scholarly journals Induction of fibroblast apolipoprotein E expression during apoptosis, starvation-induced growth arrest and mitosis

2004 ◽  
Vol 378 (3) ◽  
pp. 753-761 ◽  
Author(s):  
Carmel M. QUINN ◽  
Katarina KÅGEDAL ◽  
Alexei TERMAN ◽  
Uri STROIKIN ◽  
Ulf T. BRUNK ◽  
...  
Keyword(s):  
Apolipoprotein E ◽  
Human Fibroblasts ◽  
Growth Arrest ◽  
Plasma Lipoproteins ◽  
In Vivo Studies ◽  
Serum Starvation ◽  
Morphological Alterations ◽  
Mrna Induction ◽  
Apoe Mrna

Apolipoprotein E (apoE) mediates the hepatic clearance of plasma lipoproteins, facilitates cholesterol efflux from macrophages and aids neuronal lipid transport. ApoE is expressed at high levels in hepatocytes, macrophages and astrocytes. In the present study, we identify nuclear and cytosolic pools of apoE in human fibroblasts. Fibroblast apoE mRNA and protein levels were up-regulated during staurosporine-induced apoptosis and this was correlated with increased caspase-3 activity and apoptotic morphological alterations. Because the transcription of apoE and specific pro-apoptotic genes is regulated by the nuclear receptor LXR (liver X receptor) α, we analysed LXRα mRNA expression by quantitative real-time PCR and found it to be increased before apoE mRNA induction. The expression of ABCA1 (ATP-binding cassette transporter A1) mRNA, which is also regulated by LXRα, was increased in parallel with apoE mRNA, indicating that LXRα probably promotes apoE and ABCA1 transcription during apoptosis. Fibroblast apoE levels were increased under conditions of serum-starvation-induced growth arrest and hyperoxia-induced senescence. In both cases, an increased nuclear apoE level was observed, particularly in cells that accumulated lipofuscin. Nuclear apoE was translocated to the cytosol when mitotic nuclear disassembly occurred and this was associated with an increase in total cellular apoE levels. ApoE amino acid sequence analysis indicated several potential sites for phosphorylation. In vivo studies, using 32P-labelling and immunoprecipitation, revealed that fibroblast apoE can be phosphorylated. These studies reveal novel associations and potential roles for apoE in fundamental cellular processes.

scholarly journals In Vitro Generation of Oocyte Like Cells and Their In Vivo Efficacy: How Far We have been Succeeded

Cells ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 557 ◽  
Author(s):  
Dinesh Bharti ◽  
Si-Jung Jang ◽  
Sang-Yun Lee ◽  
Sung-Lim Lee ◽  
Gyu-Jin Rho
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem ◽  
In Vivo Studies ◽  
Special Focus ◽  
Morphological Alterations ◽  
Cell Sources ◽  
Induced Pluripotent

In the last few decades, stem cell therapy has grown as a boon for many pathological complications including female reproductive disorders. In this review, a brief description of available strategies that are related to stem cell-based in vitro oocyte-like cell (OLC) development are given. We have tried to cover all the aspects and latest updates of the in vitro OLC developmental methodologies, marker profiling, available disease models, and in vivo efficacies, with a special focus on mesenchymal stem cells (MSCs), induced pluripotent stem cells (iPSCs), and embryonic stem cells (ESCs) usage. The differentiation abilities of both the ovarian and non-ovarian stem cell sources under various induction conditions have shown different effects on morphological alterations, proliferation- and size-associated developments, hormonal secretions under gonadotropic stimulations, and their neo-oogenesis or folliculogenesis abilities after in vivo transplantations. The attainment of characters like oocyte-like morphology, size expansion, and meiosis initiation have been found to be major obstacles during in vitro oogenesis. A number of reports have either lacked in vivo studies or have shown their functional incapability to produce viable and healthy offspring. Though researchers have gained many valuable insights regarding in vitro gametogenesis, still there are many things to do to make stem cell-derived OLCs fully functional.

