Lipid-rich Cells Research Articles

From Skin and Gut to the Brain: The Infectious Journey of the Human Commensal Fungus Malassezia and Its Neurological Consequences.

The human mycobiome encompasses diverse communities of fungal organisms residing within the body and has emerged as a critical player in shaping health and disease. While extensive research has focused on the skin and gut mycobiome, recent investigations have pointed toward the potential role of fungal organisms in neurological disorders. Among those fungal organisms, the presence of the commensal fungus Malassezia in the brain has created curiosity because of its commensal nature and primary association with the human skin and gut. This budding yeast is responsible for several diseases, such as Seborrheic dermatitis, Atopic dermatitis, Pityriasis versicolor, Malassezia folliculitis, dandruff, and others. However recent findings surprisingly show the presence of Malassezia DNA in the brain and have been linked to diseases like Alzheimer's disease, Parkinson's disease, Multiple sclerosis, and Amyotrophic lateral sclerosis. The exact role of Malassezia in these disorders is unknown, but its ability to infect human cells, travel through the bloodstream, cross the blood-brain barrier, and reside along with the lipid-rich neuronal cells are potential mechanisms responsible for pathogenesis. This also includes the induction of pro-inflammatory cytokines, disruption of the blood-brain barrier, gut-microbe interaction, and accumulation of metabolic changes in the brain environment. In this review, we discuss these key findings from studies linking Malassezia to neurological disorders, emphasizing the complex and multifaceted nature of these cases. Furthermore, we discuss potential mechanisms through which Malassezia might contribute to the development of neurological conditions. Future investigations will open up new avenues for our understanding of the fungal gut-brain axis and how it influences human behavior. Collaborative research efforts among microbiologists, neuroscientists, immunologists, and clinicians hold promise for unraveling the enigmatic connections between human commensal Malassezia and neurological disorders.

MIER2/PGC1A elicits sunitinib resistance via lipid metabolism in renal cell carcinoma

IntroductionRenal cell carcinoma (RCC) is one of the most common malignant tumors of the urinary system and accounts for more than 90 % of all renal tumors. Resistance to targeted therapy has emerged as a pivotal factor that contributes to the progressive deterioration of patients with advanced RCC. Metabolic reprogramming is a hallmark of tumorigenesis and progression, with an increasing body of evidence indicating that abnormal lipid metabolism plays a crucial role in the advancement of renal clear cell carcinoma. ObjectivesClarify the precise mechanisms underlying abnormal lipid metabolism and drug resistance. MethodsBioinformatics screening and analyses were performed to identify hub gene. qRT-PCR, western blot, chromatin immunoprecipitation (ChIP) assays, and other biological methods were used to explore and verify related pathways. Various cell line models and animal models were used to perform biological functional experiments. ResultsIn this study, we identified Mesoderm induction early response 2 (MIER2) as a novel biomarker for RCC, demonstrating its role in promoting malignancy and sunitinib resistance by influencing lipid metabolism in RCC. Mechanistically, MIER2 facilitated P53 deacetylation by binding to HDAC1. Acetylation modification augmented the DNA-binding stability and transcriptional function of P53, while deacetylation of P53 hindered the transcriptional process of PGC1A, leading to intracellular lipid accumulation in RCC. Furthermore, Trichostatin A (TSA), an inhibitor of HDAC1, was found to impede the MIER2/HDAC1/P53/PGC1A pathway, offering potential benefits for patients with sunitinib-resistant renal cell cancer. ConclusionOur findings highlight MIER2 as a key player in anchoring HDAC1 and inhibiting PGC1A expression through the deacetylation of P53, thereby inducing lipid accumulation in RCC and promoting drug resistance. Lipid-rich RCC cells compensate for energy production and sustain their own growth in a glycolysis-independent manner, evading the cytotoxic effects of targeted drugs and ultimately culminating in the development of drug resistance.

Identification of a myofibroblast differentiation program during neonatal lung development.

Alveologenesis is the final stage of lung development in which the internal surface area of the lung is increased to facilitate efficient gas exchange in the mature organism. The first phase of alveologenesis involves the formation of septal ridges (secondary septae) and the second phase involves thinning of the alveolar septa. Within secondary septa, mesenchymal cells include a transient population of alveolar myofibroblasts (MyoFBs) and a stable but poorly described population of lipid-rich cells that have been referred to as lipofibroblasts or matrix fibroblasts (MatFBs). Using a unique Fgf18CreER lineage trace mouse line, cell sorting, single-cell RNA sequencing and primary cell culture, we have identified multiple subtypes of mesenchymal cells in the neonatal lung, including an immature progenitor cell that gives rise to mature MyoFB. We also show that the endogenous and targeted ROSA26 locus serves as a sensitive reporter for MyoFB maturation. These studies identify a MyoFB differentiation program that is distinct from other mesenchymal cell types and increases the known repertoire of mesenchymal cell types in the neonatal lung.

