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Background and Aims:To study the effect of rose extract on CD4 T lymphocytes, and assess the cytokines response after cell treatment. In our previous study on endothelial cells, the rose extract reduced the secretion of inflammatory markers significantly. Methods:The red rose extract used in this study was prepared and stored until use at -20 degree C. T cells were seeded in 96-well plates at 313500 cells/well in 100microl of cell culture medium in duplicate, one half of the wells were used for biomarkers screening in the culture medium, and the other half for cytotoxicity assay. 24h after plating, the cells were treated in duplicate with 100microl of red rose extract diluted at 0.5%, 0.1%, 0.05%, 0.01% and 0.005% (v/v) in cell culture medium or with culture medium only as control for 72h. Some other wells were for untreated cells, and cells treated with rose extract at 0.005% for 48h incubation time. After 48h and 72h, the corresponding wells were used for the cytotoxicity assay and from the duplicate wells, the cell culture media were collected and stored at -80degree C until the biomarkers screening assay. Results:Cytotoxicity assay revealed insignificant changes. IFN-gamma, MCP-1, GRO, RANTES and TIMP, Angiopoietin 1 and MMP-9 were elevated. Except MMP-9 which had fold changes >2 other cytokines were minimally elevated at various concentrations and timing of rose extract treatment. None of the cytokines were less than 0.8 fold. Conclusions:Unlike in the endothelial cells, there is mild elevation in few inflammatory markers on T lymphocytes treatment by rose extract. Further studies need to be performed to estimate the clinical relevance.

Background: CC-Chemokine Receptor 5 (CCR5) and Chemokine C-X-C-Motif Receptor 4 (CXCR4) are expressed in various tissues, and they are potential molecules involved in multiple pathways. CCR5 and CXCR4 targets are associated with immune regulation in patients in multiple tissues and numerous clinical conditions. The study was performed searching for a novel therapy for immune regulation on these CCR5 and CXCR4 receptors with rose extract. Methods: The crushed red rose extract was prepared, and it was processed for analysis. The HUVEC cells were obtained for seeding in the cell culture. The cells were tested in normal physiological conditions and varying degrees of hypoxia. The cells were treated with extract for 72 hours, and the resultant secreted supernatants were analyzed for expression of CCR5 and CXCR4 by the Elisa technique. Results: The CCR5 levels were significantly elevated at normoxia compared to untreated controls. The surge of CCR5 was persistent in 12% hypoxia, and at higher degrees of hypoxia, the levels were mildly lower than the untreated levels. The CXCR4 levels were not changed in normoxia, and even with significant hypoxia, the levels were similar or mildly reduced compared to untreated values. Conclusion: The rose extract has the potentials to induce the secretion of soluble CCR5 from the HUVEC cells, and it can prevent the reduction of soluble CXCR4 levels during the hypoxic challenge of the endothelial cells. This in-turn can modulate the receptor levels on the endothelial cells, which has clinical applications.

Background:Coronary artery diseases and autoimmune disorders are common in clinical practice. In this study, a novel method of immune-modulation to modify the endothelial function was studied to modulate the features of the endothelial cells, and thereby to reduce coronary artery disease and other disorders modulated by endothelium. Methods:HUVEC cells were seeded in the cell culture, and streptococcus pyogenes were added to the cell culture, and the supernatant was studied for the secreted proteins. In the second phase, the bacterial lysate was synthesized, and the lysate was added to cell culture; and the proteins in the supernatant were studied at various time intervals. Results:When streptococcus pyogenes alone was added to culture, E Cadherin, Angiostatin, EpCAM and PDGF-AB were some of the biomarkers elevated significantly. HCC1, IGFBP2 and TIMP were some of the biomarkers which showed a reduction. When the lysate was added, the cell-culture was maintained for a longer time, and it showed the synthesis of immune regulatory cytokines. Heatmap analysis showed a significant number of proteins/cytokines concerning the immune/pathways, and toll-like receptors superfamily were modified. BLC, IL 17, BMP 7, PARC, Contactin2, IL 10 Rb, NAP 2 (CXCL 7), Eotaxin 2 were maximally increased. By principal component analysis, the results observed were significant. Conclusion:There is potential for a novel method of immunomodulation of the endothelial cells, which have pleiotropic functions, using streptococcus pyogenes and its lysates.

Background: The purpose was to develop a novel hypothetical method to increase the size of coronary arteries.Methods: During long-term observation an increase in coronary size has been seen in three unexpected scenarios. Changes in coronary artery sizes were observed in patients with mitral stenosis (n=69) by angiogram prior to percutaneous balloon mitral valvuloplasty or valve replacement surgery for severe mitral stenosis. The coronaries of patients with patent ductus arteriosus who underwent surgical closure in the past (n=14) were examined by echocardiogram. Patients with renal failure on long-term dialysis through peripheral arterio-venous fistula without left ventricular hypertrophy (n=17) were studied by echocardiography. Normal age, weight and sex matched coronary sizes served as controls in the study. All these observations were made over a period of 12 years.Results: The sizes of coronaries in patients with mitral stenosis, patients who underwent closure for patent ductus arteriosus, and in patients on hemodialysis through arteriovenous fistulas were higher than normal controls (p<0.05, for all). A hypothetical model to increase the coronary sizes could be developed based on the analysis of the differential equations of Poiseuille’s. A method is proposed of creating a peripheral arterio-venous fistula, which could be closed later electively by a percutaneous method/surgery. The closure time needs to be determined by experimental studies. The other methods could be a continuous exercise program or by the use of beta-blockers.Conclusion: A novel hypothetical method of peripheral arteriovenous fistula formation is decsribed which could potentially increase the size of the coronaries, which could be closed later.