Incidence of Hashimoto Thyroiditis Among Libyans: A Retrospective Epidemiological Study

Background and aims. Hashimoto's disease is an autoimmune disorder in which the body produces antibodies that attack the thyroid gland, leading to chronic inflammation, destruction of the gland, and hypothyroidism. This study aimed to assess the epidemiology of this disease among Libyan patients. Methods. A cross-sectional retrospective study conducted from June 2012 to April 2020 in order to examine the anti TPO level among Libyan population. Data was collected from eastern and western part of Libya, and were analyzed from available sample for 244 apparently patients with thyroid disorders collected from different private clinic’s laboratories. The analysis for serum anti-TPO was done by electrochemiluminescence protein binding assay (ECLIA) using Roche diagnostics and Cobas e411 analyzer. Results. The current results showed that females predominate the study, and most of them were in the age group of (>40) years old. About 49.18% of these cases were suffering from Hashimoto's disease (High ATPO level). The mean value of anti-TPO status among females was (0.5±2) nmol/L, while among males it was (0.45±3) nmol/L. Significantly, more women (81.66%) had Anti- TPO Above (34 IU/ml), compared to (18.33%) of male participants. Conclusion. Hashimoto disease is common among patients with thyroid dysfunction especially females. Our findings suggest that different interventional strategies are needed to reduce the chances of developing Hashimoto’s and its associated negative health outcomes in Libya.

Improving the accuracy of unbound fraction measurement of drug-protein binding by preconditioning the RED membrane inserts

Aim: The objective of this study was to evaluate the rapid equilibrium dialysis (RED) device in protein binding assays in diluted protein matrices and to improve the accuracy of the unbound fraction ( fu) measurement. Methodology: Protein binding assays of drug compounds in bovine serum albumin solutions and human plasma with different folds of dilution were performed using the RED device with and without preconditioning of the dialysis membrane inserts, and the results were compared with those using other approaches in this study. Results & conclusion: Preconditioning of the RED membrane inserts improved the fu data accuracy of drug-protein binding assay in matrices with relatively low protein contents and such impact could be compound dependent.

Synthesis of novel cytotoxic tetracyclic acridone derivatives and study of their molecular docking, ADMET, QSAR, bioactivity and protein binding properties

Acridone based synthetic and natural products with inherent anticancer activity advancing the research and generating a large number of structurally diversified compounds. In this sequence we have designed, synthesized a series of tetracyclic acridones with amide framework viz., 3-(alkyloyl/ aryloyl/ heteroaryloyl/ heteroaryl)-2,3-dihydropyrazino[3,2,1-de]acridin-7(1H)-ones and screened for their in vitro anti-cancer activity. The in vitro study revealed that compounds with cyclopropyl-acetyl, benzoyl, p-hydroxybenzoyl, p-(trifluoromethyl)benzoyl, p-fluorobenzoyl, m-fluorobenzoyl, picolinoyl, 6-methylpicolinoyl and 3-nicotinoyl groups are active against HT29, MDAMB231 and HEK293T cancer cell lines. The molecular docking studies performed for them against 4N5Y, HT29 and 2VWD revealed the potential ligand–protein binding interactions among the neutral aminoacid of the enzymes and carbonyl groups of the title compounds with a binding energy ranging from − 8.1394 to − 6.9915 kcal/mol. In addition, the BSA protein binding assay performed for them has confirmed their interaction with target proteins through strong binding to BSA macromolecule. The additional studies like ADMET, QSAR, bioactivity scores, drug properties and toxicity risks ascertained them as newer drug candidates. This study had added a new collection of piperazino fused acridone derivatives to the existing array of other nitrogen heterocyclic fused acridone derivatives as anticancer agents.