scholarly journals THER-15. DEVELOPING TUMOR-HOMING CYTOTOXIC HUMAN INDUCED NEURAL STEM CELL THERAPY FOR BRAIN METASTASES

2019 ◽  
Vol 1 (Supplement_1) ◽  
pp. i13-i14
Author(s):  
Alison Mercer-Smith ◽  
Wulin Jiang ◽  
Juli Bago ◽  
Simon Khagi ◽  
Carey Anders ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Brain Metastases ◽  
Drug Carrier ◽  
Human Fibroblasts ◽  
Genetically Engineered ◽  
In Vivo Studies ◽  
Tumor Homing ◽  
The Brain

Abstract INTRODUCTION: Non-small cell lung cancer (NSCLC) and breast cancer are the most common cancers that metastasize to the brain. New therapies are needed to seek out and eradicate metastases. Genetically engineered neural stem cells (NSCs) have shown unique tumor-homing capacity, allowing them to deliver cytotoxic proteins directly to tumors. An ideal NSC drug carrier would be readily available and autologous. We have transdifferentiated human fibroblasts into induced NSCs (hiNSCs) that home to tumors and engineered the hiNSCs to release the cytotoxic protein TRAIL. Here we used intracerebroventricular (ICV) injections to deliver hiNSCs to metastatic foci. METHODS: We performed an in vitro efficacy co-culture assay, used in vivo studies to determine the migration, persistence, and efficacy of therapeutic hiNSCs against H460 NSCLC and triple-negative breast cancer MB231-Br tumors in the brain. Following the establishment of tumors in the brains of nude mice, hiNSCs were injected directly into the tumor or the ventricle contralateral to the site of tumor. The migration and persistence of hiNSCs was investigated by following the bioluminescence of the hiNSCs. The therapeutic efficacy of the hiNSCs was determined by following the bioluminescece of the tumor. RESULTS/CONCLUSION: Co-culture results demonstrated that hiNSC therapy reduced the viability of H460 and MB231-Br up to 75% and 99.8% respectively compared to non-treated controls. ICV-administered hiNSC serial imaging show that cells persisted for more than one week. Fluorescent analysis of tissue sections showed that hiNSCs co-localized with lateral and a contralateral tumors within 7 days. Using H460 and MB231-Br models, kinetic tracking of intracranial tumor volumes showed intratumoral or ICV-injected therapeutic hiNSCs reduced the growth rate of brain tumors by 31-fold and 3-fold, respectively. This work demonstrates for the first time that we can effectively deliver personalized cytotoxic tumor-homing cells through the ventricles to target brain metastases.

MULTIPHOTON MICROSCOPY: A NEW APPROACH, IN PHYSIOLOGICAL STUDIES AND PATHOLOGICAL DIAGNOSIS FOR OPHTHALMOLOGY

2009 ◽  
Vol 02 (01) ◽  
pp. 45-60 ◽  
Author(s):  
CHIU-MEI HSUEH ◽  
WEN LO ◽  
SUNG-JAN LIN ◽  
TSUNG-JEN WANG ◽  
FUNG-RUNG HU ◽  
...  
Keyword(s):  
Second Harmonic ◽  
Healing Process ◽  
Multiphoton Microscopy ◽  
In Vivo Studies ◽  
Wound Healing Process ◽  
Label Free ◽  
Multiphoton Imaging ◽  
Morphological Alterations ◽  
Corneal Scar

Multiphoton microscopy (MPM), with the advantages of improved penetration depth, decreased photo-damage, and optical sectioning capability, has become an indispensable tool for biomedical imaging. The combination of multiphoton fluorescence (MF) and second-harmonic generation (SHG) microscopy is particularly effective in imaging tissue structures of the ocular surface. This work is intended to be a review of advances that MPM has made in ophthalmic imaging. The MPM not only can be used for the label-free imaging of ocular structures, it can also be applied for investigating the morphological alterations in corneal pathologies, such as keratoconus, infected keratitis, and corneal scar. Furthermore, the corneal wound healing process after refractive surgical procedures such as conductive keratoplasty (CK) can also be studied with MPM. Finally, qualitative and quantitative SHG microscopy is effective for characterizing corneal thermal denaturation. With additional development, multiphoton imaging has the potential to be developed into an effective imaging technique for in vivo studies and clinical diagnosis in ophthalmology.

Procoagulant activities of plasma factor VIIc and factor Xc are positively and independently associated with concentrations of the high-density lipoprotein apolipoprotein, apo A-II

2008 ◽  
Vol 100 (09) ◽  
pp. 391-396 ◽  
Author(s):  
Maria Atta ◽  
David Crook ◽  
Faria Shafique ◽  
Desmond G. Johnston ◽  
Ian F. Godsland
Keyword(s):  
High Density Lipoprotein ◽  
Total Cholesterol ◽  
Multivariable Analysis ◽  
Density Lipoprotein ◽  
Hdl Cholesterol ◽  
High Density ◽  
Plasma Lipoproteins ◽  
In Vivo Studies ◽  
Plasma Factor

SummaryThe pro- and antiatherogenic roles of apolipoproteins B andA-I, respectively, are well-established although the importance of apolipoprotein A-II remains unclear. There is extensive evidence for the involvement of plasma lipoproteins in haemostatic function. However, in-vivo studies of relationships between haemostatic variables and apolipoprotein concentrations are very limited. Plasma fibrinogen, factors VIIc and Xc (FVIIc and FXc, respectively), apolipoproteins (apo) A-I, A-II and B, triglycerides, total, low-density and high-density lipoprotein (HDL) cholesterol, and cholesterol in HDL subfractions 2 and 3 were measured in 186 apparently healthy Caucasian men (aged 26–78 years; body mass index 19.9–37.8 kg/m2).Associations between haemostatic, apolipoprotein,lipid and lipoprotein variables were explored in uni- and multivariable analyses. Fibrinogen did not correlate with any of the lipid-related variables. FVIIc and FXc were significant positive univariate correlates of total cholesterol (correlation coefficients 0.26,p<0.001 and 0.19,p<0.05,respectively) triglycerides (0.37, p<0.001 and 0.36, p<0.001), and apoB (0.21, p<0.01 and 0.17, p<0.05) and apoA-II (0.19, p<0.05 and 0.29, p<0.001). HDL2 subfraction cholesterol correlated negatively with FVIIc and FXc (-0.20, p<0.01 and –0.22, p<0.01, respectively). In multivariable analysis, only the associations of FVIIc and FXc with total cholesterol, triglycerides and apoA-II remained statistically significant. In conclusion, total cholesterol and triglycerides were the major independent lipid correlates of FVIIc and FXc. The independent and positive associations of apoA-II with FVIIc and FXc suggest a prothrombotic involvement for this apolipoprotein

scholarly journals A two-step homogeneous assay for apolipoprotein E-containing high-density lipoprotein-cholesterol

2018 ◽  
Vol 56 (1) ◽  
pp. 123-132 ◽  
Author(s):  
Yuji Takahashi ◽  
Yasuki Ito ◽  
Toshihiro Sakurai ◽  
Norio Wada ◽  
Atsushi Nagasaka ◽  
...  
Keyword(s):  
High Density Lipoprotein ◽  
Apolipoprotein E ◽  
High Density Lipoprotein Cholesterol ◽  
Density Lipoprotein ◽  
High Density ◽  
Lipoprotein Cholesterol ◽  
In Vivo Studies ◽  
Homogeneous Assay

