from cells is, however, largely unknown. The shikonin production system in L. erythrorhizon is an excellent model for studying the mechanism by which lipophilic compounds are secreted from plant cells, because of the abundant amounts of these compounds produced by L. erythrorhizon, the 0 to 100% inducibility of their production, the light-specific inhibition of production, and the visibility of these products as red pigments. To date, many factors regulating shikonin biosynthesis have been identified, but no mechanism that regulates shikonin secretion without inhibiting biosynthesis has been detected. This study showed that inhibitors of membrane traffic strongly inhibit shikonin secretion without inhibiting shikonin production, suggesting that the secretion of shikonin derivatives into the apoplast utilizes pathways common to the ADP-ribosylation factor/GTPase-activating protein (ARF/GAP) system and actin filament polymerization, at least in part. These findings provide clues about the machinery involved in secreting lipid-soluble metabolites from cells.

The sebaceous gland is a neuro-immuno-endocrine organ responsible for maintaining regular skin functions. Overdose exposure of UV and visible light (e.g., blue light) can cause sebocyte gland function disorders or even different diseases (e.g., chronic actinic dermatitis). Studying the mechanism of light-induced damage in sebaceous glands has been challenging, since ex vivo culture of sebaceous glands is difficult due to its short life in culture medium. To address this issue, a versatile 3D artificial sebocyte gland model was established using the inertial focusing effect for studying the impact of light damage and screening potential drugs. The artificial sebocyte gland exhibited specific biological function and structure similar to natural sebocyte glands. Using this artificial sebocyte gland, the interactions between the artificial organ and blue light or UV were studied. The results indicated that UV and blue light upregulated lipid secretion and downregulated cell viability within the sebocytes. Light damage intensified oxidative stress and promoted pro-inflammation cytokines (i.e., IL-1β and TNF-α) production in the artificial sebocytes. Additionally, the therapeutic effects of cannabidiol, a clinically tested drug for treating acne, was also indicated on restoring light damaged sebaceous gland functions. These results indicate that the 3D artificial sebocyte gland could be a versatile, fast, and low-cost platform for skincare studies or drug screening., (© 2023 Wiley-VCH GmbH.)

Microbial oils are sustainable alternatives to petroleum for the production of chemicals and fuels. Oleaginous yeasts are promising source of oils and Yarrowia lipolytica is the most studied and engineered one. Nonetheless the commercial production of biolipids is so far limited to high value products due to the elevated production and extraction costs. In order to contribute to overcoming these limitations we exploited the possibility of secreting lipids to the culture broth, uncoupling production and biomass formation and facilitating the extraction. We therefore considered two synthetic approaches, Strategy I where fatty acids are produced by enhancing the flux through neutral lipid formation, as typically occurs in eukaryotic systems and Strategy II where the bacterial system to produce free fatty acids is mimicked. The engineered strains, in a coupled fermentation and extraction process using alkanes, secreted the highest titer of lipids described so far, with a content of 120% of DCW. [ABSTRACT FROM AUTHOR]

Bayberry ( Myrica pensylvanica ) fruits are covered with a remarkably thick layer of crystalline wax consisting of triacylglycerol (TAG) and diacylglycerol (DAG) esterified exclusively with saturated fatty acids. As the only plant known to accumulate soluble glycerolipids as a major component of surface waxes, Bayberry represents a novel system to investigate neutral lipid biosynthesis and lipid secretion by vegetative plant cells. The assembly of Bayberry wax is distinct from conventional TAG and other surface waxes, and instead proceeds through a pathway related to cutin synthesis (Simpson and Ohlrogge, 2016). In this study, microscopic examination revealed that the fruit tissue that produces and secretes wax (Bayberry knobs) is fully developed before wax accumulates and that wax is secreted to the surface without cell disruption. Comparison of transcript expression to genetically related tissues (Bayberry leaves, M. rubra fruits), cutin-rich tomato and cherry fruit epidermis, and to oil-rich mesocarp and seeds, revealed exceptionally high expression of 13 transcripts for acyl-lipid metabolism together with down-regulation of fatty acid oxidases and desaturases. The predicted protein sequences of the most highly expressed lipid-related enzyme-encoding transcripts in Bayberry knobs are 100% identical to the sequences from Bayberry leaves, which do not produce surface DAG or TAG. Together, these results indicate that TAG biosynthesis and secretion in Bayberry is achieved by both up and down-regulation of a small subset of genes related to the biosynthesis of cutin and saturated fatty acids, and also implies that modifications in gene expression, rather than evolution of new gene functions, was the major mechanism by which Bayberry evolved its specialized lipid metabolism. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner. [ABSTRACT FROM AUTHOR]

