miR-181b and miR-204 suppress the VSMC proliferation and migration by downregulation of HCK.

BACKGROUND: VSMC proliferation and migration pathways play important roles in plaque formation in the vessel stenosis and re-stenosis processes. The microRNAs affect the expression of many genes that regulate these cellular processes. The aim of this study was to investigate the effects of miR-181b, miR-204, and miR-599 on the gene and protein expression levels of hematopoietic cell kinase (HCK) in VSMCs. METHODS: miR-181b, miR-204 were predicted for the suppression of HCK in the chemokine signaling pathway using bioinformatics tools. Then, the VSMCs were transfected by PEI-containing microRNAs. The HCK gene and protein expression levels were evaluated using RT-qPCR and Western blotting techniques, respectively. Moreover, the cellular proliferation and migration were evaluated by MTT and scratch assay methods. RESULTS: The miR-181b and miR-204 decreased significantly the HCK gene and (total and phosphorylated) protein expression levels. Also, the miR-599 did not show any significant effects on the HCK gene and protein levels. The data also showed that miR-181b, miR-204, and miR-599 prevent significantly the proliferation and migration of VSMCs. CONCLUSION: The downregulation of HCK by miR-181b and miR-204 suppressed the VSMC proliferation and migration.

Postinfarction intramyocardial dissection, an interesting case report and systematic review.

Intramyocardial dissection (IMD) with ventricular septal rupture (VSR) following myocardial infarction (MI) is a rare subacute form of cardiac rupture. The evidence available in this regard is scarce. We aimed to share our experience and conduct a systematic review of previous cases. We searched the literature and performed a systematic review of previous cases. A total of 37 cases of IMD with VSR were included (1 our original and 36 literature cases). Mean age was 68 ± 8 years and 20 (54.1%) patients were male. Anterior and inferior MI were observed in 14 (37.8%) and 23 (62.2%) cases, respectively. The dissected area was the septum, RV, both septum and RV, or LV apex in 21 (56.8%), 9 (24.3%), 5 (13.5%), and 2 (5.4%), respectively. Apicoseptal and inferoseptal VSR were observed in 15 (40.5%) and 22 (59.5%) cases, respectively. At least one occluded artery was observed in 29 (90.6%) of cases. Reperfusion therapy was done for 15 (40.5%) cases before the VSR occurred. Surgery, percutaneous, and medical therapy were done for 26 (70.3%), 3 (8.1%), and 7 (18.9%) cases, respectively. The mortality rate was significantly higher in the medical versus surgical-treated group (85.7% versus 42.3%, P = .027). There was a trend to higher mortality in the group with dissection of both septum and RV (P = .15). We concluded that echocardiography has a critical role in diagnosing this complication. Surgery is mandatory in IMD with VSR.

The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages.

Atherosclerosis is developed due to the formation of atheroma plaques in the coronary arteries. In this process, M1 macrophages and vascular smooth muscle cells (VSMCs) are the main functional cells. Inflammatory mediators such as histamine may inflame M1 macrophages. The aim of this study was to determine the effect of M1 macrophage secretion contents on the gene and protein expression levels of focal adhesion kinase (FAK), vasodilator-stimulated phosphoprotein (VASP), and thrombospondin1 (THBS1). Whole blood samples from the six healthy subjects (stenosis<5%), and six patients (stenosis>70%) were prepared and peripheral blood mononuclear cells (PBMCs) were isolated. Then monocytes were differentiated into M1 macrophages using 100 ng/mL granulocyte-macrophage colony stimulating factor (GM-CSF). The differentiated M1 macrophages were treated with histamine (10-6 M), and their secretion contents were harvested and added to the culture medium of VSMCs. The FAK, VASP, and THBS1 gene expression and protein levels were measured using RT-qPCR and western blot techniques in VSMCs, respectively. The FAK and THBS1 gene expression levels significantly increased in VSMCs after adding secretion contents obtained from histamine-treated M1 macrophages (p=0.023 and 0.05, respectively), while significant results were not observed for VASP gene (p=0.45). In converse with the phosphorylated VASP (pVASP) (p<0.34), the phosphorylated FAK (pFAK) and THBS1 protein levels increased in VSMCs (p<0.001). We concluded that in inflammatory conditions, the immune events could affect the macrophages by histamine. The activated macrophages could locally activate signaling pathways via FAK and THBS1 genes that are effective in the proliferation and migration of VSMCs.

Associations between miR-661, miR-1202, lncRNA-HOTAIR, lncRNA-GAS5 and MMP9 in differentiated M2-macrophages of patients with varicose veins.

BACKGROUND: The venous hypertension is suggested as the main cause of varicose disease. Some mediators and growth factors are known as the responsible of cellular events for the progression of venous perturbations. The aim of this study was to investigate non-coding (nc) RNA and MMP9 expression levels in macrophages differentiated from monocytes of patients with varicose veins. METHODS: The monocytes were isolated from the whole blood samples by RosetteSep kit and were differentiated to macrophages M2 using M-CSF factor. The based on ncRNA-gene network, lncRNA-GAS5, lncRNA-HOTAIR, miRNA-661, miRNA-1202, and MMP9 were selected. The gene expression levels were measured by RT-qPCR technique. RESULTS: Data showed that the MMP9 gene expression increased (P=0.003) while the GAS5, miRNA-661, and miRNA-1202 expression levels reduced significantly in the differentiated macrophages of patients (P=0.035, P=0.009, and P=0.015, respectively). Furthermore, the MMP9 gene expression levels were conversely related to the GAS5, HOTAIR, miRNA-661 and miRNA-1202 expression levels. CONCLUSIONS: The results suggested that the lncRNA-GAS5, miRNA-661, miRNA-1202 and MMP9 are involved in varicose disease.

Frequent Arterial Emboli in a Patient with Acute Pulmonary Thromboembolism and Patent Foramen Ovale.

Although patent foramen ovale (PFO) is a relatively common condition, the risk of paradoxical embolism is less than 2% of all arterial ischemia. We present the case of a 52-year-old man diagnosed with pulmonary thromboembolism complicated with 2 events of paradoxical emboli in the left upper and right lower limbs secondary to PFO. We also discuss some uncertainties behind the management of PFO patients after an episode of venous thromboembolism.

Interleukin 6 may be related to indoleamine 2,3-dioxygense function in M2 macrophages treated with small dense LDL particles.

Macrophages are known as important immune cells involved in the improvement of atherosclerosis plaques. The M2 macrophages are beneficial because scavenging the non-functional components in vessel sub-endothelial space. In this study, we investigated the effects of small dense LDL (sdLDL) on the changes of indoleamine 2,3-dioxygense (IDO) and interleukin (IL6) in the differentiated M2 macrophages. The patients were selected from who underwent coronary artery angiography. The monocytes were isolated from the whole blood samples of healthy (<5% stenosis) and patient (>70% stenosis; SVD, 2VD and 3VD) subjects and, were differentiated into M2 macrophages. The IDO gene expression, activity and IL6 values were measured by RT-qPCR, colorimetry and ELISA techniques, respectively. In contrast with healthy group, the IDO gene expression and activity were significantly reduced in SVD and 2VD groups (P<0.05). Furthermore, they were conversely associated to secretion of IL6. In conclusion, the data suggested that inflammatory responses in M2 macrophages differentiated from monocytes of patients after treatment of sdLDL may be related to the reduced IDO function.

The sdLDL Reduces MRC1 Expression Level and Secretion of Histamin e in Differentiated M2-macrophages from Patients with Coronary Artery Stenosis.

BACKGROUND: The macrophage polarization is proposed to be involved in initial events and remodeling of atherosclerosis plaques. Mannose receptor, C type 1 (MRC1) is a trans-membrane glycoprotein participating in phagocytosis and, is highly expressed in the M2 macrophages. OBJECTIVE: The aim of this study was to investigate the effects of sdLDL (small dense LDL) on the MRC1 gene expression level and secretion of histamine in the differentiated M2 macrophages from monocytes of patients with coronary artery stenosis and healthy subjects. METHOD: The monocytes were isolated from healthy subjects (< 5% stenosis) and patients (> 70% stenosis, SVD (Single Vessel Disease), 2VD (Two-Vessel Disease) and 3VD (Three-Vessel Disease)) by RosetteSep kit and, were differentiated into M2 macrophages by macrophage colonystimulating factor (M-CSF). The sdLDL particles were obtained by PEG-combined precipitation method. The MRC1 gene expression and histamine levels were measured by RT-qPCR and ELISA techniques, respectively. RESULTS: The MRC1 gene expression level was significantly increased in M2 macrophages of healthy subjects (P=0.05) while it reduced in SVD (P=0.05), 2VD (P=0.01) and 3VD (P=0.9) patients after treatment with sdLDL. The histamine value secreted from M2 macrophages (7-day) was higher (>3-fold, P=0.02) in patients as compared to healthy controls. CONCLUSION: The results showed that the sdLDL particles reduce the MRC1 gene expression levels in the differentiated M2 macrophages from patients with coronary artery disease. Furthermore, they had high inflammatory capacity for the secretion of histamine.

Matrix Gla protein (MGP) promoter polymorphic variants and its serum level in stenosis of coronary artery.

Although the role of matrix Gla protein (MGP) is not completely known but, its expression within subendothelial macrophages and vascular smooth muscle cells is suggested to be involved in vascular calcification. In this study, we investigated the associations between the serum MGP levels and the MGP promoter high minor allele frequency (MAF) variants with the development of stenosis in coronary arteries. Moreover, we evaluated the allele changes within predicted transcription factor elements with bioinformatics tools. 182 subjects were recruited from who underwent coronary angiography. The MGP promoter rs1800801, rs1800802 and rs1800799 genotypes and haplotypes were detected by ARMS-RFLP PCR techniques. The serum MGP concentration was measured using ELISA method. Jaspar profiles were used for scoring the polymorphic variations within the transcription factor elements. The genotype and two-allelic haplotype distributions were not significant between the patient and control groups (P > 0.05). The serum MGP levels had not significant differences between the genotypes (P > 0.1) and haplotypes (P > 0.4). Based on the prediction studies, we did not observe significant differences between the polymorphic scores in the predicted elements (P > 0.05). We concluded that the genotype and haplotype distributions of the MGP promoter high-MAF polymorphisms, as confirmed in the prediction studies and the serum MGP level are not significantly associated with the coronary artery disease. Based on the study results, the MGP protein did not play an important role in the development of stenosis of coronary arteries.