CD49f and CD146: A Possible Crosstalk Modulates Adipogenic Differentiation Potential of Mesenchymal Stem Cells.

The lack of appropriate mesenchymal stem cells (MSCs) selection methods has given the challenges for standardized harvesting, processing, and phenotyping procedures of MSCs. Genetic engineering coupled with high-throughput proteomic studies of MSC surface markers arises as a promising strategy to identify stem cell-specific markers. However, the technical limitations are the key factors making it less suitable to provide an appropriate starting material for the screening platform. A more accurate, easily accessible approach is required to solve the issues. This study established a high-throughput screening strategy with forward versus side scatter gating to identify the adipogenesis-associated markers of bone marrow-derived MSCs (BMSCs) and tonsil-derived MSCs (TMSCs). We classified the MSC-derived adipogenic differentiated cells into two clusters: lipid-rich cells as side scatter (SSC)-high population and lipid-poor cells as SSC-low population. By screening the expression of 242 cell surface proteins, we identified the surface markers which exclusively found in lipid-rich subpopulation as the specific markers for BMSCs and TMSCs. High-throughput screening of the expression of 242 cell surface proteins indicated that CD49f and CD146 were specific for BMSCs and TMSCs. Subsequent immunostaining confirmed the consistent specific expression of CD49f and CD146 and in BMSCs and TMSCs. Enrichment of MSCs by CD49f and CD146 surface markers demonstrated that the simultaneous expression of CD49f and CD146 is required for adipogenesis and osteogenesis of mesenchymal stem cells. Furthermore, the fate decision of MSCs from different sources is regulated by distinct responses of cells to differentiation stimulations despite sharing a common CD49f+CD146+ immunophenotype. We established an accurate, robust, transgene-free method for screening adipogenesis associated cell surface proteins. This provided a valuable tool to investigate MSC-specific markers. Additionally, we showed a possible crosstalk between CD49f and CD146 modulates the adipogenesis of MSCs.

Regulation of low-density lipoprotein on lipid metabolism in macrophages of large yellow croaker (Larimichthys crocea)

Low-density lipoprotein (LDL) is the main carrier of cholesterol transport in plasma, which participates in regulating lipid homeostasis. Studies in mammals have shown that high levels of LDL in plasma absorbed by macrophages trigger the formation of lipid-rich foam cells, leading to the development of atherosclerotic plaques. Although lipid-rich atherosclerosis-like lesions have been discovered in the aorta of several fish species, the physiological function of LDL in fish macrophages remains poorly understood. In the present study, LDL was isolated from the plasma of large yellow croaker (Larimichthys crocea), and mass spectrometry analysis identified two truncated forms of apolipoprotein B100 in the LDL protein profile. Transcriptomic analysis of LDL-stimulated macrophages revealed that differentially expressed genes (DEGs) were enriched in various pathways related to lipid metabolism, as confirmed by the fact that LDL increased total cholesterol and cholesteryl esters content. Meanwhile, the gene and protein expression levels of perilipin2 (PLIN2), a DEG enriched in the PPAR signaling pathway, were upregulated in response to LDL stimulation. Importantly, knocking down plin2 significantly attenuates LDL-induced cholesterol accumulation and promotes cholesterol efflux. Furthermore, the transcription factor PPARγ, which is upregulated in response to LDL stimulation, can enhance the promoter activity of plin2. In conclusion, this study suggests that LDL may upregulate plin2 expression through PPARγ, resulting in cholesterol accumulation in fish macrophages. This study will facilitate the investigation of the function of LDL in regulating lipid homeostasis in macrophages and shed light on the evolutionary origin of LDL metabolism in vertebrates.

Heterologous Expression of CFL1 Confers Flocculating Ability to Cutaneotrichosporon oleaginosus Lipid-Rich Cells.

