Journal: The American journal of clinical nutrition

Article Title: Vitamin B-12 and liver activity and expression of methionine synthase are decreased in fetuses with neural tube defects.

doi: 10.1093/ajcn/nqy340

Figure Lengend Snippet: FIGURE 1 Nutritional and metabolic markers and activities and expression of enzymes from the homocysteine remethylation pathway in liver tissue from NTD (open bars) (n = 14) and non-NTD fetuses (black bars) (n = 16). (A) Nutritional markers, folate (B9), vitamin B-12 (B12), and holoTC (nmol/g protein). (B) SAM and SAH (nmol/g protein). (C) Enzymatic activities of CBS, MS, MTHFR, and BHMT (nmol · h–1 · mg protein–1). (D) Gene expression of MTR and MTHFR. The expressions of the 2 genes were normalized using POLR2A as reference gene. Values are means ± SEMs; ∗P < 0.05, ∗∗P < 0.005, ∗∗∗P < 0.0005. BHMT, betaine-homocysteine S-methyltransferase; CBS, cystathionine β-synthase; holoTC, holotranscobalamin; MS, methionine synthase; MTHFR, methylenetetrahydrofolate reductase; MTR, methionine synthase; NTD, neural tube defect; POL2RA, RNA polymerase2 subunitA; SAH, S-adenosyl homocysteine; SAM, S-adenosyl methionine.

Article Snippet: The membranes were incubated overnight at 4◦C with primary antibodies diluted in trisbuffered saline containing 5% nonfat dried milk at the appropriate dilution (BHMT, Abcam, Ab15144; cystathionine β-synthase, Novus Biologicals, H00000875-M01; MS, Abcam, Ab9209; MTHFR, Novus Biologicals, H00004524-M03; glyceraldehyde-3-phosphate dehydrogenase, Abcam, Ab9485).

Techniques: Expressing, Gene Expression

Journal: The American journal of clinical nutrition

Article Title: Vitamin B-12 and liver activity and expression of methionine synthase are decreased in fetuses with neural tube defects.

doi: 10.1093/ajcn/nqy340

Figure Lengend Snippet: FIGURE 2 Western blot (WB) analyses of MS, CBS, MTHFR, and BHMT in fetal liver tissues from NTD and non-NTD fetuses (controls). (A) Examples of western blots. GAPDH was used as reference protein. (B) Densitometric analysis of western blots of MS, MTHFR, BHMT, and CBS normalized with GAPDH, in fetal liver tissues from NTD (open bars) (n = 14) and non-NTD fetuses (black bars) (n = 16). Values are means ± SEMs; ∗∗P < 0.005. BHMT, betaine-homocysteine methyl- transferase; CBS, cystathionine β-synthase; GAPDH, glyceraldehyde-3- phosphate dehydrogenase; MS, methionine synthase; MTHFR, methylenete- trahydrofolate reductase; NTD, neural tube defect.

Article Snippet: The membranes were incubated overnight at 4◦C with primary antibodies diluted in trisbuffered saline containing 5% nonfat dried milk at the appropriate dilution (BHMT, Abcam, Ab15144; cystathionine β-synthase, Novus Biologicals, H00000875-M01; MS, Abcam, Ab9209; MTHFR, Novus Biologicals, H00004524-M03; glyceraldehyde-3-phosphate dehydrogenase, Abcam, Ab9485).

Techniques: Western Blot

Journal: The American journal of clinical nutrition

Article Title: Vitamin B-12 and liver activity and expression of methionine synthase are decreased in fetuses with neural tube defects.

doi: 10.1093/ajcn/nqy340

Figure Lengend Snippet: FIGURE 3 MTR gene transcript level in liver fetal tissue according to the genotypes (AA, n = 21 and AG + GG, n = 9) of polymorphism rs1805087 (A). The lowest level of transcripts is observed for the AG + GG genotype. (B) MS activity in liver fetal tissue according to the polymorphism rs1805087 of MTR. The lowest activity is observed in the AG + AG genotypes of rs1805087. The dotted line represents the threshold of the values reported in control fetuses. Ct, cycle threshold; MS, methionine synthase; MTHFR, methylenetetrahydrofolate reductase; MTR, methionine synthase; NTD, neural tube defect; POL2RA, RNA polymerase2 subunitA; wa, weeks of amenorrhea.

