Hydroxy decenoic acid down regulates <it>gtfB</it> and <it>gtfC</it> expression and prevents <it>Streptococcus mutans</it> adherence to the cell surfaces

<p>Abstract</p> <p>Background</p> <p>10<b>-</b>Hydroxy-2-decenoic acid, an unsaturated fatty acid is the most active and unique component to the royal jelly that has antimicrobial properties. <it>Streptococcus mutans</it> is associated with pathogenesis of oral cavity, gingivoperiodontal diseases and bacteremia following dental manipulations. In the oral cavity, <it>S. mutans</it> colonize the soft tissues including tongue, palate, and buccal mucosa. When considering the role of supragingival dental plaque in caries, the proportion of acid producing bacteria (particularly <it>S. mutans</it>), has direct relevance to the pathogenicity of the plaque. The genes that encode glucosyltransferases (<it>gtf</it>s) especially <it>gtfB</it> and <it>gtfC</it> are important in <it>S. mutans</it> colonization and pathogenesis. This study investigated the hydroxy-decenoic acid (HDA) effects on <it>gtfB</it> and <it>gtfC</it> expression and <it>S. mutans</it> adherence to cells surfaces.</p> <p>Methods</p> <p><it>Streptococcus mutans</it> was treated by different concentrations of HPLC purified HDA supplied by Iran Beekeeping and Veterinary Association. Real time RT-PCR and western blot assays were conducted to evaluate <it>gtfB</it> and <it>gtfC</it> genes transcription and translation before and after HDA treatment. The bacterial attachment to the cell surfaces was evaluated microscopically.</p> <p>Results</p> <p>500 μg ml^-1 of HDA inhibited <it>gtfB</it> and <it>gtfC</it> mRNA transcription and its expression. The same concentration of HDA decreased 60% the adherence of <it>S. mutans</it> to the surface of P19 cells.</p> <p>Conclusion</p> <p>Hydroxy-decenoic acid prevents <it>gtfB</it> and <it>gtfC</it> expression efficiently in the bactericide sub-concentrations and it could effectively reduce <it>S. mutans</it> adherence to the cell surfaces. In the future, therapeutic approaches to affecting <it>S. mutans</it> could be selective and it’s not necessary to put down the oral flora completely.</p

Functional convergence of Akt protein with VEGFR-1 in human endothelial progenitor cells exposed to sera from patient with type 2 diabetes mellitus

WOS: 000431155100013PubMed ID: 28732797Diabetes mellitus type 2 predisposes patients to various microvascular complications. In the current experiment, the potent role of diabetes mellitus was investigated on the content of VEGFR-1,-2, Tie-1 and-2, and Akt in human endothelial progenitor cells. The gene expression profile of mTOR and Hedgehog signaling pathways were measured by PCR array. The possible crosstalk between RTKs, mTOR and Hedgehog signaling was also studied by bioinformatic analysis. Endothelial progenitor cells were incubated with serum from normal and diabetic for 7 days. Compared to non treated cells, diabetic serum-induced cell apoptosis (similar to 2-fold) and prohibited cell migration toward bFGF (p < 0.001). EUSA analysis showed that diabetes exposed cells had increased abundance of Tie-1,-2 and VEGFR-2 and reduced amount of VEGFR-1 (p < 0.0001) in diabetic cells. Western blotting showecIA marked reduction in the protein level of Akt after cells exposure to serum from diabetic subjects (p < 0.0001). PCR array revealed a significant stimulation of both mTOR and Hedgehog signaling pathways in diabetic cells (p < 0.05). According to data from bioinformatic datasets, we showed VEGFR-1,-2 and Tie-2, but not Tie-1, are master regulators of angiogenesis. There is a crosstalk between RTK5 and mTOR signaling by involving P62, GABARAPLI, and HTT genes. It seems that physical interaction and co-expression of Akt decreased the level of VEGFR-1 in diabetic cells. Regarding data from the present experiment, diabetic serum contributed to uncontrolled induction of both mTOR and Hedgehog signaling in endothelial progenitor cells. Diabetes mellitus induces mTOR pathway by involving receptor tyrosine kinases while Hedgehog stimulation is independent of these receptors. (C) 2017 Elsevier Inc. All rights reserved.Research Council, Tabriz University of Medical Sciences [578.104.5]This work is supported by the Research Council, Tabriz University of Medical Sciences under grant number 578.104.5

Construction and Development of a Cardiac Tissue-Specific and Hypoxia-Inducible Expression Vector

Purpose: Cardiovascular gene therapy is a sophisticated approach, thanks to the safety of vectors, stable transgene expression, delivery method, and different layers of the heart. To date, numerous expression vectors have been introduced in biotechnology and biopharmacy industries in relation to genetic manipulation. Despite the rapid growth of these modalities, they must be intelligently designed, addressing the cardiac-specific transgene expression and less side effects. Herein, we conducted a pilot project aiming to design a cardiac-specific hypoxia-inducible expression cassette. Methods: We explored a new approach to design an expression cassette containing cardiac specific enhancer, hypoxia response elements (HRE), cardiac specific promoter, internal ribosome entry site (IRES), and beta globin poly A sequence to elicit specific and inducible expression of the gene of interest. Enhanced green fluorescent protein (eGFP) was sub-cloned by BglII and NotI into the cassette. The specificity and inducible expression of the cassette was determined in both mouse myoblast C2C12 and mammary glandular tumor 4T1 as ‘twin’ cells. eGFP expression was evaluated by immunofluorescence microscope and flow cytometry at 520 nm emission peak. Results: Our data revealed that the designed expression cassette provided tissue specific and hypoxia inducible (O2<1%) transgene expression. Conclusion: It is suggested that cardiac-specific enhancer combined with cardiac-specific promoter are efficient for myoblast specific gene expression. As well, this is for the first time that HRE are derived from three well known hypoxia-regulated promoters. Therefore, there is no longer need to overlap PCR process for one repeated sequence just in one promoter

Low-level laser irradiation at a high power intensity increased human endothelial cell exosome secretion via Wnt signaling

WOS: 000435589100024PubMed ID: 29603107The distinct role of low-level laser irradiation (LLLI) on endothelial exosome biogenesis remains unclear. We hypothesize that laser irradiation of high dose in human endothelial cells (ECs) contributes to the modulation of exosome biogenesis via Wnt signaling pathway. When human ECs were treated with LLLI at a power density of 80 J/cm(2), the survival rate reduced. The potential of irradiated cells to release exosomes was increased significantly by expressing genes CD63, Alix, Rab27a, and b. This occurrence coincided with an enhanced acetylcholine esterase activity, pseudopodia formation, and reduced zeta potential value 24 h post-irradiation. Western blotting showed the induction of LC3 and reduced level of P62, confirming autophagy response. Flow cytometry and electron microscopy analyses revealed the health status of the mitochondrial function indicated by normal Delta I activity without any changes in the transcription level of PINK1 and Optineurin. When cells exposed to high power laser irradiation, p-Akt/Akt ratio and in vitro tubulogenesis capacity were blunted. PCR array and bioinformatics analyses showed the induction of transcription factors promoting Wnt signaling pathways and GTPase activity. Thus, LLLI at high power intensity increased exosome biogenesis by the induction of autophagy and Wnt signaling. LLLI at high power intensity increases exosome biogenesis by engaging the transcription factors related to Wnt signaling and autophagy stimulate.Tabriz University of Medical Sciences, Tabriz, IranThis study was supported by a grant from Tabriz University of Medical Sciences, Tabriz, Iran