FUNCTIONAL ANALYSIS OF 5'-FLANKING REGION OF CYTOCHROME P450 GENES THROUGH MOLECULAR CLONING AND TRANSFECTION IN VITRO AND IN VIVO

Cytochrome P450 (CYP) enzymes are an important class of heme-containing proteins that catalyze oxidation reactions leading toward the removal of a wide variety of endogenous and exogenous substrates including prescription drugs. The activities of CYP enzymes are regulated primarily at the transcription level involving the regulatory sequences at the 5'-flanking region of the CYP genes. The objective of this dissertation study was to characterize the function of the 5'-flanking sequences of selected CYP genes primarily responsible for drug metabolism. Various sequences from the 5'-flanking regions of different CYP genes (CYP1A2, CYP2C9, CYP2C18, CYP2D6, CYP2E1, and CYP3A4) were cloned in expression vectors and tested for their activity in driving reporter gene expression in mouse livers and in transfected HepG2, 293, and BL-6 cells under optimized conditions. It was demonstrated that among the tested 5'-flanking regions of CYP genes, the CYP2D6 promoter showed the highest activity both in vivo and in vitro. The activities of various 5'-flanking regions of CYP genes in sustaining transgene expression were then tested in mouse liver and compared to those of other promoter sequences. As a result, the CYP2D6 promoter showed the highest activity and its activity was comparable to that of many established promoters. The mechanism underlying CYP promoter activities in vivo and in vitro were then studied using the CYP2C9 promoter as a model. Activities of various 5'-flanking sequences of CYP2C9 were evaluated by using deletion mutations of plasmid constructs in combination with transfection in mouse livers and in HepG2 cells. Finally, the role of PXR and CAR nuclear receptors in regulating CYP2C9 activation was investigated. The results show that both CAR and PXR are essential for CYP2C9 activation and that the regulatory elements reside in the proximal 1-2 kb region upstream of the CYP2C9 gene

EXPLORANDO O PAPEL DA UNIVERSIDADE PRÍNCIPE SATTAM BIN ABDULAZIZ NA ABORDAGEM DE CAUSAS E CONSEQUÊNCIAS DE FRUSTRAÇÃO ENTRE ESTUDANTES: UM ESTUDO DE CAMPO

Frustration among university students is a prevalent and often overwhelming emotion resulting from a variety of academic, personal, and social pressures, significantly impacting their overall college experience. This study aims to investigate the role of Prince Sattam bin Abdulaziz University in addressing the causes and consequences of students’ frustration. We employed a descriptive research methodology and surveyed 87 faculty members from the colleges of Wadi Al-Dawasir and Al-Sukayyia using an 18-item questionnaire. Data were analyzed using appropriate statistical methods. The results indicated that, from the perspective of faculty members, the role of Prince Sattam bin Abdul Aziz University in addressing the causes and consequences of student frustration was moderate. The dimensions of this role were ranked as follows: first, the university's role in raising students' awareness about the causes and consequences of frustration; second, the university's educational role; and third, the university's preventive role in alleviating student frustration. The study proposes a conceptual framework to activate the preventive, awareness-raising, and educational roles of Prince Sattam bin Abdulaziz University in addressing the issue of student frustration.A frustração entre os estudantes universitários é uma emoção predominante e muitas vezes avassaladora, resultante de uma variedade de pressões acadêmicas, pessoais e sociais, impactando significativamente a sua experiência universitária em geral. Este estudo tem como objetivo investigar o papel da Universidade Príncipe Sattam bin Abdulaziz na abordagem das causas e consequências da frustração dos estudantes. Empregamos uma metodologia de pesquisa descritiva e entrevistamos 87 professores das faculdades de Wadi Al-Dawasir e Al-Sukayyia usando um questionário de 18 itens. Os dados foram analisados utilizando métodos estatísticos apropriados. Os resultados indicaram que, do ponto de vista dos docentes, o papel da Universidade Príncipe Sattam bin Abdul Aziz na abordagem das causas e consequências da frustração dos estudantes foi moderado. As dimensões deste papel foram ordenadas da seguinte forma: primeiro, o papel da universidade na sensibilização dos estudantes sobre as causas e consequências da frustração; segundo, o papel educativo da universidade; e terceiro, o papel preventivo da universidade no alívio da frustração dos estudantes. O estudo propõe uma estrutura conceitual para ativar os papéis preventivos, de conscientização e educacionais da Universidade Príncipe Sattam bin Abdulaziz na abordagem da questão da frustração dos estudantes