Lipid extraction from microbial and microalgae biomass requires the separation of oil-rich cells from the production media. This downstream procedure represents a major bottleneck in biodiesel production, increasing the cost of the final product. Flocculation is a rapid and cheap system for removing solid particles from a suspension. This natural characteristic is displayed by some microorganisms due to the presence of lectin-like proteins (called flocculins/adhesins) in the cell wall. In this work, we showed, for the first time, that the heterologous expression of the adhesin Cfl1p endows the oleaginous species Cutaneotrichosporon oleaginosus with the capacity of cell flocculation. We used Helm's test to demonstrate that the acquisition of this trait allows for reducing the time required for the separation of lipid-rich cells from liquid culture by centrifugation without altering the productivity. This improves the lipid production process remarkably by providing a more efficient downstream.

CT of hemorrhagic adrenal adenomas: radiologic-pathologic correlation.

To describe the appearance of chronically hemorrhagic adenomas on adrenal protocol CT and correlate imaging with pathologic findings. Retrospective case series of adult patients with resected adrenal adenomas showing internal hemorrhage at histology. Seven of nine patients underwent pre-operative adrenal protocol CT and 2/7 underwent unenhanced CT with portal venous phase CT. Two abdominal radiologists in consensus assessed the CT images for the presence of calcifications, macroscopic fat, cystic/necrotic appearance, and the presence, pattern, and percent nodule volume of areas < 10 HU on unenhanced CT. Absolute washout was calculated using a large ROI, and ROIs on the highest and lowest attenuating regions on the portal venous phase. Mean adenoma length was 4.9cm. All adenomas had areas measuring < 10 HU on unenhanced CT, ranging from < 20 to > 80% nodule volume. Calcifications were present in 4/9 adenomas and gross fat in 4/9 on CT. Of the seven cases with adrenal protocol CT, the absolute washout was < 60% in 5/7 using the large ROI, 5/7 using the low attenuation ROI, and 7/7 using the high attenuation ROI. At histology, all nine cases had microscopic evidence of hemorrhage, lipid rich adenoma cells, and fibrosclerosis. Myelolipomatous changes were identified in 4/9 cases, with the remaining five cases showing lipomatous metaplasia without a myeloid component. Chronically hemorrhagic adrenal adenomas demonstrated variable areas < 10 HU on unenhanced CT corresponding to lipid rich adenoma cells. Absolute washout was most often < 60%, hypothesized to be due to fibrosclerosis within the adenomas.

