Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: BAP decreases food intake and body weight and improves glucose homeostasis in HFD-fed male CD-1 mice . (A) The chemical structure of BAP. (B) Experimental outline for the male mice cohorts. 7-week-old male CD-1 mice were acclimated with a standard chow diet in the animal facility for 1 week and were fed a HFD for 4 weeks to induce an obese phenotype (DIO). All mice received 3 days of emulsion feeding acclimation, followed by 2 or 4 weeks feeding of BAP (300 mg/kg/day) or vehicle emulsion. ipGTT was conducted in a subset of mice 3 days before the experimental endpoint. C–F, Changes in (C) body weight as a percentage of starting weight, (D) daily food intake, (E) average daily food intake per cage of 2 mice, and (F) average daily water intake per cage of 2 mice fed with BAP or vehicle emulsion for 2 weeks ( n = 10 for body weight measurements, n = 5 for food and water intake measurements as per cage of 2 mice). G-J, Changes in (G) body weight as a percentage of starting weight, (H) daily food intake, (I) average daily food intake per cage of 2 mice, and (J) average daily water intake per cage of 2 mice fed with BAP or vehicle emulsion for 4 weeks ( n = 16 for body weight measurements, n = 8 for food and water intake measurements per cage of 2 mice). K-T, Before the experimental endpoint, a subset of mice from each cohort were randomly selected for ipGTT ( n = 8). Glucose tolerance and area of curve (AOC = area of curve) measurements of CD-1 mice after (K) 11 days or (P) 25 days of BAP or vehicle emulsion feeding ( n = 8). Serum concentration of insulin and leptin of CD-1 mice fed with BAP or vehicle emulsion for (L) 2 weeks or (Q) 4 weeks ( n = 8). Serum levels of (M, R) insulin and (N, S) leptin were correlated with the final body weight of each mouse using simple linear regression with 95% confidence bands around the best-fit line ( n = 8). Serum concentration of GLP-1 and GIP of CD-1 mice fed with BAP or vehicle emulsion for (O) 2 weeks or (T) 4 weeks ( n = 8). Data were analyzed using two-way ANOVA with Bonferroni post-hoc test for all daily measurements of body weight percentage and food intake (C, D, G, H) and glucose tolerance (K, P). Unpaired two-tailed t -test was used for 2 group comparisons (E, F, I, J, K, L, O, P, Q, T) . Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns. Δ represent the net differences in body weight percentage between BAP and vehicle groups. Days or time points that show significant differences between BAP and vehicle groups for body weight (C, G) , food intake (D) and blood glucose (P) are presented with ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.005, ∗∗∗∗ P < 0.0001.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Emulsion, Concentration Assay, Two Tailed Test

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: BAP increases WAT thermogenic markers and decreases hepatic MASLD markers in male CD-1 mice . A-D, The effect of daily BAP feeding on the mRNA expression of thermogenic markers, Ucp1, Dio2, Cidea, Pgc1a , in eWAT of male CD-1 mice after (A) 2 weeks or (C) 4 weeks of 300 mg/kg/day BAP ( n = 8). Relative mRNA levels of Ucp1 were correlated with the body weight of each mouse from the (B) 2 and (D) 4-week male cohorts using simple linear regression with 95% confidence bands around the best-fit line ( n = 8). E-H, Changes in eWAT Lep mRNA expression in male CD-1 mice following (E) 2 or (G) 4 weeks of 300 mg/kg/day BAP feeding ( n = 8). Relative Lep mRNA expression in eWAT was correlated with serum leptin levels of male (F) 2 week and (H) 4-week 300 mg/kg/day BAP cohorts ( n = 8). I, The effects of 4-week BAP on the mRNA expression of thermogenic markers in BAT of male CD-1 mice. J,K, The effect of daily BAP feeding on mRNA expression of liver function markers, Alpl, ColA1, Got1, Got2, Tnfa, Ugt1a1 , in the liver of male CD-1 mice after (J) 2 weeks or (K) 4 weeks of 300 mg/kg/day BAP ( n = 8). L,M, (L) The effect of daily BAP feeding on the mRNA expression of kidney function markers, Kim-1, Timp2, Igfbp7,Il-18 , in the kidney, and (M) serum uric acid levels, in CD-1 mice after 2 weeks of 75, 150, 300 mg/kg/day BAP ( n = 8). Data was analyzed using unpaired two-tailed t -test. Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Expressing, Two Tailed Test

