Journal: iScience

Article Title: Transcellular biosynthesis of leukotriene B 4 orchestrates neutrophil swarming to fungi

doi: 10.1016/j.isci.2022.105226

Figure Lengend Snippet: Transcellular synthesis is sufficient to restore swarming responses against C. albicans (A) A schematic diagram indicates the important steps in LTB 4 synthesis, the block points for alox5−/− and lta 4 h−/− cell types, how they can collaborate by transcellular synthesis to create LTB 4 , and how LTB 4 signals through BLT1 receptors present on both cell types to drive swarming. (B) Bone marrow cells of the indicated genotypes were pre-incubated with 10 μM of the BLT1 antagonist U-75302 or vehicle for 30 min and then added to the swarming assay. The area of the swarm was quantified at 1 and 2 h. N = 48 swarms across three independent experiments. (C) The area of fungal growth at the end of the assay (16 h) was also quantified. N≥286 swarms across three independent experiments. Mean and SD are shown. n.s. is non-significant and ∗∗∗∗p≤0.0001 by Kruskal-Wallis with Dunn’s post-test. See also Figure S2 .

Article Snippet: Rabbit polyclonal Alexa fluor 488 anti-human BLT1 , Bioss , Cat# bs-2654R-A488; RRID: AB_2924305.

Techniques: Blocking Assay, Incubation

Journal: iScience

Article Title: Transcellular biosynthesis of leukotriene B 4 orchestrates neutrophil swarming to fungi

doi: 10.1016/j.isci.2022.105226

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal Alexa fluor 488 anti-human BLT1 , Bioss , Cat# bs-2654R-A488; RRID: AB_2924305.

Techniques: Recombinant, Spectrophotometry, Gas Chromatography, Enzyme-linked Immunosorbent Assay, Software, Mass Spectrometry

Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: Alpha-Lipoic Acid-Mediated Inhibition of LTB 4 Synthesis Suppresses Epithelial-Mesenchymal Transition, Modulating Functional and Tumorigenic Capacities in Non-Small Cell Lung Cancer A549 Cells

doi: 10.1016/j.curtheres.2024.100765

Figure Lengend Snippet: Reverse transcription-quantitative PCR oligonucleotides.

Article Snippet: Blocking of nonspecific binding was performed with BSA (bovine serum albumin) for 30 minutes and subsequently the sections were incubated at 4°C overnight [ ] with the primary anti-LTA4H (D-6) antibody (mouse anti-human, 1:100, sc-390567, Santa Cruz Biotechnology, INC, Dallas, Texas) or with the anti-LTB4-R1 (BLT1) antibody (rabbit anti-human, 1:200, BS-2654R, Bioss Antibodies, Waltham, Massachusetts).

Techniques:

Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: Alpha-Lipoic Acid-Mediated Inhibition of LTB 4 Synthesis Suppresses Epithelial-Mesenchymal Transition, Modulating Functional and Tumorigenic Capacities in Non-Small Cell Lung Cancer A549 Cells

doi: 10.1016/j.curtheres.2024.100765

Figure Lengend Snippet: Primary antibody for western blotting.

Article Snippet: Blocking of nonspecific binding was performed with BSA (bovine serum albumin) for 30 minutes and subsequently the sections were incubated at 4°C overnight [ ] with the primary anti-LTA4H (D-6) antibody (mouse anti-human, 1:100, sc-390567, Santa Cruz Biotechnology, INC, Dallas, Texas) or with the anti-LTB4-R1 (BLT1) antibody (rabbit anti-human, 1:200, BS-2654R, Bioss Antibodies, Waltham, Massachusetts).

Techniques: Western Blot

Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: Alpha-Lipoic Acid-Mediated Inhibition of LTB 4 Synthesis Suppresses Epithelial-Mesenchymal Transition, Modulating Functional and Tumorigenic Capacities in Non-Small Cell Lung Cancer A549 Cells

doi: 10.1016/j.curtheres.2024.100765

Figure Lengend Snippet: Protein expression of the 5-lipoxygenase pathway and the BLT1 receptor in A549 non-small lung cancer cells. (A) mRNA levels of cPLA 2 , 5-LOX, FLAP, LTA 4 H and BLT1 normalized with housekeeping (pumilio) in the cell line. (B) The protein lysate from A549 cells were analyzed by immunoblot and the membranes were incubated with primary antibodies against cPLA 2 , 5-LOX, FLAP, LTA 4 H, BLT1 and β-actin. ( C) Semi-quantification of the relative protein expression of cPLA 2 , 5-LOX, FLAP, LTA 4 H and BLT1, normalized with β-actin. The values 110, 78, 15, 70, 37, and 43 correspond to mass units (kDa). cPLA 2 , 5-LOX, LTA 4 H, BLT1; n = 3; FLAP; n = 4

