Saffron (Crocus sativus L.) Flower Water Extract Disrupts the Cecal Microbiome, Brush Border Membrane Functionality, and Morphology In Vivo (Gallus gallus)

Saffron (Crocus sativus L.) is known as the most expensive spice. C. sativus dried red stigmas, called threads, are used for culinary, cosmetic, and medicinal purposes. The rest of the flower is often discarded, but is now being used in teas, as coloring agents, and fodder. Previous studies have attributed antioxidant, anti-inflammatory, hepatoprotective, neuroprotective, anti-depressant, and anticancer properties to C. sativus floral bio-residues. The aim of this study is to assess C. sativus flower water extract (CFWE) for its effects on hemoglobin, brush boarder membrane (BBM) functionality, morphology, intestinal gene expression, and cecal microbiome in vivo (Gallus gallus), a clinically validated model. For this, Gallus gallus eggs were divided into six treatment groups (non-injected, 18 Ω H2O, 1% CFWE, 2% CFWE, 5% CFWE, and 10% CFWE) with n~10 for each group. On day 17 of incubation, 1 mL of the extracts/control were administered in the amnion of the eggs. The amniotic fluid along with the administered extracts are orally consumed by the developing embryo over the course of the next few days. On day 21, the hatchlings were euthanized, the blood, duodenum, and cecum were harvested for assessment. The results showed a significant dose-dependent decrease in hemoglobin concentration, villus surface area, goblet cell number, and diameter. Furthermore, we observed a significant increase in Paneth cell number and Mucin 2 (MUC2) gene expression proportional to the increase in CFWE concentration. Additionally, the cecum microbiome analysis revealed C. sativus flower water extract altered the bacterial populations. There was a significant dose-dependent reduction in Lactobacillus and Clostridium sp., suggesting an antibacterial effect of the extract on the gut in the given model. These results suggest that the dietary consumption of C. sativus flower may have negative effects on BBM functionality, morphology, mineral absorption, microbial populations, and iron status.

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Quinoa Fiber and Quercetin Alter the Composition and Function of the Cecal Microbiome and Improve Brush Border Membrane Functionality and Morphology In-Vivo (Gallus gallus)

Current Developments in Nutrition ◽

10.1093/cdn/nzab037_002 ◽

2021 ◽

Vol 5 (Supplement_2) ◽

pp. 292-292

Author(s):

Nikita Agarwal ◽

Noa Khen ◽

Nikolai Kolba ◽

Elad Tako

Keyword(s):

Gene Expression ◽

Brush Border ◽

Brush Border Membrane ◽

Iron Status ◽

Gallus Gallus ◽

Intestinal Morphology ◽

Soluble Fiber ◽

Mineral Solubility ◽

And Function

Abstract Objectives Assessment and comparison of the effects of various concentrations of soluble extracts of quinoa fiber (Chenopodium quinoa Willd.) and quercetin-3-glucoside on the zinc and iron status, brush border membrane (BBM) functionality, intestinal morphology, and cecal bacterial populations in-vivo (Gallus gallus). Methods The study utilized Gallus gallus intra-amniotic feeding, a clinically validated method to assess the effects of quinoa, quercetin, and control using seven groups (no injection, 18 Ω H2O, 5% inulin, 1% quercetin 3-glucoside, 2.5% quinoa fiber, 5% quinoa fiber, 1% quercetin 3-glucoside + 5% quinoa fiber). Upon hatch, the cecum, duodenum, pectoral muscle, liver, and blood samples were collected for the estimation of the relative abundance of the gut microbiome, mRNA gene expression Zn and Fe-related transporter proteins and brush border membrane functionality and morphology, glycogen, relative expression of lipid-related genes and hemoglobin levels, respectively. Results The results demonstrated an increase (P < 0.05) in villi height, weight, and surface area in the groups administered with quercetin, and a dose-dependent increase was observed with quinoa soluble fiber treatment. Additionally, an increase in ferroportin and duodenal cytochrome B (DcytB) was observed in the group injected with both quinoa and quercetin. Similarly, zinc transporter 7 (ZnT7) and sucrose-isomaltase (SI) gene expression was upregulated in this group. Further, the administration of quinoa soluble fiber altered the composition and function of the cecal microbiome. Conclusions The evidence suggests that quinoa and quercetin have a synergistic effect, together they are found to improve BBM morphology and functionality, affect the intestinal microbiome, increase short-chain fatty acid production, and thereby improving mineral solubility. Quinoa fibers, a polyphenol-rich superfood, may help fight micronutrient deficiencies in target populations. Funding Sources N/A.