The mechanism of lipotropic action provided by betaine in early diet-induced liver steatosis was investigated. Male rats received a high-fat liquid diet (HF) for 3 wk. HF intake resulted in a significant accumulation of triacyglycerol and cholesterol in the liver. Phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) was decreased, while protein expression of liver-X-receptor α (LXRα) and nuclear sterol regulatory element binding protein-1c (nSREBP-1c) was enhanced. Betaine supplement at 1% in HF inhibited hepatic triacyglycerol accumulation, but elevated serum cholesterol and triacyglycerol levels. Phosphorylation of AMPK and ACC was normalized, and induction of LXRα and nSREBP-1c was prevented. HF intake down-regulated betaine–homocysteine methyltransferase and increased homocysteine levels, both of which were blocked by betaine. Betaine supplementation elevated hepatic apolipoprotein B and serum phosphatidylcholine levels. The results suggest that both inhibition of lipid synthesis and enhancement of its secretion are involved in the anti-steatotic activity of betaine. [ABSTRACT FROM AUTHOR]

The scenario of this work was to boost concurrent lipid accumulation and secretion by Coccomyxa subellipsoidea with glucose coupling glycerol as accelerator. Results stated that glucose coupling glycerol (carbon content ratio 5:5) with batch-feeding achieved total lipid accumulation and lipids-secreted by 4.79 g/L and 2.4 g/L, representing 1.61- to −4.16 and 3.69- to 17.1-fold more than sole glucose and sole glycerol manipulation. When response to fed-batch fermentation with glucose coupling glycerol supplementation, lipid accumulation and lipids-secreted peaked 20.4 g/L and 9.11 g/L which performed 4.26- and 3.8-fold increments than batch-feeding manipulation. Whether intracellular lipidic or extracellular lipidic fatty acid profiles (mainly C16-C18) were commendable for high-quality biodiesel-making. Glucose coupling glycerol mobilized multiple channels including up-regulating PPP and glycolysis, weakening TCA cycle, and dampening MVA biosynthesis to flock acetyl-CoA into lipid accumulation. Such glucose coupling glycerol manipulation concurrently lowered PC/PE ratio to intensify cytomembrane fluidity and permeability consequently compelled lipid secretion. Findings provided exemplifications in facilitating concurrent lipid accumulation and secretion for economical-feasible alga-based biodiesel. [Display omitted] • Glu coupling Gly fed-batch fermentation boosted algal lipid synthesis and secretion. • Glu coupling Gly achieved commendable fatty acids for high-quality biodiesel-making. • Glu coupling Gly activated glycolysis/weakened TCA/dampened MVA for lipid synthesis. • Glu coupling Gly depressed PC/PE to strengthen membrane permeability to spill lipid. [ABSTRACT FROM AUTHOR]

The milk-producing alveolar epithelial cells secrete milk that remains after birth the principal source of nutrients for neonates. Milk secretion and composition are highly regulated processes via integrated actions of hormones and local factors which involve specific receptors and downstream signal transduction pathways. Overall milk composition is similar among mammalian species, although the content of individual constituents such as lipids may significantly differ from one species to another. The milk lipid fraction is essentially composed of triglycerides, which represent more than 95 % of the total lipids in human and commercialized bovine milk. Though sterols, including cholesterol, which is the major milk sterol, represent less than 0.5 % of the total milk lipid fraction, they are of key importance for several biological processes. Cholesterol is required for the formation of biological membranes especially in rapidly growing organisms, and for the synthesis of sterol-based compounds. Cholesterol found in milk originates predominantly from blood uptake and, to a certain extent, from local synthesis in the mammary tissue. The present review summarizes current knowledge on cellular mechanisms and regulatory processes determining intra- and transcellular cholesterol transport in the mammary gland. Cholesterol exchanges between the blood, the mammary alveolar cells and the milk, and the likely role of active cholesterol transporters in these processes are discussed. In this context, the hormonal regulation and signal transduction pathways promoting active cholesterol transport as well as potential regulatory crosstalks are highlighted. [ABSTRACT FROM AUTHOR]

Background and Aims Recently, molecular approaches have been used to investigate the phylogeny of Oncidiinae. This has resulted in the transfer of taxa previously considered to be species of Oncidium Sw. into Gomesa R. Br. and the re-circumscription of both genera. In this study, the structure of the floral elaiophore (oil gland) is described and compared for Gomesa echinata (Barb. Rodr.) M.W. Chase & N.H. Williams, G. ranifera (Lindl.) M.W. Chase & N.H. Williams, Oncidium amazonicum (Schltr.) M.W. Chase & N.H. Williams and O. oxyceras (Königer & J.G. Weinm.) M.W. Chase & N.H. Williams in order to determine whether phylogenetic revision is supported by differences in its anatomy. Methods The floral elaiophore structure was examined and compared at three developmental stages (closed bud, first day of anthesis and final stage of anthesis) for all four species using light microscopy, fluorescence microscopy, scanning electron microscopy, transmission electron microscopy and histochemistry. Key results In all species investigated, the floral elaiophore occurs on the labellar callus and is of the epithelial type, comprising cuboidal to palisade-like, secretory epidermal cells and a layer of sub-epidermal cells, both tissues enclosing ground parenchyma supplied with collateral vascular bundles and containing idioblasts, often with raphides or phenolic contents. A bi-layered cuticle comprising an outer, lamellate and an inner, reticulate layer is present, and sub-cuticular accumulation of secreted material results in distension of the cuticle. Secretion-filled cavities are present at anthesis in the elaiophore cell walls and, in most species, the outer, tangential walls of the elaiophore have small, peg-like projections that protrude into the cytoplasm. In all taxa examined, the elaiophore organelle complement, especially the smooth endoplasmic reticulum (SER), is typical of lipid-secreting cells. Conclusions In terms of location, morphology, anatomy and ultrastructure, the floral elaiophores of both Gomesa and Oncidium species examined are very similar, and distinction between these genera is not possible based on elaiophore features alone. Furthermore, many of these elaiophore characters are shared with representatives of other clades of Oncidiinae, including the Ornithocephalus clade. Consequently, elaiophores are considered homoplasious and of limited value in investigating the phylogeny of this subtribe. [ABSTRACT FROM AUTHOR]