In-vitro Cytotoxicity and In-silico Insights of the Multi-target Anticancer Candidates from Haplophyllum tuberculatum

This study aimed to investigate the anticancer activity of Haplophyllum tuberculatum(Forsk.) aerial parts ethanol extract and fractions and reveal the potential anticancer targets, binding modes, pharmacokinetics, and toxicity properties of its phytoconstituents. MTT assay was used to investigate the anticancer activity. TargetNet, ChemProt version 2.0, and CLC-Pred web servers were used for virtual screening, and Cresset Flare software was used for molecular docking with the 26 predicted targets. Moreover, pkCSM, swiss ADME, and eMolTox web servers were used to predict pharmacokinetics and safety. Ethanolic extracts of H. tuberculatum on HepG2 and HeLa cell lines showed promising activities with IC50 values 54.12 and 48.1 µg/mL, respectively. Further, ethyl acetate fraction showed the highest cytotoxicity on HepG2 and HeLa cell lines with IC50 values 41.7 and 52.31 µg/mL. Of 70 compounds screened virtually, polygamain, justicidin A, justicidin B, haplotubine, kusunokinin, and flindersine were predicted as safe anticancer drugs candidates. They showed the highest binding scores with targets involved in cell growth, proliferation, survival, migration, tumor suppression, induction of apoptosis, metastasis, and drug resistance. Our findings revealed the potency of H. tuberculatum as a source of anticancer candidates that further studies should support

EFFECT OF BETA VULGARIS L. ON CHOLESTEROL RICH DIET-INDUCED HYPERCHOLESTEROLEMIA IN RATS

Abstract The lyophilized aqueous extract of Beta vulgaris L. (beet root) (BVE) was investigated for its possible antihypercholesterolemic and antioxidant potential in cholesterol rich diet-induced hypercholesterolemia in Wistar albino rats. Hypercholesterolemia was induced in rats by feeding 1% cholesterol rich diet for 10 weeks. Lipid profile and glucose were estimated in serum. Malondialdehyde (MDA) and non-protein sulfhydryls (NP-SH) levels were measured in liver and heart. Hypercholesterolemic rats showed a significant increase in total cholesterol and triglycerides and a significant decrease in high-density lipoprotein-cholesterol (HDL-C) levels. BVE at the doses of 250 and 500 mg/kg body weight for 70 consecutive days showed a significant decrease in total cholesterol and triglycerides and significant increase in HDL-C. Furthermore, hypercholesterolemic rats showed free radical generation (lipid peroxidation), evident by a significant increase in MDA level and a significant reduction in NP-SH content in both liver and heart homogenates. BVE treatment significantly decreased MDA level and significantly replenished the reduced NP-SH content in both liver and heart tissue. The acute toxicity test of BVE showed no mortality or morbidity in rats. The findings indicate that BVE has a significant antihypercholesterolemic and antioxidant potential and/or free radical scavenging properties in hypercholesterolemic, rats possibly exerted by the phytoconstituents present in the beet root. Rezumat Studiul experimental evaluează acţiunea antihipercolesterolemiantă şi antioxidantă a extractului apos liofilizat al rădăcinii plantei Beta vulgaris (Chenopodiaceae). Studiul a fost realizat pe şobolani albi de laborator, cărora li s-a indus experimental hipercolesterolemia. A fost evaluat profilul lipidic şi glucidic al animalelor, concentraţia serică a malonildialdehidei. De asemenea, au fost evaluate (ĭn ţesutul hepatic şi cardiac) grupările sulfhidril non-proteice. Rezultatele obţinute indică proprietăţile antihipercolesterolemiante şi antioxidante, datorate fitoconstituenţilor prezenţi în rădăcina plantei studiate

Survival Rate of Patients with Cardiothoracic Injuries in Road Traffic Accidents, and their Relationship with ISS, GCS and blood transfusions.