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: BAP decreases body weight without changing food intake in HFD-fed female CD-1 mice . A, Experimental outline for the female mice cohorts. 7-week-old female CD-1 mice were acclimated with a standard chow diet in the animal facility for 1 week and were then fed a HFD for 16 weeks to induce an obese phenotype. All mice received 3 days of emulsion feeding acclimation, and a subset of mice were switched to MFD during this time while the rest remained on HFD. All mice were then randomly assigned to be fed BAP (200 mg/kg/day) or vehicle emulsion for 4 weeks. B-D, Changes in (B) body weight as a percentage of starting weight, (C) average food intake, and (D) daily food intake per cage of 2 mice of female CD-1 mice fed 200 mg/kg/day BAP or vehicle emulsion for 4 weeks, with or without MFD intervention ( n = 12 for HFD groups, n = 8–9 for MFD groups). E-J , Relative serum concentration of metabolic hormones of (E) female CD-1 mice fed with BAP (200 mg/kg/day, 4 weeks) or vehicle emulsion without MFD intervention ( n = 11). Values were normalized to vehicle. Body weight was correlated to serum concentration of (F) ghrelin and (G) leptin in female mice using simple linear regression with 95% confidence bands around the best-fit line ( n = 11). H, I, Changes in the mRNA expression of (H) thermogenic markers and (I) Lep in WAT of female mice after 4 weeks of 200 mg/kg/day BAP ( n = 11). J, K, Changes in the mRNA expression of (J) thermogenic markers in BAT and (K) liver health markers in the liver of female mice after 4 weeks of 200 mg/kg/day BAP ( n = 11). Data were analyzed using three-way ANOVA with Bonferroni post-hoc test for daily measurements of body weight percentage and food intake in the HFD vs MFD experiments (B, D) . Two-way ANOVA with Bonferroni post-hoc test was used for average body percentage weight comparison (B). Unpaired t-tests were used to compare serum hormones and tissue markers between BAP and vehicle-fed groups (E, H–K). Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns. Δ represent the net differences in body weight percentage between the BAP group and the vehicle control group.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Emulsion, Concentration Assay, Expressing, Comparison, Control

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: BAP differentially alters RER and the mRNA expression of WAT function markers in female and male CD-1 mice . A, Experimental outline for the metabolic cage experiment. 7-week-old female and male CD-1 mice were acclimated with a standard chow diet in the animal facility for 1 week and were then fed with HFD for 8 and 4 weeks, respectively, to induce an obese phenotype. All mice were moved into single-caged metabolic cage 1 weeks for cage acclimation and received 3 days of handling/emulsion feeding acclimation. All mice were then randomly assigned to be fed BAP (Female: 200 mg/kg/day, Male: 300 mg/kg/day) or vehicle emulsion for 2 weeks within the metabolic cage. B-E, Changes in (B) body weight as a percentage of starting weight, (C) fat tissue mass, (D) daily food intake, and (E) energy expenditure (EE) in female CD-1 mice fed 200 mg/kg/day BAP or vehicle emulsion for 2 weeks ( n = 8). F, Regression plot of energy expenditure from female mice fed BAP or vehicle emulsion over the second week of the experiment (Day 8–14), n = 8. G, A representative fragment of the effects of BAP on respiratory exchange ratio (RER) in female CD-1 (left). The differences between total area under the curve (AUC) value of the RER curve over entire 2-week BAP exposure between BAP and vehicle fed group (right). H, Linear regression plot of body weight percentage and RER AUC in female CD-1 mice. I, The effects of BAP on locomotor activity over the 2-week exposure in female. J, K , The effects of 13 days of BAP or vehicle emulsion feeding on (J) interscapular temperature in female mice with (K) representative images. L-N, Changes in mRNA expression of markers associated with (L) browning, (M) catecholamine signaling, and (N) fatty acid oxidation in the inguinal WAT (iWAT) of female CD-1 mice after 2 weeks of BAP or vehicle feeding. O–R, Changes in (O) body weight as a percentage of starting weight, (P) fat tissue mass, (Q) daily food intake, and (R) energy expenditure (EE) in male CD-1 mice fed 300 mg/kg/day BAP or vehicle emulsion for 2 weeks ( n = 8). S, Regression plot of energy expenditure from male mice fed BAP or vehicle emulsion over the second week of the experiment (Day 8–14), n = 7–8. T, A representative fragment of the effects of BAP on RER in male CD-1 (left). The differences between total AUC value of the RER curve over entire 2-week BAP exposure between BAP and vehicle fed group (right). U, The effects of BAP on locomotor activity over the 2-week exposure in male. V , The effects of 13 days of BAP or vehicle emulsion feeding on interscapular temperature in the male mice. W, X, Changes in mRNA expression of markers associated with (W) browning and (X) fatty acid oxidation in the epididymal WAT (eWAT) of male CD-1 mice after 2 weeks of BAP feeding. Y, Z , A summary figure of (Y) the effects of BAP on WAT function markers expression and (Z) the overall in vivo effects of BAP in female and male CD-1 mice. Data were analyzed using two-way ANOVA with Bonferroni post-hoc test for all daily measurements of body weight percentage and food intake (B, D, E, O, Q, R). Unpaired two-tailed t -test was used for 2 group comparisons (C, G, I, J, L-N, P, T-X). ANCOVA analysis controlled for body weight (F, S). Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns. Δ represent the net differences in body weight percentage between the BAP group and the vehicle control group.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Expressing, Emulsion, Activity Assay, In Vivo, Two Tailed Test, Control