Background and Aims A significant number of species assigned to the Neotropical orchid sub-tribe Oncidiinae reward insect pollinators with oil produced in floral glands termed elaiophores. The latter may be glabrous (epithelial elaiophores) or hirsute (trichomal elaiophores). Although the detailed anatomy and ultrastructure of epithelial elaiophores have been studied for a number of genera, such as Oncidium Sw., Gomesa R. Br. and Trichocentrum Poepp. & Endl., hitherto, trichomal elaiophores have been investigated only for a single species of Oncidiinae, Ornithocephalus ciliatus Lindl. Furthermore, this is the only representative of the Ornithocephalus clade to be investigated to date. Here, an examination is made of the elaiophore anatomy and ultrastructure of a further four species currently assigned to this clade (Ornithocephalus gladiatus Hook., Phymatidium falcifolium Lindl., Zygostates grandiflora (Lindl.) Mansf. and Zygostates lunata Lindl.) and the results compared with those obtained for other Oncidiinae. Methods Elaiophore structure was examined for all species at three stages of flower development: closed bud, first day of anthesis and final stage of anthesis, using light microscopy, fluorescence microscopy, scanning electron microscopy, transmission electron microscopy and histochemistry. Key Results Elaiophores of O. gladiatus occur upon the lateral lobes of the labellum and display characters intermediate between those of typical epithelial and trichomal elaiophores, in that they are largely glabrous, consisting mainly of cuboidal epidermal cells, but bear short, unicellular hairs proximally. By contrast, the elaiophores of all the other species investigated occur on the callus and are of the trichomal type. In P. falcifolium, these unicellular hairs are capitate. In all species, oil secretion commenced at the closed floral bud stage. Ultrastructurally, the mainly trichomal elaiophores of the four representatives of the Ornithocephalus clade closely resembled the epithelial elaiophores of other Oncidiinae, in that their cells displayed an organelle complement typical of lipid-secreting cells. However, in some taxa, a number of noteworthy characters were present. For example, the elaiophore cuticle of O. gladiatus and P. falcifolium was bi-layered, the outer layer being lamellate, the inner reticulate. The cuticle of Z. grandiflora and Z. lunata was also lamellate, but here, a reticulate layer was absent. Accumulation of secreted oil resulted in the localized distension of the cuticle. Cuticular cracks and pores, however, were absent from all species. The walls of the secretory cells of Z. grandiflora were also atypical in that they had short protuberances or ingrowths, and contained cavities which are thought to be involved in the secretory process. Conclusions Of the species investigated, most displayed similar anatomical organization, their trichomal elaiophores occurring on the labellar callus. They, thus, differ from many other members of the Oncidiinae, where epithelial elaiophores are found either on the callus, or on the lateral lobes of the labellum. However, ultrastructurally, all elaiophores, whether those of representatives of the Ornithocephalus clade, or those of other oil-secreting Oncidiinae, possessed a similar complement of organelles, regardless of whether the elaiophores were trichomal or epithelial. In view of the latter, and the similar chemical composition of oils derived from all Oncidiinae investigated to date, it is probable that position and type of elaiophore, and possibly the structure of the overlying cuticle, play an important role in pollinator selection in these oil-secreting orchids. [ABSTRACT FROM AUTHOR]

Abstract: Fish oil (FO) has traditionally been used as the dominating lipid component in fish feed. However, FO is a limited resource and the price varies considerably, which has led to an interest in using alternative oils, such as vegetable oils (VOs), in fish diets. It is far from clear how these VOs affect liver lipid secretion and fish health. The polyunsaturated fatty acids (PUFAs), eicosapentanoic acid (EPA) and docosahexanioc acid (DHA), reduce the secretion of lipoproteins rich in triacylglycerols (TAGs) in Atlantic salmon, as they do in humans. The mechanism by which n-3 fatty acids (FAs) in the diet reduce TAG secretion is not known. We have therefore investigated the effects of rapeseed oil (RO) and n-3 rich diets on the accumulation and secretion of 3H-glycerolipids by salmon hepatocytes. Salmon, of approximately 90 g were fed for 17 weeks on one of four diets supplemented with either 13.5% FO, RO, EPA-enriched oil or DHA-enriched oil until a final average weight of 310 g. Our results show that the dietary FA composition markedly influences the endogenous FA composition and lipid content of the hepatocytes. The intracellular lipid level in hepatocytes from fish fed RO diet and DHA diet were higher, and the expressions of the genes for microsomal transfer protein (MTP) and apolipoprotein A1 (Apo A1) were lower, than those in fish fed the two other diets. Secretion of hepatocyte glycerolipids was lower in fish fed the EPA diet and DHA diet than it was in fish fed the RO diet. Our results indicate that EPA and DHA possess different hypolipidemic properties. Both EPA and DHA inhibit TAG synthesis and secretion, but only EPA induces mitochondrial proliferation and reduce intracellular lipid. Expression of the gene for peroxisome proliferator-activated receptor alpha (PPARα) was higher in the DHA dietary group than it was in the other groups. [Copyright &y& Elsevier]