S3004 Giant Hepatic Hemangioma: Dulling the Blade of Ockham’s Razor

Introduction: Ockham’s razor suggests the simplest diagnosis is likely correct. Hickman’s dictum says multiple diagnoses may occur in one patient. We present a patient with abdominal pain due to giant hepatic hemangioma, as well as secondary hypertension due to adrenal mass masked by giant hepatic hemangioma. Case Description/Methods: A 38-year-old female with abnormal uterine bleeding due to cervical ectropion developed right-sided abdominal pain and hypertension two years prior to presentation. She reported increased facial hair and acne which improved on spironolactone. Other medications included amlodipine, irbesartan, and progestin intrauterine device. Physical exam showed normal vital signs, facial hirsutism and acne, and abdominal distension with tenderness to palpation in the right upper and lower quadrants. Laboratory studies including serum adrenocorticotropic hormone, cortisol, aldosterone, and renin, and plasma and urine norepinephrine, epinephrine, and dopamine levels were unremarkable (Table 1). Magnetic resonance imaging with intravenous gadobutrol showed a 13.9 x 14.1 x 20.2 centimeter (cm) mass replacing the right hepatic lobe, compatible with hemangioma (Figure 1A), normal left adrenal gland, and patent renal vessels. The right adrenal gland was not visualized. She underwent right hepatic resection of hemangioma with enucleation (Fig. 1B). This allowed visualization and palpation of the right adrenal gland, revealing a 3 cm mass on the inferior portion (Fig. 1C). Right adrenalectomy was performed. Pathology revealed a well-demarcated vascular neoplasm consistent with liver hemangioma. Right adrenalectomy specimen revealed nodular cortical hyperplasia (Fig. 1D), correlating with aldosterone hypersecretion. Following surgery, she was normotensive without medication, and hirsutism resolved. Hepatic hemangioma alone is not known to cause hypertension. Her hypertension was attributed to adrenal hyperplasia, with potential elevation of serum aldosterone masked by spironolactone. Discussion: Our patient presented with concurrent onset of abdominal pain and hypertension, secondary to two distinct, unrelated entities – hepatic hemangioma and unilateral adrenal hyperplasia. This unique case of a giant hepatic hemangioma obscuring radiographic views of an adrenal mass reminds us that patients can have as many diseases as they please.Figure 1.: A, MRI of the abdomen with intravenous gadobutrol shows 13.9 x 14.1 x 20.2 cm hemangioma replacing the right lobe. B, Giant liver hemangioma specimen. C, Intraoperative photo of right adrenal mass. D, Adrenal cortical hyperplasia with enlarged lipid-rich cells (black open arrow) and lipid-depleted cells (black solid arrow. Hematoxylin and Eosin stain, magnification 200X. Table 1. - Laboratory Data Serologies Patient Values Reference Range White blood cell count (K/μL) 4.38 3.98-10.04 Hematocrit (%) 10.9 11.2-15.7 Hemoglobin (g/dL) 33.3 34.1-44.9 Platelet count (K/μL) 185 173-369 Sodium (mmol/L) 139 136-145 Potassium (mmol/L) 3.7 3.4-5.1 Chloride (mmol/L) 101 98-107 Total CO2 (mmol/L) 26 22-29 Urea nitrogen (mg/dL) 14 6-20 Creatinine (mg/dL) 0.60 0.51-0.95 Glucose (mg/dL) 88 74-99 Alkaline phosphatase (U/L) 66 35-105 Alanine aminotransferase (U/L) 19 0-33 Aspartate aminotransferase (U/L) 16 0-32 Bilirubin, total (mg/dL) 0.5 0.0-1.2 Bilirubin, direct (mg/dL) < 0.2 0.0-0.3 Activated partial thromboplastin time (sec) 31.2 25.3-37.3 Prothrombin time (sec) 14.9 11.6-15.2 Hemoglobin A1c (%) 5.6 4.0-6.0 Albumin (g/dL) 4.2 3.5-5.2 Calcium (mmol/L) 2.30 2.15-2.55 Magnesium (mmol/L) 0.81 0.66-1.07 Phosphorus (mg/dL) 3.1 2.5-4.5 Thyroid stimulating hormone (mcIU/mL) 1.24 0.27-4.2 Cortisol, serum 5.9 5.0-25.0 Follicle stimulating hormone (U/L) 3.5 < 21 Luteinizing hormone (U/L) 3.9 < 77 Estradiol, serum (pg/mL) 212.1 15-350 Dehydroepiandrosterone sulfate (mcg/mL) 0.85 0.35-4.30 Norepinephrine, plasma (pg/mL) 286 112-750 Epinephrine, plasma (pg/mL) 21 0-50 Dopamine, plasma (pg/mL) < 25 0-29 Fractionated normetanephrine (pg/mL) 70 18-112 Fractionated metanephrine (pg/mL) 14 12-61 Aldosterone, serum (ng/dL) 6.6 < 44.5 Renin, plasma (ng/mL/h) 0.8 0.6-4.3 Adrenocorticotropic hormone (pg/mL) 8.2 5.0-46.0 Alpha fetoprotein (ng/mL) 3.7 0.6-6.6 Sex hormone binding globulin (nmol/L) 100 18-114 Testosterone, total (ng/dL) < 20 < 81 Vitamin D, 1,25-dihydroxy (pg/mL) 64 20-79 Parathyroid hormone, intact (pg/mL) 48.1 15-65 Serotonin, whole blood (ng/mL) 87 < 330 Chromogranin A (ng/mL) 38 < 93 Human chorionic gonadotropin (IU/L) < 1 0-5 Epinephrine, urine (mcg/24h) 8.3 < 21 Norepinephrine, urine (mcg/24h) 61 15-80 Dopamine, urine (mcg/24h) 250 65-400

Total ice content and lipid saturation determine adipose tissue cryolipolysis by injection of ice-slurry.

Cryolipolysis uses tissue cooling to solidify lipids, preferentially damaging lipid-rich cells. Topical cooling is popular for the reduction of local subcutaneous fat. Injection of biocompatible ice-slurry is a recently introduced alternative. We developed and verified a quantitative model that simulates the heat exchange and phase changes involved, offering insights into ice-slurry injection for treating subcutaneous fat. Finite element method was used to model the spatial and temporal progression of heat transfer between adipose tissue and injected ice-slurry, estimating dose-response relationships between properties of the slurry and size of tissue affected by cryolipolysis. Phase changes of both slurry and adipose tissue lipids were considered. An in vivo swine model was used to validate the numerical solutions. Oils with different lipid compositions were exposed to ice-slurry in vitro to evaluate the effects of lipid freezing temperature. Microscopy and nuclear magnetic resonance (NMR) were performed to detect lipid phase changes. A ball of granular ice was deposited at the injection site in subcutaneous fat. Total injected ice content determines both the effective cooling region of tissue, and the duration of tissue cooling. Water's high latent heat of fusion enables tissue cooling long after slurry injection. Slurry temperature affects the rate of tissue cooling. In swine, when 30 ml slurry injection at -3.5°C was compared to 15 ml slurry injection at -4.8°C (both with the same total ice content), the latter led to almost twice faster tissue cooling. NMR showed a large decrease in diffusion upon lipid crystallization; saturated lipids with higher freezing temperatures were more susceptible to solidification after ice-slurry injection. Total injected ice content determines both the volume of tissue treated by cryolipolysis and the cooling duration after slurry injection, while slurry temperature affects the cooling rate. Lipid saturation, which varies with diet and anatomic location, also has an important influence.