Article Snippet: Blocking of nonspecific binding was performed with BSA (bovine serum albumin) for 30 minutes and subsequently the sections were incubated at 4°C overnight [ ] with the primary anti-LTA4H (D-6) antibody (mouse anti-human, 1:100, sc-390567, Santa Cruz Biotechnology, INC, Dallas, Texas) or with the anti-LTB4-R1 (BLT1) antibody (rabbit anti-human, 1:200, BS-2654R, Bioss Antibodies, Waltham, Massachusetts).

Techniques: Expressing, Western Blot, Incubation

Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: Alpha-Lipoic Acid-Mediated Inhibition of LTB 4 Synthesis Suppresses Epithelial-Mesenchymal Transition, Modulating Functional and Tumorigenic Capacities in Non-Small Cell Lung Cancer A549 Cells

doi: 10.1016/j.curtheres.2024.100765

Figure Lengend Snippet: Localization of LTA 4 H and BLT1 in samples from patients with lung adenocarcinoma. Images indicate the localization of LTA 4 H and BLT1 in samples from patients with lung adenocarcinoma. (A) Immunohistochemistry analysis of LTA 4 H. Scale bar: 50 µm. (B) Immunohistochemistry analysis of BLT1. Scale bar: 50 µm.

Article Snippet: Blocking of nonspecific binding was performed with BSA (bovine serum albumin) for 30 minutes and subsequently the sections were incubated at 4°C overnight [ ] with the primary anti-LTA4H (D-6) antibody (mouse anti-human, 1:100, sc-390567, Santa Cruz Biotechnology, INC, Dallas, Texas) or with the anti-LTB4-R1 (BLT1) antibody (rabbit anti-human, 1:200, BS-2654R, Bioss Antibodies, Waltham, Massachusetts).

Techniques: Immunohistochemistry

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: Top 20 ranked compounds with higher libdock scores than Rapamycin.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: ADME (Adsorption, Distribution, Metabolism, Excretion) properties of compounds.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Adsorption, Solubility

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: Toxicities of compounds.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: The structures of Rapamycin and novel compounds selected from virtual screening. ( A ) ZINC000013374324; ( B ) ZINC000012495776; ( C ) Rapamycin.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: CDOCKER interaction energy of compounds with mTORC1.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: Hydrogen bond interaction parameters for each compound and mTORC1 residues.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: ( A ) ZINC000013374324-mTORC1 complex. Schematic drawing of interactions between ligands and mTORC1, the surface of the binding area was added, blue represented positive charge, red represented negative charge, and ligands were shown in the sticks, the structure around the ligand-receptor junction was shown in thinner sticks. ( B ) ZINC000012495776 -mTORC1 complex. Schematic drawing of interactions between ligands and mTORC1, the surface of the binding area was added, blue represented positive charge, red represented negative charge, and ligands were shown in the sticks, the structure around the ligand-receptor junction was shown in thinner sticks. ( C ) Rapamycin-mTORC1 complex. Schematic drawing of interactions between ligands and mTORC1, the surface of the binding area was added, blue represented positive charge, red represented negative charge, and ligands were shown in the sticks, the structure around the ligand-receptor junction was shown in thinner sticks.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Binding Assay

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: The inter-molecular interaction of the predicted binding modes of ( A ) ZINC000013374324 to mTORC1; ( B ) ZINC000012495776 to mTORC1.; ( C ) Rapamycin to mTORC1.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Binding Assay

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: Pi-Sigma interaction, Pi-Pi interaction, Pi-Alkyl interaction and Alkyl interaction parameters for each compound and mTORC1 residues.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: The molecular docking by Schrodinger. Ligands were docked into the defined binding pocket. Red represents positive charge; blue represents negative charge. ( A ) ZINC000013374324 to mTORC1; ( B ) ZINC000012495776 to mTORC1.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Binding Assay

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: The inter-molecular interaction by Schrodinger of the predicted binding modes of ( A ) ZINC000013374324 to mTORC1; ( B ) ZINC000012495776 to mTORC1.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Binding Assay