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Effects of short-term saffron (Crocus sativus L.) intake on the in vivo activities of xenobiotic metabolizing enzymes in healthy volunteers

Food and Chemical Toxicology ◽

10.1016/j.fct.2019.05.013 ◽

2019 ◽

Vol 130 ◽

pp. 32-43 ◽

Cited By ~ 2

Author(s):

Elias Begas ◽

Maria Bounitsi ◽

Thomas Kilindris ◽

Evangelos Kouvaras ◽

Konstantinos Makaritsis ◽

...

Keyword(s):

Healthy Volunteers ◽

Crocus Sativus ◽

Short Term ◽

Metabolizing Enzymes ◽

Xenobiotic Metabolizing Enzymes ◽

Crocus Sativus L

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Simultaneous gene quantitation of multiple genes in individual bovine nuclear transfer blastocysts

Reproduction ◽

10.1530/rep.1.0966 ◽

2007 ◽

Vol 133 (1) ◽

pp. 231-242 ◽

Cited By ~ 43

Author(s):

Craig Smith ◽

Debbie Berg ◽

Sue Beaumont ◽

Neil T Standley ◽

David N Wells ◽

...

Keyword(s):

Gene Expression ◽

Nuclear Transfer ◽

Inner Cell Mass ◽

Cell Mass ◽

Ectopic Expression ◽

Cell Number ◽

Transcript Levels ◽

Multiple Genes

During somatic cell nuclear transfer (NT), the transcriptional status of the donor cell has to be reprogrammed to reflect that of an embryo. We analysed the accuracy of this process by comparing transcript levels of four developmentally important genes (Oct4,Otx2,Ifitm3,GATA6), a gene involved in epigenetic regulation (Dnmt3a) and three housekeeping genes (β-actin, β-tubulinandGAPDH) in 21 NT blastocysts with that in genetically half-identicalin vitroproduced (IVP,n=19) andin vivo(n=15) bovine embryos. We have optimised an RNA-isolation and SYBR-green-based real-time RT-PCR procedure allowing the reproducible absolute quantification of multiple genes from a single blastocyst. Our data indicated that transcript levels did not differ significantly between stage and grade-matched zona-free NT and IVP embryos except for Ifitm3/Fragilis, which was expressed at twofold higher levels in NT blastocysts.Ifitm3expression is confined to the inner cell mass at day 7 blastocysts and to the epiblast in day 14 embryos. No ectopic expression in the trophectoderm was seen in NT embryos. Gene expression in NTand IVP embryos increased between two- and threefold for all eight genes from early to late blastocyst stages. This increase exceeded the increase in cell number over this time period indicating an increase in transcript number per cell. Embryo quality (morphological grading) was correlated to cell number for NT and IVP embryos with grade 3 blastocysts containing 30% fewer cells. However, only NT embryos displayed a significant reduction in gene expression (50%) with loss of quality. Variability in gene expression levels was not significantly different in NT, IVP orin vivoembryos but differed among genes, suggesting that the stringency of regulation is intrinsic to a gene and not affected by culture or nuclear transfer.Oct4levels exhibited the lowest variability. Analysing the total variability of all eight genes for individual embryos revealed thatin vivoembryos resembled each other much more than did NT and IVP blastocysts. Furthermore,in vivoembryos, consisting of 1.5-fold more cells, generally contained two- to fourfold more transcripts for the eight genes than did their cultured counterparts. Thus, culture conditions (in vivoversusin vitro) have greater effects on gene expression than does nuclear transfer when minimising genetic heterogeneity.

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Effect of Cordyceps militaris Hot Water Extract on Immunomodulationassociated Gene Expression in Broilers, Gallus gallus

The Journal of Poultry Science ◽

10.2141/jpsa.0180067 ◽

2019 ◽

Vol 56 (2) ◽

pp. 128-139 ◽

Cited By ~ 1

Author(s):

Yeong-Hsiang Cheng ◽

Yi-Chun Hsieh ◽

Yu-Hsiang Yu

Keyword(s):

Gene Expression ◽

Water Extract ◽

Gallus Gallus ◽

Cordyceps Militaris ◽

Hot Water ◽

Hot Water Extract

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AV-65, a Novel Inhibitor of the Wnt/β-Catenin Signaling Pathway, Inhibits the Proliferation of Myeloma Cells.

Blood ◽

10.1182/blood.v114.22.2866.2866 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2866-2866

Author(s):

Hisayuki Yao ◽

Eishi Ashihara ◽

Rina Nagao ◽

Shinya Kimura ◽

Hideyo Hirai ◽

...

