Nur77 and FHL2: Novel players in vascular and immune disease

The nuclear receptor Nur77 is an early response gene that is induced by diverse extracellular signals in a wide range of tissues and cultured cells. It has been implicated in the regulation of genes involved in metabolic disease, adipogenesis, inflammation, and vascular disease. FHL2/DRAL/SLIM3 is a LIM-only protein that has been shown to interact with many proteins and acts as a co-activator or co- repressor depending on the cell-type and cellular context. It is a crucial adaptor protein and plays a pivotal role in a range of physiological and pathological processes, including proliferation, migration, differentiation and apoptosis. The aim of this thesis is to increase our understanding of fundamental pathways critical in vascular diseases including atherosclerosis, restenosis, coagulation, and immune diseases including asthma, airway inflammation and schistosomiasis. To achieve this goal, we performed numerous distinct studies on the role of nuclear receptor Nur77 and LIM-only protein FHL2 using mouse models and several cell types. In this thesis, functional properties of Nur77 and FHL2, as well as their impact on multiple signaling pathways in vascular and immune disease were studied. These research efforts are ultimately directed at designing novel therapeutic strategies that may aid in mitigating the development of vascular and immune disease

Load Frequency Control of Three area Multi-Unit Deregulated Power System with FOSMC and Performance analysis using Regulation constant

This paper presents three area multi-unit Deregulated Power System (DPS) for Load Frequency Control (LFC) using Fractional Order Sliding Mode Controller (FOSMC) along with Thyristor Controlled Phase Shifters (TCPS) and Superconducting Magnetic Energy Storage (SMES) combination. The FOSMC can be used to overcome nonlinearities and uncertainties of the system for bilateral and unilateral transactions under different Step Load Perturbations (SLPs). The deregulated power system performance is analyzed for different Regulation constants (R) such as 1.8, 2.4 and 3.0. For stabilization of oscillations in frequency and to stabilize the deregulated power system dynamically for different SLPs, TCPS is incorporated with the tie line in series and SMES is used as an energy storage unit. The dynamic responses of LFC problems have been simulated and analyzed with MATLAB/Simulink-based computer simulations. Further simulation results have also been tabulated as a comparative performance with respect to peak overshoot and settling time.&nbsp

SELF-NANO EMULSIFYING DRUG DELIVERY SYSTEM (SNEDDS) FOR ORAL DELIVERY OF ATORVASTATIN- FORMULATION AND BIOAVAILABILITY STUDIES

The aim of the study was to develop a self-nano emulsifying drug delivery system (SNEDDS) for the  atorvastatin which belongs to BCS class II lipid lowering agent with poor water solubility and dissolution rate. The solubility of atorvastatin in individual micro emulsion components viz. oil and surfactants was determined. The surfactants were screened for emulsification ability. Based on the solubility determinations and emulsification properties oleic acid as oil; surfactants Brij 80 and Tween 80 were selected for further study. The solubility of atorvastatin in different ratios of selected oil and surfactants was determined. The composition of oil: Surfactant with maximum solubility for atorvastatin was used for SNEDDS formulation. Ternary phase diagrams were used to evaluate the micro emulsification existence area. The micro emulsions were evaluated for emulsion droplet size, self-emulsification time, phase separation, in vitro dissolution and stability. SNEDDS formulations found to be self-emulsified in 70 to 120 seconds without precipitation and their mean droplet sizes was 150 to 230 nm. Among the optimized formulations, formulation BF4 showed highest in vitro drug release. Formulation BF4 was composed of 20% oleic acid, 60% tween 80 and 20% brij 30 showed significant increases in the dissolution rate (99.65%) in 90 minutes and intestinal absorption (86.67%) than marketed product with 56.86% release and 45.34% oral absorption. Stability studies were conducted according to the Q1 ICH guidelines and found stable at different conditions

A COST-EFFECTIVE ANALYSIS OF TIOTROPIUM WITH FORMOTEROL AND BUDESONIDE WITH FORMOTEROL BASED ON EFFICACY AND QUALITY OF LIFE IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE PATIENTS