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: BAP modulates hypothalamic feeding neuropeptide expression . A, D, The effect of oral BAP feeding on the mRNA expression of hypothalamic feeding neuropeptides, Npy, Agrp, Pomc , in the hypothalamus, including the (A) male groups that underwent 2 ( n = 10) or 4 weeks ( n = 8) of 300 mg/kg/day BAP, and (D) female 4 weeks 200 mg/kg/day BAP while on HFD or MFD ( n = 8–10). B, C, Final body weight as a percentage of starting weight was correlated to hypothalami Npy mRNA levels of the (B) 2-week and (C) 4-week male cohorts using simple linear regression with 95% confidence bands around the best-fit line ( n = 10–8). E, Changes in Npy mRNA expression in mHypoE-46 ( n = 4), mHypoE-44 ( n = 4), mHypoA-Bmal1-WT/F ( n = 3), and hiPSC-HLN ( n = 4) treated with 100 μM BAP for 16 h. F, Changes in Npy expression after 2-to-24-hour treatment with100 μM BAP in mHypoE-46 neuronal cell line ( n = 4). G, Changes in Npy expression after treatment of mHypoE-46 ( n = 3) cell line with increasing concentrations of BAP for 16 h. H, Changes in Pomc mRNA expression in mHypoA-Bmal1-WT/F ( n = 3), mHypoA-POMC/GFP1 ( n = 3), and primary hypothalamic cultures were generated from 8 weeks old male ( n = 7) and female ( n = 6) CD-1 mice treated with 100 μM BAP for 16 h. I, J, Changes in (I) normalized fluorescence intensity of NPY in mHypoE-46 cells treated with 100 μM BAP or DMSO for 24 h with (J) representative ICC images. Data was analyzed using unpaired two-tailed t -test when comparing neuropeptide expressions between vehicle and BAP groups (A, D, E, H, I). Two-way ANOVA with Bonferroni post-hoc test was used for the analysis of the timecourse (F), and One-way ANOVA with Bonferroni post-hoc test was used for the dose curve analysis (G). Values were normalized to vehicle. Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Expressing, Generated, Fluorescence, Two Tailed Test