Electrocardiographic manifestations in acute methanol poisoning cannot predict mortality.

The aim of this retrospective observational case series was to determine electrocardiographic (ECG) manifestations in patients poisoned with methanol and see whether they could predict mortality. We also wanted to see whether there was an association between ECG changes and time elapsed between ingestion and treatment, age, sex, seizure, coma (Glasgow Coma Scale ≤8), arterial blood gas (ABG) parameters, and serum potassium levels on hospital admission. The study included 42 patients aged 31.14±12.5 years. Twenty-five survived and 17 died. Almost all patients had one or more abnormal ECG findings, including heart rate, rhythm, and conduction abnormalities. However, we found no significant difference between survivors and non-survivors. QTc interval did not correlate with time elapsed between ingestion and treatment, age, sex, seizure and coma, HCO3(-), or serum potassium level. Similarly, T waves showed no correlation with serum potassium. ECG abnormalities did not correlate with coma or seizure. Even though cardiotoxicity in methanol poisoning is high, none of the ECG abnormalities found in our study predicted mortality. This however does not rule out the need to routinely run ECG for cardiotoxicity in every single patient poisoned by methanol.

No association between MGP rs1800802 polymorphism and stenosis of the coronary artery.

BACKGROUND AND OBJECTIVES: Matrix Gla protein (MGP) was originally isolated from bone but it is known to be expressed in several tissues including kidney, lung, heart, cartilage and vascular smooth muscle cells (VSMC) of the blood vessel wall. Since it inhibits calcification in subendothelial space of vessels thus, we evaluated the association of rs1800802(T > C) polymorphism and stenosis of the coronary artery. DESIGN AND SETTING: Cross-sectional case-control. SUBJECTS AND METHODS: One hundred eighty two subjects recruited on the basis of study protocol from who underwent coronary angiography. The controls (n=70) had normal coronary arteries (up to 5% stenosis). The patients (n=112) subdivided into three subgroups; single-vessel disease (SVD), two-vessel disease (2VD) and three-vessel disease (3VD) based on the number of stenosed coronary vessels (at least 50% stenosis). rs1800802 (T > C) polymorphism was determined by PCR-RFLP technique. RESULTS: Genotype distribution was not significant between control and patient groups. In addition, there were no significant differences between rs1800802 (T > C) frequency and gender (P=.092), and also patient subgroups (one-, two- and three vessel disease) (P=.840). CONCLUSION: We concluded that rs1800802 (T > C) polymorphism within the MGP promoter is not related to stenosis of the coronary artery.

Central retinal artery occlusion in Crohn disease.

Crohn disease is an inflammatory bowel disease frequently associated with extraintestinal manifestations. Ocular manifestations are uncommon but may cause significant morbidity, including blindness. We report the first case of a 9-year-old boy with biopsy-proven Crohn disease who developed a cilioretinal artery-sparing central retinal artery occlusion. After 2 months of follow-up, the patient developed optic atrophy with no change in visual acuity.

Association between rs4673 (C/T) and rs13306294 (A/G) haplotypes of NAD(P)H oxidase p22phox gene and severity of stenosis in coronary arteries.

Phagocytic NADH/NADPH oxidase is an important enzyme producing reactive oxygen species within subendothelial space of vessels. Findings have shown that p22phox subunit is an essential element related to the enzyme activity. Since some p22phox polymorphisms are thought to have functional roles in the enzyme thus, we studied the association between rs4673 (C242T) and rs13306294 (A/G) haplotypes and the severity of stenosis in coronary arteries. One hundred eighty-two subjects undergoing coronary angiography were recruited on the base of study design. Patients (n=114) had at least a stenosed coronary artery (>50% stenosis) and subdivided into three subgroups; SVD (n=28), 2VD (n=31) and 3VD (n=55) while controls (n=68) had the normal coronary arteries (<5% stenosis). The direct haplotyping technique of SNPs was performed using ARMS-RFLP-PCR method. Furthermore, alphabet-based tools predicted the changes of secondary structure at the rs4673 position. All haplotypes being proposed theoretically were found in the study population. The distribution of two-allele haplotypes had no significant difference between patients and controls (P=0.1). Although the rs4673 allele frequency was not significant between the groups (P>0.5), chi square test and multinomial regression analysis showed an observed high risk for rs13306294 A allele among patients. The bioinformatics tools predicted that the p22phox secondary structure is not changed due to the substitution of Tyr→His at the rs4673 position. We concluded that the polymorphisms have no allele linkage on the chromosome. In addition, the rs13306294 A allele is a potential factor of stenosis of coronary arteries that increases susceptibility for the extent of disease.