COMPARATIVE BIOAVAILABILITY (BIOEQUIVALENCE) STUDY FOR FIXED DOSE COMBINATION TABLET CONTAINING AMLODIPINE, VALSARTAN, AND HYDROCHLOROTHIAZIDE USING A NEWLY DEVELOPED HPLC-MS/MS METHOD

Objective: The aim of the study was to evaluate the bioequivalence between a newly developed generic tablet containing fixed-dose combination of amlodipine besylate, valsartan and hydrochlorothiazide (10/160/25 mg), and the reference brand product Exforge HCT® tablet; using a newly developed HPLC-MS/MS method for simultaneous determination of these drugs in human plasma.Methods: The brand (reference) and the test (generic) products were administered to thirty-nine healthy subjects. A fasting, laboratory blind, single-dose, two-treatment, two-period, two-sequence, randomized crossover design was conducted with 14 d washout period between dosing. Serial blood samples were withdrawn from each subject immediately before dosing (zero time), and then at 0.33, 0.66, 1.0, 1.33, 1.66, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 14, 16, 24, 48 and eventually at 72 h post dosing. Plasma samples were analyzed for simultaneous determination of amlodipine, valsartan and hydrochlorothiazide by a newly developed HPLC coupled with MS/MS detector. The linearity of the method was established for plasma concentration ranges of 0.2-12 ng/ml, 50-8000 ng/ml, and 2-250 ng/ml for amlodipine, valsartan, and hydrochlorothiazide, respectively.Results: Plasma concentration-time data of each individual were analyzed by non-compartmental method to measure the pharmacokinetics parameters; Cmax, Tmax, AUC0-t, AUC0-¥, lZ, T1/2. For amlodipine truncated AUC72hr was calculated. The 90% confidence interval for the pharmacokinetic parameters used for bioequivalence evaluation (Cmax and AUC) for amlodipine, valsartan, and hydrochlorothiazide were well within FDA acceptable ranges of 80-125%.Conclusion: It is concluded that the newly devolved generic product is bioequivalent with the brand product Exforge HCT® tablet. Thus, both products are clinically interchangeable.Keywords: Amlodipine, Valsartan, Hydrochlorothiazide, Pharmacokinetics, Bioequivalence, HPLC-MS/M

Chronic Toxicity Assessment of Histological Changes and Micronuclei in Fish Cyprinus carpio L. After Exposed to Copper

This study was conducted to assess the histological changes of (gill, liver, kidney and muscle) and The micronucleus test were applied in circulating erythrocytes of in freshwater fish common carp, Cyprinus carpio after chronic exposure to copper. For chronic tests, the fish were exposed to different concentrations (0.5, 0.9 and 1.2 mg/L) of copper for 3 and 6 weeks. Control fish were maintained in parallel with the experimental groups. Several histological alterations were observed in the gills, including the epithelium of gill filaments and secondary lamellae, degeneration and congestion of secondary lamellae and short villi. The liver showed dilation in cells hepatic, degenerative and necrosis of hepatocyte cell with mild inflammatory cell and accumulation of cholesterol inside the cell. Regarding in kidney, Renal tissue showed congestion and haemorrhage with certain degeneration and necrosis of renal tubules tissue. In the muscle, showed mild hyalinization of the skeletal muscles fibres with the loss of interstitial fibres in between the muscles fibres and focal degeneration and necrosis with mild inflammatory cell infiltration. Micronucleus test was applied to evaluate the genotoxic effects of heavy metals on Cyprinus carpio. Results of micronucleus test showed a progressive increase in the percentage of micronuclei (P≥0.001) with increases in the intensity of exposure of  copper. The obtained results showed that fish common carp, Cyprinus carpio erythrocytes are good models for cytotoxicity studies

Application of Gaussia luciferase in bicistronic and non-conventional secretion reporter constructs