Background Apolipoprotein E-containing high-density lipoprotein shows antiatherogenic properties in vitro. There is a need for a homogeneous assay to determine the concentration of apolipoprotein E-containing high-density lipoprotein for in vivo studies. Methods In the proposed homogeneous assay, lipoproteins other than apolipoprotein E-containing high-density lipoprotein were eliminated in the first step. Apolipoprotein E-containing high-density lipoprotein-cholesterol was measured in the second step. The control study used a 13% polyethylene glycol precipitation assay (control assay). Results The homogeneous assay showed good performance in validation studies. In subjects with normal liver function ( n = 78), a significant correlation was found between the control assay and the homogeneous assay ( r = 0.824). Serum apolipoprotein E-containing high-density lipoprotein cholesterol concentrations, determined by the control assay and the homogeneous assay, respectively, were 0.05 (0.04–0.10) (median [25th–75th percentile]) mmol/L and 0.10 (0.06–0.13) mmol/L for healthy individuals ( n = 12), and 0.03 (0.01–0.13) mmol/L and 0.02 (0.01–0.02) mmol/L for patients with cholestasis ( n = 6). The results indicate that the homogeneous assay recovers cholesterol contained in physiological apolipoprotein E-containing high-density lipoprotein, but not in pathological apolipoprotein E-containing high-density lipoprotein from cholestatic patients. Conclusions The proposed two-step homogeneous assay enables selective measurement of physiological apolipoprotein E-containing high-density lipoprotein cholesterol in common autoanalysers. This assay might uncover a role for apolipoprotein E-containing high-density lipoprotein in physiological conditions.

scholarly journals Prolonged Glucocorticoid Exposure Does Not Accelerate Telomere Shortening in Cultured Human Fibroblasts

Genes ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1425
Author(s):  
Anthony S. Zannas ◽  
Oksana Kosyk ◽  
Calvin S. Leung
Keyword(s):  
Telomere Length ◽  
Psychosocial Stress ◽  
Prolonged Exposure ◽  
Disease Risk ◽  
Human Fibroblasts ◽  
Stress Hormones ◽  
In Vivo Studies ◽  
Biological Aging ◽  
Telomere Shortening

Psychosocial stress, especially when chronic or excessive, can increase disease risk and accelerate biological aging. Although the underlying mechanisms are unclear, in vivo studies have associated exposure to stress and glucocorticoid stress hormones with shorter telomere length. However, the extent to which prolonged glucocorticoid exposure can shorten telomeres in controlled experimental settings remains unknown. Using a well-characterized cell line of human fibroblasts that undergo gradual telomere shortening during serial passaging in culture, we show that prolonged exposure (up to 51 days) to either naturalistic levels of the human endogenous glucocorticoid cortisol or the more potent synthetic glucocorticoid dexamethasone is not sufficient to accelerate telomere shortening. While our findings await extension in other cell types and biological contexts, they indicate that the in vivo association of psychosocial stress with telomere shortening is unlikely to be mediated by a direct and universal glucocorticoid effect on telomere length.

Glycosphingolipids and plasma lipoproteins: a review

1984 ◽  
Vol 62 (6) ◽  
pp. 385-397 ◽  
Author(s):  
Subroto Chatterjee ◽  
Peter O. Kwiterovich Jr.
Keyword(s):  
Human Fibroblasts ◽  
Ldl Receptor ◽  
Plasma Lipoproteins ◽  
In Vivo Model ◽  
Renal Tubular Cells ◽  
Cultured Human Fibroblasts ◽  
Vitro Model ◽  
Renal Tubular

Glycosphingolipids (GSL) are complex, sugar-containing lipids that are transported in plasma on lipoproteins, particularly low density (beta) lipoproteins (LDL). LDL are taken up and metabolized by cells through a LDL receptor-mediated pathway. Cells from receptor-negative familial hypercholesterolemic (FH) homozygotes lack a functional LDL receptor and the cellular uptake of LDL occurs through a LDL receptor-independent pathway. We have studied the relation between LDL and GSL metabolism using an in vitro model of cultured human fibroblasts and an in vivo model of renal tubular cells shed in the urine. Using biochemical, morphological, and immunocytological techniques, we have demonstrated that LDL taken up through the LDL receptor-independent pathway results in the accumulation of GSL in cells in vitro and in vivo. The storage of GSL is localized to intracytoplasmic vesicles and such storage can be modulated by changing the concentration of LDL in the culture medium or in plasma. The possible effect of LDL on certain steps in the intracellular metabolism of GSL in normal and receptor-negative homozygous FH cells is reviewed.

scholarly journals High density lipoproteins of blood plasma as a transport form of actinomycin D

2019 ◽  
Vol 17 (6) ◽  
pp. 64-69
Author(s):  
L. M. Polyakov ◽  
R. A. Knyazev ◽  
A. V. Ryabchenko ◽  
N. V. Trifonova ◽  
M. V. Kotova
Keyword(s):  
Actinomycin D ◽  
Physicochemical Characteristics ◽  
The Body ◽  
Plasma Lipoproteins ◽  
In Vivo Studies ◽  
High Density Lipoproteins ◽  
Transport Form ◽  
Apolipoprotein A

Introduction.The development of new and highly effective antitumor therapy is one of the priorities of pharmacology. The paper presents one of the solutions to the problem related to the development of transport forms of antitumor drugs.The aimof the study was to study the ability of various fractions of plasma lipoproteins (VLDLP, LDL, HDL) to interact with actinomycin D and show the role of HDL as a transport form of actinomycin D in the body cells.Material and methods. The studies were conducted using unlabeled and tritium-labeled actinomycin D, preparative ultracentrifugation of the rat plasma lipoprotein fractions, chromatography, and in vivo experiments with intravenous administration of HDL complexes with labeled actinomycin D.Results.The important role of HDL in the formation of complexes with actinomycin D in comparison with LDL and LPA was shown. The basic physicochemical characteristics of the interaction of HDL and apolipoprotein A-I with actinomycin were obtained. The constants of the association were of the order of 105 M-1, and the number of binding sites for the drug was 26 for HDL and 12 for apolipoprotein A-I. In vivo studies on rats, the highest radioactivity after intravenous injection of HDL complexes with tritium-labelled actinomycin D was observed in the adrenal glands, then in the liver and kidneys. The uptake of tritium-labelled actinomycin D was twice lower in the lungs, adipose tissue, thymus and spleen. The low uptake of the label was observed in the myocardial tissue.Conclusion.The results obtained demonstrate the feasibility of using HDL as a transport form of actinomycin D in body cells.

Apolipoprotein E-Mimetic Therapeutic Peptides Mediate CLL Cell Cytotoxicity

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 359-359
Author(s):  
Dale J. Christensen ◽  
Karen M. Bond ◽  
Alicia D. Volkheimer ◽  
Jessica Oddo ◽  
Youwei Chen ◽  
...  
Keyword(s):  
B Cells ◽  
Apolipoprotein E ◽  
Colorimetric Assay ◽  
In Vivo Studies ◽  
Regulatory Subunit ◽  
Community Control ◽  
Mimetic Peptides ◽  
Therapeutic Peptides

Abstract Background and Significance : Even though we have treatments for CLL, it remains an incurable leuke mia. We need new and better treatments for this disease. The Akt kinase is usually constitutively acti vated (phosphorylated) in CLL, and it acts to maintain CLL cell viability. Chromosome deletion at 11q22–23 is frequently seen in CLL, and this cytogenetic abnormality portends a bad prognosis. The PPP2R1B gene that encodes the Ab constant regulatory subunit of the tumor suppressor protein phos phatase 2a (PP2a) is within the deleted segment in CLL patients with deleted 11q22–23. The resulting underexpression noted in those with deleted 11q22–23 leads to decreased PP2a activity in CLL cells. PP2a is important in deactivation of Akt, the mitogen activated protein kinases (MAPK) p38, JNK, ERK, and nuclear factor kB (through IkK). We have developed apolipoprotein E-mimetic therapeutic peptides that potently decrease phosphorylation of Akt, decrease TNF and nitric oxide (NO) and NO synthase (NOS) expression, and display anti-inflammatory activity in vitro and in vivo. NOS is overexpressed in CLL, and its product NO inhibits CLL cell apoptosis. Methods : Patients were from the V.A. and Duke University Medical Centers, and normal controls were from the community. Control normal PBMC were isolated by ficoll-Hypaque centrifugation, and CD19+ CLL or normal B cells were purified using negative selection with antibodies. We determined cytotoxicity using the MTS colorimetric assay. The apoE-mimetic peptides were prepared by chemical synthesis. Results : The apoE-mimetic COG compounds are peptides of 10 to 34 amino acids derived from the ligand-binding region of the apolipoprotein E holoprotein. Samples from 6 early stage CLL patients and 11 normal individuals were examined. Five of 6 patients were Rai stage 0 at presentation, and one was stage 1. They had been followed 5.6 yr (median; range 1.9–10.6 yr). Five of 6 were CD38 negative, and 2 of 6 were Zap-70 positive. Of 5analyzed, all had mutated IgVH gene. Five of 6 patients had not been treated. Of 6 peptides examined, all displayed some cytotoxicity for CLL cells in vitro. Peptide COG 112 was the most potent, while the control peptide with an inverted sequence (COG 056) had very little or no activity (Table). CLL PBMC Agent ED50 (nM) ED50 range (n) ED50 (nM) ED50 range (n) COG 112 (active) 215 64 to 351 (6) 5,150 1,100 to &gt;12,500 (6) COG 056 (control) 13,546 2771 to 20,418 (6) &gt;25,000 20,470 to &gt;25,500 (6) COG 112 induced cell death in a dose-dependent fashion in all patients’ samples.The ED50 of COG 112 for normal B cells was very high (&gt; 24,400 nM). COG 112 was approximatel 24 to 116 fold more potent for killing of CLL cells compared to normal PBMC or purified B cells. In vivo studies in normal mice using COG 112 revealed no toxicity even with doses of 100 mg/kg. Conclusions : ApoE mimetic peptides kill CLL cells in vitro with high efficacy and potency (ED50s in the low nanomolar range). The cytotoxicity is very specific for CLL cells compared to normal PBMC and B cells (24 to 116 fold more potent for CLL cells). Preliminary studies show that the peptide is nontoxic in vivo in normal mice. In vivo trials with apoE peptides in patients with CLL should help determine the toxicity and efficacy in patients.

scholarly journals Sustained Release of Levobupivacaine, Lidocaine, and Acemetacin from Electrosprayed Microparticles: In Vitro and In Vivo Studies

2020 ◽  
Vol 21 (3) ◽  
pp. 1093
Author(s):  
Jian-Ming Chen ◽  
Kuan-Chieh Liu ◽  
Wen-Ling Yeh ◽  
Jin-Chung Chen ◽  
Shih-Jung Liu
Keyword(s):  
Pain Relief ◽  
Release Kinetics ◽  
Human Fibroblasts ◽  
Fracture Site ◽  
In Vivo Studies ◽  
Infrared Spectrometry ◽  
Plga Microparticles ◽  
Low Toxicity

In this study, we explored the release characteristics of analgesics, namely levobupivacaine, lidocaine, and acemetacin, from electrosprayed poly(D,L-lactide-co-glycolide) (PLGA) microparticles. The drug-loaded particles were prepared using electrospraying techniques and evaluated for their morphology, drug release kinetics, and pain relief activity. The morphology of the produced microparticles elucidated by scanning electron microscopy revealed that the optimal parameters for electrospraying were 9 kV, 1 mL/h, and 10 cm for voltage, flow rate, and travel distance, respectively. Fourier-transform infrared spectrometry indicated that the analgesics had been successfully incorporated into the PLGA microparticles. The analgesic-loaded microparticles possessed low toxicity against human fibroblasts and were able to sustainably elute levobupivacaine, lidocaine, and acemetacin in vitro. Furthermore, electrosprayed microparticles were found to release high levels of lidocaine and acemetacin (well over the minimum therapeutic concentrations) and levobupivacaine at the fracture site of rats for more than 28 days and 12 days, respectively. Analgesic-loaded microparticles demonstrated their effectiveness and sustained performance for pain relief in fracture injuries.

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