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: Pharmacophore predictions using Schrodinger. Red represents hydrogen acceptor; blue represents hydrogen donor, green represents the hydrophobic center, and yellow represents Aromatic Ring. ( A ) ZINC000013374324 to mTORC1; ( B ) ZINC000012495776 to mTORC1.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques:

Journal: Aging (Albany NY)

Article Title: Computational research of mTORC1 inhibitor on cerebral ischemia-reperfusion injury

doi: 10.18632/aging.203371

Figure Lengend Snippet: The establishment of an enzymatic reaction system of different concentrations of selected molecules and the determination of mTOR protein activity. ( A ) Aurantiamide Acetate; ( B ) Ltb4 Ethanol Amide.

Article Snippet: mTOR protein (bought from Wuhan Huamei Biological Company), Atg13 (bought from Shanghai Kemin Biological Technology Co., Ltd.), ZINC000013374324: Aurantiamide Acetate (CAS No.: 56121-42-7; bought from MedChemExpress) and ZINC000012495776: Ltb4 Ethanol Amide (CAS No.: 877459-63-7; bought from Good Laboratory Practice Bioscience).

Techniques: Activity Assay

Journal: JCI Insight

Article Title: Excessive localized leukotriene B 4 levels dictate poor skin host defense in diabetic mice

doi: 10.1172/jci.insight.120220

Figure Lengend Snippet: (A) WT CT, STZ-treated WT, STZ-treated Alox5–/–, and STZ-treated Ltb4r1–/– mice were infected s.c. with MRSA, and infection areas were measured for 9 days. (B) Bacterial CFUs in the skin at day 9 after infection. Data are mean ± SEM of 5–10 mice. *P < 0.05 vs. WT CT mice. #P < 0.05 vs. STZ-treated WT mice. (C) CT and STZ-treated mice were infected s.c. with MRSA. STZ-treated mice were treated daily with topical ointments, vehicle-control, 0.001% BLT1 antagonist (U-75302), or 0.001% BLT2 antagonist (LY255283), and infection area was measured as in A. (D) Bacterial CFUs from mice in C at day 9 after infection. Data are mean ± SEM of 4–6 mice. *P < 0.05 vs. CT mice. #P < 0.05 vs. STZ-treated mice treated with vehicle control ointment. (E) Mice were infected s.c. with bioluminescent-expressing MRSA. Infection areas were measured every other day in CT and STZ-treated mice treated daily with vehicle control or 0.001% BLT1 antagonist (U-75302) ointments. (F) Bioluminescence imaging (BLI) to quantify bacterial burden in the skin at days 1, 3, 4, 7, and 9 after infection. (G) Representative images of bioluminescent MRSA infection in control and diabetic mice that were treated or not with BLT1 antagonist scanned by BLI. (H) Lesion size of nondiabetic NOD (ctNOD) and diabetic NOD (dbNOD) mice infected with MRSA by s.c. injection. Infected dbNOD mice were treated daily with vehicle-control or 0.001% BLT1 antagonist (U-75302) ointments as in C. Data are mean ± SEM of 5–10 mice. *P < 0.05 vs. CT mice. #P < 0.05 vs. STZ-treated mice treated with vehicle control. (I) Gram stains of CT and diabetic mice that were infected and treated with the BLT1 antagonist with MRSA for 1 and 9 after infection. Top panels show 100× and bottom panels show 1,000× magnification with an inset of a cropped zoomed view of 5 ,000× magnification. Arrows indicate bacteria.

Article Snippet: The following antibodies were utilized: BLT1-PE (BD Biosciences; catalog 552836; clone 203/14F11), BLT2-AF647 (Bioss; catalog bs-2655R-A647; polyclonal), Ly6G-AF488 (BioLegend; catalog 127626; clone 1A8),Ly6G-PerCPCy5.5 (BioLegend; catalog 127616; clone 1A8), F4/80-PE (BioLegend; catalog 123110; clone BM8), F4/80-AF700 (BioLegend; catalog 123130; clone BM8), CD45-Pacific blue (BioLegend; catalog 103126; clone 30-F11), mouse CD16/32 Fc blocking antibody (BioLegend; catalog 101320; clone 93), and Zombie UV viability dye (BioLegend; catalog 423107).

Techniques: Infection, Expressing, Imaging, Injection