Mammary epithelial cells from lactating cows were cultured onto inserts coated with type I collagen. Every second day, the rates of fatty acid synthesis and secretion were determined by measuring the amount of [14C]-labeled sodium acetate incorporated into lipids over a 4-h period. The [14C]-containing lipids were identified by thin layer chromatography fractionation. In parallel, the integrity of the cell layer was evaluated by measurement of transepithelial electrical resistance. The integrity increased progressively to reach a maximum after 8 d of culture. Cells incorporated acetate into lipids; 1.34% of acetate was incorporated into lipids produced by freshly isolated cells. This percentage decreased to 0.5% after 2 d of culture. Moreover, this capacity decreased with the duration in culture; on d 8, the rate of incorporation dropped to about 3% of that on d 2. In the cell extracts, the [14C]-labeled lipids were mainly triglycerides, although the proportion of diglycerides and phospholipids progressively increased as a part of total newly synthesized lipids. The proportion of triglycerides decreased 0.66 times between d 2 and 8 when the proportion of diglycerides and phospholipids increased 1.33 and 2.18 times, respectively. About 28% of the newly synthesized lipids were secreted within 4 h of incubation. Around 65 to 85% of these labeled lipids were found in the apical compartment, suggesting a partially vectorial secretion. But 58 to 80% of labeled lipids found in the apical and basolateral medium were free fatty acids. Functional tight junctions and incorporation of labeled fatty acids into triglycerides are not compatible with an inferred status of complete dedifferentiation of the cell layer. Moreover, triglyceride secretion seems compromised, probably due to the lack of an appropriate cell environment and cell shape. [ABSTRACT FROM AUTHOR]

Lichens are symbiotic associations of fungi and algae (or cyanobacteria). Fungal cells produce large amounts of lipid, assisted by algae, and secrete them out of the cells. Some of the secreted lipids crystallize in the thallus of the lichen. The crystalline materials sometimes occupy 30% of total dry weight of the thallus. This unusual amount of lipid crystal led to our interest in investigating the mechanism of lipid secretion. To begin the cell biological study of lipid secretion and to know the significance of the existence of such crystals in the thallus, it is essential to identify the crystal. The lipid crystal extracted from the thallus of a lichen, Myelochroa leucotyliza, was studied by EM observations, TLC analysis, and EM and X-ray diffraction methods. Atranorin is the predominant component of the crystalline materials in the lipids extracted. [Copyright &y& Elsevier]

• ALA-PDT inhibits lipid secretion of primary human sebocytes and induces atrophy of sebaceous gland. • ALA-PDT suppresses the expression of SREBP-1 in sebocytes. • ALA-PDT suppresses sebum secretion in the treatment of acne vulgaris through down-regulating the AMPK/SREBP-1 pathway, providing a potential mechanism of ALA-PDT for acne vulgaris. Acne vulgaris is a chronic inflammatory skin disease around pilosebaceous unit. 5-Aminolaevulinic acid photodynamic therapy (ALA-PDT) is an effective therapy for severe acne vulgaris. However, its specific treatment mechanism remains unclear. In the present study, we investigated the potential mechanism of how ALA-PDT induced lipid secretion inhibition in acne vulgaris. Primary human sebocytes and sebaceous gland of golden hamster were treated with/without ALA-PDT. Cell viability was evaluated by Live/Dead Cell assay. Fluorescence microscope was used to observe lipids secretion in sebocytes after Nile red staining. The expression of SREBP-1 after ALA-PDT was evaluated by qRT-PCR. Regulation of ALA-PDT on AMPK/SREBP-1 was evaluated by western blot. The results showed that ALA-PDT suppressed lipid secretion of primary human sebocytes. In addition, ALA-PDT could inhibit the expression of SREBP-1 in vitro. We also found that ALA-PDT activated AMPK pathway, down-regulating the expression of SREBP-1 in sebocytes after ALA-PDT. These findings elucidate that ALA-PDT suppresses lipid secretion through AMPK/SREBP-1 pathway in treatment of acne vulgaris. [ABSTRACT FROM AUTHOR]

Aims/Background: Biliary phospholipid secretion involves predominant segregation of canalicular phosphatidylcholine into bile. We tested the hypothesis that micellar concentrations of the major physiologic bile salt taurocholate can preferentially solubilize phosphatidylcholine from the canalicular rat liver plasma membrane. Methods: Subcellular fractions from rat liver and kidney were isolated with standardized procedures, incubated in vitro with taurocholate or 3-[(3-cholamidopropyl)dimethylammonio]-propane-1-sulphonate (CHAPS) and released phospholipids determined after centrifugation. Results: After incubation of canalicular (cLPM) and basolateral (blLPM) rat liver plasma membrane vesicles with 6 and 8 mM taurocholate, the proportion of phosphatidylcholine released was about two-fold higher as compared with its relative contribution to the overall lipid composition of the membranes. Quantitatively, this taurocholate-induced preferential phosphatidylcholine release was about four-fold higher in cLPM (117 nmol) as compared with blLPM (28 nmol). Comparison of membranes from different organs showed that increased sphingomyelin content reduced taurocholate-induced phosphatidylcholine release. Furthermore, phosphatidylcholine release from cLPM did not fit an inverse exponential relationship between membrane sphingomyelin content and phosphatidylcholine release from different starting material, indicating that cLPM is especially prone to taurocholate-induced phosphatidylcholine release. In contrast, in rat liver microsomes and kidney brush border membranes, taurocholate released phospholipids in proportion of their membrane contents, indicating an unspecific membrane solubilizing effect only. Similarly, CHAPS had an unselective lipid solubilizing effects in cLPM and blLPM. Conclusion: These results support the concept that the very last step of canalicular phospholipid secretion is mediated in vivo by bile salt-induced vesiculation of phosphatidylcholine-enriched microdomains from the outer leaflet of cLPM. [ABSTRACT FROM AUTHOR]

Various solutions of labeled precursors were absorbed by the cotyledons of etiolated Euphorbia lathyris L. seedlings. Incorporation of C into triterpenes from [2-C]mevalonic acid, [1-C]acetate, [3-C]pyruvate, [U-C]glyoxylate, [U-C]glycerol, [U-C]serine, [U-C]xylose, [U-C]glucose, and [U-C]sucrose was obtained. The [] triterpenes synthesized from [C] sugars were mainly of latex origin. [C]mevalonic acid was only involved in terpenoid synthesis outside the laticifers. Exogenously supplied glyoxylate, serine, and glycerol were hardly involved in lipid synthesis at all. The C-distribution over the various triterpenols was consistent with the mass distribution of these constituents in gas liquid chromatography when [C]sugars, [C]acetate, and [C]pyruvate were used. These precursors were supplied to the seedlings in the presence of increasing amounts of unlabeled substrates. The amount of substrate directly involved in lipid synthesis as well as the absolute triterpenol yield was calculated from the obtained [C]triterpenols. The highest yield was obtained in the sucrose incorporated seedlings, being 25% of the daily increase of latex triterpenes in growing seedlings. [ABSTRACT FROM AUTHOR]

The effects of pretreatment for 4 days with the hepatic microsomal enzyme inducers phenobarbital (8 mg/100 g body weight), spironolactone (20 mg/100 g body weight) and pregnenolone-16α-carbonitrile (7 mg/100 g body weight) on bile flow and bile lipid secretion have been compared in rats. Similar to phenobarbital, spironolactone and pregnenolone-16α-carbonitrile increased bile flow but did not alter bile salt excretion, indicating that these agents increased bile salt independent bile formation. This finding could be substantiated for spironolactone by studies of the relationship between bile salt excretion and bile flow during bile salt infusions. Whereas phenobarbital decreased cholesterol and phospholipid secretion to 39 and 49%, respectively, spironolactone and pregnenolone-16α-carbonitrile more than doubled cholestal excretion without influencing phospholipid output. As a consequence, marked differences in the effect on cholesterol saturation were observed: a decrease by phenobarbital and an increase following spironolactone and pregnenolone-16α-carbonitrile. The present studies demonstrate that different types of enzyme inducers may share certain effects on bile formation and differ in others. [ABSTRACT FROM AUTHOR]

