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Production of Encecalin in Cell Cultures and Hairy Roots of Helianthella quinquenervis (Hook.) A. Gray

Production of Encecalin in Cell Cultures and Hairy Roots of Helianthella quinquenervis (Hook.) A. Gray

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Plant cell and organ cultures of Helianthella quinquenervis, a medicinal plant whose roots are used by the Tarahumara Indians of Chihuahua, Mexico, to relieve several ailments, were established to identify and quantify some chromenes with biological activity, such as encecalin, and to evaluate their potential for biotechnological production.

 

    • Gas chromatography-mass spectrometry (GC-MS) analysis corroborated the presence of quantifiable amounts of encecalin in H. quinquenervis cell cultures (callus and cell suspensions).

 

    • In addition, hairy roots were obtained through three transformation protocols (prick, 45-s sonication and co-culture), using wild type Agrobacterium rhizogenes A4.

 

    • After three months, cocultivation achieved the highest percentage of transformation (66%), and a comparable production (FW) of encecalin (110 μg/g) than the sonication assay (120 μg/g), both giving far higher yields than the prick assay (19 μg/g).

 

    • Stable integration of rolC and aux1 genes in the transformed roots was confirmed by polymerase chain reaction (PCR).

 

Hairy roots from cocultivation (six months-old) accumulated as much as 1086 μg/g (FW) of encecalin, over three times higher than the cell suspension cultures. The production of encecalin varied with growth kinetics, being higher at the stationary phase.

 

This is the first report of encecalin production in hairy roots of H. quinquenervis, demonstrating the potential for a future biotechnological production of chromenes.

 

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Keywords: Agrobacterium rhizogenes A4; Helianthella quinquenervis; chromenes; encecalin; medicinal plant; secondary metabolites; sonication.

 

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Diogenes Kit

NAT1000 1KIT
EUR 270

Hydrogen Peroxide Assay Kit

NAT1002 1KIT
EUR 308.4

ProtoBlock System

NAT1004 1KIT
EUR 122.4

450ML PBS 10X

NAT1006 450ML
EUR 96

4L PBS 10X

NAT1008 4L
EUR 238.8

ProtoGlow ECL 200ml Kit

NAT1010 EACH
EUR 354

ProtoGlow ECL 500ml Kit

NAT1012 EACH
EUR 627.6

100G Acrylamide

NAT1014 100G
EUR 120

500G Acrylamide

NAT1016 500G
EUR 261.6

1KG Acrylamide

NAT1018 1KG
EUR 412.8

25G AquaPor LE

NAT1020 25G
EUR 132

100G AquaPor LE GTAC

NAT1022 100G
EUR 250.8

500G AquaPor LE GTAC

NAT1024 500G
EUR 878.4

25G AquaPor ES

NAT1026 25G
EUR 152.4

100G AquaPor ES

NAT1028 100G
EUR 350.4

25G AquaPor LM

NAT1030 25G
EUR 288

100G AquaPor LM

NAT1032 100G
EUR 860.4

25G AquaPor HR

NAT1034 25G
EUR 236.4

100G AquaPor HR

NAT1036 100G
EUR 592.8

25G AquaPor 3:1

NAT1038 25G
EUR 250.8

100G AquaPor 3:1

NAT1040 100G
EUR 570

25G Bis (Methylene Bis-Acrylamide)

NAT1042 25G
EUR 100.8

25G DATD Ultrapure

NAT1044 25G
EUR 168

250G Glycine

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1KG Glycine Ultrapure

NAT1048 1KG
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250G Tris

NAT1050 250G
EUR 102

1KG Tris Ultrapure

NAT1052 1KG
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100G Tricine

NAT1054 100G
EUR 120

100G Ion Exchange Resin (mixed bed)

NAT1056 100G
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25G Riboflavin

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25ML TEMED

NAT1060 25ML
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25G Ammonium Persulfate

NAT1062 25G
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100G Ammonium Persulfate

NAT1064 100G
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1G Dithiothreitol (DTT)

NAT1066 1G
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5G Dithiothreitol (DTT)

NAT1068 5G
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Intrinsic Type 1 Interferon (IFN1) Profile of Uncultured Human Bone Marrow CD45 low CD271 + Multipotential Stromal Cells (BM-MSCs): The Impact of Donor Age, Culture Expansion and IFNα and IFNβ Stimulation

 

Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence.

 

Considering that BM-HSCs and BM-MSCs share the same BM niche, we investigated IFN1 expression profile in human BM-MSCs in relation to donor age, culture-expansion and IFN1 (α and β) stimulation.