Complex meristematic activity induced by Eucecidoses minutanus on Schinus engleri turns shoots into galls.

Gall-inducing organisms change the development of their host plant organs, resulting in ontogenetic patterns not observed in the non-galled plants. Distinct taxa induce galls on Schinus spp., manipulating meristematic patterns in the host plant in distinct ways. Here we report ontogenetic novelties induced in the lateral buds of S. engleri by Eucecidoses minutanus, a Cecidosidae, whose galls have been poorly understood. The anatomy, histochemistry, and histometry of galls in distinct phases of development, non-galled buds, and stems of Schinus engleri were analyzed in parallel with the instars of E. minutanus to detail the morphogenetic changes in the host with each larval stage. Ontogenetic phases of the galls were intricately associated with larval development. First and second-instar larvae induced pericycle and pith cells to dedifferentiate into the gall inner meristem, where hyperplasia and cell hypertrophy characterized the growth and development phase of the gall. The innermost layers were lipid-rich nutritive cells that lined the larval chamber. Additional vascular bundle rows were produced in young galls. Third and fourth instar-larvae were associated with the gall maturation phase: centripetal lignification of the outer parenchyma cell layers, epidermal stratification, and activation of a cambium-like meristem (CLM). The CLM activity resulted in new layers of nutritive cells that differentiated inward as the first layers of nutritive cells were consumed by E. minutanus larvae, and, also, in more parenchyma cell layers that formed outward. All tissues between the innermost layer of nutritive tissue that surround the gall chamber and the outermost layer of the dermal system that externally covers the gall form the gall wall, and increased in thickness until the end of gall maturation. E. minutanus induces a structurally complex globoid stem gall, modifying all host plant tissues and stimulating a novel meristematic pattern in S. engleri. The gall developmental stages are each related to specific gall-inducing instars, as gall development progresses according to the development of E. minutanus.

Evaluation of Plaque Burden and Lipid Content in Atherosclerotic Plaques.

Atherosclerosis is a chronic inflammatory disease characterized by the formation of lipid-rich, fibrous plaques within the arterial wall of medium and large arteries. Plaques prone to rupture are typically rich in lipids and pro-inflammatory markers. Cells within the plaque can take up lipids via different mechanisms leading to the formation and accumulation of lipid-rich foam cells, a key hallmark of the disease. Evaluation of plaque burden and lipid content is hence important to determine disease progression and severity. This chapter describes the most commonly used staining methods that enable visualization and analysis of mouse atherosclerotic plaques. These methods include en face preparation of mouse aorta, and staining sections of arteries using hematoxylin and eosin, Oil Red O, and Masson's Trichrome.

Development of mutant microalgae that accumulate lipids under nitrate-replete conditions

Microalgal biofuels have attracted global attention as an alternative energy source to fossil fuels. Microalgae generally accumulate triacylglycerols (lipids) under nitrogen-deficient conditions; however, cell growth is highly suppressed under these conditions, subsequently reducing lipid productivity. Therefore, microalgae that can accumulate lipids even under nitrogen-replete conditions are needed to realize the one-step nutrient-replete cultivation with improved lipid productivity. Thus, the aim of this study was to develop a method for screening mutant microalgae showing high lipid accumulation in the presence of a nitrogen source. Mutant cells were generated by irradiating the oleaginous green microalga Chlamydomonas sp. KOR1 with carbon-ion beams, followed by cultivation under nitrate-replete conditions, and then subjected to fluorescence-activated cell sorting-based screening for detecting lipid-rich cells. By repeatedly performing the sequential procedures of cultivation and selection, the strains KAC1710 and KAC1801, which showed high lipid accumulation under nitrate-replete conditions, were obtained. In the presence of abundant nitrate, these mutants formed a large number of lipid droplets in the cells, and achieved 1.5-fold (KAC1710) and 2.1-fold (KAC1801) higher lipid content than KOR1. Metabolome analysis revealed that the pool size of pyruvate was significantly increased in KAC1710 and KAC1801, consistent with the activation of lipid synthesis under nitrate-replete conditions. In conclusion, this study reports the use of a novel nitrogen-based screening method for developing mutant microalgae that accumulate lipids in the presence of a nitrogen source, and found that an increase in pyruvate pool size is an important metabolic phenomenon associated with lipid production.

