Journal: Journal of pharmaceutical sciences

Article Title: Physiologically-based pharmacokinetic modeling of nanoparticles

doi: 10.1016/j.xphs.2018.10.037

Figure Lengend Snippet: Selected PBPK model structures for nanoparticles.

Article Snippet: The iridium-derived model parameters were not able to predict the kinetics of silver nanoparticles. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Reference Nanopartic Dosing route Species Modeling entity Tissues considered Clearance mechanism Tissue models Kagan, Gershkovich et al. 2014 106 AmBisome® IV Mice, rats Liposomal amphotericin B (AmB) and free AmB Plasma, liver, spleen, kidneys, GI tract, lungs, heart, and the remainders Clearance of free AmB was defined in the liver, the kidneys, and the remainder compartment In the spleen and kidneys: In others: Li, Johanson et al. 2014 21 PEG-coated polyacrylamide nanoparticle IV Rats Nanoparticle Arterial blood, venous blood, lungs, bone marrow, brain, heart, kidneys, liver, spleen, and the remainders Biodegradation was considered to be negligible.

Techniques:

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific deletion of Fto alleviated T4-induced AF. ( A ) Representative traces of AF induced by electrical stimulation in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups. ( B ) Quantification of AF inducibility ( n = 10). ( C ) AF duration was quantified only in animals with inducible episodes. The sample size for each group is n = 10, of which 2 (Myh6-cre − ; Fto fl/fl ) and 7 (Myh6-cre + ; Fto fl/fl ) animals did not have inducible AF and are therefore not included in (C) . ( D ) Quantification of HW/TL ( n = 10). ( E ) Representative maps of atrial activation time and dispersion of conduction in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups ( n = 6). ( F ) Quantification of active time ( n = 6). ( G ) Quantification of dispersion of conduction ( n = 6). ( H ) Quantification of LACV ( n = 6). ( I ) Quantification of ERP ( n = 6). ( J ) Representative images of LAD and Masson staining in atria. ( K ) Quantification of fibrosis area ( n = 6). ( L ) Quantification of LAD ( n = 6). AF, atrial fibrillation; β-Actin, beta-actin; ERP, effective refractory period; Fto, fat mass and obesity-associated protein; HR, heart rate; HW/TL, heart weight-to-tibial length ratio; LACV, left atrial conduction velocity; LAD, left atrial diameter. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant. Statistical analysis was performed with ordinary one-way ANOVA ( D , F , G , H , I , K , and L ), Fisher's exact test ( B ) and unpaired Student's t -test ( C ).

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Saline, Activation Assay, Dispersion, Staining

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific Fto knockout attenuated Lox expression in T4-treated mice. ( A and B ) GO functional enrichment analysis of differentially expressed genes. ( C ) Venn diagram showing overlap of differentially expressed genes between two groups. ( D ) Representative immunoblots of Fto and Lox protein in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups ( n = 6). ( E ) Quantification of Lox mRNA levels ( n = 6). ( F ) Quantification of Fto protein levels ( n = 6). ( G ) Quantification of Lox protein levels ( n = 6). β-Actin, beta-actin; Fto, fat mass and obesity-associated protein; GO, gene ontology; Lox, lysyl oxidase. **** P < 0.0001, comparison made using ordinary one-way ANOVA.

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Knock-Out, Expressing, Functional Assay, Western Blot, Saline, Comparison