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: Bioinformatic analysis of transcriptome changes in BAP-treated mHypoE-46 neurons and hypothalami of BAP-fed CD-1 male mice . A, RNA-seq heatmap of the top 150 DEGs based on padj. value in mHypoE-46 neurons treated with 4 and 16 h of 100 μM BAP ( n = 3). Over-represented pathways were determined using DAVID ORA analysis and were listed on the right. B,C, Top 10 GO pathways up- and down-regulated by (B) 4 or (C) 16 h of BAP treatment in mHypoE-46 cells, determined by threshold-free AUROC method using ErmineR GO analysis. d-f, Top enriched pathways identified by GSEA in (D) mHypoE-46 neurons treated with 4 or 16 h of BAP ( n = 3), and (E) whole hypothalami of male CD-1 mice after 2 or 4 weeks of 300 mg/kg/day BAP feeding ( n = 8). (F) Individual enrichment plots of pathways that are changed by BAP at 4 h in mHypoE-46 neurons and are related to receptor/signal transductions. G, iLINCS analyses of the top perturbagens targets that are concordant with the transcriptomic signature of BAP-treated (100 μM, 4 or 16 h) mHypoE-46 neurons. Targets that belong to the receptor tyrosine kinases family are shown in pink.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: RNA Sequencing

Journal: Molecular Metabolism

Article Title: The plant hormone, 6-benzylaminopurine, ameliorates obesity in male and female mice while on a high-fat diet

doi: 10.1016/j.molmet.2026.102366

Figure Lengend Snippet: Delineating the molecular pathways modulated by BAP in hypothalamic neurons and 3T3-L1 cells . A-D, The effect of 5 or 15 min 100 μM BAP treatment on the phosphorylation of (A) ERK1/2 (Thr202/Tyr204) ( n = 9), (B) JNK (Thr183/Tyr185) ( n = 3), (C) p38 (Thr180/Tyr182) ( n = 5), and (D) AKT (Ser473) ( n = 6) in mHypoE-46 cells. E, Changes in the phosphorylation levels of p70-S6K (Thr389) in mHypoE-46 neurons treated with 100 μM BAP over a 90 min timeourse ( n = 4). F, Treatment outline for dual EGFR/ErbB2 inhibitor afatinib pretretment experiment. G, Changes of ERK1/2 phosphorylation in mHypoE-46 cells pretreated with 10 μM or 20 μM afatinib or DMSO for 1 h, followed by 5 min exposure with 100 μM BAP or DMSO. H, Treatment outline for MEK/ERK inhibitor PD0325901 pre/cotreatment experiments. I, Changes of Npy and Egr1 mRNA expression in mHypoE-46 neurons pre/cotreated with DMSO or 10 μM PD0325901 for 1 h, followed by 16 h with 50 μM BAP or DMSO. J, Venn diagram that cross-references DEGs at 4 and 16 h based on the RNA-seq of BAP-treated mHypoE-46 and a list of putative TFs of Npy generated using JASPAR and ENCODE ChIP-seq database. K, The effect of 16 h of 100 μM BAP treatment on the mRNA expression of Egr1 in hypothalamic neuron primary culture from male and female CD-1 mice, and hypothalamic neuronal models from mice and humans ( n = 3–6). L, The effect of BAP feeding on the mRNA expression of Egr1 in male and female CD-1 mice after 2 or 4 weeks of BAP feeding (300 mg/kg/day for males, 200 mg/kg/day for females) while on HFD ( n = 8–13). M, Heatmap of the fold change of Egr1 and Npy mRNA expression in BAP-treated hypothalamic neuronal models ( n = 3–6), primary cultures ( n = 4), and hypothalami of BAP-fed mice ( n = 8–13), relative to their respective vehicle controls. N, Changes in EGR1 protein expression in mHypoE-46 neurons treated with 100 μM BAP in a 4 h timecourse. O, ChIP analysis of relative EGR1 or IgG binding to mouse Npy 5′UTR after 1 h or 4 h of 100 μM BAP or DMSO treatment in mHypoE-46 neurons ( n = 3–4). P. Summary schematic of the current understanding of the effects of BAP exposure on molecular signal transduction in mHypoE-46 hypothalamic neurons. Green and red represent signaling components that are activated or inhibited by BAP, respectively. Black starts representing putative binding targets of BAP identified by SwissTargetPrediction. Q-T, The effects of 2 weeks oral BAP feeding on Egr1 mRNA expression in the (Q) eWAT and (S) iWAT of male and female CD-1 mice, respectively ( n = 8). Body weight change as a percentage of starting weight was correlated to WAT Egr1 mRNA levels of 2-week BAP and vehicle fed (R) male and (T) female CD-1 mice using simple linear regression with 95% confidence bands around the best-fit line ( n = 8). U, Differentiation and treatment outline for the 3T3-L1 adipocyte cells. V–Y, Changes in mRNA expression of (V) Egr1 , (W) Ucp1 , (X) Pgc1a , and (Y) Pdk4 in 3T3-L1 cells pretreated with 10 μM PD0325901, followed by 24 h cotreatment with 100 μM BAP or DMSO ( n = 4). Data were analyzed using unpaired two-tailed t -test for 2 components comparisons (A-D, J, L, O, Q, S), and Two-way ANOVA with Benjamini test was used to analysis with two variables (E, G, I, N, V–Y) . One-way ANOVA with Bonferroni post-hoc test was used for ChIP analysis (O). Values were normalized to vehicle/DMSO controls. A representative western blot image was included below each western blot analysis. Values are presented as mean ± SEM with P values. P -values greater than 0.2 are presented as ns.