Background: Secreted luciferases are highly useful bioluminescent reporters for cell-based assays and drug discovery. A variety of secreted luciferases from marine organisms have been described that harbor an N-terminal signal peptide for release along the classical secretory pathway. Here, we have characterized the secretion of Gaussia luciferase in more detail. / Results: We describe three basic mechanisms by which GLUC can be released from cells: first, classical secretion by virtue of the N-terminal signal peptide; second, internal signal peptide-mediated secretion and third, non-conventional secretion in the absence of an N-terminal signal peptide. Non-conventional release of dNGLUC is not stress-induced, does not require autophagy and can be enhanced by growth factor stimulation. Furthermore, we have identified the golgi-associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1) as a suppressor of release of dNGLUC. / Conclusions: Due to its secretion via multiple secretion pathways GLUC can find multiple applications as a research tool to study classical and non-conventional secretion. As GLUC can also be released from a reporter construct by internal signal peptide-mediated secretion it can be incorporated in a novel bicistronic secretion system

Brain herniation in a patient with apparently normal intracranial pressure: a case report

Introduction Intracranial pressure monitoring is commonly implemented in patients with neurologic injury and at high risk of developing intracranial hypertension, to detect changes in intracranial pressure in a timely manner. This enables early and potentially life-saving treatment of intracranial hypertension. Case presentation An intraparenchymal pressure probe was placed in the hemisphere contralateral to a large basal ganglia hemorrhage in a 75-year-old Caucasian man who was mechanically ventilated and sedated because of depressed consciousness. Intracranial pressures were continuously recorded and never exceeded 17 mmHg. After sedation had been stopped, our patient showed clinical signs of transtentorial brain herniation, despite apparently normal intracranial pressures (less than 10 mmHg). Computed tomography revealed that the size of the intracerebral hematoma had increased together with significant unilateral brain edema and transtentorial herniation. The contralateral hemisphere where the intraparenchymal pressure probe was placed appeared normal. Our patient underwent emergency decompressive craniotomy and was tracheotomized early, but did not completely recover. Conclusions Intraparenchymal pressure probes placed in the hemisphere contralateral to an intracerebral hematoma may dramatically underestimate intracranial pressure despite apparently normal values, even in the case of transtentorial brain herniation

P04.74 Preclinical evaluation of combinations targeting the DNA damage response in 2D and 3D models of glioblastoma stem cells

Background Despite surgical resection followed by DNA-damaging adjuvant therapies, glioblastoma remain incurable. Increasing evidence demonstrates that aberrations within the DNA damage response (DDR) of cancer stem cells contribute to treatment resistance. We have previously shown that the Fanconi Anaemia (FA) pathway, a key DDR process, remains inactive in normal brain but is re-activated in glioblastoma, making it an appealing foundational target for cancer-specific combination therapies. Since intratumoural heterogeneity in glioblastoma and inherent capacity for functional redundancy within DDR networks are established concepts - we aimed to determine whether combined and hypothesis-driven targeting of the FA pathway along with interconnected DDR processes could form a basis for effective multimodal therapies. Material and Methods Bioinformatic analysis of mRNA expression data (REMBRANDT database) was used to confirm the relevance of FA pathway activity in glioma. Subsequently, immunofluorescence and cell viability assays were used to validate and establish the therapeutic potential of novel FA pathway inhibitors (nFAPi) and inhibition of related DDR targets in established cell models. Finally, combinations targeting the DDR were optimised using immunoblotting, and assessed using clonogenic survival in 2D and novel 3D patient-derived glioblastoma stem cell models. Results High expression of downstream FA pathway genes is strongly associated with poor survival (-17.1% 5-year OS, n=329, Log-rank, P Conclusion Simultaneously targeting the FA pathway and interconnected DDR processes in glioblastoma represents a promising therapeutic strategy. Early mechanistic studies suggest this approach augments DNA damage and enhances IR-induced cell cycle arrest in G2/M, however further preclinical evaluation is ongoing

