Dhurgham Al-Fahad

Antimicrobial resistance (AMR) generates serious negative impacts on health-care systems worldwide, and Iraq is not an exception. To uncover the prevalence of AMR and to visualize the magnitude of the multidrug-resistant (MDR) dilemma in Iraqi hospitals, this study is carried out. A total of 11592 clinical records from ten different health-care facilities in seven Iraqi provinces are collected and analyzed. Our data show that 4984 (43.0%) of all clinical samples are negative for bacterial growth. In adults, Gram-negative bacteria (GNB) represented 48.9% and Gram-positive bacteria (GPB) represented 51.1% of clinical isolates; in children, GNB represented 60.8% and GPB represented 39.2%. Furthermore, in adults, Klebsiella pneumoniae (30.1%) and Staphylococcus aureus (40.8%) are among the most common GNB and GPB isolates, respectively. In children, K. pneumoniae (37.9%) and Staphylococcus haemolyticus (41.8%) are the most common GNB and GPB, respectively. Adults’ samples showed that Escherichia coli and Proteus mirabilis were the most resistant GNB; S. aureus and Staphylococcus epidermidis are among the most resistant GPB. In children, K. pneumoniae is found to be the most resistant GNB. This study confirms the persistence of antimicrobial resistance and multidrug-resistant gram-negative and gram-positive bacteria in adults and children alike. Ampicillin and oxacillin have been recognized as ineffective drugs in adults, and ampicillin, nafcillin, cefoxitin, and benzylpenicillin have been found to be highly resisted by pathogenic bacteria in children. The outcomes confirm the necessity of conducting AMR surveillance on a regular basis and establishing national antibiotic prescription guidelines to manage AMR development in Iraq.

Serine/threonine kinase 16 (STK 16) is involved in many facets of cellular regulation; activation of STK 16 plays a crucial role in the migration of cancer cells. Therefore, it is a novel target for the discovery of anticancer agents. Herein, virtual screening and dynamics simulation were used to screen a large library of natural compounds against STK 16 using Schrodinger suit 2021-2 and GROMACS 2021.6. The results predicted five molecules with high binding affinity against the target, with NPC132329 (Arcyriaflavin C) and NPC160898 having higher binding affinity and molecular mechanics generalized born surface area (MM/GBSA), suggesting that it is better than the standard inhibitor. The molecular dymanics (MD) simulation studies showed that the STK 16-NPC132329 complex has the lowest root mean square deviation, and STK 16-NPC160898 was the most stable compared with the standard drug and selective STK 16 inhibitor. The minimal fluctuation was observed in the STK 16-NPC132329 and STK 16-NPC160898 complexes based on the root mean square fluctuation trajectory with NPC132329 and NPC160898 forming 2 and 3 hydrogen bonds respectively with the amino acid residue of the target’s binding site. Overall, NPC132329 and NPC160898 are better STK 16 inhibitors than the standard drug and selective inhibitor, which can be further studied to discover novel anticancer drugs.

Histone deacetylases (HDACs) are enzymes that remove acetyl groups from ɛ-amino of histone, and their involvement in the development and progression of cancer disorders makes them an interesting therapeutic target. This study seeks to discover new inhibitors that selectively inhibit HDAC enzymes which are linked to deadly disorders like T-cell lymphoma, childhood neuroblastoma, and colon cancer. MOE was used to dock libraries of ZINC database molecules within the catalytic active pocket of target HDACs. The top three hits were submitted to MD simulations ranked on binding affinities and well-occupied interaction mechanisms determined from molecular docking studies. Inside the catalytic active site of HDACs, the two stable inhibitors LIG1 and LIG2 affect the protein flexibility, as evidenced by RMSD, RMSF, Rg, and PCA. MD simulations of HDACs complexes revealed an alteration from extended to bent motional changes within loop regions. The structural deviation following superimposition shows flexibility via a visual inspection of movable loops at different timeframes. According to PCA, the activity of HDACs inhibitors induces structural dynamics that might potentially be utilized to define the nature of protein inhibition. The findings suggest that this study offers solid proof to investigate LIG1 and LIG2 as potential HDAC inhibitors.

Rab5A and Akt pathways are reported to be responsible for the invasiveness of cancer cells, indicated by the fact that Rab5A activates the downstream Phosphoinositide-3-kinases (PI3K)/Akt signalling pathway, which results in promoting cancer metastasis. However, little attention has been given to the emerging role of Rab5A and Akt signalling pathways in regulating the direction of MDA-MB-231 cell migration. MDA-MB-231 breast cancer cell line was used as a model in this study because it is highly metastatic and motile. Time-lapse microscopy was used to examine the effect of Akt and Rab5A inhibitors on cell migration, proliferation and wound healing. Later, the cells were transfected with GFP-Akt-PH or GFP-Rab5A (used as a biosensor to detect Akt and Rab5A). Therefore, confocal time-lapse images were used to visualize Akt and Rab5A at the front and rear edges of the cells. The recorded data demonstrated that Akt and Rab5A inhibition reduced cell migration, proliferation and wound healing. The results of the current study also demonstrated that Akt localizes at the trailing edge while Rab5A localize more at the leading edge than the trailing edge of cells. This study suggests that Akt and Rab5A inhibition might regulate the direction of breast cancer migration.

