OBJECTIVE: To analyze the difference in the gene expression, amino acid and carnitine levels in the cervical secretions between the endometria of pre-receptive and receptive stages, with an aim to provide clues for identifying new molecular markers for endometrial receptivity. METHODS: Fifty nine infertile women treated at the Department of Reproductive Medicine of Linyi People's Hospital from January 6, 2020 to January 31, 2022 were selected as as the study subjects, which were matched with 3 pairs (6 cases) of infertile women preparing for embryo transfer based on factors such as age, body mass index, and length of infertility. Endometrial tissue samples were collected for gene transcription and expression analysis. Twenty five women who had become pregnant through assisted reproductive technology were selected as the control group, and 28 non-pregnant women receiving ovulation monitoring at the Outpatient Department were enrolled as the case group. Status of endometrial receptivity was determined by ultrasonography. In the former group, endometrial tissues were sampled for sequencing, and GO and KEGG database enrichment analysis of differentially expressed genes was carried out. In the latter group, cervical secretions were collected, and amino acid and carnitine levels were measured by mass spectrometry. Statistical analysis was carried out using rank sum test, t test and chi-square test with SPSS v25.0 software. RESULTS: No difference was found in the clinical data of the patients with regard to age, body mass index, infertility years, AMH, FSH, LH, E2, and type of infertility. Compared with the receptive endometrial tissues, there were 100 significantly up-regulated genes and 191 significantly down-regulated genes in the pre-receptive endometrial tissue, with the most significantly altered ones being HLA-DRB5 and MMP10. The biological processes, molecular functions and pathways enriched by more differentially expressed genes in GO and KEGG were mainly immune regulation, cell adhesion and tryptophan metabolism. Analysis of secretion metabolism also revealed a significant difference in the levels of amino acids and carnitine metabolites between the two groups (P < 0.05), in particular those of Alanine, Valine, 3-hydroxybutyrylcarnitine (C4OH) + malonylcarnitine (C3DC)/captoylcarnitine (C10). CONCLUSION A significant difference has been discovered in the levels of gene transcription and protein expression in the endometrial tissues from the pre-receptive and receptive stages. The levels of amino acids and carnitine, such as Alanine, Valine, 3-hydroxybutyryl carnitine (C4OH)+malonyl carnitine (C3DC)/caproyl carnitine (C10), may be associated with the receptive status of the endometrium, though this need to be verified with larger samples.
Pregnancy ; Humans ; Female ; Infertility, Female/genetics* ; Endometrium/metabolism* ; Amino Acids/metabolism* ; Gene Expression ; Carnitine ; Alanine/metabolism* ; Valine/metabolism*

In industrial fermentation of amino acids the cells are often forced to synthesize the biochemicals excessive of their physiological needs. The knowledge of metabolic networks and their regulation relevant usually come from biochemical research, especially from enzymology, not from engineering study. To enrich the knowledge of metabolic sub-network of L-valine syntheses for higher production of L-valine, Corynebacterium glutamicum AS1.495 and its genetic derivatives AA361, AAT231, AATV341 were used for metabolic flux analysis. AS1.495 is a leucine auxotrophic (Leu-), and the three derivatives carry additional mutations. AA361 contains D-aspartic acid-beta-hydroxamate supersensitive marker (Leu-, L-AAHss), AAT231 (Leu-, L-AAHss, 2-TAr) is D-aspartic acid-beta-hydroxamate supersensitive and 2-thiazole alanine resistant, and AAT341 (Leu-, L-AAHss, 2-TAr, Vd-) is a D-aspartic acid-beta-hydroxamate supersensitive, 2-thiazole alanine resistant and valine-decompose-ability imperfect (Vd-). The concentrations of extra-cellular metabolites were determined under sub-steady-state of the batch culture. The metabolic flux distribution maps of the four strains were obtained, compared and analyzed. Our analysis showed that the flux ratio of EMP and HMP from the glucose-6-phosphate had increased from 0.205 in the parental strain AS1.495 to 0.321 in the multiple-mutation strain AATV341; the flux ratio of L-valine synthesis branch and the rest branches from the pyruvate node increased from 0.188 in AS1.495 to 3.29 in AATV341; the flux of lactic acid synthesis branch decreased from 11.1 in AS1.495 to 1.16 in AATV341; the flux of L-valine synthesis branch increased from 5.37 in AS1.495 to 37.3 in AATV341; and the productivity of L-valine correspondently increased from 4 g/L in AS1.495 to 24.5 g/L in AATV341. These results indicate that the introduction of analog supersensitive marker L-AAH55 and/or analog resistant marker 2-TAr skew the metabolic flux towards the formation of L-valine. This study revealed the usefulness of the metabolic flux analysis as a tool for verification of existing production strains. The analysis may play an important role in helping us b to rationally re-design metabolism for further improvement of fermentation process.
Corynebacterium glutamicum ; classification ; genetics ; metabolism ; Fermentation ; Glucose-6-Phosphate Isomerase ; metabolism ; Industrial Microbiology ; methods ; Mutation ; Pyruvic Acid ; metabolism ; Valine ; analysis ; biosynthesis

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Hepatocytes ; cytology ; metabolism ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

Maple syrup urine disease(MSUD) is an autosomal recessive disorder involving the metabolism of the branched-chain amino acids(BCAA) such as leucine, isoleucine and valine. The disorder is due to a defect in branched-chain alpha-ketoacid dehydrogenase(BCKAD) and the classic form causes rapid progressive and overwhelming illness beginning in the first weeks of life, present with poor feeding, lethargy, change in muscle tone, acidosis, seizures and coma. The goal of therapy in acutely ill patients with MSUD is an immediate reduction in the plasma levels of the BCAAs and branched-chain ketoacids. In this report, we describe an infant with MSUD who was treated by dietary therapy alone. During the therapy, acrodermatitis enteropathica-like syndrome developed with low plasma isoleucine concentration while she was receiving a formula deficient in BCAAs.
Acer* ; Acidosis ; Acrodermatitis* ; Amino Acids, Branched-Chain ; Coma ; Diet Therapy* ; Diet* ; Humans ; Infant ; Isoleucine ; Lethargy ; Leucine ; Maple Syrup Urine Disease* ; Metabolism ; Plasma ; Seizures ; Valine

BACKGROUNDVascular smooth muscle cell proliferation is an important process in the development of atherosclerosis and is associated with other cellular processes in atherogenesis. Telmisartan is reported to have partial peroxisome proliferator-activated receptor (PPAR)-γ activating properties and has been referred to as selective PPAR modulators, but valsartan just blocks angiotensin II (AngII) type 1 (AT1) receptors. This study aimed to compare the different effects of telmisartan and valsartan on human aortic smooth muscle cells (HASMCs) proliferation.

METHODSAbility of telmisartan and valsartan to inhibit proliferation of HASMCs was evaluated by the Cell Counting Kit-8 (CCK-8) in continuous cell culture. Whether the antiproliferative effects of telmisartan and valsartan depend on their effects on AngII receptors or activating the peroxisome PPAR-γ was also investigated in this study.

RESULTSTelmisartan inhibited proliferation of HASMCs by 52.4% (P < 0.01) at the concentration of 25 µmol/L and the effect depended on the dose of telmisartan, but valsartan had little effect on HASMCs proliferation (P > 0.05) and no dose response. When tested in cells stimulated with AngII, telmisartan had the same inhibition of HASMCs by 59.2% (P < 0.05) and valsartan also inhibited it by 41.6% (P < 0.05). Telmisartan and valsartan had the same effect on down-regulating AT1 receptor expression and telmisartan was superior to valsartan up-regulating AngII type 2 (AT2) receptor expression. Antiproliferative effects of telmisartan were observed when HASMCs were treated with the PPAR-γ antagonist GW9662 but antiproliferative effects of the PPAR-γ activator pioglitazone were not observed.