 

    • Fluorescence-activated cell sorting was used to purify uncultured BM-MSCs from younger (19-41, n = 6) and older (59-89, n = 6) donors based on the CD45lowCD271+ phenotype, and hematopoietic-lineage cells (BM-HLCs, CD45+CD271) were used as controls.

 

    • Gene expression was analysed using integrated circuits arrays in sorted fractions as well as cultured/stimulated BM-MSCs and Y201/Y202 immortalised cell lines.

 

    • IFN1 stimulation led to BM-MSC growth arrest and upregulation of many IFN1-stimulated genes (ISGs), with IFNβ demonstrating stronger effects. Uncultured MSCs were characterised by a moderate-level ISG expression similar to Y201 cells.

 

Age-related changes in ISG expression were negligible in BM-MSCs compared to BM-HLCs, and intracellular reactive oxygen species (ROS) levels in BM-MSCs did not significantly correlate with donor age. Antiaging genes Klotho and SIRT6 correlated with more ISGs in BM-MSCs than in BM-HLCs.

 

In patients with osteoarthritis (OA), BM-MSCs expressed considerably lower levels of several ISGs, indicating that their IFN1 signature is affected in a pathological condition. In summary, BM-MSCs possess homeostatic IFN1 gene expression signature in health, which is sensitive to in vitro culture and external IFN1 stimulation. IFN signalling may facilitate in vivo BM-MSC responses to DNA damage and combating senescence and aberrant immune activation.

 

Keywords: aging; bone marrow; mesenchymal stromal cells; senescence; type 1 interferon.

 

Ozone disinfection kinetics of poliovirus 1 determined by cell culture assay, RT-qPCR, and Ethidium Monoazide qPCR Reduction in a Continuous Quench-Flow Reactor

 

Aims: A continuous quench-flow (CQF) reactor was developed to collect samples at the reaction times of less than one second. The reactor is applied to determine ozone disinfection kinetics of poliovirus and to study whether EMA-qPCR can assess the viral infectivity after ozone disinfection.

 

Methods: Ozone disinfection of poliovirus was conducted in the developed CQF, and the disinfection kinetics were tested in the range of 0.7 – 5.0 seconds at ozone concentration of 0.08 and 0.25 mg/L. Inactivation, damage on viral genome and damage on capsid integrity were determined by plaque assay, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and ethidium monoazide treatment coupled with RT-qPCR (EMA-qPCR), respectively.

 

 

Postsynthetic Functionalization of DNA-Nanocomposites with Proteins Yields Bioinstructive Matrices for Cell Culture Applications

 

We report on the directed postsynthetic functionalization of soft DNA nanocomposite materials with proteins. Using the example of the functionalization of silica nanoparticle-modified DNA polymer materials with agonists or antagonists of the epidermal growth factor receptor EGFR cell membrane receptor, we demonstrate that hierarchically structured interfaces to living cells can be established.

 

Due to the modular design principle, even complex DNA nanostructures can also be integrated into the materials, thereby enabling the high-precision arrangement of ligands on the lower nanometer length scale. We believe that such complex biohybrid material systems can be used for new applications in biotechnology.

 

Keywords: DNA nanocomposites; DNA origami nanostructure; epidermal growth factor receptor; phosphorylation; protein-DNA conjugates.

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cDNA - Human Adult Normal Tissue: Skeletal Muscle

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Frozen Tissue Section - Human Adult Normal: Skeletal Muscle

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cDNA - Mouse Normal Tissue: Skeletal Muscle

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Total RNA - Human Adult Normal Tissue: Skeletal Muscle

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Paraffin Tissue Section - Human Adult Normal: Skeletal Muscle

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Human Adult Skeletal Muscle (Normal) Whole tissue lysate

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Total Protein - Human Adult Normal Tissue: Skeletal Muscle

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Mouse Skeletal Muscle Cells

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Human Soft Tissue Slide (Skeletal Muscle Normal) (5 slides/pk)

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Human Skeletal Muscle Cells (HSkMC), fetal

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Monkey (Rhesus) Normal Skeletal Muscle Whole tissue lysate

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Total RNA - Monkey (Cynomolgus) Normal Tissue: Skeletal Muscle

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Mouse Normal Skeletal muscle (5 slides/pack, single section/slide)

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Rat Normal Skeletal muscle (single section per slide) (5 slides/pack)

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Skeletal Muscle Tissue Slides, Normal Human Paraffin Sections, 5 slides/pack

TS-H5019 -
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Genomic DNA - Human Adult Normal Tissue: Skeletal Muscle, from a single donor

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