scholarly journals THE TRANSDUCTION OF CELLULAR MEMBRANE PROTEINS AND STRUCTURAL CELL MEMBRANE ALTERATIONS INDUCED BY EXOGENOUS ENERGY WAVES ENERGIZING LIPID DROPLETS AS THE PROPOSED UNDERLYING MECHANISM IN VERY INTENSIVE PRESSURE PULSES TREATMENTS CLAIMS TO A CANCER CURE

2019 ◽  
Vol 7 (5) ◽  
pp. 299-310
Author(s):  
Abraham A. Embi BS ◽  
Steve Haltiwanger
Keyword(s):  
Lipid Droplets ◽  
Underlying Mechanism ◽  
Cellular Membrane ◽  
In Vitro Experiments ◽  
Intracellular Lipid ◽  
Structural Cell ◽  
Available Energy ◽  
Video Recordings ◽  
Pressure Pulses

The purpose of this manuscript is to propose a mechanism for a cancer cure claim resulting from exogenous stimulation of cancer tumors by very intense pressure pulses (VIPPs) treatments from commercially available energy hardware (CellSonic). Could it be that exogenous continuous pulsating waves alter the cellular lipid bilayer; and this in turn also influence intracellular cell signaling? In Vitro experiments are presented supporting the above-stated thesis. The evidence will show via in vitro experiments how trapping energy from bursting oxygen bubbles induces static electricity discharges up to causing luminescence of intracellular lipid droplets. Figures and video recordings documenting the above-mentioned phenomena are presented. In summary, proposed is a mechanism explaining a cancer cure claim via VIPPs. 1 AAE: Idealized, designed wrote manuscript and conducted in vitro experiments possibly demonstrating a mechanism for VIPPs cancer cure. 2 SH: Added theoretical principles in the discussion supporting the proposed VIPPs mechanism to cancer cure.

scholarly journals SUN-564 ATG7 Overexpression Results in Reduction of Hepatocellular Lipid Content in Vitro

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Federica Tavaglione ◽  
Guido Baselli ◽  
Ester Ciociola ◽  
Umberto Vespasiani Gentilucci ◽  
Luca Valenti ◽  
...  
Keyword(s):  
Liver Disease ◽  
Lipid Content ◽  
Lipid Droplets ◽  
Low Frequency ◽  
Underlying Mechanism ◽  
Loss Of Function ◽  
Wild Type ◽  
Intracellular Lipid ◽  
Heparg Cells

Abstract Abstract: Non-alcoholic fatty liver disease (NAFLD) is currently the most common liver disease worldwide, paralleling the epidemic of obesity and type 2 diabetes. Despite the high prevalence of NAFLD, only a minority of patients progress to NASH and advanced fibrosis. The reasons for this inter-individual variability are not completely understood but can be partially accounted for by genetic risk factors (1). Although several common genetic variants associated with liver disease have been identified, there is still a proportion of NAFLD heritability that remains unknown. The rare rs143545741 C>T variant in the autophagy related 7 (ATG7) gene (P426L) has been associated with a higher risk of progressive NAFLD (2). Interestingly, ATG7 encodes a E1-like ubiquitin activating enzyme which is involved in hepatic lipophagy (3). We hypothesized that the unknown heritability of NAFLD might be partially explained by rare genetic variants, therefore not identified in the GWAS studies. Moreover, we assumed that loss-of-function variants in ATG7 might confer an increased susceptibility to NAFLD by reducing autophagic catabolism of lipid droplets in the liver. To examine the underlying mechanism of the low-frequency V471A variant and the rare T86I, L127I, Q170E, and P426L variants in ATG7, we performed in vitro experiments of HepaRG cells overexpressing the human V5-tagged ATG7. We observed a reduction in intracellular lipid content in HepaRG cells overexpressing the ATG7 wild type and the 86I mutant protein (p=0.029, n=4) but not the 127I, 170E, 426L, and 471A mutant proteins. Cells with the ATG7 127I, 170E, 426L, and 471A mutants had higher intracellular lipid content compared to cells overexpressing the wild type protein (p=0.029, n=4). Our data suggested that the low-frequency V471A variant and the rare L127I, Q170E, and P426L variants in ATG7 are loss-of-function, resulting in defective lipophagy, reduced hepatocellular lipid droplets turnover, and excessive lipid accumulation. More experiments are needed to clarify the underlying mechanism of the T86I variant. In conclusion, we highlighted a role for ATG7 in reducing hepatocellular lipid content. Furthermore, we provided evidence showing non-synonymous variants in ATG7 increase the risk of NAFLD and that these variants are loss-of-function. We speculate that ATG7 might be a new susceptibility risk genetic locus for liver disease development and progression. References: (1) Eslam et al. J Hepatol. 2018;68(2):268–279. (2) Baselli et al. The Liver Meeting 2018 - AASLD. Hepatology. October 2018. Volume 68, Issue S1. (3) Martinez-Lopez and Singh. Annu Rev Nutr. 2015;35:215–37.

scholarly journals Exon 9-deleted CETP inhibits full length-CETP synthesis and promotes cellular triglyceride storage

2020 ◽  
Vol 61 (3) ◽  
pp. 422-431 ◽  
Author(s):  
Lahoucine Izem ◽  
Yan Liu ◽  
Richard E. Morton
Keyword(s):  
Lipid Metabolism ◽  
Lipid Droplets ◽  
Full Length ◽  
Intracellular Lipid ◽  
Transfer Protein ◽  
Lipid Phenotype ◽  
Exon 9 ◽  
And Storage

Cholesteryl ester transfer protein (CETP) exists as full-length (FL) and exon 9 (E9)-deleted isoforms. The function of E9-deleted CETP is poorly understood. Here, we investigated the role of E9-deleted CETP in regulating the secretion of FL-CETP by cells and explored its possible role in intracellular lipid metabolism. CETP overexpression in cells that naturally express CETP confirmed that E9-deleted CETP is not secreted, and showed that cellular FL- and E9-deleted CETP form an isolatable complex. Coexpression of CETP isoforms lowered cellular levels of both proteins and impaired FL-CETP secretion. These effects were due to reduced synthesis of both isoforms; however, the predominate consequence of FL- and E9-deleted CETP coexpression is impaired FL-CETP synthesis. We reported previously that reducing both CETP isoforms or overexpressing FL-CETP impairs cellular triglyceride (TG) storage. To investigate this further, E9-deleted CETP was expressed in SW872 cells that naturally synthesize CETP and in mouse 3T3-L1 cells that do not. E9-deleted CETP overexpression stimulated SW872 triglyceride synthesis and increased stored TG 2-fold. Expression of E9-deleted CETP in mouse 3T3-L1 cells produced a similar lipid phenotype. In vitro, FL-CETP promotes the transfer of TG from ER-enriched membranes to lipid droplets. E9-deleted CETP also promoted this transfer, although less effectively, and it inhibited the transfer driven by FL-CETP. We conclude that FL- and E9-deleted CETP isoforms interact to mutually decrease their intracellular levels and impair FL-CETP secretion by reducing CETP biosynthesis. E9-deleted CETP, like FL-CETP, alters cellular TG metabolism and storage but in a contrary manner.

scholarly journals FACILE IN VITRO GLASS SLIDE LIGHT MICROSCOPY METHOD USING TETRACYCLINE TO VISUALIZE A REPETITIVE PATTERN IN AERIAL PLANT ROOT TIPS FILAMENTOUS NETWORK

2021 ◽  
Vol 8 (12) ◽  
pp. 155-166
Author(s):  
Abraham A. Embi
Keyword(s):  
Light Microscopy ◽  
Plant Root ◽  
Plant Roots ◽  
In Vitro Experiments ◽  
Root Tips ◽  
Video Recordings ◽  
Glass Slides ◽  
Hair Roots ◽  
Correct Conclusion

The main purpose of this manuscript is to introduce a facile light microscopy methodology to visualize plant roots filaments. In a previous manuscript in vitro experiments on freshly plucked human hair roots documented the commonly used antibiotic Tetracycline (TE) deleterious effect on soft tissue, severe enough to allow for visualization of an underlying filamentous skeleton. In this manuscript, TE was also evaluated in a similar fashion of in vitro experiments, this time aerial plant roots were immersed in liquid Tetracycline. Images and video recordings are presented where plant aerial root tissue cells appeared to interact with Tetracycline, thus allowing for exposure of an underlying filamentous network. These filaments were documented undergoing biosorption of Tetracycline, thus indicating a probable cellulose base. It is emphasized that a literature search showed similar, albeit visually different displays of roots filaments obtained by using a Scanning Electron Microscopy. The method herein introduced could be an adjunct to existing established methodology in root function research. Two salient advantages are identified, firstly that the essential minimal material and equipment is limited to a light microscope, glass slides, chosen biological material, water and powder Tetracycline. Secondly, the speed in obtaining results would offer researchers a preliminary or perhaps a final correct conclusion.

