Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the prefrontal (prelimbic/infralimbic) cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. ***: p < 0.001 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the anterior cingulate cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. **: p < 0.01 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the ventral hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. *: p < 0.05; **: p < 0.02 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the dorsal hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group.

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 1. Physicochemical characterization of PEGylated HSA-NT3-BDNF (NT3 10 mg L−1, BDNF 10 mg L−1) nanoparticles. The particle size distribution was measured by intensity (A) and by number (B), the nanoparticle concentration (C), and the zeta potential (D) were measured against Ringer buffer at pH 4.9 and 0.15 M ionic strength. The intensity (percent) indicates the size frequency of the most common particles. All syntheses were performed fifteen times, and the error bars represent the mean ± standard deviation (SD). The individual curves indicate the distribution of NP diameters from 24 samples.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Concentration Assay, Zeta Potential Analyzer, Standard Deviation

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 2. Stability of PEGylated HSA-NT3-BDNF (NT 10 mg L−1) nanoparticles over 90 days after formulation. The particle size distribution was measured by intensity (A) and by number (B), the zeta potential was measured against Ringer buffer (C), and the loading of NTs was measured via ELISA (D). The intensity (percent) indicates the size frequency of the most common particles. All syntheses were performed six times, and the error bars represent the mean ± standard deviation (SD). The individual curves indicate the distribution of NP diameters from 8 samples. Note that the dispersed type of PEGylated HSA-NT3-BDNF nanoparticles was maintained for 90 days.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Formulation, Zeta Potential Analyzer, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 4. Release of NT3 (in black, circle dots) and BDNF (in gray, square) from the PEGylated HSA-NT3-BDNF nanoparticles in a cell-free system. Kinetics of neurotrophin release over 2 months. n = 3; statistics: Two-way ANOVA with Bonferroni post hoc correction was used to compare the time course between NT3 and BDNF. The relative release of the adsorbed neurotrophins was calcu lated via ELISA.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 3. Representative binding curves for the interaction between HSA and BDNF (upper curve, Part A) and NT3; (upper curve, Part B). Representative MST/TRIC time traces of 40 nM labeled BDNF (lower curve, Part A) and 20 nM labeled NT3; (lower curve, Part B) in PBS supplemented with 1 mM HSA for the NT3 assay and 0.5 mM HSA for BDNF. Cold (blue) and hot (red) regions were defined for central data analysis to calculate Fnorm. The data for both figures are from instrument N06. Diagrams exported from MOAA. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Binding Assay, Labeling

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 6. Release of NT3 (in black) and BDNF (in gray) from the PEGylated HSA-NT3-BDNF nanoparticles in vitro. Kinetics of NT3 and BDNF release in culture media supplemented with (A) the complex (10 % FBS) or (B) serum starved (1 % FBS). The time course was >72 h. (n = 3; statistics: Two-way ANOVA with Bonferroni post hoc correction to compare the time course and either complex media or serum starvation conditions). The relative release of the adsorbed neurotrophins was calculated via ELISA.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 7. Intracellular levels of BDNF and NT3 in the ARPE-19 cell line after 72 h of incubation with PEGylated HSA-NT3-BDNF.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Incubation

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 5. The influence of the neurotrophin concentrations in PEGylated HSA-NT3-BDNF nanoparticles on viability and proliferation (in relation to the untreated control). After 24 h of preincubation, ARPE-19 (A) or L-929 (B) cells were treated with 5, 10 or 20 mg L−1 neurotrophin in PEGylated HSA-NT3-BDNF (described in the picture as HSA-NT3-BDNF-PEG) nanoparticles and stored in regular cell culture conditions. After 48 h, alamarBlue™ reagent was added to the cell culture mixture according to the manufacturer's instructions. The experiment was performed in three independent replicates. One-way ANOVA (GraphPad Prism) did not reveal any significant differences.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: Control, Cell Culture

Journal: International journal of biological macromolecules

Article Title: In vitro and in vivo characterization of human serum albumin-based PEGylated nanoparticles for BDNF and NT3 codelivery.

doi: 10.1016/j.ijbiomac.2024.130726

Figure Lengend Snippet: Fig. 8. NT3 and BDNF release profiles in the retina and vitreous body of BALB/c mice.

Article Snippet: Unfiltered stock solutions (typically 250 mg L− 1) of carrier-free recombinant human BDNF (rhBDNF) (248-N4-250/CF; R&D Systems, Canada), as well as carrier-free recombinant human NT3 (rhNT3) (248- BDB-250/CF; R&D Systems), were prepared by dissolving lyophilizates of known concentrations in phosphate-buffered saline (PBS) (pH 7.4 ± 0.2, 0.15 M; Biomed, Lublin, Poland) and storing them for no longer than 2 months at − 20 ◦C.

Techniques: