Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: NOSTRIN is an emerging negative regulator of decompensated cirrhotic patients with portal hypertension.

doi: 10.1016/j.dld.2024.08.050

Figure Lengend Snippet: Fig. 3. (A, i) Pathological changes observed by H&E of human cirrhotic liver and control liver sections (magnification - 10X). The cirrhotic patient’s liver showed increased Nostrin expression and decreased peNOS expression, which was concomitant with reduced NO bioavailability. Representative images (magnification - 20X) of human cirrhotic liver tissue sections showing elevated hepatic Nostrin levels by DAB immunohistochemistry (A, ii). Representative images (magnification - 20X) of human liver tissue sections showing elevated hepatic eNOS levels in the control liver compared to the cirrhotic liver by DAB immunohistochemistry A, iii). Hepatic Nostrin protein expression by immunoblot assay with densitometry quantification (B) and mRNA expression by qPCR (C). Liver total eNOS and p-eNOS expression were evaluated by immunoblot assay with densitometry quantification (D). Hepatic total and phosphorylated eNOS/eNOS in the serine 1177 residue (peNOS) expression by immunoblot assay with densitometry quantification (D) and mRNA expression by qPCR (E). The hepatic cGMP levels were measured by ELISA (F). Data are expressed as mean ± standard error of mean (SEM). Symbols represent ∗p < 0.05; ∗∗p < 0.01 compared to control liver. β-actin was used as an internal loading control.

Article Snippet: Tissues were incubated with 5 % goat s t s ( t o r a H m ( t c s 2 f a a u R t e R A i ( A a w r i P t m 2 t p ( s t ( p R a t ( t i w i a ( H t a 2 t s a s S t l ( P 3 3 a c w t i t s p o B 3 a a erum for 1 hr and incubated overnight at 4 °C with primary anibodies against rabbit polyclonal anti-human Nostrin (1:100; cat: c-134,803, Santacruz) and rabbit polyclonal anti- human eNOS 1:100; cat: PA-1712–1, Bosterbio).

Techniques: Control, Expressing, Immunohistochemistry, Western Blot, Residue, Enzyme-linked Immunosorbent Assay

Journal: Basic Research in Cardiology

Article Title: Reduced nitric oxide bioavailability impairs myocardial oxygen balance during exercise in swine with multiple risk factors

doi: 10.1007/s00395-021-00890-8

Figure Lengend Snippet: In vitro assessment of left ventricular endothelial nitric oxide synthase (eNOS) and NO bioavailability in Normal and DM + HFD + CKD swine. Left ventricular endothelial eNOS mRNA ( a ) and protein ( b ) levels, as well as phosphorylation (p-eNOS) at the Ser1177 site ( c ) were not different between groups, while p-eNOS at the inhibitory Thr495 site was lower in DM + HFD + CKD swine ( d ). No differences between groups in eNOS glutathionylation ( e ), coupling ( f ), myocardial nitric oxide metabolites nitrite (NO 2 − ) and nitrate (NO 3 − , g ) and the ratio of phosphorylated to non-phosphorylated vasodilator activated phosphoprotein (p-VASP/VASP, h ) were observed. Values are mean ± SEM. The original western blots are available in the online supplemental file: original eNOS and p-eNOS (Ser1177) western blots are shown in Supplementary Fig. 6, p-eNOS (Thr495), VASP and p-VASP blots are shown in Supplementary Fig. 7, Glutathione/eNOS blots are shown in Supplementary Fig. 8, and eNOS monomer/dimer blots are shown in Supplementary Fig. 9. * P < 0.05 for DM + HFD + CKD versus Normal by Student’s t test

Article Snippet: Blots were probed with a mouse anti-human glutathione monoclonal antibody (1:1000, MA1-7620, Invitrogen) and a rabbit anti-human polyclonal eNOS antibody (1:1000, 9572, Cell Signalling Technology Inc.).

Techniques: In Vitro, Phospho-proteomics, Western Blot

Journal: Basic Research in Cardiology

Article Title: Reduced nitric oxide bioavailability impairs myocardial oxygen balance during exercise in swine with multiple risk factors

doi: 10.1007/s00395-021-00890-8

Figure Lengend Snippet: In vitro assessment of left ventricular NO bioavailability and oxidative stress in Normal and DM + HFD + CKD swine. Left ventricular myocardial nitrite (NO 2 − ) and nitrate (NO 3 − ) levels plotted as a function of eNOS protein levels ( a ), left ventricular 8-isoprostane levels ( b ), NADPH-oxidase 2 (NOX2) mRNA expression levels ( c ) left ventricular total antioxidant capacity ( d ), and in vitro small coronary artery endothelium-dependent vasodilation to bradykinin (BK) in the absence/presence of ROS scavengers Tempol and MPG ( e and f ) in Normal and DM + HFD + CKD swine. Values are mean ± SEM. * P < 0.05 and (*) P = 0.086 for DM + HFD + CKD versus Normal by logistic regression analysis ( a ), Student’s t test ( b–d ) and two-way ANOVA for repeated measures ( f )

Article Snippet: Blots were probed with a mouse anti-human glutathione monoclonal antibody (1:1000, MA1-7620, Invitrogen) and a rabbit anti-human polyclonal eNOS antibody (1:1000, 9572, Cell Signalling Technology Inc.).

Techniques: In Vitro, Expressing

Journal: Basic Research in Cardiology

Article Title: Reduced nitric oxide bioavailability impairs myocardial oxygen balance during exercise in swine with multiple risk factors

doi: 10.1007/s00395-021-00890-8

Figure Lengend Snippet: Proposed mechanisms contributing to coronary microvascular dysfunction in DM + HFD + CKD swine. NOS, nitric oxide synthase; PDE5, phosphodiesterase 5; NO, nitric oxide, eNOS, endothelial nitric oxide synthase; cGMP, cyclic guanosine monophosphate; TNFα, tumor necrosis factor α; ROS, reactive oxygen species

Article Snippet: Blots were probed with a mouse anti-human glutathione monoclonal antibody (1:1000, MA1-7620, Invitrogen) and a rabbit anti-human polyclonal eNOS antibody (1:1000, 9572, Cell Signalling Technology Inc.).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Characterization of Cardiovascular Alterations Induced by Different Chronic Cisplatin Treatments

doi: 10.3389/fphar.2017.00196

Figure Lengend Snippet: Representative images of histology and immunohistochemistry of the effect of cisplatin treatment on rat hearts. Rats were injected intraperitoneally for 5 weeks with saline (0.9% NaCl, left column) or cisplatin (3 mg/kg week −1 , right column). Histological samples embedded in paraffin and stained with Masson’s trichrome (A,B) . Samples processed for immunohistochemistry with anti connexin-43 antibody (black arrows) (C,D) and with anti eNOS (E,F) . Heterogenicity in fiber staining is clearly seen. Bar: 100 μm.

Article Snippet: Sections were then incubated overnight at 4°C with the following antibodies: monoclonal mouse anti-human connexin-43 (1:800; Santa Cruz Biotechnology, Santa Cruz, CA, USA); polyclonal rabbit anti-human eNOS (1:50; Novus Biologicals).

Techniques: Immunohistochemistry, Injection, Saline, Staining

Journal: Frontiers in Pharmacology

Article Title: Characterization of Cardiovascular Alterations Induced by Different Chronic Cisplatin Treatments

doi: 10.3389/fphar.2017.00196

Figure Lengend Snippet: (A) Representative immunoblots for connexin-43 protein expression in whole cardiac left ventricle. Diagram bars show the results of densitometric analysis of connexin-43 in whole cardiac left ventricle. Homogenized samples from heart show essentially the connexin-43 in its phosphorylated form (strongest bands for the quantification) (B) Quantitative analysis of connexin-43 mRNA levels in whole cardiac left ventricle. (C) Representative immunoblots for eNOS protein expression. Diagram bars show the results of densitometric analysis of eNOS in whole cardiac left ventricle. (D) Quantitative analysis of eNOS mRNA levels in whole cardiac left ventricle Data are presented as means ± SEM of observations obtained for 5–8 tissue samples from 5 to 8 animals per treatment. A one-way ANOVA followed by Bonferroni/Dunn post hoc test was used for statistical analysis ( ∗ P < 0.05, CPT 2 mg/kg vs. control, ∗∗∗ P < 0.001, CPT 2 mg/kg vs. control, +++ P < 0.001, CPT 3 mg/kg vs. control, $ P < 0.05, CPT 3 mg/kg vs. CPT 2 mg/Kg, $$ P < 0.01, CPT 3 mg/kg vs. CPT 2 mg/Kg).

Article Snippet: Sections were then incubated overnight at 4°C with the following antibodies: monoclonal mouse anti-human connexin-43 (1:800; Santa Cruz Biotechnology, Santa Cruz, CA, USA); polyclonal rabbit anti-human eNOS (1:50; Novus Biologicals).

Techniques: Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Characterization of Cardiovascular Alterations Induced by Different Chronic Cisplatin Treatments

doi: 10.3389/fphar.2017.00196

Figure Lengend Snippet: Representative images of histology and immunohistochemistry of the effect of cisplatin treatment on rat aorta. Rats were injected intraperitoneally for 5 weeks with saline (0.9% NaCl, left column) or cisplatin (3 mg/kg week −1 , right column). Histological samples embedded in paraffin and stained with Masson’s trichrome (A,B) . Samples processed for immunohistochemistry with anti eNOS antibody (C,D) . Bar: 100 μm.

Article Snippet: Sections were then incubated overnight at 4°C with the following antibodies: monoclonal mouse anti-human connexin-43 (1:800; Santa Cruz Biotechnology, Santa Cruz, CA, USA); polyclonal rabbit anti-human eNOS (1:50; Novus Biologicals).

Techniques: Immunohistochemistry, Injection, Saline, Staining

Journal: Frontiers in Pharmacology

Article Title: Characterization of Cardiovascular Alterations Induced by Different Chronic Cisplatin Treatments

doi: 10.3389/fphar.2017.00196

Figure Lengend Snippet: (A) Representative immunoblots for eNOS protein expression in aorta. Diagram bars show the result of densitometric analysis of eNOS in aorta. (B) Quantitative analysis of eNOS mRNA levels in aorta. (C) Representative immunoblots for plasminogen activator inhibitor-1 (PAI-1) protein expression in aorta. Diagram bars show the result of densitometric analysis of PAI-1 in aorta. Data are presented as means ± SEM of observations obtained for 5–8 tissue samples from 5 to 8 animals per treatment. A one-way ANOVA followed by Bonferroni/Dunn post hoc test was used for statistical analysis.

Article Snippet: Sections were then incubated overnight at 4°C with the following antibodies: monoclonal mouse anti-human connexin-43 (1:800; Santa Cruz Biotechnology, Santa Cruz, CA, USA); polyclonal rabbit anti-human eNOS (1:50; Novus Biologicals).

Techniques: Western Blot, Expressing