140 EXPRESSION OF PERILIPIN IN PORCINE OOCYTES AND CUMULUS CELLS AT DIFFERENT STAGES OF IN VITRO MATURATION

2016 ◽  
Vol 28 (2) ◽  
pp. 200
Author(s):  
A. Oh ◽  
J.-X. Jin ◽  
S. Lee ◽  
G. A. Kim ◽  
B. C. Lee
Keyword(s):  
Messenger Rna ◽  
Lipid Droplets ◽  
In Vitro Maturation ◽  
Statistical Significance ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Intracellular Lipid ◽  
Germinal Vesicle Breakdown ◽  
Interacting Protein

Perilipin, one of the perilipin adipophilin tail-interacting protein of 47 kDa (PAT) family, has been found to coat the surface of intracellular lipid droplets. It limits the interaction of lipases with intracellular lipid droplets and is involved in the formation and regulation of lipids in various kinds of cells. However, little is known about the effect of perilipin on porcine oocytes and cumulus cells. Therefore, this study aimed to detect the expression of perilipin1 (PLIN1), perilipin2 (PLIN2), perilipin3 (PLIN3), and perilipin4 (PLIN4) in porcine oocytes and cumulus cells at 4 stages of in vitro maturation (IVM) by quantitative real-time PCR (RT-qPCR). Germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), and metaphase II (MII) stage were found to occur predominantly at 18 to 24 h, after 24 h, 30 to 36 h, and 36 to 44 h of IVM, respectively. Cumulus-oocyte complexes (COC) were cultured in IVM medium and oocytes and cumulus cells were isolated after different durations of IVM (20, 26, 32, and 44 h). The data were analysed by one-way ANOVA using GraphPad Prism 5.0 (GraphPad Software, Sn Diego, CA, USA) and the threshold for statistical significance was set at P < 0.05. Messenger RNA expression of PLIN1 was not detected in either oocytes or cumulus cells during all periods of IVM. PLIN2, on the other hands, showed a significant lower expression in GVBD, MI, and MII oocytes compared with the GV oocytes, but showed no difference in cumulus cells. PLIN3 expression was significantly decreased in oocytes of MI stage, whereas PLIN3 from cumulus cells was expressed significantly lower in GVBD and MII stage compared with GV stage. Expression of PLIN4 was significantly decreased in only cumulus cells of GVBD and MI stage. These findings suggest that PLIN2 may have important roles in lipid metabolism during porcine oocyte maturation, whereas PLIN4 may play a major role in cumulus cells. PLIN3 can be hypothesised as a common lipid droplet-associated protein in both oocytes and cumulus cells. Further studies should be conducted to characterise the expression and distribution of PLIN1, PLIN2, PLIN3, and PLIN4 in porcine oocytes and cumulus cells. This study was supported by Ministry of Trade, Industry and Energy (#10048948), Korea IPET (#311011-05-4-SB010), Research Institute for Veterinary Science, and the BK21 plus program.

scholarly journals A single nucleotide polymorphism of PLIN2 is associated with nonalcoholic steatohepatitis and causes phenotypic changes in hepatocyte lipid droplets

2019 ◽  
Author(s):  
Claire S Faulkner ◽  
Collin White ◽  
Loretta L Jophlin
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Nonalcoholic Steatohepatitis ◽  
Lipid Droplets ◽  
Nucleotide Polymorphism ◽  
Intracellular Lipid ◽  
Single Nucleotide ◽  
Phenotypic Changes ◽  
Intracellular Lipid Accumulation ◽  
And Gender

ABSTRACTPerilipin 2 (PLIN2) is a lipid droplet-associated protein which regulates cellular lipid storage and is highly expressed in the liver. Previous studies of a missense single nucleotide polymorphism (SNP) in PLIN2, Ser251Pro, (i.e. rs3556875) have shown this SNP to cause decreased lipolysis and increased intracellular lipid accumulation. To explore if this SNP is associated with nonalcoholic steatohepatitis (NASH), we genotyped 116 adults with NASH and 67 age- and gender-matched controls. rs3556875 was significantly associated with NASH with an allelic odds ratio of 2.98 (95% confidence interval 1.12 - 7.31, p=0.02) and in a dominant inheritance model. In an in vitro model of hepatic steatosis, expression of the Pro251 variant protein led to phenotypic changes in intracellular lipid droplets, yielding smaller diameter and up to 7 fold more lipid droplets per cell when compared to wild type PLIN2. In conclusion, the Ser251Pro SNP of PLIN2 may convey risk for NASH and causes phenotypic changes in hepatocyte lipid droplets.