Article Snippet: The membranes were incubated overnight at 4◦C with primary antibodies diluted in trisbuffered saline containing 5% nonfat dried milk at the appropriate dilution (BHMT, Abcam, Ab15144; cystathionine β-synthase, Novus Biologicals, H00000875-M01; MS, Abcam, Ab9209; MTHFR, Novus Biologicals, H00004524-M03; glyceraldehyde-3-phosphate dehydrogenase, Abcam, Ab9485).

Techniques: Activity Assay, Control

Journal: Frontiers in pharmacology

Article Title: Hydrogen Sulfide Promotes Thyroid Hormone Synthesis and Secretion by Upregulating Sirtuin-1.

doi: 10.3389/fphar.2022.838248

Figure Lengend Snippet: FIGURE 1 | H2S generation and related enzymes in human thyroid. (A) H2S generation in human primary thyrocytes was measured by an in situ fluorescent H2S probe. (B) H2S levels in supernatants were detected by H2S-selective sensor in human primary thyrocytes with different culture days. (C) The enzymes that generated H2S were detected by immunohistochemistry staining in human thyroid tissues. All of them were located in the cytoplasm of thyrocytes. CBS (a), CSE (b), 3-MPST (c). (D) The enzymes that generated H2S in human primary thyrocytes were detected by real-time PCR. The level of CBS mRNA expression is the highest. *p < .05, **p < .01. CBS, cystathionine β-synthase; CSE, cystathionine γ-lyase; 3-MPST, 3-mercaptosulfurtransferase; H2S, hydrogen sulfide. Data are expressed as the mean ± SEM, and all experiments were performed independently three times.

Article Snippet: After blocking with 3% bovine serum albumin (BSA, Sigma–Aldrich), tissue slides were incubated with the following primary antibodies at 4°C overnight: CBS antibody (1:200), CSE antibody (1:25), and 3-MPST antibody (1:25) (all from Santa Cruz, California, United States).

Techniques: In Situ, Generated, Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction, Expressing

Journal: Journal of medicinal food

Article Title: Rice Protein Exerts Endogenous Antioxidant Capacity via Methionine Sulfoxide Reductase and the Nrf2 Antioxidant System Independent of Age.

doi: 10.1089/jmf.2019.4504

Figure Lengend Snippet: FIG. 2. Effects of RP on hepatic expression levels of MsrA, MsrB2, and MsrB3 in growing and adult rats. (A) Hepatic protein expression levels and mRNA levels of total MsrA. (B) Nuclear and cytosolic MsrA protein contents. (C) Hepatic protein expression levels and mRNA levels of total MsrB2. (D) Hepatic protein expression levels and mRNA levels of total MsrB3. (E) Nuclear MsrB2 protein contents. (F) Nuclear MsrB3 protein contents. (G) Cytosolic MsrB2 protein levels. (H) Cytosolic MsrB3 protein levels. Values are the means – SEM (n = 6). *P < .05, in comparison with CAS-A. **P < .05, in comparison with CAS-G. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MsrA, methionine sulfoxide reductase A; MsrB2, methionine sulfoxide reductase B2; MsrB3, methionine sulfoxide reductase B3.

Article Snippet: Protein extraction and Western blotting analysis The total, cytoplasmic, and nuclear proteins were used for Western blot analysis, which were prepared as described in our previous studies.16,22,23,28 The primary antibodies of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), Cullin 3 (Cul3), MsrA, MsrB2, MsrB3, glutamate cysteine ligase catalytic subunit (GCLC), glutamate cysteine ligase modulatory subunit (GCLM), glutathione synthase (GS), GR, GSTA1, SOD, CAT, heme oxygenase 1 (HO-1), NAD(P)H: quinone oxidoreductase 1 (NQO1), H1.2, and GAPDH were purchased from Proteintech (Wuhan, China), and methionine adenosyltransferase Ia (MAT1A), S-adenosylhomocysteine hydrolase (SAHH), cystathionine b-synthase (CBS), cystathionine c-lyse (CTH), GPx1/2, and second antibody were Table 1.

Techniques: Expressing, Comparison