CONCLUSIONSTelmisartan, but not valsartan, inhibits HASMCs proliferation and has dose-dependent response without stimulation of AngII. AT2 receptor up-regulation of telmisartan contributes to its greater antiproliferative effects than valsartan. Its PPAR-γ activation does not play a critical role in inhibiting HASMCs proliferation.

Benzimidazoles ; pharmacology ; Benzoates ; pharmacology ; Cell Proliferation ; drug effects ; Humans ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; cytology ; drug effects ; PPAR gamma ; metabolism ; Receptor, Angiotensin, Type 1 ; metabolism ; Receptor, Angiotensin, Type 2 ; metabolism ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

Change in fibrin stabilizing activity of factor XIII A subunit (FXIII-A) caused by a specific mutation, Val34Leu, is recently implicated to incidences of pathophysiology of thrombosis. In an effort to understand the effect of Val34Leu on enhanced catalytic role of FXIII-A, wild type human factor XIII A (HFXIII-A) and mutant HFXIII-A: HFXIII-A (V34L), HFXIII-A (V35L) and HFXIII-A (V34L/V35L) cDNA were expressed in E.coli system where the purified recombinant FXIII-A (rFXIII-A) showed a similar specific transglutaminase activity comparable to the human native FXIII-A from platelet. Using these rFXIII-A mutants, the activation kinetics by thrombin and the enzymatic properties of the activated rFXIII-A were characterized. rFXIII-A (V34L) and rFXIII-A (V34L/V35L) mutants were activated by thrombin much faster than those of wild type rFXIII-A and V35L variant. However, the activated rFXIII-A and mutants showed the identical catalytic efficiency as measured by in vitro assay. These results suggest that ready activation caused by a specific mutation of neighboring thrombin cleavage site(s) in the activation peptide of FXIII-A like V34L resulted in the real-time amount of the activated factor XIII-A that could influence the outcome of fibrin stabilization in vivo such as alpha2- plasmin inhibitor crosslinking to fibrin, a reaction known to be dependent on the initial concentration of active factor-XIII-A.
Blood Coagulation Tests ; Catalysis ; Enzyme Activation/genetics ; Escherichia coli/genetics ; Factor XIII/*genetics/*metabolism ; Fibrin/metabolism ; Human ; Immunoblotting ; Leucine/genetics ; Mutagenesis, Site-Directed ; *Polymorphism (Genetics) ; Recombinant Proteins/genetics/metabolism ; Thrombin/metabolism ; Valine/genetics

OBJECTIVETo assess the effect of valsartan eluting-stents on restenosis and collagen deposition in neointima hyperplasia in rabbits.

METHODSValsartan eluting-stents and the carrier eluting-stents were made with patented multi-layers coating techniques. Bare stents (n = 8), carrier eluting-stents (n = 8) and valsartan eluting-stents (n = 10) were implanted into rabbit abdominal aortas, respectively. Quantitive angiography (QA) was performed before, immediately post and 3 months after stents implantations to determine the diameter of aortas. Rabbits were killed 3 months post stents implantation and the cross sections of the stented vessels were analyzed for neointimal formation: luminal area (LA), neointimal area (NIA), inner elastic lumina area (IELA), the maximal inner-membrane thickness (MIT) and percent stenosis. MASSON and picrosirius red staining were performed to observe the collagen deposition in neointima analyzed.

RESULTSThe mean aortic diameters measured by QA at different time points were similar between the groups. LA was significantly larger (5 016 269 microm(2) +/- 207,934 microm(2) vs. 4,345,548 microm(2) +/- 125,822 microm(2) and 4,302,061 microm(2) +/- 167,952 microm(2), P < 0.01 vs. valsartan stents) while NIA (441,577 microm(2) +/- 74,099 microm(2) vs. 1,119,635 microm(2) +/- 163,503 microm(2) and 1,135,636 microm(2) +/- 136,555 microm(2)) and MIT (116 microm +/- 12 microm vs. 240 microm +/- 30 microm and 192 microm +/- 21 microm) as well as percent stenosis (8% +/- 2% vs. 20% +/- 2% and 21% +/- 2%) were significantly reduced in valsartan eluting-stents group compared to bare and carrier stents groups. MASSON and picrosirius red staining revealed rich type III collagen deposition in neointima and spare type I collagen patched around stents struts in bare and carrier stents groups and collagen deposition was rarely seen in neointima and stents struts in valsartan eluting-stents group.

CONCLUSIONValsartan eluting-stents inhibited neointimal hyperplasia by decreasing collagen deposition.

Animals ; Collagen ; metabolism ; Coronary Restenosis ; metabolism ; pathology ; therapy ; Coronary Vessels ; pathology ; Drug-Eluting Stents ; Female ; Graft Occlusion, Vascular ; metabolism ; pathology ; Hyperplasia ; Male ; Rabbits ; Tetrazoles ; therapeutic use ; Tunica Intima ; pathology ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

OBJECTIVETribbles, a protein family controlling mitogen-activated protein kinase cascades, might contribute to the remodeling process in dilated cardiomyopathy. We investigated the gene expression of Tribble 3 (TRB(3)), cardiac function and collagen changes in rats with diabetic cardiomyopathy (DCM) and the modulating effects of valsartan on them.

METHODSMale Wistar rats were fed with high cholesterol diet throughout the study period, streptozocin (30 mg/kg, i.p) was given at the 28th day, valsartan (30 mg.kg(-1).d(-1), n = 13) or placebo (n = 11) was administered at the 35th day to rats with fasting blood glucose > or = 11.1 mmol/L per gavage for another 12 weeks. Control rats (n = 8) were fed with regular chow. Fasting blood glucose was monitored throughout the study, left ventricular function was determined by echocardiography, myocardial collagen content quantified after Masson-staining and myocardial mRNA expression of TRB(3) detected by quantification real-time RT-PCR at the end of study.

RESULTSCardiac function was significantly improved (EF: 74% +/- 10% vs. 66% +/- 7%, P < 0.05), myocardial collagen content decreased (13.23 +/- 3.14 vs. 16.92 +/- 3.18, P < 0.05) in rats with DCM treated with valsartan. Moreover, TRB(3) mRNA was significantly increased in rats with DCM compared to control rats (0.0198 +/- 0.0082 vs. 0.1108 +/- 0.0933, P < 0.05) and the increase could be significantly attenuated by valsartan (0.0367 +/- 0.0234, P < 0.05 vs. DCM). A significant positive correlation was observed between myocardial TRB(3) mRNA and myocardial collagen content (r = 0.67, P < 0.05) and between TRB(3) mRNA and fasting blood glucose (r = 0.69, P < 0.05) in rats with DCM.

CONCLUSIONOur results show for the first time that myocardial TRB(3) mRNA is upregulated in rats with DCM and which could be down-regulated by valsartan.

Animals ; Cardiomyopathy, Dilated ; metabolism ; physiopathology ; Diabetes Mellitus, Experimental ; Gene Expression ; Male ; Protein Kinases ; metabolism ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

