Metabolism, transformation and dynamic changes of alkaloids in silkworm during feeding mulberry leaves

<p>Metabolism, transformation and dynamic changes of DNJ, 2-O-<i>α</i>-D-Gal-DNJ, fagomine, isofagomine and 4-O-<i>β</i>-d-Glc-fagomine from mulberry leaves in silkworms at different instars were observed. UPLC-Q/TOF-MS and UPLC-TQ/MS methods were adopted for qualitative and quantitative analysis respectively. Three species mulberry leaves were used to feed the silkworm as controls. By analyzing and comparing the content changes of DNJ, fagomine and their derivatives in silkworms and silkworm excrement at different instar, we revealed the dynamic changes, confirmed the enrichment effect of the polyhydroxy alkaloids by silkworm, and inferred the conversion process behind this effect. The experimental results indicated that DNJ and its derivatives turned into some intermediate substances in the metabolic process, and finally they converted back and the content increased. Fagomine and its derivatives interconverted into each other in the process, 4-O-<i>β</i>-d-Glc-fagomine transformed into fagomine, while fagomine transformed into isofagomine.</p

Cultural policy of Prague

This thesis follows mainly approach of the city of Prague to culture and cultural policy. Part one contains mainly the definition of culture, specifics of Czech environment a culture as public service. It follows approach since 2000 to today, from the view of strategic documents and its effects on culture, culture funding and advocacy

Metabolomic Study of Biochemical Changes in the Plasma and Urine of Primary Dysmenorrhea Patients Using UPLC–MS Coupled with a Pattern Recognition Approach

Primary dysmenorrhea (PD) is characterized by painful menstrual cramps without any organic pathology and has a prevalence of up to 90% in adolescents. Recent advances in its etiology and pathogenesis are providing more speculative hypotheses focused on integral systems. Using a targeted tandem mass spectrometry (MS/MS)-based metabolomic platform, we explored the changes of metabolic profiling in plasma/urine simultaneously between PD patients and healthy controls before and after a 3-month herbal medicine (namely Shaofu Zhuyu formula concentrated-granule, SFZYFG) therapy. To detect and identify potential biomarkers associated with PD and SFZYFG treatment, we also performed a combined UPLC–QTOF-MS/MS-based metabolomic profiling of the plasma/urine samples, indicating a further deviation of the patients’ global metabolic profile from that of controls. The total thirty-five metabolites (nineteen in plasma and sixteen in urine), up-regulated or down-regulated (<i>p</i> < 0.05 or 0.01), were identified and contributed to PD progress. These promising identified biomarkers underpinning the metabolic pathway including sphingolipid metabolism, steroid hormone biosynthesis, and glycerophospholipid metabolism are disturbed in PD patients, which were identified by using pathway analysis with MetPA. Twenty-four altered metabolites and fourteen biochemical indicators were restored back to the control-like level after the treatment of SFZYFG and could be potential biomarkers for monitoring therapeutic efficacy. These findings may be promising to yield a valuable insight into the pathophysiology of PD and to advance the approaches of treatment, diagnosis, and prevention of PD and related syndromes

Altered Cytokine Gene Expression in Peripheral Blood Monocytes across the Menstrual Cycle in Primary Dysmenorrhea: A Case-Control Study

<div><p>Primary dysmenorrhea is one of the most common gynecological complaints in young women, but potential peripheral immunologic features underlying this condition remain undefined. In this paper, we compared 84 common cytokine gene expression profiles of peripheral blood mononuclear cells (PBMCs) from six primary dysmenorrheic young women and three unaffected controls on the seventh day before (secretory phase), and the first (menstrual phase) and the fifth (regenerative phase) days of menstruation, using a real-time PCR array assay combined with pattern recognition and gene function annotation methods. Comparisons between dysmenorrhea and normal control groups identified 11 (nine increased and two decreased), 14 (five increased and nine decreased), and 15 (seven increased and eight decreased) genes with ≥2-fold difference in expression (<em>P</em><0.05) in the three phases of menstruation, respectively. In the menstrual phase, genes encoding pro-inflammatory cytokines (IL1B, TNF, IL6, and IL8) were up-regulated, and genes encoding TGF-β superfamily members (BMP4, BMP6, GDF5, GDF11, LEFTY2, NODAL, and MSTN) were down-regulated. Functional annotation revealed an excessive inflammatory response and insufficient TGF-β superfamily member signals with anti-inflammatory consequences, which may directly contribute to menstrual pain. In the secretory and regenerative phases, increased expression of pro-inflammatory cytokines and decreased expression of growth factors were also observed. These factors may be involved in the regulation of decidualization, endometrium breakdown and repair, and indirectly exacerbate primary dysmenorrhea. This first study of cytokine gene expression profiles in PBMCs from young primary dysmenorrheic women demonstrates a shift in the balance between expression patterns of pro-inflammatory cytokines and TGF-β superfamily members across the whole menstrual cycle, underlying the peripheral immunologic features of primary dysmenorrhea.</p> </div