scholarly journals Universal phase behaviors of intracellular lipid droplets

2019 ◽  
Author(s):  
Shunsuke F. Shimobayashi ◽  
Yuki Ohsaki
Keyword(s):  
Phase Transition ◽  
Phase Diagram ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Droplet Surface ◽  
Molecular Organization ◽  
Cholesteryl Esters ◽  
Intracellular Lipid

Lipid droplets are cytoplasmic micro-scale organelles involved in energy homeostasis and handling of cellular lipids and proteins. The core structure is mainly composed of two kinds of neutral lipids, triglycerides and cholesteryl esters, which are coated by a phospholipid monolayer and proteins. Despite the liquid crystalline nature of cholesteryl esters, the connection between the lipid composition and physical states is poorly understood. Here, we present the first universal intracellular phase diagram of lipid droplets, semi-quantitatively consistent with the in vitro phase diagram, and reveal that cholesterol esters cause the liquid-liquid crystal phase transition under near-physiological conditions. The internal molecules of the liquid crystallized lipid droplets are aligned radially. We moreover combine in vivo and in vitro studies, together with the theory of confined liquid crystals, to suggest that the radial molecular alignments in intracellular lipid droplets are caused by an anchoring force at the droplet surface. Our findings on the phase transition of lipid droplets and resulting molecular organization contribute to a better understanding of their biological functions and diseases.

scholarly journals Retinyl esters form lipid droplets independently of triacylglycerol and seipin

2021 ◽  
Vol 220 (10) ◽  
Author(s):  
Martijn R. Molenaar ◽  
Kamlesh K. Yadav ◽  
Alexandre Toulmay ◽  
Tsjerk A. Wassenaar ◽  
Muriel C. Mari ◽  
...  
Keyword(s):  
Lipid Bilayers ◽  
Lipid Droplet ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Yeast Cells ◽  
In Vitro Experiments ◽  
Steryl Esters ◽  
Retinyl Esters

Lipid droplets store neutral lipids, primarily triacylglycerol and steryl esters. Seipin plays a role in lipid droplet biogenesis and is thought to determine the site of lipid droplet biogenesis and the size of newly formed lipid droplets. Here we show a seipin-independent pathway of lipid droplet biogenesis. In silico and in vitro experiments reveal that retinyl esters have the intrinsic propensity to sequester and nucleate in lipid bilayers. Production of retinyl esters in mammalian and yeast cells that do not normally produce retinyl esters causes the formation of lipid droplets, even in a yeast strain that produces only retinyl esters and no other neutral lipids. Seipin does not determine the size or biogenesis site of lipid droplets composed of only retinyl esters or steryl esters. These findings indicate that the role of seipin in lipid droplet biogenesis depends on the type of neutral lipid stored in forming droplets.

Neurofilaments and Paired Helical Filaments

1985 ◽  
Vol 43 ◽  
pp. 740-743
Author(s):  
J. Metuzals
Keyword(s):  
Animal Model ◽  
Posttranslational Modifications ◽  
Posttranslational Modification ◽  
Giant Axon ◽  
Paired Helical Filaments ◽  
Squid Giant Axon ◽  
In Vitro Experiments ◽  
Thin Sections ◽  
Structural Relationships

It has been demonstrated that the neurofibrillary tangles in biopsies of Alzheimer patients, composed of typical paired helical filaments (PHF), consist also of typical neurofilaments (NF) and 15nm wide filaments. Close structural relationships, and even continuity between NF and PHF, have been observed. In this paper, such relationships are investigated from the standpoint that the PHF are formed through posttranslational modifications of NF. To investigate the validity of the posttranslational modification hypothesis of PHF formation, we have identified in thin sections from frontal lobe biopsies of Alzheimer patients all existing conformations of NF and PHF and ordered these conformations in a hypothetical sequence. However, only experiments with animal model preparations will prove or disprove the validity of the interpretations of static structural observations made on patients. For this purpose, the results of in vitro experiments with the squid giant axon preparations are compared with those obtained from human patients. This approach is essential in discovering etiological factors of Alzheimer's disease and its early diagnosis.