We investigated the association between superoxide dismutase (SOD) Ala16Val polymorphism and the levels of oxidized LDL lipoprotein-C (ox-LDL-C) in two age-different Greek cohorts. Four hundred fifteen middle-aged (n=147 females: 43.2+/-13 years, n=268 males: 43.3+/-14 years) Caucasian Greek subjects consisted the middle aged cohort. One hundred seventy five elderly (n=88 females: 79.9+/-4 years; n=87 males: 80.6+/-4 years) were selected from the elderly cohort. Genotype data were obtained for all of them. Multiple linear regression analysis, stratified by gender and adjusted for age, smoking habits and body mass index as covariates, showed higher ox-LDL-C levels for the middle aged men with the Val/Val genotype, compared to the other allele (Ala/Ala and Ala/Val) carriers (65.9+/-25.7 vs. 55.7+/-20.5 mg/dl; standardized beta coefficient=0.192, P=0.012). On the contrary, elderly women with the Val/Val genotype occurred with lower ox-LDL-C levels compared to the Ala/Ala or Ala/Val genotype (74.2+/-22.1 vs. 86.5+/-26.6 mg/dl; standardized beta coefficient= -0.269, P=0.015). The same trend was also recorded in elderly men, however without reaching statistical significance (standardized b coefficient= -0.187, P=0.077). Moreover, elderly men and women with the Ala/Ala or Ala/Val genotype presented higher triglycerides levels compared to Val/Val (women: 145.2+/-68.7 vs. 114.3+/-34.3 mg/dl, P= 0.027; men: 147.8+/-72.4 vs. 103.7 +/-38.0 mg/dl, P=0.002). Additionally, middle aged men with the Val/Val genotype had higher HDL-C levels compared to the Ala allele carriers. The results suggest that SOD Ala16Val polymorphism is an age-dependent modulator of ox-LDL-C levels in middle-aged men and elderly women.
Adult ; Aged ; Aged, 80 and over ; Aging/*genetics ; Alanine/*genetics ; Female ; Genotype ; Humans ; Lipoproteins, LDL/*metabolism ; Male ; Middle Aged ; Polymorphism, Single Nucleotide/*genetics ; Regression Analysis ; Sex Characteristics ; Superoxide Dismutase/*genetics ; Valine/*genetics

BACKGROUND: Many studies document the presence of abnormalities in amino acids metabolism in chronic uremia. These abnormalities have been attributed to low protein intake, deficiency of excretory and metabolic functions of the diseased kidneys, toxic effects of uremia on the intermediary metabolism of amino acids and in dialysis patients, loss of protein and amino acids by the dialytic procedure. METHODS: This study was designed to compare anthropometric measurement, biochemical characteristics and plasma amino acid concentration between patients with end stage renal disease on maintenance hemodialysis (HD) and normal controls. A cross sectional study of overnight fasting plasma amino acids and plasma albumin, prealbumin, triglyceride (TG), cholesterol, transferrin concentration were performed on 20 hemodialysis patients and 20 normal controls, matched by age and sex. RESULTS: The concentrations of prealbumin (25.60+/-7.05 mg/dL vs 35.08+/-8.11 mg/dL, p<0.005), transferrin (158.30+/-39.66 mg/dL vs 275.50+/-55.46 mg/dL, p<0.001) were found to be lower in HD patients. No differences in albumin, cholesterol and TG were observed between the two groups. Several amino acids (taurine, cystine, phosphoserine) were found to be higher in the HD patients, while the concentrations of other five amino acids (serine, alanine, valine, leucine, tyrosine) were lowered in HD patients. No differences in nine amino acids (asparagine, glutamine, proline, glycine, methionine, isoleucine, lysine, histidine, arginine) were observed between the two groups. CONCLUSION: Our results suggest that chronic renal failure patients have malnutrition and amino acids abnormalities. To correct the amino acids abnormalities and improve nitrogen utilization in hemodialysis patients, correction of acidosis and supplementation of the diet with serine should be considered.
Acidosis ; Alanine ; Amino Acids ; Cholesterol ; Cystine ; Dialysis ; Diet ; Fasting ; Glutamine ; Glycine ; Histidine ; Humans ; Isoleucine ; Kidney ; Kidney Failure, Chronic ; Leucine ; Lysine ; Malnutrition ; Metabolism ; Methionine ; Nitrogen ; Nutritional Status* ; Plasma* ; Prealbumin ; Proline ; Renal Dialysis* ; Serine ; Serum Albumin ; Transferrin ; Triglycerides ; Uremia ; Valine