DAVID analysis of differentially expressed genes in women with primary dysmenorrhea on the first day of menstruation.

<p>DAVID analysis of differentially expressed genes in women with primary dysmenorrhea on the first day of menstruation.</p

DAVID analysis of differentially expressed genes in women with primary dysmenorrhea on the fifth day of menstruation.

<p>DAVID analysis of differentially expressed genes in women with primary dysmenorrhea on the fifth day of menstruation.</p

Multivariate analysis of cytokine gene expression profiles from controls (NM1-NM3) and dysmenorrheic (DM1–DM6) samples on the first day of menstruation.

<p>A) Heat map showing hierarchical clustering of individual arrays by gene expression. B) 3D PCA score plot showing separate clustering of expression profiles corresponding to DM vs. NM. C) OPLS-DA model results for DM vs NM group. D) S-plot of OPLS-DA model for DM vs. NM group.</p

PLS-DA score plot (3D) of PBMC cytokine gene expression between controls and primary dysmenorrhea groups.

<p>DS, the secretory phase in the primary dysmenorrhea group (seventh day before menstruation); DM, the menstrual phase in the primary dysmenorrhea group (first day of menstruation); DP, the proliferative phase in the primary dysmenorrhea group (fifth day of menstruation); NS, NM, NP, the secretory, menstrual and proliferative phase, respectively, in unaffected controls.</p

DataSheet1_Multi-omics analysis reveals the pathogenesis of db/db mice diabetic kidney disease and the treatment mechanisms of multi-bioactive compounds combination from Salvia miltiorrhiza.docx

Diabetic kidney disease (DKD) is a common diabetic complication. Salvia miltiorrhiza has significant therapeutic effects on diabetes complications, although the mechanism remains unclear. Here, biochemical indicators and pathological changes were used to screen out the optimal Salvia miltiorrhiza multi-bioactive compounds combination. Metabolomics, transcriptomics and proteomics were used to explore the pathogenesis of DKD. RT-PCR and parallel reaction monitoring targeted quantitative proteome analysis were utilized to investigate treatment mechanisms of the optimal Salvia miltiorrhiza multi-bioactive compounds combination. The db/db mice showed biochemical abnormalities and renal lesions. The possible metabolic pathways were steroid hormone biosynthesis and sphingolipid metabolism. The 727 differential genes found in transcriptomics were associated with biochemical indicators via gene network to finally screen 11 differential genes, which were mainly key genes of TGF-β/Smad and PI3K/Akt/FoxO signaling pathways. Salvia miltiorrhiza multi-bioactive compounds combination could significantly regulate the Egr1, Pik3r3 and Col1a1 genes. 11 differentially expressed proteins involved in the two pathways were selected, of which 9 were significantly altered in db/db mice compared to db/m mice. Salvia miltiorrhiza multi-bioactive compounds combination could callback Q9DBM2, S4R1W1, Q91Y97, P47738, A8DUK4, and A2ARV4. In summary, Salvia miltiorrhiza multi-bioactive compounds combination may ameliorate kidney injury in diabetes through regulation of TGF-β/Smad and PI3K/Akt/FoxO signaling pathways.</p

A simplified representation of biological cross-talk between multiple TNFα/IL-1-induced actions and the TGF-β superfamily member signaling pathway.

<p>TGF-β family members may interfere with the multiple roles of TNF-α/IL-1via HO-1, p300, PPARα, and PPARγ.</p