Drosophila melanogaster as a model host to study arbovirus–vector interaction

ABSTRACTArboviruses cause the most devastating diseases in humans and animals worldwide. Several hundred arbovirus are transmitted by mosquitoes, sand flies or ticks and are responsible for more than million deaths annually. Development of a model system is essential to extrapolate the molecular events occurring during infection in the human and mosquito host. Virus overlay protein binding assay (VOPBA) combined with MALDI TOF/TOF MS revealed that Dengue-2 virus (DENV-2) exploits similar protein molecules in Drosophila melanogaster and Aedes aegypti for its infection. Furthermore, the virus susceptibility studies revealed that DENV-2 could propagate in D. melanogaster, and DENV-2 produced in fruit fly is equally infectious to D. melanogaster and Ae. aegypti. Additionally, real time PCR analysis revealed that RNAi coupled with JAK-STAT and Toll pathway constitutes an effector mechanism to control the DENV-2 infection in flies. These observations point out that D. melanogaster harbors all necessary machineries to support the growth of arboviruses. With the availability of well-established techniques for genetic and developmental manipulations, D. melanogaster, offers itself as the potential model system for the study of arbovirus-vector interactions.

Novel isoniazid derivative as promising antituberculosis agent

Background: A major focus of tuberculosis drug discovery is aimed at the development of novel antibiotics with activity against drug-resistant strains of Mycobacterium tuberculosis. Results: We have synthesized ten isoniazid derivatives and investigated for antibacterial activity toward M. tuberculosis H37Rv and isoniazid-resistant strain SRI 1369. It was revealed that only one compound, isonicotinic acid (1-methyl-1H-pyrrol-2-ylmethylene)-hydrazide (1), is active toward isoniazid-resistant strain with minimum inhibitory concentration value of 0.14 μM. This compound is not cytotoxic toward human liver cells (HepG2; IC50 >100 μM), demonstrates good permeability in Caco-2 cells. Accordingly to the results of plasma protein binding assay, unbound fraction of compound 1, which potentially exhibits pharmacologic effects, is 57.9%. Conclusion: Therefore, isonicotinic acid (1-methyl-1H-pyrrol-2-ylmethylene)-hydrazide is a promising compound for further preclinical studies.

Competitive Protein Binding Assay of Naproxen by Human Serum Albumin Functionalized Silicon Dioxide Nanoparticles

We have developed a new competitive protein binding assay (CPBA) based on human serum albumin functionalized silicon dioxide nanoparticles (nano-SiO2-HSA) that can be used for naproxen determination in urine. Compared with a conventional multi-well reaction plate, nano-SiO2 with a high surface-area-to-volume ratio could be introduced as a stationary phase, markedly improving the analytical performance. Nano-SiO2-HSA and horseradish peroxidase-labeled-naproxen (HRP-naproxen) were prepared for the present CPBA method. In this study, a direct competitive binding to nano-SiO2-HSAwas performed between the free naproxen in the sample and HRP-naproxen. Thus, the catalytic color reactions were investigated on an HRP/3,3′5,5′-tetramethylbenzidine (TMB)/H2O2 system by the HRP-naproxen/nano-SiO2-HSA composite for quantitative measurement via an ultraviolet spectrophotometer. A series of validation experiments indicated that our proposed methods can be applied satisfactorily to the determination of naproxen in urine samples. As a proof of principle, the newly developed nano-CPBA method for the quantification of naproxen in urine can be expected to have the advantages of low costs, fast speed, high accuracy, and relatively simple instrument requirements. Our method could be capable of expanding the analytical applications of nanomaterials and of determining other small-molecule compounds from various biological samples.

Annexin II as a Dengue Virus Serotype 2 Interacting Protein Mediating Virus Interaction on Vero Cells

Recent evidence has demonstrated that dengue virus requires active filopodia formation for a successful infection. However, the cellular factor involved in the interaction has not been fully elucidated. We used a combination of virus overlay protein binding assay and LC-MS/MS, and identified annexin II as a dengue virus serotype 2 (DENV2) interacting protein on Vero cells, upon filopodia induction. Flow cytometry analysis showed annexin II on the Vero cells surface increased when DENV2 was added. The amount of annexin II in the plasma membrane fraction was reduced as the infection progressed. Antibody-mediated inhibition of infection and siRNA-mediated knockdown of annexin II expression significantly reduced DENV2 infection and production levels. Collectively, we demonstrated that annexin II is one of the host factor involved in DENV2 binding on Vero cells.