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific Fto overexpression promoted AF susceptibility via m 6 A-dependent mechanisms. ( A ) Representative immunoblots of Fto and Lox proteins in mice injected with AAV9-cTnT- Fto wt -oe, AAV9-cTnT- Fto mut -oe, and AAV9-cTnT-nc ( n = 6). ( B and C ) Quantification of Fto and Lox protein levels ( n = 6). ( D ) Representative intracardiac bipolar electrograms showing induction of AF from mice injected with AAV9-cTnT- Fto wt -oe, AAV9-cTnT- Fto mut -oe, and AAV9-cTnT-nc. ( E ) Quantification of AF inducibility ( n = 10). ( F ) AF was inducible only in the AAV9-cTnT- Fto wt -oe group (7 out of 10 animals). Therefore, data are presented only for this group. AF was not inducible in any animal from the AAV9-cTnT-nc ( n = 10) or AAV9-cTnT- Fto mut -oe ( n = 10) groups; hence, no AF duration data are available for these groups, and they are not represented in this graph. ( G ) Representative maps of atrial activation time and dispersion of conduction. ( H ) Quantification of active time ( n = 6). ( I ) Quantification of dispersion of conduction ( n = 6). ( J ) Quantification of LACV ( n = 6). ( K ) Quantification of ERP ( n = 6). ( L ) Representative images of LAD and Masson staining. ( M ) Quantification of LAD ( n = 6). ( N ) Quantification of percentage of fibrotic area ( n = 6). AF, atrial fibrillation; β-Actin, beta-actin; ERP, effective refractory period; Fto, fat mass and obesity-associated protein; ; LACV, left atrial conduction velocity; LAD, left atrial diameter; Lox, lysyl oxidase. ** P < 0.01; *** P < 0.001; **** P < 0.0001. Statistical analysis was performed with ordinary one-way ANOVA ( B , C , H , I , J , K , M , and N ), Fisher's exact test ( E ).

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Over Expression, Western Blot, Injection, Activation Assay, Dispersion, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: The expression of HKla is upregulated in cardiomyocytes of mice subjected to TAC. (A) The ratios of HW/BW and HW/TL were measured in both the sham and TAC groups ( n = 6 mice/group). (B) Histological analysis of myocardial tissue. (C) Levels of markers for cardiac hypertrophy and fibrosis were measured in mice from the TAC and sham groups. (D) Echocardiography was performed to measure LVEDD, LVESD, LVPWT, heart rate, LVEF and LVFS in both the TAC and sham groups. (E) The levels of LDHA, LDHB and P300 were measured in the hearts of mice from the TAC and sham groups. (F, G) Western blot analysis was conducted to examine the levels of Pan Kla, H3K18la, H3K9la, H4K5la, H4K12la and H3K14la in hearts from mice in the TAC and sham groups. (H, I) The levels of H3K18ac, H4K5ac and pan Kac were measured in cardiomyocytes from both the TAC and sham groups ( n = 3, ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared to the sham group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Expressing, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: Ang II contributes to cardiomyocyte hypertrophy and promotes HKla. (A) After 24 h of Ang II stimulation, the cell surface area of NMCMs increased. (B) Representative western blots of ANP, BNP and β‐MHC in cultured NMCMs treated with Ang II (1 μM) for 24 h. (C) NMCMs were exposed to Ang II (1 μM) for 24 h, and intracellular lactate levels were evaluated using a colorimetric lactate measurement kit. (D) Expression levels of LDHA, LDHB and P300 proteins were detected in NMCMs using western blotting. (E, F) Western blot analysis was performed to measure the expression levels of H3K18la, H3K9la, H4K5la, H4K12la, H3K14la and Pan HKla in NMCMs ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, vs. control group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Western Blot, Cell Culture, Expressing, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: Lactate enhances HKla and exacerbates cardiac hypertrophy. (A) NMCMs were exposed to different concentrations of L‐lactate for 48 h, and intracellular lactate levels were evaluated using a colorimetric lactate measurement kit. (B) Effects of increasing lactate on the surface areas of cardiomyocytes. C‐H NMCMs were cultured with various concentrations of L‐lactate (0, 1, 5 and 10 mM) for 48 h. Afterward, cells at different concentrations were harvested for western blot analysis. (C) Effects of increasing lactate on the levels of cardiac hypertrophy markers. (D) Expression levels of LDHA, LDHB and P300 proteins were detected in NMCMs using western blotting. (E, F) Western blot analysis was performed to measure the expression levels of H3K18la, H3K9la, H4K5la, H4K12la, H3K14la and Pan HKla in NMCMs. (G, H) Expression levels of H3K18ac, H4K5ac and Pan Kac were detected in NMCMs using western blotting ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. indicated group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Cell Culture, Western Blot, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: Glucose increases lactate levels, which upregulate HKla expression. (A) Lactate levels in NMCMs cultured with different concentrations of glucose were measured by a lactate colorimetric kit. (B) Effects of glucose increase on cardiomyocyte surface areas. (C‐H) NMCMs were cultured for 48 h with different concentrations of glucose (0, 1, 5 and 25 mM). Cells at varying concentrations were harvested and utilized for western blot analysis. (C) Effects of increasing glucose on the levels of cardiac hypertrophy markers. (D) LDHA, LDHB and P300 protein expression levels in NMCMs were assessed using western blotting. (E, F) Western blot analysis was performed to measure the expression levels of H3K18la, H3K9la, H4K5la, H4K12la, H3K14la and Pan HKla in NMCMs. (G, H) Expression levels of H3K18ac, H4K5ac and Pan HKla were detected in NMCMs using western blotting. (I) The glycolysis ability of cardiomyocytes was analysed by Seahorse ECAR assay ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, vs. indicated group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Expressing, Cell Culture, Western Blot, ECAR Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: 2‐DG can decrease HKla levels and inhibits the advancement of cardiac hypertrophy. (A) NMCMs were exposed to different concentrations of 2‐DG for 48 h, and intracellular lactate levels were evaluated using a colorimetric lactate measurement kit. (B) The impact of 2‐DG increase on the surface area of cardiomyocytes. (C–H) NMCMs were cultured with various concentrations of 2‐DG (0, 1, 5 and 10 mM) for 48 h, and cells were then collected for western blot analysis. (C) The impact of increasing 2‐DG on the levels of cardiac hypertrophy marker genes. (D) Western blot analysis detected the expression of LDHA, LDHB and P300 proteins in NMCMs. (E, F) Western blot analysis was conducted to examine the expression of H3K18la, H3K9la, H4K5la, H4K12la, H3K14la and Pan HKla in NMCMs. (G, H) Western blot analysis detected the expression of H3K18ac, H4K5ac and Pan Kac levels in NMCMs. (I) The glycolysis ability of cardiomyocytes was analysed by Seahorse ECAR assay ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. indicated group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Cell Culture, Western Blot, Marker, Expressing, ECAR Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lactate regulates pathological cardiac hypertrophy via histone lactylation modification

doi: 10.1111/jcmm.70022

Figure Lengend Snippet: Oxamate can reduce HKla levels and inhibit the progression of cardiac hypertrophy. (A) Lactate levels in NMCM cultured with different concentrations of oxamate were measured by a lactate colorimetric kit. (B) The effect of increased oxamate concentration on the surface area of cardiomyocytes was detected. (C–H) The NMCMs model was cultured for 48 h using different concentrations of oxamate (0, 5, 10 and 20 mM), and cells were then collected for western blot analysis. (C) The impact of an increase in oxalate concentration on the levels of marker genes for cardiac hypertrophy. (D) The protein expression levels of LDHA, LDHB and P300 in NMCMs exposed to different concentrations of oxamate were analysed by western blotting. (E and F) Western blot analysis was performed to assess the expression level of H3K18la, H3K9la, H4K5la, H4K12la, H3K14la and Pan HKla in NMCMs. (G and H) Expression of H3K18ac, H4K5ac and Pan Kac levels were detected in NMCMs by western blotting ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. indicated group).

Article Snippet: After removing the cell suspension by aspiration, the NMCMs were transferred to complete medium and incubated in a 37°C incubator with 5% CO 2 for 24 h. Finally, the cardiomyocyte hypertrophy model was established by stimulating NMCMs with 1 μM Ang II (MCE, China) for 24 h.

Techniques: Cell Culture, Concentration Assay, Western Blot, Marker, Expressing