Daunorubicin and Platelet Function

1981 ◽  
Vol 45 (01) ◽  
pp. 038-042 ◽  
Author(s):  
M E Pogliani ◽  
R Fantasia ◽  
G Lambertenghi-Deliliers ◽  
E Cofrancesco
Keyword(s):  
Electron Microscope ◽  
Cellular Membrane ◽  
Hemorrhagic Diathesis ◽  
Clot Retraction ◽  
Freezing And Thawing ◽  
Platelet Factor ◽  
High Doses ◽  
Very High ◽  
Platelet Functions

SummaryThe influence of Daunorubicin on some platelet functions in vitro was investigated, using different concentrations of the drug (0.01-0.02-0.04 μg/ml). Daunorubicin was shown to inhibit Collagen and Thrombin induced platelet aggregation and the intensity of inhibition depended on both drug concentration and the time of preincubation.Daunorubicin was also shown to inhibit the release reaction, the platelet prostaglandin pathway and the availability platelet factor 3; the drug at concentrations for clinical use does not damage the platelet membrane, as is the case with the freezing and thawing test, in platelet uptake of 14C-serotonin and as confirmed by the electron microscope. When very high doses (0.16 mg) of Daunorubicin are used, lysis of the platelets can be observed and this is confirmed under the electron microscope by the presence of empty platelets with fractures at the level of the cytoplasmic membrane.Finally, Daunorubicin causes irreversible inhibition of reptilase clot-retraction, even if this is less severe than with Vincristine. Working with gel-filtered platelets, it would appear that the inhibition exercised by the drug on platelet reactions is not caused through modifications in Ca++ metabolism.The authors suggest that Daunorubicin, at the dosages used clinically, induces in vitro thrombocytopathy without damaging the cellular membrane as confirmed by the electron microscope.This impairment of platelet functions could play a part in hemorrhagic diathesis observed during Daunorubicin therapy.

Nano-Curcumin Regulates p53 Phosphorylation and PAI-1 Expression during Bleomycin Induced Injury in Alveolar Basal Epithelial Cells

2020 ◽  
Vol 16 (1) ◽  
pp. 85-89
Author(s):  
Mahesh M. Gouda ◽  
Ashwini Prabhu ◽  
Varsha Reddy S.V. ◽  
Rafa Jahan ◽  
Yashodhar P. Bhandary
Keyword(s):  
Epithelial Cells ◽  
Lung Injury ◽  
Molecular Mechanisms ◽  
A549 Cells ◽  
Plasminogen Activator Inhibitor ◽  
Underlying Mechanism ◽  
Protective Role ◽  
Alveolar Epithelial Cells ◽  
Cellular Damage

Background: Bleomycin (BLM) is known to cause DNA damage in the Alveolar Epithelial Cells (AECs). It is reported that BLM is involved in the up-regulation of inflammatory molecules such as neutrophils, macrophages, chemokines and cytokines. The complex underlying mechanism for inflammation mediated progression of lung injury is still unclear. This investigation was designed to understand the molecular mechanisms associated with p53 mediated modulation of Plasminogen Activator Inhibitor-I (PAI-I) expression and its regulation by nano-curcumin formulation. Methods: A549 cells were treated with BLM to cause the cellular damage in vitro and commercially available nano-curcumin formulation was used as an intervention. Cytotoxic effect of nano-curcumin was analyzed using Methyl Thiazolyl Tetrazolium (MTT) assay. Protein expressions were analyzed using western blot to evaluate the p53 mediated changes in PAI-I expression. Results: Nano-curcumin showed cytotoxicity up to 88.5 % at a concentration of 20 μg/ml after 48 h of treatment. BLM exposure to the cells activated the phosphorylation of p53, which in turn increased PAII expression. Nano-curcumin treatment showed a protective role against phosphorylation of p53 and PAI-I expression, which in turn regulated the fibro-proliferative phase of injury induced by bleomycin. Conclusion: Nano-curcumin could be used as an effective intervention to regulate the severity of lung injury, apoptosis of AECs and fibro-proliferation during pulmonary injury.

Sign in / Sign up

Export Citation Format

Share Document