Article Snippet: For the animal experiments, 7-week-old male and female CD-1 mice were purchased from Charles River Laboratories (Strain code: 482) and were acclimatized to our facilities (Biological Sciences Facility and Division of Comparative Medicine, University of Toronto) 1 week prior to the commencement of our experiments.

Techniques: Phospho-proteomics, Expressing, RNA Sequencing, Generated, ChIP-sequencing, Binding Assay, Transduction, Two Tailed Test, Western Blot

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Multifunctional ACE2-nanobody fusion design for pan-specific neutralization and cardiovascular protection in SARS coronavirus infection

doi: 10.1128/aac.00024-26

Figure Lengend Snippet: Cardiovascular effects of the lead construct 72opt-ACE2-R28 in normotensive animals. Male CD-1 mice were injected intravenously via the tail vein with 72opt-ACE2-R28 at a single dose of 10 mg/kg or PBS as vehicle control. Each group consisted of seven animals (mean ± SEM). ( A ) Systolic blood pressure was measured at the tail using a high-throughput blood pressure apparatus at predetermined time intervals post-injection. ( B ) Body weights were monitored daily and compared to the initial body weights of the animals at the time of injection. ( C ) Total protein concentration in urine samples was measured 7 days post-injection.

Article Snippet: Male CD-1 mice (19 g–21 g, 4 weeks old) were obtained from Charles River Laboratories (Saint-Constant, QC, Canada).

Techniques: Construct, Injection, Control, High Throughput Screening Assay, Protein Concentration

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Multifunctional ACE2-nanobody fusion design for pan-specific neutralization and cardiovascular protection in SARS coronavirus infection

doi: 10.1128/aac.00024-26

Figure Lengend Snippet: Cardiovascular effects of lead construct 72opt-ACE2-R28 in hypertensive animals. ( A ) Progression of systolic blood pressure over the 2 weeks of establishing hypertensive male CD-1 mice via implanted mini-pumps used to inject Ang-II daily at a rate of 1.44 mg/kg/day. Animals were split into two groups ( n = 8 and n = 7) for follow-up administration of test articles. Measurements were taken every 3 days (mean ± SEM). The control group included four animals without the Ang-II mini-pump. ( B ) Effect of 72opt-ACE2-R28 on decreasing systolic blood pressure in established hypertensive mice. On the 14th day after the beginning of daily Ang-II infusion, 72opt-ACE2-R28 was injected at a single dose of 10 mg/kg into the tail vein of one hypertensive group ( n = 8), while the other hypertensive group ( n = 7) received IV saline injections. Daily Ang-II infusion continued at the same rate of 1.44 mg/kg/day during treatment. SBP was measured at the tail using the high-throughput blood pressure apparatus at predetermined time intervals post-injection. Statistical analysis of SBP reduction gave a P -value <0.005 (#) at 4 h versus 0 h in the treated group, and P -values <0.05 (*) at 4 h and 24 h in the treated versus saline groups, based on a two-way ANOVA test performed in Prism 10.

Article Snippet: Male CD-1 mice (19 g–21 g, 4 weeks old) were obtained from Charles River Laboratories (Saint-Constant, QC, Canada).

Techniques: Construct, Control, Injection, Saline, High Throughput Screening Assay