Journal: Frontiers in Immunology

Article Title: The phytohormone abscisic acid enhances remyelination in mouse models of multiple sclerosis

doi: 10.3389/fimmu.2024.1500697

Figure Lengend Snippet: ABA promotes lipid droplet load in macrophages in vitro. (A) Schematic representation showing the experimental set-up: mouse BMDMs were left untreated or treated with 100µg/ml myelin for 24 hours (Mye 24h ) or 72 hours (Mye 72h ). Myelin exposure was performed in the presence or absence of ABA. Created with biorender.com. (B) Representative images of ORO staining of BMDMs exposed to vehicle (PBS) or ABA and treated with myelin for 24 or 72 hours. Scale bars, 50 µm. (C) Quantification of lipid load (defined as percent ORO+ area of total cell area)(n = 3). (D) Mean fluorescence intensity of BODIPY in BMDMs (n = 5) exposed to vehicle or ABA and treated with myelin for 24 or 72 hours, as measured by flow cytometry. Data is represented as relative lipid droplet load compared to vehicle. (E) Internalization of pHrodo™-labelled myelin by BMDMs exposed to vehicle or ABA for 24 hours. Data are measured by flow cytometry and depicted relative to the vehicle treated group. (F) Quantification of total cholesterol (TC), free cholesterol (FC), and esterified cholesterol (EC) in BMDMs (n = 6) exposed to vehicle or ABA and treated with myelin for 0, 24 or 72 hours. (G-I) mRNA expression of Igf1, Tgfβ and Tnf α in BMDMs (n = 4). Each dot represents one well. Data are represented as mean ± SEM and statistically analyzed using a one-way ANOVA with correction for multiple testing or Student’s t-test. *p<0.05, **p<0.01.

Article Snippet: Chemokines and cytokines were measured in plasma using a Bio-Plex assay for tumor necrosis factor α (TNFα) (12002444; Bio-Rad), interferon γ (IFNγ) (12002438; Bio-Rad), interleukin 6 (IL6) (12002241; Bio-Rad), interleukin 10 (IL10) (12002242; Bio-Rad), C-C motif chemokine 5 (CCL5) (12002256; Bio-Rad) and monocyte chemoattractant protein 1 (MCP1) (12002441; Bio-Rad) according to the manufacturer’s instructions.

Techniques: In Vitro, Staining, Fluorescence, Flow Cytometry, Expressing

Journal: Materials

Article Title: Combined Effects of HA Concentration and Unit Cell Geometry on the Biomechanical Behavior of PCL/HA Scaffold for Tissue Engineering Applications Produced by LPBF

doi: 10.3390/ma16144950

Figure Lengend Snippet: SEM images of surfaces of DO and RD PCL/HA scaffolds with 5 wt.% ( a , b ), 30 wt.% ( c , d ) and 50 wt.% ( e , f ) HA. Inset in ( f ) shows details of a hydroxyapatite particle.

Article Snippet: Three different powder mixtures obtained by combining polycaprolactone (PCL, Eurocoating S.p.a., Pergine Valsugana, Italy) and hydroxyapatite (HA, Boc Sciences, Inc., Shirley, NY, USA) in 5, 30 and 50 wt.% were used for scaffold manufacturing by laser powder bed fusion (LPBF) technology, using a Formiga P110 Velocis (EOS GmbH, Munich, Germany) manufacturing system.

Techniques:

Journal:

Article Title: ACF consists of two subunits, Acf1 and ISWI, that function cooperatively in the ATP-dependent catalysis of chromatin assembly

doi:

Figure Lengend Snippet: Purification of the native form of Acf1 leads to the isolation of ACF comprising Acf1 (p170 and p185) and ISWI. (A) Scheme for the purification of native form of Acf1 from Drosophila embryos. (B) Hydroxyapatite chromatography. The peak gradient fractions from the Source 15Q (Pharmacia Biotech) column were applied to a Bio-Gel HT hydroxyapatite (Bio-Rad) column, and protein was eluted with a linear potassium phosphate gradient. The column fractions were subjected to Western blot analysis with antibodies against Drosophila Acf1 (p170/p185), ISWI, topoisomerase II, and dCAF-1 p55 in conjunction with 125I-labeled protein A. With the Acf1 Western blot, the p170 and p185 forms of Acf1 were not clearly resolved. Also, the slower migrating species that cross-reacts with the Acf1 antiserum is not recognized by the affinity-purified antibodies (e.g., see Fig. ​Fig.2).2). (C) POROS heparin chromatography. The peak hydroxyapatite fractions were applied to a POROS heparin (PerSeptive Biosystems) column, and protein was eluted with a linear NaCl gradient. The column fractions were subjected to Western blot analysis, as in B. The control sample is an ACF-containing fraction from the Source 15Q chromatography step. The p170 and p185 forms of Acf1 were not clearly resolved. (D) Glycerol gradient sedimentation. The peak POROS heparin fractions were subjected to 15%–40% (vol/vol) glycerol gradient sedimentation. The glycerol gradient fractions were subjected to Western blot analysis, as in B and C. The p170 and p185 forms of Acf1 were not clearly resolved. (E) Micrococcal nuclease digestion analysis. ACF activity in the glycerol gradient fractions was tested by micrococcal nuclease digestion analysis. Chromatin assembly reactions contained 10 μl of each 400 μl fraction and were carried out as described in Materials and Methods. The samples were then partially digested with two different concentrations of micrococcal nuclease. The resulting DNA fragments were deproteinized, resolved by 1.5% agarose gel electrophoresis, and visualized by staining with ethidium bromide. The mass markers (M) are the 123-bp DNA ladder (GIBCO-BRL). The peak of ACF activity is seen in fractions 7–9. (F) Native ACF consists of Acf1 (p185 and p170) and ISWI. Glycerol gradient fractions were subjected to 6% polyacrylamide–SDS gel electrophoresis, and proteins were visualized by silver staining. The sizes of molecular mass markers and the ACF subunits are indicated. The traces of dCAF-1 p55/NURF-55 that were seen in Western blots of the glycerol gradient fractions (D) could not be detected in these silver-stained SDS–polyacrylamide gels.

Article Snippet: The dialyzed Bio-Gel HT hydroxyapatite fractions were applied directly to a POROS 20 HE1 heparin (PerSeptive Biosystems) resin [column volume = 0.2 ml, column dimensions (diam.

Techniques: Purification, Isolation, Chromatography, Western Blot, Labeling, Affinity Purification, Sedimentation, Activity Assay, Agarose Gel Electrophoresis, Staining, SDS-Gel, Electrophoresis, Silver Staining

Journal: Nanotheranostics

Article Title: Dual Potential of Cetuximab Conjugated Hydroxyapatite Zirconium Nanoparticle as Nanocarrier for Radioenhancer in X-Ray Dynamic Therapy and 177 Lu-based Radioimmunotherapy of Lung Cancer

doi: 10.7150/ntno.101699

Figure Lengend Snippet: (a) XRD diffractogram of HApZr-CTX compared to ICDD 00-009-0432 of standard hydroxyapatite, (b) DLS-PSA analysis of HApZr-APTES and (c) HApZr-CTX dispersion in water, (d) hydrodynamic size stability of HApZr-APTES and HApZr-CTX dispersion in water for 14 days in 10°C storage, (e) TEM image of bare HApZr nanoparticle showed bar type nanoparticle with average size of 33 nm, (f) TEM image of agglomerated HApZr-CTX nanoparticle with size around 500 nm, note that thick shadow was alleged as CTX bound to agglomerated HApZr nanoparticle, (g) enlarged inset of HApZr-CTX nanoparticle from figure .f showed smaller HApZr-CTX nanoparticle with particle size around 50 nm

Article Snippet: CTX conjugated hydroxyapatite is yet to be investigated as a nanocarrier platform for radioenhancer and therapeutic radionuclide delivery.

Techniques: Dispersion

Journal: Nanotheranostics

Article Title: Dual Potential of Cetuximab Conjugated Hydroxyapatite Zirconium Nanoparticle as Nanocarrier for Radioenhancer in X-Ray Dynamic Therapy and 177 Lu-based Radioimmunotherapy of Lung Cancer

doi: 10.7150/ntno.101699

Figure Lengend Snippet: Cellular imaging of A549 cells using CLSM stained with DAPI (blue) and FITC conjugated in HApZr-CTX (green) in various concentrations compared to untreated cells as a control condition. Scale bar: 100 µm

Article Snippet: CTX conjugated hydroxyapatite is yet to be investigated as a nanocarrier platform for radioenhancer and therapeutic radionuclide delivery.

Techniques: Imaging, Staining, Control