Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: Experimental groups of rats used in the study.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Isolation

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: Suppression of mTOR pathway by RAPA or pp242 following SCI. Western blot analysis of proteins isolated proximal to the lesion site ( A – F ). Representative immunoblots of experimental and control groups are shown for each analyzed protein ( A , D ). Stain-free technology was used for visualization of total protein content ( B , E ) to ensure equal protein loading and normalization of relative band intensities of individual proteins. Representative total protein stains correspond to the sample lanes in representative blots of individual analyzed proteins. The phosphorylation level of proteins was assessed by calculating ratios of normalized phosphorylated and unphosphorylated protein band intensities. All rats after SCI had a higher p-Akt/Akt ratio compared to uninjured rats ( A – C ), and RAPA caused an increase in p-Akt/Akt ( C ). All rats after SCI had a higher p-4E-BP1/4E-BP1 ratio compared to uninjured rats ( D – F ), and RAPA caused an increase, while pp242 showed a comparable phosphorylation rate to that measured in healthy tissue ( F ). Immunohistochemical staining of spinal cord sections with phospho-S6 ribosomal protein (p-S6) and hematoxylin ( G – J ). Sections from rats treated with RAPA ( H ) and pp242 ( I ) had a significantly lower number of p-S6 positive cells per section area ( J ) compared to vehicle-treated controls ( G ). Scale bars: 500 µm; G.1, H.1, and I.1 images are 1:4 magnifications of corresponding areas of the spinal cord. Arrows point to examples of DAB-stained cells. Data are shown as means ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Western Blot, Isolation, Staining, Immunohistochemical staining

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: Separation of mTOR pathway by RAPA or pp242 enhances autophagy in SCI. LC3b-II content was assessed by Western blot analyses ( A – C ). Representative immunoblot lanes of each sample group are presented ( A ). Protein band intensities were normalized to total protein content visualized by stain-free technology ( B ). Representative total protein stains are corresponding to the sample lanes in representative immunoblots. LC3b-II levels were higher in all injured rats compared to non-injured rats; however, significantly higher levels were present in animals treated with RAPA or pp242 compared to vehicle-treated controls ( C ). Immunohistochemical analysis of LC3b in spinal cord sections from rats treated with vehicle control ( D ), RAPA ( E ), or pp242 ( F ) revealed a significant increase in LC3b expression in both RAPA- and pp242-treated groups ( G ). Scale bars: 500 µm; D.1, E.1, and F.1 images are 1:4 magnifications of corresponding areas of the spinal cord. Arrows point to examples of DAB-stained cells. Data are shown as means ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Western Blot, Staining, Immunohistochemical staining, Expressing

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: Suppression of mTOR pathway by RAPA or pp242 alters cytokine production in SCI. Analysis of cytokine levels in spinal cord tissue ( A ) and serum ( B ) after SCI. Calculated concentration data were normalized to the average concentration of the vehicle control group for each cytokine. Data are presented as means ± SEM; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Concentration Assay

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: Inhibition of mTOR pathway by RAPA ( B ) leads to greater axonal sprouting assessed by the number of GAP43 + puncta compared to inhibition by pp242 ( C ) and controls ( A ), as shown in the graph ( D ). Scale bars: 500 µm; A.1, B.1, and C.1 images are 1:5 magnifications of corresponding areas of the spinal cord, whereas A.2, B.2, and C.2 are 1:4 magnifications of the indicated areas of A.1, B.1, and C.1, respectively. Arrows point to examples of GAP43 + puncta. Data are shown as means + SEM; * p < 0.05, *** p < 0.001.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Inhibition

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: mTOR inhibition caused an improvement in locomotor function and a reduction in lesion size in acute SCI. A reduction in lesion size ( A ) was observed in the RAPA and pp242 groups. Significantly higher Basso, Beattie, and Bresnahan (BBB) scores ( B ) were observed following the treatment with rapamycin or pp242 after 7 days of SCI. Data are shown as means ± SEM; ** p < 0.01, *** p < 0.001.

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Inhibition

Journal: Biomedicines

Article Title: Involvement of mTOR Pathways in Recovery from Spinal Cord Injury by Modulation of Autophagy and Immune Response

doi: 10.3390/biomedicines9060593

Figure Lengend Snippet: mTOR pathway and its inhibition by rapamycin (RAPA) and pp242 after spinal cord injury (SCI). RAPA, attached to FKBP12, inhibits mTOR complex 1 (mTORC1), which leads to decreased phosphorylation of downstream proteins S6, eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), autophagy activation through ULK1 and transcription factor EB (TFEB), and reduced inflammation. A negative feedback loop through disinhibition of insulin receptor substrate-1 (IRS) is also induced with RAPA inhibition, which leads to activation of AKT, through phosphatidylinositol 4,5-bisphosphate (PIP2) conversion to phosphatidylinositol (3,4,5)-trisphosphate (PIP3) by phosphoinositide 3-kinase (PI3K). Akt inhibits tuberous sclerosis complex (TSC), which leads to disinhibition of the small GTPase RHEB (Ras homolog–mTORC1 binding) and RHEB activates mTORC1. Inhibition of mTORC1 and mTOR complex 2 (mTORC2) by pp242 leads to inhibition of AKT and, therefore, more effective inhibition of the mTOR pathway. Panels on the right display observed changes in protein modifications and physiological responses during inhibition of the mTOR pathway after SCI compared to vehicle treated controls. Phosphatase and tensin homolog (PTEN); ribosomal protein S6 kinase 1 (S6K1).

Article Snippet: Rats were randomized into four groups postoperatively: RAPA (5 mg/kg, ChemScience, Quebec, CA), pp242 (5 mg/kg, APExBIO, Huston, TX, USA), vehicle-treated SCI control group (2% DMSO, 5% triton, saline), and a no-lesion and no-treatment control group ( ).

Techniques: Inhibition, Binding Assay, Activation Assay

Journal: bioRxiv

Article Title: Genomic analysis reveals shared genes and pathways in human and canine angiosarcoma

doi: 10.1101/570879

Figure Lengend Snippet: Including: A) somatic mutations in the seven significantly mutated genes B) somatic mutations in the PI3K pathway and tumor location. ‘Other PI3K’ includes mutations in PIK3CB , PIK3C2G , PIK3C3 , PIK3R1 , and PIK3R5 ). SNV, single nucleotide variant; Indel, insertion/deletion; SCNA, somatic copy number aberrations. SCNA frequencies are calculated based on oaCGH data from 69 canine angiosarcoma samples.

Article Snippet: Liver tumors had slightly fewer overall alterations in the PI3K gene family (3/8, 37.5%), and were significantly less likely to have PIK3CA mutations (1/8, 12.5%) than heart tumors (p chi-sq =0.009).

Techniques: Variant Assay