Epi-Drugs in Heart Failure

Unveiling the secrets of genome's flexibility does not only foster new research in the field, but also gives rise to the exploration and development of novel epigenetic-based therapies as an approach to alleviate disease phenotypes. A better understanding of chromatin biology (DNA/histone complexes) and non-coding RNAs (ncRNAs) has enabled the development of epigenetic drugs able to modulate transcriptional programs implicated in cardiovascular diseases. This particularly applies to heart failure, where epigenetic networks have shown to underpin several pathological features, such as left ventricular hypertrophy, fibrosis, cardiomyocyte apoptosis and microvascular dysfunction. Targeting epigenetic signals might represent a promising approach, especially in patients with heart failure with preserved ejection fraction (HFpEF), where prognosis remains poor and breakthrough therapies have yet to be approved. In this setting, epigenetics can be employed for the development of customized therapeutic approaches thus paving the way for personalized medicine. Even though the beneficial effects of epi-drugs are gaining attention, the number of epigenetic compounds used in the clinical practice remains low suggesting that more selective epi-drugs are needed. From DNA-methylation changes to non-coding RNAs, we can establish brand-new regulations for drug targets with the aim of restoring healthy epigenomes and transcriptional programs in the failing heart. In the present review, we bring the timeline of epi-drug discovery and development, thus highlighting the emerging role of epigenetic therapies in heart failure

Epigenetic remodeling in heart failure with preserved ejection fraction

PURPOSE OF REVIEW In this review, we critically address the role of epigenetic processing and its therapeutic modulation in heart failure with preserved ejection fraction (HFpEF). RECENT FINDINGS HFpEF associates with a poor prognosis and the identification of novel molecular targets and therapeutic approaches are in high demand. Emerging evidence indicates a key involvement of epigenetic signals in the regulation of transcriptional programs underpinning features of HFpEF. The growing understanding of chromatin dynamics has led to the development of selective epigenetic drugs able to reset transcriptional changes thus delaying or preventing the progression toward HFpEF. Epigenetic information in the setting of HFpEF can be employed to: (i) dissect novel epigenetic networks and chromatin marks contributing to HFpEF; (ii) unveil circulating and cell-specific epigenetic biomarkers; (iii) build predictive models by using computational epigenetics and deep machine learning; (iv) develop new chromatin modifying drugs for personalized management of HFpEF. SUMMARY Acquired epigenetic signatures during the lifetime can contribute to derail molecular pathways involved in HFpEF. A scrutiny investigation of the individual epigenetic landscape will offer opportunities to develop personalized epigenetic biomarkers and therapies to fight HFpEF in the decades to come

Genetic Study for G-Protein Coupled Receptor from Saccharomyces Cerervisiae and From Sera of Patients with Heart Thrombosis

Aim of the present study is Identification of specific gene for GPCR using specific primers .and identification of difference in PCR analysis in patients with heart thrombosis  and compared with healthy, Sequencing of PCR product regarding GPCR compared for all three subject, Identification  the similarity of human GPCR with local strain of yeast fifty healthy control and fifty patients with thrombosis which diagnosed medically with cardiac specific troponin t, troponin 1 levels and electro myocardiogram ECG. The aged for all subjects ranged (39-75) years   patients were lying in cardiac care unit at Ibn- al- Nafees teaching hospital and  Sheikh  Zayed  teaching  hospital. Genomic DNA of whole blood was extracted from buffy coat and cell cultured handbook protocol using Bioneer- kit and  Genomic DNA fungus/yeast  kit was used in isolation  and purification of DNA. patients divided  into three  groups according  to their age: group A (60-75) years , group B (50-59) years ,  group C (39-49) years the  results of genomic  DNA  isolation  from blood cells extracted in pure  form which ensured by the absorbance ratio (260/280 ) was (1.6 – 1.9 ) with a concentration of 50µg/ml and one DNA band with high resolution in gel electrophoresis. The result of  genomic DNA extracted from  the local strain of  S. cerevisiae showed that DNA   extracted with  high   purity   because   the  absorbance  ratio  (260 /280 )was (1.7 to 2.0) with a concentration of 60 µg/ml  and presence one DNA band with high resolution  in gel electrophoresis. primers were designed  depending on the sequence of the gene responsible for the production of GPCR on the chromosome 11 , GPCR  contain  three exons which covered  with six primers to detect a defect in gene sequence among. Results of gel electrophoresis are showed that primer  GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. with molecular weight of this band is 1000 bp. The  GPRX2  primer  used to amplify second exon in the GPCR gene ,the molecular weight of amplified bands are  400 bp were present in all control samples and three groups of thrombosis patientsand yeast. GPRX2A primer that designed to amplify  part two from second exon of GPCR gene by PCR gave one band for all samples which include control and patient, the molecular weight of this band is 500 bp. PCR analysis  showed one amplify band for all control and patients group with molecular weight 500 bp  for GPRX3 primer and 400 bp for GPRX3A ,300 bp for GPRX3B. The specific primers which  designed to covering  GPCR gene used to amplification  genomic DNA  of the local strain  S.cerevisiae by PCR technique. Results showed all six primers which gave  one band with difference molecular weight  for each primer. All samples demonstrate identity planned sizes to control and local strain of S. cerevisiae samples except patient number 8 and 9 in the group(C) that  showed non specialist bands in specific primer  with  first  exon (GPRX1) .So the genetic sequence analysis of these two case based on the sequence  of the  remainder exons to detect the genetic defect in these case. The sequence of the first part for the second exon (X2) was identity standard sequence found on the NCBI web site for case( 8). The  case (9) showed  identity with  the sequence present in the human gene bank but some difference in the first of sequence which neglected because it is in the place link of primer. The  results for  case  8  showed some mutation for Exon X2(part2). but case (9) demonstrate one  deletion and one substitution. The results, also, illustrated that the ether48 thrombosis patients didn't appeared any mutation despite the positive results for ( Troponin) that gives strong indication of thrombosis. The conclusion Primer  GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. The molecular weight of this band is 1000 bp. The amplified band with molecular weight 400 bp were present in all control samples and three groups of thrombosis patients with primer GPRX2  and 500 bp with primer GPRX2A.PCR analysis  showed one amplify band for all control and patients group with molecular weight 500 bp for GPRX3 primer.,400 bp with GPRX3A and 300 bp for primer GPRX3B.Similarity between results given by healthy group and local strain of yeast. Genetic study  showed that there are  only two case of  patients  eight and nine demonstrated mutation in nucleic location on exon two and three from GPCR gene Keyword: GPCR, Genetic Study, Saccharomysis  cerevisiae, Patient with Thrombosis