Severe thoracic trauma is one of the major causes of injury-related mortality. In the United States, thoracic trauma results in one-fourth of all trauma deaths. Globally, cardiothoracic trauma is also a major contributor to mortality. The most common cardiothoracic injuries include rib fractures, thoracic vertebral fractures, haemothorax, pneumothorax, flail chest, and lung contusions. The purpose of the present study was to determine the survival rate of patients with cardiovascular injuries in road traffic accidents and its relationship with ISS, GCS and blood transfusions at King Khalid Hospital. This study is a useful addition to the literature, as research in this topic is lacking. A total of 189 patients were transported to the hospital with cardiothoracic injuries during the study period. Data was gathered regarding age, gender, nationality, vehicle user type, anatomical region injured, Intensive Care Unit (ICU) admission, Glasgow Coma Scale (GCS), Injury Severity Score (ISS), blood transfusion, treatment and mortality rate. The neurological status was assessed using the GCS score. Injury Severity Scores were calculated to categorize the injury severity. The mean patient age was 31.81 years, with a peak age of between 21–30 years. Males predominated (93.7%) with a male to female ratio of 15:1. Most of the patients were Saudi nationals (61.3%). Overall mortality was 7.9%. Factors that were significantly associated with mortality were head and neck involvement, ICU admission, age (above 60), treatment delivered, and blood transfusions. Cardiothoracic trauma is associated with a high mortality rate, which may depend on the clinical presentation such as GCS, ISS, degree of shock, pattern of injuries, and associated injuries. Immediate management is vital for patients with life-threatening cardiothoracic trauma, as mortality is high if the diagnosis is missed, wrong or left untreated

Reinventing Patient Support and Continuity of Care Using Innovative Physician-staffed Hotline: More than 60,000 Patients Served Across 15 Medical and Surgical Specialties During the First Wave of COVID-19 Lockdown in Qatar

Rising disease prevalence early during the COVID-19 pandemic in the State of Qatar led to stoppage of all non-emergency health care services. To maintain continuity of care and information exchanges for non-emergency patients, a physician-operated telephone hotline was set up that involved triage followed by immediate consultation with a specialized physician. We describe the initiation and evaluate the operations of the Urgent Consultation Centre (UCC) hotline manned by 150 physicians and aimed at urgent non-life-threatening consultations at Hamad Medical Corporation, the public health provider in Qatar. UCC established a hotline to triage inbound patient calls related to 15 medical and surgical specialties. For calls between April-August 2020, we describe call volume, distribution by specialty, outcomes, performance of UCC team, as well as demographics of callers. During the study period, UCC received 60229 calls (average 394 calls/day) from Qatari nationals (38%) and expatriates (62%). Maximum total daily calls peaked at 1670 calls on June 14, 2020. Call volumes were the highest from 9 AM to 2 PM. Response rate varied from 89% to 100%. After an initial telephone triage, calls were most often related to and thus directed to internal medicine (24.61%) and geriatrics (11.97%), while the least percentage of calls were for pain management and oncology/hematology (around 2% for each). By outcome of consultation, repeat prescriptions were provided for 60% of calls, new prescriptions (15%), while referrals were to outpatient department (17%), emergency department/pediatric emergency center (5%), and primary health care centres (3%). We conclude that during a pandemic, physician-staffed telephone hotline is feasible and can be employed in innovative ways to conserve medical resources, maintain continuity of care, and serve patients requiring urgent care

Novel quinolines carrying pyridine, thienopyridine, isoquinoline, thiazolidine, thiazole and thiophene moieties as potential anticancer agents