Macrophages in Atherosclerosis, First or Second Row Players?

Macrophages represent a cell type that has been widely described in the context of atherosclerosis since the earliest studies in the 17th century. Their role has long been considered to be preponderant in the onset and aggravation of atherosclerosis, in particular by participating in the establishment of a chronic inflammatory state by the release of pro-inflammatory cytokines and by uncontrolled engorgement of lipids resulting in the formation of foam cells and later of the necrotic core. However, recent evidence from mouse models using an elegant technique of tracing vascular smooth muscle cells (VSMCs) during plaque development revealed that resident VSMCs display impressive plastic properties in response to an arterial injury, allowing them to switch into different cell types within the plaque, including mesenchymal-like cells, macrophage-like cells and osteochondrogenic-like cells. In this review, we oppose the arguments in favor or against the influence of macrophages versus VSMCs in all stages of atherosclerosis including pre-atherosclerosis, formation of lipid-rich foam cells, development of the necrotic core and the fibrous cap as well as calcification and rupture of the plaque. We also analyze the relevance of animal models for the investigation of the pathophysiological mechanisms of atherosclerosis in humans, and discuss potential therapeutic strategies targeting either VSMCs or macrophage to prevent the development of cardiovascular events. Overall, although major findings have been made from animal models, efforts are still needed to better understand and therefore prevent the development of atherosclerotic plaques in humans.

Exosomes and Atherogenesis.

Myocardial infarction and ischemic stroke are the leading causes of mortality worldwide. Atherosclerosis is their common pathological foundation. It is known that atherosclerosis is characterized by endothelial activation/injury, accumulation of inflammatory immune cells and lipid-rich foam cells, followed by the development of atherosclerotic plaque. Either from arterial vessel wall or blood circulation, endothelial cells, smooth muscle cells, macrophages, T-lymphocytes, B-lymphocytes, foam cells, and platelets have been considered to contribute to the pathogenesis of atherosclerosis. Exosomes, as natural nano-carriers and intercellular messengers, play a significant role in modulation of cell-to-cell communication. Under physiological or pathological conditions, exosomes can deliver their cargos including donor cell-specific proteins, lipids, and nucleic acids to target cells, which in turn affect the function of the target cells. In this review, we will describe the pathophysiological significance of various exosomes derived from different cell types associated with atherosclerosis, and the potential applications of exosome in clinical diagnosis and treatment.

FABP5 Is a Sensitive Marker for Lipid-Rich Macrophages in the Luminal Side of Atherosclerotic Lesions.

Lipid-rich macrophages in atherosclerotic lesions are thought to be derived from myeloid and vascular smooth muscle cells. A series of studies with genetic and pharmacological inhibition of fatty acid binding protein 4 (FABP4) and FABP5 and bone marrow transplant experiments with FABP4/5 deficient cells in mice have demonstrated that these play an important role in the development of atherosclerosis. However, it is still uncertain about the differential cell-type specificity and distribution between FABP4- and FABP5-expressing cells in early- and late-stage atherosclerotic lesions. In this study, we first explored spatial distribution of FABP4/5 in atherosclerotic lesions in apolipoprotein E deficient (ApoE-/-) mice. FABP4 was only marginally detected in early and advanced lesions, whereas FABP5 was abundantly expressed in these lesions. In advanced lesions, the FABP5-positive area was mostly restricted to the foam cell layer adjacent to the lumen above collagen and elastic fibers with a high signal/noise ratio. Oil red O (ORO) staining revealed that FABP5-positive cells were lipid-rich in early and advanced lesions. Together, most of lipid-rich FABP5-positive cells reside adjacent to the lumen above collagen and elastic fibers. We next studied involvement of FABP5 in lesion formation of atherosclerosis using ApoE-/- FABP5-/- mice. However, deletion of FABP5 did not affect the development of atherosclerosis. These findings, along with previous reports, suggest a novel notion that FABP5 is a sensitive marker for bone marrow-derived lipid-rich macrophages in the luminal side of atherosclerotic lesions, although its functional significance remains elusive.

