Andrea Paravić Radičević

Pliva (Croatia)

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Vesna Eraković

University of Rijeka

Martina Bosnar

Pliva (Croatia)

Ines Glojnarić

Pliva (Croatia)

Ivan Faraho

Pliva (Croatia)

Kosjenka Frka Borić

GlaxoSmithKline (Croatia)

Daniela Belamarić

Pliva (Croatia)

Daslav Hranueli

University of Zagreb

Mila Vrančić

GlaxoSmithKline (Croatia)

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Snježana Čužić

Pliva (Croatia)

Mihailo Banjanac

Pliva (Croatia)

Cooperative Institutions

Pliva (Croatia)

GlaxoSmithKline (Croatia)

Galapagos (Croatia)

GlaxoSmithKline (United Kingdom)

University of Zagreb

Johannes Gutenberg University Mainz

University Medical Center of the Johannes Gutenberg University Mainz

Rudjer Boskovic Institute

University of Rijeka

Agency for Medicinal Products and Medical Devices of Croatia

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Mammalian Genome Recombineering: Yeast, Still a Helper Microorganism of Choice?

DOAJ (DOAJ: Directory of Open Access Journals) (2008)

Mila Vrančić Sandra Gregorić Andrea Paravić Radičević Krešimir Gjuračić

Summary Functional studies of complex mammalian genomes have been revolutionized by the development of the recombineering methodology. Recombineering represents the sum of in vivo recombinant DNA techniques used for the production and manipulation of targeting vectors by the process of homologous recombination within the host microorganism. Although this methodology had initially been developed in yeast, the term recombineering was coined after successful introduction of similar techniques in bacterial cells. Since then, due to simplicity of amplification, manipulation and purification of mammalian targeting vectors, Escherichia coli has become the dominant helper microorganism in functional genomics studies. However, some types of experiments in functional genomics still employ yeast as a unique host for the manipulation of megabase-sized mammalian genomic regions.

Recombineering

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Enhancement of Wound Healing in Normal and Diabetic Mice by Topical Application of Amorphous Polyphosphate. Superior Effect of a Host–Guest Composite Material Composed of Collagen (Host) and Polyphosphate (Guest)

Polymers (2017)

Wernér E.G. Müller Dinko Relkovic Maximilian Ackermann Shunfeng Wang Meik Neufurth

The effect of polyphosphate (polyP) microparticles on wound healing was tested both in vitro and in a mice model in vivo. Two approaches were used: pure salts of polyphosphate, fabricated as amorphous microparticles (MPs, consisting of calcium and magnesium salts of polyP, "Ca⁻polyp-MPs" and "Mg⁻polyp-MPs"), and host⁻guest composite particles, prepared from amorphous collagen (host) and polyphosphate (guest), termed "col/polyp-MPs". Animal experiments with polyP on healing of excisional wounds were performed using both normal mice and diabetic mice. After a healing period of 7 days "Ca⁻polyp-MP" significantly improved re-epithelialization in normal mice from 31% (control) to 72% (polyP microparticle-treated). Importantly, in diabetic mice, particularly the host⁻guest particles "col/polyp-MP", increased the rate of re-epithelialization to ≈40% (control, 23%). In addition, those particles increased the expression of COL-I and COL-III as well as the expression the α-smooth muscle actin and the plasminogen activator inhibitor-1. We propose that "Ca⁻polyp-MPs", and particularly the host⁻guest "col/polyp-MPs" are useful for topical treatment of wounds.

Polyphosphate

10.3390/polym9070300

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Citations (34)

Precision-cut lung slices from bleomycin treated animals as a model for testing potential therapies for idiopathic pulmonary fibrosis

Pulmonary Pharmacology & Therapeutics (2019)

Matea Cedilak Mihailo Banjanac Daniela Belamarić Andrea Paravić Radičević Ivan Faraho

Nintedanib

Pirfenidone

TIMP1

10.1016/j.pupt.2019.02.005

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Citations (32)

The histone-deacetylase-8 selective inhibitor PCI-34051 enhances IFN-λ production in vitro and reduces inflammation in mouse models of rhinoviral infection and rhinovirus-induced exacerbations of asthma in vivo

European Respiratory Journal (2013)

Martina Bosnar Gabrijela Ergović Boška Hrvačić Kosjenka Frka Borić Andrea Paravić Radičević

An impaired immune response to viral infection, in particular reduced production of type I and III interferons (IFN), is considered to be an important mechanism of greater susceptibility of asthmatic patients to respiratory infections. Therefore, novel therapies aimed at replacing or augmenting deficient IFN production are being investigated. According to the literature, histone-deacetylase-8 (HDAC8) is a negative regulator of innate antiviral responses since it functions as a repressor for IFN-β gene expression. Consequently, HDAC8 silencing increases IFN-β mRNA levels. The aim of this work was to validate HDAC8 as a pharmacological target by using PCI-34051, HDAC8 selective inhibitor, as a tool compound. We have shown that PCI-34051, HDAC8 selective inhibitor (IC 50 10nM) with >200-fold selectivity over the other HDAC isoforms, enhanced IFN-λ production in human bronchial cells from normal and asthmatic donors after rhinovirus (RV1B and RV16) infection. In vivo , PCI-34051 reduced cell counts in bronchoalveolar lavage fluid (BALF) and viral load in lung tissue of mice infected with RV-1B. Finally, the compound was also effective at reducing BALF cell counts and cytokine concentration in a mouse model of rhinovirus-induced exacerbation of allergic airway inflammation. Taken together, these findings indicate that inhaled selective HDAC8 inhibitors could be useful as a novel treatment for rhinovirus-induced exacerbations of asthma.

Rhinovirus

HDAC8

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Persistence of the Chromosome End Regions at Low Copy Number in Mutant Strains of Streptomyces rimosus and Streptomyces lividans

DOAJ (DOAJ: Directory of Open Access Journals) (2005)

Dalia Denapaite Andrea Paravić Radičević Branka Čajavec Iain S. Hunter Daslav Hranueli

Summary Streptomycetes are important antibiotic producing bacteria that often exhibit genetic instability. One or both ends of the linear Streptomyces chromosome are lost spontaneously, resulting in viable mutant strains sometimes lacking hundreds of genes. We examined some strains of Streptomyces rimosus and Streptomyces lividans, which had been classified as »deletion mutants« and appeared to have lost chromosome end sequences. We discovered that the »deleted« sequences were still present in vegetative mycelium at a very low copy number so that they were normally not detected. The copy number in S. rimosus was estimated as 0.1–1.0 10 –3 /chromosome. Streptomyces spores contain the disappearing chromosome end sequences at a higher copy number than the vegetative mycelium, promoting their inheritance via spore preparations. This, in effect, represents a separation between germ line and deleted vegetative genomes, which has not been recognised before in Streptomyces, and has practical implications both for strain preservation and genetic studies.

Streptomycetaceae

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2D versus 3D cell culture platforms for screening antitumoral activity of novel chemical entities

Marija Trzun Andrea Paravić Radičević Kosjenka Frka Borić Dubravko Jelić Ines Vujasinović

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Correlation of ex vivo cytokine secretion profiles with scoring indices in ulcerative colitis

European Journal of Clinical Investigation (2023)

Marija Podolski Lidija Požgaj Ivan Faraho Adriana Petrinić Grba Daniela Belamarić

Abstract Background In ulcerative colitis, the complexity of mucosal cytokine secretion profiles and how they correlate with endoscopic and clinical scores is still unclear. Methods In this study, we collected fresh biopsies from UC patients to investigate which cytokines are produced in ex vivo culture conditions, a platform increasingly used for testing of novel drugs. Then, we correlated cytokine production with several scoring indices commonly used to assess the severity of the disease. Results Increased levels of IL‐1β, IL‐2, IL‐4, IL‐6, IL‐8, IL‐10, IL‐12, IL‐13, TNFα and IFNɣ were produced by biopsies of UC patients compared to non‐IBD controls. Our results show a better correlation of cytokine levels with Mayo Endoscopic Subscore (MES) and Mayo score, than the more complex Ulcerative Colitis Endoscopic Index of Severity (UCEIS). Out of 10 measured cytokines, eight correlated with MES, six with Mayo score and only three with UCEIS, due to the partial increase in cytokine secretion observed in donors with UCEIS = 7–8. When we analysed individual subscores within the UCEIS, Vascular Network subscore showed a correlation similar to MES (7/10 cytokines), while Bleeding as well as Erosions and Ulcers subscores correlated with only 3/10 cytokines, similarly to the total UCEIS. Conclusions Our findings suggest that choosing biopsies from donors with MES = 2–3 and UCEIS = 2–6 from areas with no bleeding and no superficial and/or deep ulcers could enable a deeper insight into the cytokine profile of the inflamed tissue and represent a better tool for studying potential therapeutic targets and evaluation of novel therapies.

Ex vivo

10.1111/eci.14070

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Citations (1)