Objectives: The primary objective of the study was to conduct the cost-effective analysis of tiotropium with formoterol versus budesonide with formoterol in the management of Stage-I chronic obstructive pulmonary disease (COPD) based on efficacy and quality of life (QoL). The study also aims to analyze the improvement of the QoL through pulmonary rehabilitation. The secondary objective was to evaluate the incidence of depression and anxiety in COPD patients.Methods: A prospective interventional study, with a sample size of 74 patients diagnosed with COPD Stage-I was conducted for 6 months. The study population was divided based on simple randomization into two main groups, one receiving tiotropium with formoterol and the other receiving budesonide with formoterol. Based on spirometry, their pulmonary function test (PFT) was recorded. The patients QoL was recorded using the WHOQoL-BREF questionnaire.Results: Data of 74 patients were collected for both the zero-degree and first-degree follow-up. The patients demonstrated good compliance with the transinhaler of the prescribed drugs. A significant improvement after providing pulmonary rehabilitation in PFT, i.e., forced expiratory volume1/ forced vital capacity values (p=0.000**) and the WHOQoL scoring (P = 0.001**) was observed in patients receiving tiotropium with formoterol. Tiotropium with formoterol was found to be more cost-effective treatment than budesonide with formoterol.Conclusion: The study showed that transinhalation of 9 mcg/12 mcg tiotropium/formoterol once a day is a better cost-effective treatment than 200 mcg/6 mcg transinhalation of budesonide/formoterol twice a day

Functional outcome of proximal 1/3rd, distal 1/3rd and diaphysial tibial fractures in adults operated with expert tibial nailing

Background: Fractures of the tibial shaft are increasing due to high velocity trauma and industrialisation. Not only they are common but also difficult to treat. Until recently surgeons had to rely on non-operative treatment, V nailing, plates and screws and external fixator but they had their drawbacks like prolonged immobilisation infection, delayed union and non-union. Numerous modifications in nail and screw design have led to development of the expert tibial nail. Multidirectional interlocking screws ensure that alignment can be well maintained and stability preserved in short proximal or distal tibial segments.Methods: 30 patients were admitted and operated during September 2014 to September 2016 in Mamata general hospital Khammam. All patients were evaluated with Klemm Borner’s criteria and complications following surgery.Results: 87% of patients achieved good or excellent results, fair results were obtained in 3 (10%) patient and poor result in one (3%) patient. 2 (6%) patients had malunion, 2 (6%) patients had delayed union, 1 (3%) patient had deep infection led to implant failure.Conclusions: Intramedullary nailing is a safe and effective technique for the treatment of tibial metaphyseal fractures. It avoids the additional soft-tissue dissection associated with traditional open procedures as well as the complications associated with external fixators. Expert tibial nail can give excellent functional and clinical results. Complications such as failure of the bone-implant construct or post-operative malallignment are avoidable if careful pre-operative planning is allied with meticulous surgical technique

HEPATOPROTECTIVE EFFECT OF PHYLLANTHUS NIRURI ALKALOID FRACTION IN D–GALACTOSAMINE INDUCED HEPATITIS IN RATS

Objective: The current study was designed to explore the hepatoprotective activity of Phyllanthus niruri (PN) alkaloid fraction in induced hepatitis in rats.Methods: The rats are divided into 5 groups as per the experimental design. Pre alkaloid treatment of PN was given to the hepatitis rats for 21 d. After completion of the treatment, the serum was collected, and biochemical analysis was carried out. The hepatic markers like alanine aminotransferase (ALT), aspartate aminotransferase (AST) activities, bilirubin (BL) urea (UR), creatinine (CR) levels are estimated in serum.Results: ALT, AST activities, BL, CR levels are increased, and UR levels are decreased in hepatitis rats. However with PN alkaloid fraction pre-treatment all these hepatic markers significantly (P<0.01) came back to near normal levels. Histopathological studies also prove that PN protected the hepatic tissue from hepatitis.Conclusion: The outcome of the present study showed that the alkaloid fraction of PN possesses hepatoprotective effect in hepatitis rats.Keywords: Phyllanthus niruri, Hepatitis, Liver marker

Validated HPLC method for quantification of copanlisib in mice plasma: application to a pharmacokinetic study