Cobalt Chloride Induces Macrophage Foam Cell Formation: A Chemical Hypoxia Model for Anti-Atherosclerotic Drug Screening.

Macrophages would engulf circulating oxidized (ox)- low-density lipoprotein and form lipid droplet-laden foam cells. Macrophage foam cells are considered an important therapeutic target of atherosclerosis. The aim of the study was to investigate a hypoxic foam cell model for anti-atherosclerotic drug screening using the chemical hypoxia-mimicking agent cobalt chloride (CoCl2). The oil red O stating results showed that treatment with CoCl2 could induce lipid accumulation and lead to cell transformation to spindle-shaped and lipid-rich foam cells in RAW 264.7 macrophages. Incubation with 150 μM CoCl2 for 24 h significantly increased the area of intracellular lipid droplets in macrophages, compared with the control group. Our findings indicate that CoCl2-triggered macrophage foam cells should be a potential in vitro hypoxia model for atherosclerosis drug discovery.

A benzothiadiazole-quinoline hybrid sensor for specific bioimaging and surgery procedures in mice

The work describes the design, synthesis, characterization and biological application (bioimaging) of a new hybrid and fluorescent 2,1,3-benzothiadiazole-quinoline sensor. Both photophysical data of the designed compound and DFT calculations have indicated its high stability and have pointed to its possible efficient application as a bioprobe for bioimaging experiments. The new sensor has been designed to display lipophilic character being capable of selectively stain lipid bodies independent whether live and fixed cancer cells were used. The fluorescent sensor has also been tested as a specific bioimaging marker for C. elegans (complex multicellular model) and has showed far better selectivity in the worm than the commercially available BODIPY. A live mice has been submitted to a surgery procedure and the adipocytes (lipid-rich) cells have been differentiated from standard cells. Experiments using high fat fed mice have enabled a high-resolution 3D image reconstruction. Also, in another surgery procedure, hepatocyte cells could have been successfully followed in the blood flow using the designed sensor in a multistaining procedure alongside with a commercially available blue emitting nuclei marker.

Data set for reporting of carcinoma of the adrenal cortex: explanations and recommendations of the guidelines from the International Collaboration on Cancer Reporting

Complete resection of adrenal cortical carcinoma (ACC) with or without adjuvant therapy offers the best outcome. Recurrence is common, and in individual cases, the long-term outcome is difficult to predict, making it challenging to personalize treatment options. Current risk stratification approaches are based on clinical and conventional surgical pathology assessment. Rigorous and uniform pathological assessment may improve care for individual patients and facilitate multi-institutional collaborative studies. The International Collaboration on Cancer Reporting (ICCR) convened an expert panel to review ACC pathology reporting. Consensus recommendations were made based on the most recent literature and expert opinion. The data set comprises 23 core (required) items. The core pathological features include the following: diagnosis as per the current World Health Organization classification, specimen integrity, greatest dimension, weight, extent of invasion, architecture, percentage of lipid-rich cells, capsular invasion, lymphatic invasion, vascular invasion, atypical mitotic figures, coagulative necrosis, nuclear grade, mitotic count, Ki-67 proliferative index, margin status, lymph node status, and pathological stage. Tumors were dichotomized into low-grade (<20 mitoses per 10mm2) and high-grade (>20 mitoses per 10mm2) ones. Additional noncore elements that may be useful in individual cases included several multifactorial risk assessment systems (Weiss, modified Weiss, Lin-Weiss-Bisceglia, reticulin, Helsinki, and Armed Forces Institute of Pathology scores/algorithms). This data set is now available through the ICCR website with the hope of better standardizing pathological assessment of these relatively rare but important malignancies.

Mitochondrial Fatty Acid Oxidation Is Not Vital for Melanoma Cell Proliferation and Migration

Abstract Objectives Melanoma, the 5th most common cancer in the US, is the most aggressive form of skin cancer. Excess adiposity is associated with an increased risk of melanoma in males, and a high-fat diet promotes melanoma tumor growth in mice. Our group found that lipid droplet (LD) content increases with melanoma cell aggressiveness and that fatty acid uptake inhibition reduces cell proliferation and migration; however, the function of these lipids in melanoma is unknown. Since lipids can be used to produce energy by fatty acid oxidation (FAO), we sought to determine whether melanoma cells were using FAO to maintain aggressiveness. Methods Gene expression microarray was used to identify differences in gene expression, which was confirmed by RT-qPCR. WM983A, WM983B, 1205Lu and A375 human melanoma cells were treated with 5 µM Etomoxir and cell proliferation, migration, and respiration were quantified using the Quant-iT™ PicoGreen™ dsDNA Assay Kit, Boyden Chamber transwell migration, and Seahorse XFe96 Flux Analyzer, respectively. Results The most significantly altered genes expressed between four invasive, lipid-rich and four non-invasive, lipid-poor melanoma cells pertained to fatty acid metabolism. Carnitine palmitoyltransferase 1A (CPT1a), the rate limiting enzyme in mitochondrial FAO (mFAO), mRNA was increased in lipid-rich cells compared to the lipid-poor cells (p &amp;lt; 0.05, n = 6–7), indicating that melanoma cells may use LD lipids for mFAO. To determine the importance of mFAO to melanoma cells, we inhibited mFAO with etomoxir, an irreversible CPT1 inhibitor. Treatment of lipid-rich 1205Lu and A375 cells or lipid-poor WM983A and WM983B cells with etomoxir for 4 days had no effect on cell proliferation. Extracellular flux analysis of cells with or without etomoxir showed no difference in ATP production, indicating that melanoma cells do not use mFAO to generate energy under normal conditions. In motile cells, etomoxir reduced lipid-rich A375 cell migration by 8.8% (p = 0.05, n = 2). Conclusions Lipids play a role in melanoma aggressiveness; however, our results indicate that mFAO of lipids is not vital to melanoma cell proliferation or migration. Further studies are required to understand the implication of excess adiposity and circulating lipids in melanoma development and progression. Funding Sources Institutional Department Fund

SUN-157 Delayed Diagnosis of Cushings Syndrome: Hiding in Plain Sight!

BACKGROUND: Endogenous Cushing’s syndrome (CS) is rare, with an incidence of 0.7-2.4/million people/year.1 It should be considered in individuals with diabetes (DM), hypertension (HTN), osteoporosis or electrolyte abnormalities.2 We present a patient with DM2 and persistent hypokalemia for 10 years found to have ACTH-independent Cushing’s disease as the cause of metabolic syndrome. Case: 62 y.o. M admitted with abdominal pain, a history of DM2 (2012) on Metformin 500 mg BID, HTN (2010) and on ramipril 10 mg qd, with chronic lymphedema on furosemide 20 mg qd. He reported 3-inch height loss. On exam, had facial plethora, moon facies, supraclavicular fullness, thick violaceous abdominal striae and kyphosis. Past history was significant for abdominal/leg cellulitis, muscle weakness, difficult to heal wounds, easy bruising and recurrent hospitalizations for hypokalemia, despite being on KCl up to 80 mEq/d for 10 years. Labs showed AM cortisol at 22.6 ug/dL, ACTH <5 pg/mL, 24-hour urine free cortisol of 523 ug/d (normal < 60 ug/d). AM cortisol after 1 mg overnight dexamethasone suppression was 36.8 g/dL, ACTH <5 pg/mL. CT abdomen showed right adrenal nodule, 4.0 x 3.5 cm with density of 22 HU. MRI showed lipid-poor adenoma measuring 3.9 x 3.5 cm, raising concern for adrenocortical malignancy. Patient underwent right adrenalectomy. Pathology was consistent with benign adenoma showing no nuclear pleomorphism, lipid rich cells containing eosnophillic cytoplasm. Mib-1 stain <1% cells and positive inhibin. He was maintained on steroids post op due to concern about adrenal insufficiency. Hypokalemia, DM and lymphedema resolved completely 4 months post op with weight loss of ~30 pounds. HbA1c improved to 5.1%, metformin was stopped and he was maintained on Carvedilol 6.125 mg BID for HTN. He was diagnosed with osteoporosis with T score -4.0 at mean femoral neck, -2.9 for mean total hip with non-diagnostic spine. He had multi-level chronic compression fractures of the mid-thoracic spine. Conclusion: Delayed diagnosis of CS, as occurred in our patient, can result in detrimental consequences such as life threatening electrolyte abnormalities, cardiovascular events, fractures and premature death.1 Identifying CS can be challenging as clinical presentation is variable. 2,3 Early recognition, diagnosis and control of CS is crucial to decrease morbidity and mortality. Our patient demonstrated rapid resolution of DM, hypokalemia and lymphedema after surgery; however, prolonged exposure to endogenous cortisol resulted in compression fractures and osteoporosis requiring follow up treatment.