In the migration and metastasis of cancer cells, it is necessary to rotate the focal adhesion (FA). MAP4K4 plays a vital role in the formation of cytoskeletal regeneration, but its role in regulating FA dynamics and cancer cell migration is not well understood. This present aimed to investigate the role of MAP4K4 in regulating FA dynamics and cell migration in the human breast cancer cell line. For this purpose, different variants, including MAP4K4 (wild type), partial active mutation kinase (MAP4K4-T178D), mutant with inactivated or reduced activity kinase (MAP4K4-T178A) and inactive kinase mutation (MAP4K4-K54R) was used in the evaluation. GFP-paxillin was also used as a marker in basal breast cancer cells (MDA-MB-231) in determining FA dynamics. Time-lapse and confocal microscopes were used to record FA dynamics and cell migration. The present study's findings showed that cells expressing MAP4K4-K54R, MAP4K4-T178D and MAP4K4-T178A type slowly down the FA turnover rates and had much larger FAs than those expressing WT MAP4K4 in MDA-MB-231 breast cancer cell line. Furthermore, inhibiting MAP4K4 strongly inhibited FA formation and reduced cell migration speed. In conclusion, MAP4K4 regulates FA dynamics and cancer cell migration, most probably through activating FA proteins and cytoskeleton.

Background: Focal adhesions (FAs) are highly dynamic complex structures that assembled and disassembled on an ongoing basis. The balance between the two processes mediates various aspects of cell behavior, ranging from cell adhesion to cell migration. Assembly and disassembly processes of FAs are regulated by a variety of cellular signaling proteins and adaptors. ...

Background: The assembly and disassembly of the focal adhesions (FA) components occurs throughout life cycle of adhesion, with conservation of balance between removal and recruitment rate during temporal stages. Previous studies have demonstrated that phosphotidyilinositols play a role in regulating FA turnover. However, a little attention has been given to quantify the dynamics changes of ...

The increase in multidrug-resistant pathogens in urinary tract infections (UTIs) among communities and hospitals threatens our ability to treat these common pathogens. Uropathogenic Escherichia coli (UPEC) strains are the most frequent uropathies linked to the development of UTIs. This work aims to introduce bioactive natural products via virtual screening of small molecules from a public database to prevent biofilm formation by inhibiting FimH, a type 1 fimbriae that plays a crucial role in UPEC pathogenicity. A total of 30926 small molecules from the NPASS database were subjected to screening via molecular docking. Followed by performing in silico ADME studies, seven molecules showed promising docking results ranging from −6.8 to −8.7 kcal/mol. As a result of the docking score findings, 100 ns Molecular dynamics (MD) simulations were performed. Based on MM-PBSA analysis, NPC313334 ligand showed high binding affinity −42 and stability with the binding pocket of FimH protein during molecular dynamic simulations. DFT calculations were also performed on the ligands to calculate the HOMO-LUMO energies of the compounds in order to an idea about their structure and reactivity. This research suggests that NPC313334 may be a possible antibacterial drug candidate that targets FimH to reduce the number of UPEC-related urinary tract infections. Communicated by Ramaswamy H. Sarma

Phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5) P3) and Phosphatidylinositol 4,5-trisphosphate (PtdIns(4,5) P2] form an insignificant amount of phospholipids but play important roles in controlling membrane-bound signalling. Little attention has been given to visualize and monitor changes or differences in the local generation of PtdIns(4,5) P2 and PtdIns(3,4,5) P3 in the cell membranes of MDA-MB-231 breast cancer cell lines. PLCδ1-PH-GFP and Btk-PH-GFP were used as biosensors to detected PtdIns(4,5) P2 and PtdIns(3,4,5)P3 respectively. These biosensors and antibodies were transfected, immuostained and then visualized by confocal microscopy on different cell surfaces. Our results showed that PLCδ1-PH-GFP/mCherry was localized at the cell membrane, while Btk-PH-GFP/mCherry was sometimes localized at the cell membrane but there was also a large amount of fluorescence present in the cytosol and nucleus. Our results also showed that the cells that expressed low levels of Btk-PH-GFP the fluorescence was predominantly localised to the cell membrane. While the cells that expressed high levels of Btk-PH-GFP the fluorescence was localization in the cytosol and cell membrane. Our results demonstrated that both anti-PtdIns(4,5)P2 and anti-PtdIns(3,4,5)P3 antibodies were localized everywhere in cell. Our results suggest that PLCδ1-PH-GFP and Btk-PH-GFP/mCherry have more specificity, reliability, suitability and accuracy than antibodies in binding with and detecting PtdIns(4,5)P2 and PtdIns(3,4,5)P3 and in studying the molecular dynamics of phospholipids in live and fixed cells.