Keyword(s):

Gene Expression ◽

Mouse Model ◽

Signaling Pathway ◽

Small Molecule ◽

Conflicts Of Interest ◽

Flow Cytometric Analysis ◽

Cancer Cell Line ◽

Dependent Manner ◽

Dose Dependent

Abstract Abstract 2866 Poster Board II-842 Although new molecular targeting agents against multiple myeloma (MM) have been developed, MM still remains an incurable disease. It is important to continue to investigate new therapeutic agents based on the biology of MM cells. β-catenin is the downstream effector of Wnt signaling and it regulates genes implicated in malignant progression. We have demonstrated that blockade of Wnt/β-catenin signaling pathway inhibits the progression of MM by using RNA interference methods with an in vivo mouse model (Ashihara E, et al. Clin Cancer Res 15:2731, 2009.). In this study, we investigated the effects of AV-65, a novel inhibitor of the Wnt/β-catenin signaling pathway, on MM cells. The system to identify a series of small molecule compounds using a biomarker driven approach has been established. A gene expression biomarker signature reporting on the inhibition of Wnt/β-catenin signaling was generated upon treatment of a colon cancer cell line with β-catenin siRNA. This gene expression signatiure was used to screen a small molecule compound library to identify compounds which mimic knockdown of β-catenin and thus potentially inhibit the Wnt/β-catenin signaling pathway. One compound series, LC-363, was discovered from this screen and validated as novel Wnt/β-catenin signaling inhibitors (Strovel JW, et al. ASH meeting, 2007.). We investigated the inhibitory effects of AV-65, one of LC-363 compounds, on MM cell proliferation. AV-65 inhibited the proliferation of MM cells in a time- and a dose-dependent manner and the values of IC50 at 72 hrs were ranging from 11.7 to 82.1 nM. AV-65 also showed an inhibitory effect on the proliferation of RPMI8226/LR-5 melphalan-resistant MM cells (provided from Dr. William S. Dalton). In flow cytometric analysis, apoptotic cells were increased by AV-65 treatment in a time- and a dose-dependent manner. Western blotting analysis showed that β-catenin was ubiquitinated and that the expression of nuclear β-catenin diminished (Figure 1). Moreover, AV-65 suppressed T-cell factor transcriptional activities, resulting in the decrease of c-myc expression. Taken together, AV-65 promotes the degradation of β-catenin, resulting in the induction of apoptosis of MM cells. We next investigated the in vivo effects of AV-65 using an orthotopic MM-bearing mouse model. AV-65 inhibits the growth of MM cells and significantly prolongs the survival rates (Figure 2). In conclusion, AV-65 inhibited the proliferation of MM cells via inhibition of the Wnt/β-catenin signaling pathway. AV-65 is a promising therapeutic agent for treatment of MM. Disclosures: No relevant conflicts of interest to declare.

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The Immunomodulatory Effects of AlbuminIn VitroandIn Vivo

Advances in Pharmacological Sciences ◽

10.1155/2011/691928 ◽

2011 ◽

Vol 2011 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Derek S. Wheeler ◽

John S. Giuliano ◽

Patrick M. Lahni ◽

Alvin Denenberg ◽

Hector R. Wong ◽

...

Keyword(s):

Gene Expression ◽

Lethal Dose ◽

Intraperitoneal Administration ◽

Peritoneal Macrophages ◽

Luciferase Reporter Assay ◽

Luciferase Reporter ◽

Reporter Assay ◽

Dose Dependent

Albumin appears to have proinflammatory effectsin vitro. We hypothesized that albumin would induce a state of tolerance to subsequent administration of lipopolysaccharide (LPS)in vitroandin vivo. RAW264.7 and primary peritoneal macrophages were treated with increasing doses of bovine serum albumin (BSA) and harvested for NF-κB luciferase reporter assay or TNF-αELISA. In separate experiments, RAW264.7 cells were preconditioned with 1 mg/mL BSA for 18 h prior to LPS (10 μg/mL) treatment and harvested for NF-κB luciferase reporter assay or TNF-αELISA. Finally, C57Bl/6 mice were preconditioned with albumin via intraperitoneal administration 18 h prior to a lethal dose of LPS (60 mg/kg body wt). Blood was collected at 6 h after LPS administration for TNF-αELISA. Albumin produced a dose-dependent and TLR-4-dependent increase in NF-κB activation and TNF-αgene expressionin vitro. Albumin preconditioning abrogated the LPS-mediated increase in NF-κB activation and TNF-αgene expressionin vitroandin vivo. The clinical significance of these findings remains to be elucidated.

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Apocarotenoid gene expression in saffron (Crocus sativus L.)

Scientific Research and Essays ◽

10.5897/sre2015.6277 ◽

2015 ◽

Vol 10 (15) ◽

pp. 482-488 ◽

Cited By ~ 1

Author(s):

I Mir J ◽

Ahmed N ◽

H Khan M ◽

A Mokhdomi T

Keyword(s):

Gene Expression ◽

Crocus Sativus ◽

Crocus Sativus L

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282 GENE EXPRESSION PATTERNS AND QUALITY OF BOVINE EMBRYOS PRODUCED UNDER DIFFERENT OXYGEN CONCENTRATIONS

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab282 ◽

2007 ◽

Vol 19 (1) ◽

pp. 256

Author(s):

W. J. Son ◽

M. K. B. ◽

Y. J. Jeong ◽

S. Balasubramanian ◽

S. Y. Choe ◽

...