Purification of G-Protein Coupled Receptor from Whole Cell of Local Strain of Saccharomyces cerevisiae

The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtrationchromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the alreadyused in published researches, which depend on the costly affinity chromatography and other expensive methods ofpurification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR).The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated onYeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C° for 24 h .Loop fully of the yeast culture wastransferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C°for 24h , after that itwas stored at 4C° ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae wasidentified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cellof S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negativelycharged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtrationchromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fractionwas measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISAKit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weightof GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plottedbetween log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR thatextracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ionexchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient(0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration ofsodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured inthe fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M ofNaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don'tgive any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step ofpurification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical withthe peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed inthese fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S.cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.Key words: GPCR purification, S.Cerevisea, whole cel

Inflammation in Metabolic Cardiomyopathy

Overlapping pandemics of lifestyle-related diseases pose a substantial threat to cardiovascular health. Apart from coronary artery disease, metabolic disturbances linked to obesity, insulin resistance and diabetes directly compromise myocardial structure and function through independent and shared mechanisms heavily involving inflammatory signals. Accumulating evidence indicates that metabolic dysregulation causes systemic inflammation, which in turn aggravates cardiovascular disease. Indeed, elevated systemic levels of pro-inflammatory cytokines and metabolic substrates induce an inflammatory state in different cardiac cells and lead to subcellular alterations thereby promoting maladaptive myocardial remodeling. At the cellular level, inflammation-induced oxidative stress, mitochondrial dysfunction, impaired calcium handling, and lipotoxicity contribute to cardiomyocyte hypertrophy and dysfunction, extracellular matrix accumulation and microvascular disease. In cardiometabolic patients, myocardial inflammation is maintained by innate immune cell activation mediated by pattern recognition receptors such as Toll-like receptor 4 (TLR4) and downstream activation of the NLRP3 inflammasome and NF-κB-dependent pathways. Chronic low-grade inflammation progressively alters metabolic processes in the heart, leading to a metabolic cardiomyopathy (MC) phenotype and eventually to heart failure with preserved ejection fraction (HFpEF). In accordance with preclinical data, observational studies consistently showed increased inflammatory markers and cardiometabolic features in patients with HFpEF. Future treatment approaches of MC may target inflammatory mediators as they are closely intertwined with cardiac nutrient metabolism. Here, we review current evidence on inflammatory processes involved in the development of MC and provide an overview of nutrient and cytokine-driven pro-inflammatory effects stratified by cell type

Modeling and Simulation Affect of Binder Addition Rate on Ammonia Emission During Granulation Process

This paper presents the modeling and simulation of binder addition in Fluidized bed granulator in order to quantify the amount of ammonia (NH3) gas release. The model development is based on experimental observations using MATLAB 12b with the ability of combining expert knowledge with mapping of experimental data sets is expected to establish a robust model which then taken up for process optimization and control the ammonia emission.

IN VITRO ACTIVITY OF VARIOUS POTENCIES OF HOMEOPATHIC DRUG THUJA AGAINST MOLDS INVOLVED IN MYCOTIC KERATITIS

Objective: Isolation and characterisation of clinically isolated fungi from mycotic keratitis and exploration of the in vitro efficacy of various potencies of homeopathic preparations of Thuja occidentalis on the ocular fungal isolates. Methods: Clinical samples were collected from fungal keratitis patients attending a tertiary eye care hospital in Coimbatore, Tamilnadu state, India. The scrapings were also subjected to Gram staining and 10% KOH mount to detect the presence of fungal hyphae. The fungal isolates were subjected to lacto phenol cotton blue (LCB) mount employing cello tape flag method. Homeopathic drug Thuja occidentalis with various potencies viz., Q, 30 C, 200 C, 1 M, 10 M and 50 M were investigated for the growth inhibition of various fungal isolates by plate assay method. Further, a follow up analyses with varying dilutions of Q and 10 M homeopathic potencies of Thuja was carried out for the determination of minimum inhibitory concentration of both microdilution and minimum fungicidal concentration for the Biopolaris isolates. Results: Out of 35 samples analysed, Fusarium spp. (n=5), Aspergillus flavus (n=6), Bipolaris spp. (n=3), Exserohilum spp. (n=3) and Curvularia spp (n=3) were identified. All the potencies of Thuja had good inhibitory activity against Bipolaris spp., followed by Curvularia spp., Exserohilum spp. and Aspergillus flavus. Statistical analysis revealed significant inhibition of all the test isolates by Thuja Q and 50 M. Significant growth inhibition was exhibited by Thuja 30, 200 C, I M for Bipolaris, Exserohilum & A. flavus isolate and Thuja 10 M for all the isolates tested except Fusaria. It was revealed that for the Biopolaris isolates BS1, BS2, BS3 Thuja 10 M and Thuja Q had MIC and MFC of 0.125/10[20,00]0& 0.0625/10 and 0.25/10[20][00]0 & 0.125/10, respectively. Conclusion: The present investigation concludes that homeopathic drug Thuja has good inhibitory activity against the fungi causing keratitis, irrespective of the potencies. It is evident that no definite co-relation exists between various potencies of the same homeopathic drug with regard to their antimycotic properties

Bilateral Anterior Ischaemic Optic Neuropathy in a Child on Continuous Peritoneal Dialysis : Case report and literature review

Non-arteritic anterior ischaemic optic neuropathy (NAION) is a serious complication of continuous peritoneal dialysis (CPD) which can lead to poor vision and blindness. We report a five-year-old girl who had undergone a bilateral nephrectomy at the age of one year and was on home CPD. She was referred to the Paediatric Ophthalmology Unit of Sultan Qaboos University Hospital, Muscat, Oman, in 2013 with acute bilateral vision loss, preceded by a three-day history of poor oral intake. At presentation, the patient had severe systemic hypotension. An ophthalmological examination revealed severe bilateral visual impairment and NAION. She was treated with intravenous methylprednisolone and normal saline boluses. At a five-month follow-up, the visual acuity of the right eye had improved but vision in the left eye remained the same. Acute bilateral blindness due to NAION while on CPD is a rare condition in childhood. Paediatricians should be aware of this complication in order to ensure prompt management.Keywords:

Strengthening and Retrofitting of Reinforced Concrete Hollow Columns using High Strength Ferrocement Fibers Composites

Eight RC circular hollow columns (external diameter = 220 mm, internal diameter = 100 mm, length = 1000 mm and the hollow part = 700mm) casted and strengthened with ferrocement fibers composites to illustrate the behavior of these columns under concentric and eccentric axial compression force. Two columns where used as reference columns, which were repaired after failure to be tested as retrofitted columns. Six specimens were strengthened with one and two WWM layers as required. The variables considered included number of the WWM layers (N), the loading configuration and the eccentricity value (e) of loading. The ferrocement thickness was constant at 20 mm in all retrofitted and strengthened specimens. The test results revealed that the maximum increase in the ultimate concentric loads were 67% by strengthening the reference column with two layers of WWM, and the maximum increase in the ultimate eccentric load of columns was 78% by increasing of the WWM from one to two layers. For a constant number of WWM layers, the change from concentric to eccentric force caused a decrease in the ultimate load value attaining 73.5% for one- layer WWM strengthened columns. The failure of columns occurred by yielding of steel reinforcement followed by concrete crushing (i.e. tension failure)

Epigenetic Signatures in Arterial Hypertension: Focus on the Microvasculature

Systemic arterial hypertension (AH) is a multifaceted disease characterized by accelerated vascular aging and high cardiometabolic morbidity and mortality. Despite extensive work in the field, the pathogenesis of AH is still incompletely understood, and its treatment remains challenging. Recent evidence has shown a deep involvement of epigenetic signals in the regulation of transcriptional programs underpinning maladaptive vascular remodeling, sympathetic activation and cardiometabolic alterations, all factors predisposing to AH. After occurring, these epigenetic changes have a long-lasting effect on gene dysregulation and do not seem to be reversible upon intensive treatment or the control of cardiovascular risk factors. Among the factors involved in arterial hypertension, microvascular dysfunction plays a central role. This review will focus on the emerging role of epigenetic changes in hypertensive-related microvascular disease, including the different cell types and tissues (endothelial cells, vascular smooth muscle cells and perivascular adipose tissue) as well as the involvement of mechanical/hemodynamic factors, namely, shear stress