Copanlisib is a pan phosphatidylinositol 3-kinase (PI3K) inhibitor approved for follicular lymphoma. In this paper, we present the data of development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of copanlisib in mice plasma as per the FDA regulatory guideline. The method involves the extraction of copanlisib along with internal standard (IS, enasidenib) from mice plasma (100 µL) using ethyl acetate as an extraction solvent. The chromatographic resolution of copanlisib and the IS was achieved on a Hypersil Gold C18 column maintained at 40 °C using a binary gradient mobile phase [10 mM ammonium formate (pH 4.0) and acetonitrile]. The flow-rate was 0.8 mL/min. For the detection of copanlisib and the IS, the photo-diode array detector was set at λmax 310 nm. Copanlisib and the IS eluted at 6.60 and 7.80 min, respectively with a total run time of 10 min. The calibration curve was linear over a concentration range of 50 to 5000 ng/mL for copanlisib (r2 0.998). The results of intra- and inter-day accuracy and precision studies were within the acceptable limits. Copanlisib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycle and long-term storage at -80 °C. The application of the validated method was shown in a mice pharmacokinetic study

Thoracic aortic aneurysm development in patients with bicuspid aortic valve: what is the role of endothelial cells?

Bicuspid aortic valve (BAV) is the most common type of congenital cardiac malformation. Patients with a BAV have a predisposition for the development of thoracic aortic aneurysm (TAA). This pathological aortic dilation may result in aortic rupture, which is fatal in most cases. The abnormal aortic morphology of TAAs results from a complex series of events that alter the cellular structure and extracellular matrix (ECM) composition of the aortic wall. Because the major degeneration is located in the media of the aorta, most studies aim to unravel impaired smooth muscle cell (SMC) function in BAV TAA. However, recent studies suggest that endothelial cells play a key role in both the initiation and progression of TAAs by influencing the medial layer. Aortic endothelial cells are activated in BAV mediated TAAs and have a substantial influence on ECM composition and SMC phenotype, by secreting several key growth factors and matrix modulating enzymes. In recent years there have been significant advances in the genetic and molecular understanding of endothelial cells in BAV associated TAAs. In this review, the involvement of the endothelial cells in BAV TAA pathogenesis is discussed. Endothelial cell functioning in vessel homeostasis, flow response and signalling will be highlighted to give an overview of the importance and the under investigated potential of endothelial cells in BAV-associated TAA. Stem cells & developmental biolog

The Inflammatory Profile of CTEPH-Derived Endothelial Cells Is a Possible Driver of Disease Progression

Chronic thromboembolic pulmonary hypertension (CTEPH) is a form of pulmonary hypertension characterized by the presence of fibrotic intraluminal thrombi and causing obliteration of the pulmonary arteries. Although both endothelial cell (EC) dysfunction and inflammation are linked to CTEPH pathogenesis, regulation of the basal inflammatory response of ECs in CTEPH is not fully understood. Therefore, in the present study, we investigated the role of the nuclear factor (NF)-κB pro-inflammatory signaling pathway in ECs in CTEPH under basal conditions. Basal mRNA levels of interleukin (IL)-8, IL-1β, monocyte chemoattractant protein-1 (MCP-1), C-C motif chemokine ligand 5 (CCL5), and vascular cell adhesion molecule-1 (VCAM-1) were upregulated in CTEPH-ECs compared to the control cells. To assess the involvement of NF-κB signaling in basal inflammatory activation, CTEPH-ECs were incubated with the NF-κB inhibitor Bay 11-7085. The increase in pro-inflammatory cytokines was abolished when cells were incubated with the NF-κB inhibitor. To determine if NF-κB was indeed activated, we stained pulmonary endarterectomy (PEA) specimens from CTEPH patients and ECs isolated from PEA specimens for phospho-NF-κB-P65 and found that especially the vessels within the thrombus and CTEPH-ECs are positive for phospho-NF-κB-P65. In summary, we show that CTEPH-ECs have a pro-inflammatory status under basal conditions, and blocking NF-κB signaling reduces the production of inflammatory factors in CTEPH-ECs. Therefore, our results show that the increased basal pro-inflammatory status of CTEPH-ECs is, at least partially, regulated through activation of NF-κB signaling and potentially contributes to the pathophysiology and progression of CTEPH