Keyword(s):

Gene Expression ◽

Embryo Development ◽

Expression Patterns ◽

Cell Number ◽

Blastocyst Stage ◽

Total Cell ◽

Blastocyst Development ◽

Glut 1

Various factors are known to influence the survival and development of in vitro-produced embryos, including co-culture with somatic cells, antioxidants, and O2 tension. Studies in several species report that embryo development and quality were enhanced at low O2 concentrations. This study compared the effects of 2 O2 concentrations on IVP embryo development, embryo quality, and gene expression to those of in vivo counterparts. Cumulus–oocyte complexes were matured in vitro in TCM-199 with hormones and 10% FCS, and inseminated in TALP medium. Presumptive zygotes were cultured in SOF medium under either 5% or 20% O2 in air. In triplicate, sets of 5 embryos at the 2-cell, 4-cell, 8-cell, 16-cell, morula, and Day 7 blastocyst stages were used for analyzing the expression patterns of apoptotic (Bax and Bcl2), metabolism (Glut-1 and Glut-5), stress (Sox, Hsp70, and G6PDH), compaction (Cx43), oxidation (PRDX5, NADH, and MnSOD), and implantation (VEGF and IFN-tau) genes using real-time quantitative PCR. The expression of each gene was normalized to that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Statistical analysis was performed with Bonferroni and Duncan tests by ANOVA (P < 0.05). Cleavage rates did not differ among groups. Blastocyst and hatched blastocyst development in 5% O2 was significantly (P < 0.05) higher than in 20% O2. Total cell number of in vivo blastocysts was significantly (P < 0.05) higher than that of IVP blastocysts. ICM ratio and apoptosis of in vivo blastocysts were significantly (P < 0.05) lower than for IVP blastocysts. The relative abundances (RAs) of Glut-1, Glut-5, MnSOD, NADH, PRDX5, Cx43, Bcl2, and IFN-τ were significantly (P < 0.05) higher in in vivo embryos, whereas the RAs of Sox, G6PDH, Hsp70, Bax, and VEGF were significantly (P < 0.05) lower than for IVP counterparts. In conclusion, culture at 5% O2 concentration resulted in higher rates of development to the blastocyst stage, higher total cell numbers, and decreased apoptosis. Furthermore, differences in expression of genes including Glut-1, Glut-5, Sox, G6PDH, Hsp70, Bax, Bcl2, Cx43, PRDX5, NADH, MnSOD, VEGF, and IFN-τ may prove useful in determining optimal culture conditions. This work was supported by ARPC (204119-03-SB010), Republic of Korea.

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Adrenomedullin is a potent stimulator of osteoblastic activity in vitro and in vivo

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1997.273.6.e1113 ◽

1997 ◽

Vol 273 (6) ◽

pp. E1113-E1120 ◽

Cited By ~ 12

Author(s):

Jillian Cornish ◽

Karen E. Callon ◽

David H. Coy ◽

Ning-Yi Jiang ◽

Liqun Xiao ◽

...

Keyword(s):

Thymidine Incorporation ◽

Cell Number ◽

Osteoblastic Activity ◽

Adult Mice ◽

Fetal Rat ◽

Neonatal Mouse Calvaria ◽

Dose Dependent Increase ◽

Dose Dependent

Adrenomedullin is a 52-amino acid vasodilator peptide produced in many tissues, including bone. It has 20% sequence identity with amylin, a regulator of osteoblast growth, and circulates in picomolar concentrations. The present study assesses whether adrenomedullin also acts on osteoblasts. At concentrations of 10−12 M and greater, adrenomedullin produced a dose-dependent increase in cell number and [3H]thymidine incorporation in cultures of fetal rat osteoblasts. This effect was also seen with adrenomedullin-(15—52), -(22—52), and -(27—52), but adrenomedullin-(40—52) was inactive. These effects were lost in the presence of amylin blockers, suggesting they were mediated by the amylin receptor. Adrenomedullin also increased [3H]thymidine incorporation into cultured neonatal mouse calvaria but, unlike amylin, did not reduce bone resorption in this model. Adrenomedullin stimulated phenylalanine incorporation into both isolated osteoblasts and calvaria. When injected daily for 5 days over the calvariae of adult mice, it increased indexes of bone formation two- to threefold ( P < 0.0001) and increased mineralized bone area by 14% ( P = 0.004). It is concluded that adrenomedullin regulates osteoblast function and that it increases bone mass in vivo. The potential of this family of peptides in the therapy of osteoporosis should be further evaluated.

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