Pistil development was studied in transgenic tobacco plants in which the stigma is ablated by expression of a stigma-specific cytotoxic gene. These plants offer a tool to investigate the process of differentiation of the secretory zone, in that cell death caused by barnase activity provides a marker to follow cell fate at high resolution. After fusion of the carpel walls in the region most distal from the ovary, the epidermal cells begin to divide in both wild-type and stigmaless plants. Divisions of the L1 layer of the pistil are immediately followed by the morphogenetic events that lead to three different cell types: rounded-angular cells showing an equal number of anti- and periclinal divisions, cells that are more oblong forming the transition zone, and the square cells of the transmitting tissue dividing mostly anticlinally with respect to the original carpel wall. In the stigmaless plants, cell death caused by the expression of STIG 1-barnase begins at stage −1 and proceeds gradually, but is always associated with round epidermal cells and with angular-rounded cells underneath them. Studies at the ultrastructural level show that cell death caused by barnase activity occurs first in solitary cells and gradually extends to groups of cells. In situ hybridizations using the STIG 1 RNA probe in wild-type pistils confirm these results. Most likely, the cells in which STIG 1 is expressed are those that have just differentiated into the secretory cell type. Our results indicate that the transition zone or neck is autonomously differentiated from the secretory zone and the transmitting tissue. Furthermore, our results indicate that in both wild-type and stigmaless pistils secretion of lipids most likely occurs through the plasmodesmata. This observation suggests that bulk transport can occur via plasmodesmata. [ABSTRACT FROM AUTHOR]

So far, it has been still unknown how liamocins are biosynthesized, regulated, transported and secreted. In this study, a highly reducing polyketide synthase (HR-PKS), a mannitol-1-phosphate dehydrogenase (MPDH), a mannitol dehydrogenase (MtDH), an arabitol dehydrogenase (ArDH) and an esterase (Est1) were found to be closely related to core biosynthesis of extracellular liamocins in Aureobasidium melanogenum 6-1-2. The HR-PKS was responsible for biosynthesis of 3,5-dihydroxydecanoic acid. The MPDH and MtDH were implicated in mannitol biosynthesis and the ArDH was involved in arabitol biosynthesis. The Est1 catalyzed ester bond formation of them. A phosphopantetheine transferase (PPTase) activated the HR-PKS and a transcriptional activator Ga11 activated expression of the PKS1 gene. Therefore, deletion of the PKS1 gene, all the three genes encoding MPDH, MtDH and ArDH, the EST1, the gene responsible for PPTase and the gene for Ga11 made all the disruptants (Δpks13, Δpta13, Δest1, Δp12 and Δg11) totally lose the ability to produce any liamocins. A GLTP gene encoding a glycolipid transporter and a MDR1 gene encoding an ABC transporter took part in transport and secretion of the produced liamocins into medium. Removal of the GLTP gene and the MDR1 gene resulted in a Δgltp1 mutant and a Δmdr16 mutant, respectively, that lost the partial ability to secrete liamocins, but which cells were swollen and intracellular lipid accumulation was greatly enhanced. Hydrolysis of liamocins released 3,5-dihydroxydecanoic acid, mannitol, arabitol and acetic acid. We proposed a core biosynthesis pathway, regulation, transport and secretion of liamocins in A. melanogenum., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Cell free diagnosis of cancer is one of the crucial fields in new generation of medical technology. In this regard, cancer detection based on coastal fluids secreted from the tissues (named as secretome) has attracted a lot of attention. Lipids are important macromolecules could be found with much higher concentrations in secretome of cancer tissues vs. normal ones. On the other hand, lipids are the main dielectric components of the secretome with respect to proteins and ions. Here for the first time we introduced an electrochemical lipidomics based on electrical impedance spectroscopy (EIS) of the secretomes to detect the cancerous samples due to the lipidic content of their secretions. The EIS sensor was fabricated by multiwall carbon nanotube (MWCNT) arrays as conductive and super hydrophobic materials to have great interactive surface with the lipidic content of the solution. Results of the tests on the secretions of more than 100 human biopsied breast tissues showed the promising match between the charge transfer resistance (R CT) of samples' secretions and pathological states of the tissues with meaningful boundary (up to 8 kΩ for normal and more than 13 kΩ for cancer samples). Mass spectroscopic analyses confirmed the higher content of lipids in cancer secretomes. Electrical lipidomics of the secretome shed new lights in cell free cancer diagnosis and could be applied as a complementary clinical approach in all of biopsy based diagnoses in future. • A novel cell free approach for rapid cancer diagnosis was developed. • The sensor has been based on electrochemical recording of the lipids secreted from cancer cells. • Results showed impressive diagnostic patterns between the EIS spikes of normal and tumor secretions of human samples. [ABSTRACT FROM AUTHOR]