Combined inhibition of the Fanconi anaemia (FA) pathway and ATR promotes R-loop generation and profound radiosensitisation in glioblastoma

Glioblastoma is a deadly cancer in which treatment resistance is mediated through extensive intratumoural heterogeneity including difficult-to-treat glioblastoma stem cell (GSC) subpopulations. GSC eradication represents an attractive therapeutic goal, but these cells possess upregulated DNA damage response (DDR) processes, resulting in a chemo- and radioresistant phenotype. However, recent studies have demonstrated that elevated replication stress in GSCs may partially explain DDR upregulation and resistance, thus highlighting a potential therapeutically exploitable vulnerability. ATR and the FA-pathway are both fundamental to cellular DNA replication stress responses and maintaining replication fork stability. Since we have previously shown the FA-pathway is inactive in normal brain, but is re-activated in glioblastoma with potential to provide a cancer-specific foundation for combination DDR therapies, we explored the therapeutic potential of simultaneous inhibition of the FA-pathway (FAPi) and ATR (ATRi), in addition to other FA-pathway-based DDR inhibitor (DDRi) combinations. We find that compared with single agent treatments, combined inhibition of the FA-pathway and ATR in both 2D and 3D GSC ex vivo models promotes a substantial increase in conflicts between DNA replication and transcription (R-loops) which is further exacerbated by ionising radiation (IR). Molecular analyses of DNA damage indicate that FAPi+ATRi increases peak DNA damage post-IR treatments, with sustained elevation of DNA damage even at 24 hours post-treatment. In conclusion, simultaneously targeting the FA-pathway and ATR represents an appealing therapeutic strategy for glioblastoma. This approach promotes substantial R-loop generation, likely through exacerbating constitutively high levels of DNA replication stress previously observed in GSCs, with deleterious effects in these treatment resistant cells. Our findings underline the value of developing clinical FA pathway inhibitors and also support the application of current ATR inhibitors to molecularly-selected subsets of glioblastoma, namely, those with defects in one of 22 currently known FA-pathway genes which include BRCA1/FANCS and BRCA2/FANCD1

Survival analysis modeling with hidden censoring

There are well-established survival analysis methodologies for data sets that are complete, with accurate information on censoring. But what if they are not complete? In this article we consider how to analyze cases where “hidden censoring” occurs, where individuals have effectively left the study but the hospital is unaware of this. We develop a new Markov chain-based methodology for generating survival curves and hazard functions, and demonstrate this using a breast cancer data set from the Kurdistan region of Iraq

The management of corporate social responsibility through projects : a more economically developed country perspective

Corporate social responsibility (CSR) is an increasingly influential concept, claimed to be implemented more in the West than in the Gulf Cooperation Community (GCC) and Middle East and North Africa (MENA) region where CSR is often portrayed as less developed and mainly philanthropic. The research reported in this paper investigates capacity building for CSR in organizations and examines the research question as to whether the project management of CSR can contribute to increasing levels of awareness, engagement, and implementation. The empirical study therefore examines the management of CSR through projects and consists of data analysed from 29 interviews conducted during 2011–2016 with CSR representatives for organizations operating in the United Arab Emirates (UAE), which is a member of the GCC, located in the MENA region, and categorized as a more economically developed country. The findings reveal evidence supporting the argument that organizations with capabilities in the project management of CSR are in the position to develop capacities for strategic CSR. This empirical analysis of CSR capacity development across varying degrees of sophistication in the project management of CSR activities implies a more developed stage of CSR engagement, at least in the sample of organizations studied, than is expressed in the extant literature on CSR in the UAE, and elsewhere in the MENA region. The use of project management to organize, implement, align, and monitor CSR activity supports capacity development in CSR and can contribute to its sustainability. We recommend that future research examines the political, business, and institutional factors that could further develop CSR in organizations when managed through projects