Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Schematic drawing of PEG-ylated BDNF-PAMAM dendrimer nanoparticles synthesis process as well as BDNF delivery to differentiated 6-OHDA treated neuroblastoma SH-SY5Y cells. Inserted frame comprises molecular structures of BDNF protein, PAMAM and PEG polymer molecules with their overall charge at pH 7.4

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Polymer

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Typical size distribution of BDNF-PAMAM ( a ) and PEG-ylated BDNF-PAMAM ( b ) nanoparticles measured in the bulk by DLS (0.15 M PBS, pH 7.4) without an ultrafiltration process. All values are representative of 5 independent experiments and are expressed as mean ± SD

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques:

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: AFM analysis of PAMAM-BDNF (above part) and PEG-ylated PAMAM-BDNF (below part) nanoparticles adsorbed at mica surface at 0.15 M pH 7.4: PBS. a PAMAM-based nanoparticles at a scan area of 0.5 × 05 µm. b Structure of PAMAM-based nanoparticles after cross-section. c Histogram of adsorbed nanoparticles indicated by direct AFM enumeration, obtained for a low surface molecules concentration. The figure was created by taking into account 10 randomly chosen areas, where each micrograph of PAMAM-based monolayer at the mica surface has a size of 0.5 × 05 µm

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Concentration Assay

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Desorption characteristics of BDNF from PAMAM G5.5 dendrimers-based nanoparticles in PBS electrolyte with increasing loading of protein concentrations from 0.02 to 1 mgL −1 . BDNF detection by ELISA over 24 h incubation of ( a ) PAMAM-BDNF and ( b ) PEG-ylated PAMAM-BDNF nanoparticles

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Cytotoxicity curves for BDNF, PAMAM-BDNF and PEG-ylated PAMAM-BDNF nanoparticles in differentiated human neuroblastoma cell line SH-SY5Y treated with the 100 µmol/L 6-OHDA neurotoxin. The control (100% viability) are cells treated with 100 µmol/L 6-OHDA only. The data represent means ± SD for 30 experiments

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Control

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Cellular localisation of PAMAM-based nanoparticles in SH-SY5Y cells exposed to 6-OHDA. The cells were incubated either with BDNF-PAMAM-AF488 ( a ) or BDNF-PAMAM-AF488-PEG ( b ) nanoparticles (green color) for different time (5, 10, 30 min, 1 and 24 h) and then co-stained with WGA-Texas Red-X (red) and DAPI (blue). Confocal fluorescence images on the right panel are render series of z-stack with applied surface mode. Panel on the left presents single stack from the z-stack and only nanoparticles (green) and surface glycoproteins (red) are shown. The images were taken at 400 × magnification

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Incubation, Staining, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Desorption characteristic of BDNF from PAMAM G5.5 dendrimers-based nanoparticles for cell culture supernatant under increasing nanocarriers loading of protein concentrations from 0.02 to 2 mgL −1 . BDNF detection by ELISA over 24 h in cells incubated with BDNF, PAMAM-BDNF nanoparticles and PEG-ylated PAMAM-BDNF nanoparticles. The data represent means ± SD for five experiments

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Cell Reports Medicine

Article Title: High cellular plasticity state of medulloblastoma local recurrence and distant dissemination

doi: 10.1016/j.xcrm.2024.101914

Figure Lengend Snippet:

Article Snippet: Brain Tumor Dissociation Kit , Miltenyi Biotech , Cat# 130-095-942.

Techniques: Recombinant, Protease Inhibitor, Reverse Transcription, SYBR Green Assay, Red Blood Cell Lysis, DNA Library Preparation, Polymer, Virus, Sequencing, Software

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Blood-based detection of MMP11 as a marker of prostate cancer progression regulated by the ALDH1A1-TGF-β1 signaling mechanism

doi: 10.1186/s13046-025-03299-6

Figure Lengend Snippet: TGFβ1 regulates the members of the MMP family. A The ROC curve analysis of the potential association of the expression levels of 20 MMP genes with biochemical recurrence-free survival (BRFS) in the TCGA PRAD gene expression dataset ( n = 493). ROC analysis was conducted using easyROC web-tool . B Correlation of MMP11 and MMP26 gene expression with TGFB1 mRNA levels in the PCa gene expression datasets TCGA PRAD ( n = 493), MSKCC primary ( n = 131), Broad/Cornell ( n = 31), DKFZ ( n = 118), and SMMU ( n = 65) and non-cancerous tissues (MSKCC cohort, n = 29); * p < 0.05; n.s.- non significant. C PC3 and DU145 cells were treated with 5 ng/ml of TGFβ1 for 48 h, and MMP11 levels were analysed by qPCR. n ≥ 3; Error bars = SD; * p < 0.05; ** p < 0.01. D Analysis of the previously published dataset indicated that TGF-β signaling in stroma cells induces MMP expression in LNCaP cells. LNCaP cells overexpressing TGFB1 and co-cultured with stroma cells were compared to the control LNCaP co-cultured with stroma. * p < 0.05. E qPCR analysis of the relative MMP11 expression in LNCaP and PC3 cells upon knockdown of ALDH1A1 or ALDH1A3. Cells transfected with scrambled siRNA (siScr) were used as controls. n = 3; Error bars = SD; * p < 0.05; ** p < 0.01; n.s.- non significant. F qPCR analysis of the relative MMP11, ALDH1A1 and ALDH1A3 expression in LNCaP and PC3 cells upon MMP11 knockdown. Cells transfected with scrambled siRNA (siScr) were used as controls. n ≥ 3; Error bars = SD; * p < 0.05; ** p < 0.01; *** p < 0.001. G Relative cell radiosensitivity was analysed by 2D radiobiological colony forming assay after siRNA-mediated knockdown of MMP11 in LNCaP and PC3 cells. Cells transfected with scrambled (Scr) siRNA were used as controls. n = 3; Error bars = SD; ** p < 0.01. H PC3 cells were cultured either under 2D conventional adherent conditions, in the polymer-based microcapsules mimicking mechanical stress within tumors , or in 3D anchorage-free cultures as a model of the intermediate stage of metastasis, circulating tumor cells (CTC). In all conditions, cells were kept in the same culture media. At day 3, the cells were collected, and relative gene expression of TGFB1, MMP11, ALDHA1 and ALDHA3 was analysed by qPCR. n ≥ 3; Error bars = SD; * p < 0.05; *** p < 0.001. I ALDH1A1 regulates TGFB1 expression in androgen-sensitive cells through AR- and RA-dependent mechanisms. In contrast, the interplay between TGF-β1 and MMP11 is present in the androgen-sensitive and castration-resistant models of PCa (like in LNCaP and PC3 cells, respectively)

Article Snippet: The human PCa cell lines PC3 (derived from bone metastasis), LNCaP (derived from lymph node metastasis), LNCaP-C4-2B (further named C4-2B, a bone metastatic derivative subline of the LNCaP cell line), 22Rv1 (derived from a xenograft tumor after castration-induced regression and relapse of the parental androgen-dependent xenograft), and DU145 (derived from brain metastasis) were purchased from the American Type Culture Collection (Manassas, VA, USA), head and neck squamous cell carcinoma (HNSCC) cell line Cal33 was obtained from Deutsche Sammlung von Microorganismen und Zellkulturen DSMZ GmbH, Braunschweig, Germany.

Techniques: Expressing, Gene Expression, Cell Culture, Control, Knockdown, Transfection, Polymer

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Blood-based detection of MMP11 as a marker of prostate cancer progression regulated by the ALDH1A1-TGF-β1 signaling mechanism

doi: 10.1186/s13046-025-03299-6

Figure Lengend Snippet: TGFβ1 regulates the members of the MMP family. A The ROC curve analysis of the potential association of the expression levels of 20 MMP genes with biochemical recurrence-free survival (BRFS) in the TCGA PRAD gene expression dataset ( n = 493). ROC analysis was conducted using easyROC web-tool . B Correlation of MMP11 and MMP26 gene expression with TGFB1 mRNA levels in the PCa gene expression datasets TCGA PRAD ( n = 493), MSKCC primary ( n = 131), Broad/Cornell ( n = 31), DKFZ ( n = 118), and SMMU ( n = 65) and non-cancerous tissues (MSKCC cohort, n = 29); * p < 0.05; n.s.- non significant. C PC3 and DU145 cells were treated with 5 ng/ml of TGFβ1 for 48 h, and MMP11 levels were analysed by qPCR. n ≥ 3; Error bars = SD; * p < 0.05; ** p < 0.01. D Analysis of the previously published dataset indicated that TGF-β signaling in stroma cells induces MMP expression in LNCaP cells. LNCaP cells overexpressing TGFB1 and co-cultured with stroma cells were compared to the control LNCaP co-cultured with stroma. * p < 0.05. E qPCR analysis of the relative MMP11 expression in LNCaP and PC3 cells upon knockdown of ALDH1A1 or ALDH1A3. Cells transfected with scrambled siRNA (siScr) were used as controls. n = 3; Error bars = SD; * p < 0.05; ** p < 0.01; n.s.- non significant. F qPCR analysis of the relative MMP11, ALDH1A1 and ALDH1A3 expression in LNCaP and PC3 cells upon MMP11 knockdown. Cells transfected with scrambled siRNA (siScr) were used as controls. n ≥ 3; Error bars = SD; * p < 0.05; ** p < 0.01; *** p < 0.001. G Relative cell radiosensitivity was analysed by 2D radiobiological colony forming assay after siRNA-mediated knockdown of MMP11 in LNCaP and PC3 cells. Cells transfected with scrambled (Scr) siRNA were used as controls. n = 3; Error bars = SD; ** p < 0.01. H PC3 cells were cultured either under 2D conventional adherent conditions, in the polymer-based microcapsules mimicking mechanical stress within tumors , or in 3D anchorage-free cultures as a model of the intermediate stage of metastasis, circulating tumor cells (CTC). In all conditions, cells were kept in the same culture media. At day 3, the cells were collected, and relative gene expression of TGFB1, MMP11, ALDHA1 and ALDHA3 was analysed by qPCR. n ≥ 3; Error bars = SD; * p < 0.05; *** p < 0.001. I ALDH1A1 regulates TGFB1 expression in androgen-sensitive cells through AR- and RA-dependent mechanisms. In contrast, the interplay between TGF-β1 and MMP11 is present in the androgen-sensitive and castration-resistant models of PCa (like in LNCaP and PC3 cells, respectively)

Article Snippet: The human PCa cell lines PC3 (derived from bone metastasis), LNCaP (derived from lymph node metastasis), LNCaP-C4-2B (further named C4-2B, a bone metastatic derivative subline of the LNCaP cell line), 22Rv1 (derived from a xenograft tumor after castration-induced regression and relapse of the parental androgen-dependent xenograft), and DU145 (derived from brain metastasis) were purchased from the American Type Culture Collection (Manassas, VA, USA), head and neck squamous cell carcinoma (HNSCC) cell line Cal33 was obtained from Deutsche Sammlung von Microorganismen und Zellkulturen DSMZ GmbH, Braunschweig, Germany.

Techniques: Expressing, Gene Expression, Cell Culture, Control, Knockdown, Transfection, Polymer

Journal:

Article Title:

doi:

Figure Lengend Snippet: Elevated renal CB 1 receptor (CB 1 R) and inducible NOS (iNOS) protein expression in humans with chronic kidney disease. (a) Elevated protein expression levels of CB 1 receptors and iNOS were detected in 25 kidney biopsies from focal segmental glomerulosclerosis (FSGS), diabetic nephropathy (DN), and obesity‐related glomerulopathy (ORG) patients. (b, c) CB 1 receptors and iNOS are highly expressed in the kidney tubules in DN and ORG. Scale bar, 100 μm in all images except for the right panels in (b), where the scale is 50 μm. Data represent the mean ± SEM from six normal, eight DN, and 10 ORG kidney samples. * P < .05, significantly different from normal kidney; one‐way ANOVA, followed by a Bonferroni post hoc test

Article Snippet: Human kidney biopsies were stained with rabbit anti‐human CB 1 receptor (1:100; Alomone, Cat# ACR‐001, RRID:AB_2039795) and iNOS (1:100; Abcam, Cat# ab3523, RRID:AB_303872), followed by an HRP One‐Step Polymer Detection System: anti‐Mouse–Rabbit–Rat ZyoChem.

Techniques: Expressing

Journal:

Article Title:

doi:

Figure Lengend Snippet: MRI‐1867 and peripheral CB 1 receptor blockade effectively restore metabolic and kidney morphological changes in diet‐induced obese (DIO) mice in comparison with inducible NOS inhibition or pair feeding. Six‐week‐old male C57Bl/6J mice were fed with high‐fat diet (HFD) for 18 weeks and then treated chronically by oral gavage with either vehicle (Veh; 1% Tween 80, 4% DMSO, and 95% saline), MRI‐1867 (3 mg·kg −1 ), peripherally restricted CB 1 receptor antagonist, JD5037 (3 mg·kg −1 ), and inducible NOS inhibitor, 1400W (10 mg·kg −1 ). A parallel group of mice, which was pair fed similarly to the MRI‐1867 group, was treated with Veh. Treatment was given for 28 consecutive days. MRI‐1867 and JD5037 reduced (a) body weight (BW) and (b) food intake. Both drugs reduced (c) body fat mass and increased (d) lean mass. Histologically, MRI‐1867 and JD5037 reduced (e, h) HFD‐induced glomerular enlargement, increased (f, h) Bowman's space area, and (g, h) mesangial expansion. Scale bar, 20 μm. Data represent the mean ± SEM from 7 to 14 mice per group. * P < .05, significantly different from diet‐induced obese animals treated with Veh; # P < .05, significantly different from diet‐induced obese mice treated with MRI‐1867; one‐way ANOVA, followed by a Bonferroni post hoc test

Article Snippet: Human kidney biopsies were stained with rabbit anti‐human CB 1 receptor (1:100; Alomone, Cat# ACR‐001, RRID:AB_2039795) and iNOS (1:100; Abcam, Cat# ab3523, RRID:AB_303872), followed by an HRP One‐Step Polymer Detection System: anti‐Mouse–Rabbit–Rat ZyoChem.

Techniques: Inhibition

Journal:

Article Title:

doi:

Figure Lengend Snippet: A greater efficacy of MRI‐1867 in restoring kidney function in diet‐induced obese mice in comparison with peripheral CB 1 receptor blockade, inducible NOS inhibition, or pair feeding. MRI‐1867 or JD5037 treatment reduced (a) BUN levels and (b) the kidney‐to‐body weight ratio. Only MRI‐1867 was effective in reducing urinary (c) albumin, (d) creatinine, and (e) the albumin‐to‐creatinine (ACR) ratio, as well as the urinary secretion levels of (f) kidney injury marker 1 (KIM‐1) and (g) tissue inhibitor of metalloproteinase 1 (TIMP‐1). Data represent the mean ± SEM from 7 to 14 mice per group. * P < .05, significantly different from diet‐induced obese animals treated with Veh; # P < .05, significantly different from diet‐induced obese mice treated with MRI‐1867; one‐way ANOVA, followed by a Bonferroni post hoc test

Article Snippet: Human kidney biopsies were stained with rabbit anti‐human CB 1 receptor (1:100; Alomone, Cat# ACR‐001, RRID:AB_2039795) and iNOS (1:100; Abcam, Cat# ab3523, RRID:AB_303872), followed by an HRP One‐Step Polymer Detection System: anti‐Mouse–Rabbit–Rat ZyoChem.

Techniques: Inhibition, Marker

Journal:

Article Title:

doi:

Figure Lengend Snippet: MRI‐1867 reverses the fatty acid‐induced reduction in adiponectin signalling. Mice on standard diet (STD) or high‐fat diet (HFD) for 18 weeks were treated with vehicle (Veh) or MRI‐1867 (3 mg·kg −1 ) orally for 28 days. (a–c) MRI‐1867 restored the HFD‐induced reduction in the mRNA renal expression of adiponectin, and Adipo2 but not Adipo1 receptors. Similarly, exposing HK‐2 cells to O:P (0.5 mM, 2:1, respectively) resulted in reduced (d–f) mRNA and protein expression of adiponectin as well as reduced mRNA levels of (g) Adipo1 and (h) Adipo2 receptors. Pretreatment of the cells with MRI‐1867 (100 ng·ml −1 ) completely normalized these changes. (i) A proposed mechanism for the dual blockade of CB 1 receptors and inducible NOS by MRI‐1867 in reversing obesity‐induced chronic kidney disease (CKD) is shown. RQ, relative quantitation. In vivo data represent the mean ± SEM from 8 to 14 mice per group. * P < .05, significantly different from animals on STD; # P < .05, significantly different from animals on the same diet. In vitro data represent the mean ± SEM from five independent experiments. * P < .05, significantly different from Veh‐treated cells under normal conditions; # P < .05, significantly different from Veh‐treated cells under O:P conditions. Data were analysed by one‐way ANOVA, followed by a Bonferroni post hoc test

Article Snippet: Human kidney biopsies were stained with rabbit anti‐human CB 1 receptor (1:100; Alomone, Cat# ACR‐001, RRID:AB_2039795) and iNOS (1:100; Abcam, Cat# ab3523, RRID:AB_303872), followed by an HRP One‐Step Polymer Detection System: anti‐Mouse–Rabbit–Rat ZyoChem.

Techniques: Expressing, Quantitation Assay, In Vivo, In Vitro