From osteoarthritis to bladder cancer… hyaluronic acid is not just cosmetics!

Hyaluronic Acid (HA) is important in many biological processes such as wound repair, tissue hydration and inflammation. HA is also a potential biomarker for diseases such as osteoarthritis, liver cirrhosis and bladder cancer. So its importance as a prognostic / diagnostic / predictive biomarker is still to be elucidated… but it might be present even when you don’t expect it!

Hyaluronan (HA) is a linear polysaccharide comprised of a repeating disaccharide of N-acetylglucosamine and D-glucuronic acid. The major function of HA is to provide structural support of tissue as part of the extracellular matrix (ECM). Thus, HA is widely presented in connective tissue in higher animals. The size of HA varies from 100 kD to 10,000 kD and is responsible for different functions. [Read more…]

New Glycan Array – Sweet 17? Sweet 100!

Glycocalyx, literally meaning ‘sugar coat’, is an extracellular polymeric coating surrounding many prokaryotic and eukaryotic cells that consists of glycoproteins, glycolipids, proteoglycans and glycosaminoglycans. The constituents of the glycocalyx play an important role in the process of cell signaling, virus transfection, and immunity and glycosylation status is important in processes such as inflammation and tumour microenvironment. However, the range of detection tools available for glycobiology research is quite limited, and in most cases, studies aimed at characterising glycosylation patterns have to be done with HPLC using home-based methods. [Read more…]

13th European ISSX Meeting – Meet Jean-François in Glasgow!

WP_20150622_16_20_49_Pro twThis week, the 13th European ISSX (International Society for the Study of Xenobiotics) meeting is taking place in Glasgow (Scotland) at the University of Strathclyde.

The meeting brings scientists together during courses, keynote sessions, poster sessions and much more.

One of this blog’s authors, Jean-François, is attending as Cells & ADME-Tox specialist at tebu-bio.

If you’re there (or know anyone that is…) stop by booth 102 to say hello!

social_twitter_circle-02 Follow live tweets with #ISSX

 

A new photo sent by one of our Travel Grants winners!

Frances

Here is a photo sent by Francesca who is back from the AACR meeting  in Pennsylvania.

[Read more…]

Blood as a cells highway

MAB3838-98-12-1

IF on CTCs. HepG2 cells were stained with CD44-FITC labeled monoclonal antibody (Green). The cell nucleus were counterstained with DAPI (Blue) (Cat. No. 157MAB3838).

For years, we already knew that a lot of biomarkers are circulating in the blood (i.e. the secretome), as if it were a massive highway. Cells (apart from haematological cells, that is) also circulate.

There is much research now being focused on the role of several types of circulating cells in different diseases. Rapid advances in CRC research are pushing their utility to the in vitro diagnostic (IVD) arena, specifically in non-invasive diagnoses, companion diagnostics, and therapeutic monitoring. Up to date, several circulating rare cell (CRC) types have been described: Circulating Stem Cell (CSC), Circulating Endothelial Cell (CEC), Circulating Hematopoiectic Stem & Progenitor Cell (CHSPC), Circulating Mesenchymal Cell (CMC) and Circulating Tumour Cell (CTC).

Role of these CRCs still has to be elucidated, but there are some things we already know. For example, CSCs have a role in epithelial tumours & metastasis, as well as leukemia & lymphoma, together with CHPSCs and CTCs. CECs play a role in some tumour types. For cardiovascular, neurodegenerative and autoimmune diseases, CECs, together with CMSCs seem to be also involved.

Major challenges for the research on these CRCs is the optimisation of isolation methods, to make sure that we are indeed isolating CRCs and not other cell types. Current isolation methods depend on surface markers specific for these cell types, including EPCAM (1), CD44 (2), CD34 (3),  CD38 (4), CTNNB1 (5), etc.

Working on CRCs and their role in your disease? Leave your comments below!

 

 

References

1.- Zhao, M. et alAnal Chem. Oct 15, 2013; 85(20). doi: 10.1021/ac401985r.

2.- Qin, J. et al. Int J Clin Exp Pathol. 2014; 7(6): 3235–3244. PMCID: PMC4097256.

3.- Sho, E. et al. Arteriosclerosis, Thrombosis, and Vascular Biology, 2004; 24: 1916-1921.

4.- Ferrero, E. & Malavasi, F. J Leukoc Biol. 1999 Feb;65(2):151-61.

5.- Nowak, M. et al. Folia Histochem Cytobiol. 2007;45(3):233-8.

 

Thiostrepton: a novel lead compound against Dengue?

Dengue Virus (DenV) is transmitted by mosquito vectors. It infects 50-100 million people each year and is at the origin of Dengue Fever and the more lethal Dengue Hemorrhagic Fever (DHF) and Shock Syndrome (DSS) leading to an estimated 500,000 cases of DHF and 22 000 deaths. The World Health Organization (WHO) estimates that 40% of the world’s population is at risk of infection.

In the June issue of Journal of Biomolecular Screening, investigators at San Diego State University (Dept of Biology) and Institute Pasteur Korea (Seoul, South Korea) developed a multiplexed cell-based assay for the identification of modulators of pre-membrane processing as a target for the discovery of DenV inhibitors. (1)

The DenV pre-membrane protein (prM) is an essential chaperone for the viral envelope protein which prevents premature fusion with vesicles during viral export. Inhibition of pre-membrane protein cleavage restricts fusion and represents, thus, a novel druggable target.

The new in vitro assay developed in this study, is the first described cell-based assay that monitors DenVprM processing within the classical secretory pathway. In a pilot screen of 1,280 small molecules on that assay, Thiostrepton, a known cyclopeptide Antibiotic and FOXM1 inhibitor,  was identified as a novel positive hit in this assay (IC50=4.94 µM).

The utility of this novel assay has been proven by the identification of Thiostrepton (available at Focus Biomolecules cat. nr 10-2108) which may be a novel lead compound for the discovery of new drugs effective against Dengue Virus.

Thiostrepton Focus Biomolecules tebu-bio Catalog # 10-2108

Thiostrepton – Antibiotic and FOXM1 inhibitor. C72H85N19O18S5 – CAS No: 1393-48-2, 98% by TLC and HPLC at Focus Biomolecules.

 

1- Stolp Z.D. et al. “A Multiplexed Cell-Based Assay for the Identification of Modulators of Pre-Membrane Processing as a Target against Dengue Virus” (2015) J. Biomol. Screen. 20:616-626. DOI: 10.1177/1087057115571247.

 

Identification of a MicroRNA signature for Fibromyalgia diagnosis

journal.pone.0121903.g001

Heatmap of PBMCs miRNome from FM patients and controls. The microarray analysis was performed with Toray’s 3D-Gene Human miRNA Oligo chips (v.16.0; current version is v21.0). Patient samples are labeled (FM 1–11) and controls (C 1–10). Color palette is included to indicate signal intensity.

A recent paper by Cerdá-Olmedo, G. et al. unravels the miRNA signature in fibromyalgia (FM). Diagnosis of FM, a chronic musculoskeletal pain syndrome characterised by generalized body pain, hyperalgesia and other functional and emotional comorbidities, is a challenging process hindered by symptom heterogeneity and clinical overlap with other disorders. No objective diagnostic method exists at present.

This study aimed at identifying changes in miRNA expression profiles (miRNome) of FM patients for the development of a quantitative diagnostic method of FM. In addition, knowledge of FM patient miRNomes would lead to a deeper understanding of the etiology and/or symptom severity of this complex disease.

A broad profiling was first performed using Toray’s technology. miRNAs found were validated by qPCR in a later step. The profiling of FM patients PBMCs showed a marked downregulation of hsa-miR223-3p, hsa-miR451a, hsa-miR338-3p, hsa-miR143-3p, hsa-miR145-5p and hsa-miR-21-5p (4-fold or more).

Globally, 20% of the miRNAs analyzed (233/1212) showed downregulation of at least 2-fold in patients. This might indicate a general de-regulation of the miRNA synthetic pathway in FM. No significant correlations between miRNA inhibition and FM cardinal symptoms could be identified. However, the patient with the lowest score for mental fatigue coincided with the mildest inhibition in four of the five miRNAs associated with the FM-group.

Therefore, the authors propose a signature of five strikingly downregulated miRNAs (hsa-miR223-3p, hsa-miR451a, hsa-miR338-3p, hsa-miR143-3p and hsa-miR145-5p) to be used as biomarkers of FM. Validation in larger study groups would be required before the results can be transferred to the clinic, as the authors indicate.

Looking for miRNA signatures? Don’t hesitate to leave your comments below!

Mechanism of action for angiogenesis inhibitor Borrelidin discovered

Originally identified as an active molecule against the Borrelia species, Borrelidin has since been described as a selective inhibitor of threonyl tRNA synthetase (ThrRS). More recent research has found that Borrelidin (also called Treponemycin, Antibiotic U 78548 or C2989)  induces the collapse of newly formed capillary tubules, exhibits anti-bacterial, -malarial, -insecticidal activities, and displays a potent anti-VEGF induced angiogenic activity (IC50=0.8 nM).

[Read more…]

Biomarkers and shear stress

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Costaining of pAkt-1S473 (red) and PECAM-1 (green) after 30 min flow-adaptation and 5 min ortho- (D) or retrograde flow (E). Image was obtained using Cat. No. 039200-301-J34L for p-Akt staining.

Research nowadays aims at working on models as similar as possible to the real physiological status. This includes the modification of cell culture conditions, For example, one should perform cell culture under “real” oxygen levels (e.g. hypoxia, normoxia, physioxia). For circulating cells, shear stress is a key factor, as cells behave in a different way depending on whether they are cultured under static or dynamic conditions. [Read more…]

Neuroregenerative effects of microtubule stabilizing Epothilone B

Epothilone B is a bioactive microtubule-stabilizing small molecule. This blood-brain barrier permeable coumpound belongs to the most popular potent cytoskeletal modeling molecules for in vitro cell based assays (see the previous post dedicated to “5 Cytoskeletal Modeling Molecules“).Epothilone B structure cat. nr 10-2133 Focus Biomolecules tebu-bio
Recently, Ruschel J. et al. have demonstrated that its delayed systemic administration in rodents promoted axonal regeneration with Central Nervous System (CNS) injuries. These findings open new therapeutic areas regarding the use of microtubule-stabilizing drug compounds, like Epothilones, in CNS recovery and neuroregeneration. [Read more…]