As a part of ongoing studies in developing new anticancer agents, a class of structurally novel 1,2-dihydropyridine 4, thienopyridine 5, isoquinolines 6-20, acrylamide 21, thiazolidine 22, thiazoles 23-29 and thiophenes 33-35 bearing a biologically active quinoline nucleus were synthesized. The structure of newly synthesized compounds was confirmed on the basis of elemental analyses and spectral data. All the newly synthesized compounds were evaluated for their cytotoxic activity against the breast cancer cell line MCF7. 2,3-Dihydrothiazole-5-carboxamides 27, 25, 4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxamide (34), 1,2-dihydroisoquinoline-7-carbonitrile (7), 5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carboxamide (35), 1,2-dihydroisoquinoline-7-carbonitrile (6), 2-cyano-3-(dimethylamino)-N-quinolin-3-yl)acrylamide (21), 1,2-dihydroisoquinoline-7-carbonitriles (11) and (8) exhibited higher activity (IC50 values of 27-45 μmol L–1), compared to doxorubicin (IC50 47.9 μmol L–1). (Quinolin-3-yl)-1,2-dihydroisoquinoline-7-carbonitrile (12), 2-thioxo-2,3-dihydrothiazole-5-carboxamide (28) and (quinolin-3-yl)-1,2-dihydroisoquinoline-7-carbonitrile (15) show comparable activity to doxorubicin, while (quinolin-3-yl)-1,2-dihydroisoquinoline-7-carbonitrile (9), 2, 3-dihydrothiazole-5-carboxamide (24), thieno [3,4-c] pyridine-4(5H)-one (5), cyclopenta[b]thiophene-3-carboxamide (33) and (quinolin-3-yl)-6-stryl-1,2-dihydroisoquinoline-7-carbonitrile (10) exhibited moderate activity, lower than doxorubicin

Healthcare Management In Promoting Role Of Paramedics And Nurses In Critical Care; Review

Healthcare unit managers play a crucial role in promoting the importance of paramedics and nurses in critical care. They are responsible for ensuring the health and well-being of nurses and paramedics, as well as maintaining high-quality care. However, there is a lack of research focused on the actions and organizational strategies employed by healthcare unit managers to support nurses, as well as the specific working environments that facilitate such support. This paper utilizes a mixed methods approach, combining qualitative interviews and quantitative surveys with healthcare unit managers and nurses. The findings highlight the crucial role of healthcare unit managers\u27 accessibility to their nursing staff in promoting nurses. Additionally, the study emphasizes the importance of responsive support from the care unit managers\u27 superiors, administration, and managerial colleagues in creating favorable working conditions. The care unit manager\u27s own support was highly supported by the superior manager, and this support was positively connected with nurses

Green extraction of Milletia pinnata oil for the development, and characterization of pectin crosslinked carboxymethyl cellulose/guar gum herbal nano hydrogel

Milletia pinnata oil and Nardostachys jatamansi are rich sources of bioactive compounds and have been utilized to formulate various herbal formulations, however, due to certain environmental conditions, pure extract form is prone to degradation. Therefore, in this, study, a green hydrodistillation technology was used to extract M. pinnata oil and N. jatamansi root for the further application in development of pectin crosslinked carboxymethyl cellulose/guar-gum nano hydrogel. Both oil and extract revealed the presence of spirojatamol and hexadecanoic acid methyl ester. Varied concentrations (w/w) of cross-linker and gelling agent were used to formulate oil emulsion extract gel (OEEG1, OEG1, OEEG2, OEG2, OEEG3, OEG3, OEEG4, OEG4, OEEG5, OEG5), in which OEEG2 and OEG2 were found to be stable. The hydrogel displayed an average droplet size of 186.7 nm and a zeta potential of −20.5 mV. Endo and exothermic peaks and the key functional groups including hydroxyl, amide II, and amide III groups confirmed thermal stability and molecular structure. The smooth surface confirmed structural uniformity. Bactericidal activity against both Gram-positive (25.41 ± 0.09 mm) and Gram-negative (27.25 ± 0.01 mm) bacteria and anti-inflammatory activity (49.25%–83.47%) makes nanohydrogel a potential option for treating various infections caused by pathogenic microorganisms. In conclusion, the use of green hydrodistillation technology can be used to extract the bioactive compounds that can be used in formulation of biocompatible and hydrophobic nanohydrogels. Their ability to absorb target-specific drugs makes them a potential option for treating various infections caused by pathogenic microorganisms

Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy

Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent