Hassan A. Madkhali

Introduction: Diabetes mellitus (DM) is a chronic metabolic disorder characterized by impaired wound healing. The goal of this study was to create and assess Schiff base derivatives (SB) of L-arginine and aromatic aldehydes as possible inhibitors of nitric oxide synthase (NOS). Materials and Methods: Synthesis of SB derivatives was followed by ADMET analysis and molecular docking against the NOS protein (PDB ID: 4NOS, 2.25 Å resolution) obtained from the RCSB Protein Data Bank. Structure-activity correlations were established, and NOS inhibitory activity was evaluated in vitro. Result: FTIR and 1H NMR spectroscopy were used to characterize the five most promising derivatives (CM7, CM8, CM9, CM11, and CM14), which were produced utilizing L-arginine and aromatic natural aldehydes in ethanol/methanol solvents. High binding affinities for CM8 (-9.0 kcal/mol), CM9 (-8.8 kcal/mol), and CM11 (-9.1 kcal/mol) were found via molecular docking. Discussion: The compounds met all pharmacokinetic ADME and toxicity criteria and demonstrated NOS inhibitory action on par with the standard. In order to improve wound healing in diabetics, this study presents L-arginine Schiff base derivatives, a novel class of NOS inhibitors. Conclusion: This research introduces L-Arginine Schiff base (SB) derivatives as a potential class of Nitrous oxide inhibitors that have remained elusive until now.

Background: Platelet concentrates (PCs) are vital for treating hematologic disorders and thrombocytopenia, yet their short shelf life (3–5 days) is limited by platelet storage lesion (PSL)—a process involving biochemical and structural deterioration that reduces post-transfusion efficacy. This study aimed to characterize alterations in platelet surface receptors and RNA content during storage to better understand PSL mechanisms. Methods: Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from healthy donors and stored PCs. Flow cytometry was used to assess the expression of GPIbα, GPVI, Integrin αIIbβ3, and CD9. Thiazole orange (TO) staining evaluated RNA content to distinguish young from aged platelets, while soluble GPVI (sGPVI) levels were quantified by ELISA. Data were analyzed using one-way ANOVA and Student’s t-test (p < 0.05). Results: Baseline receptor profiles were established from fresh donor platelets. Stored PCs showed a progressive decline in GPIbα and GPVI expression from day 6, with significant reductions by day 11 (p < 0.05). αIIbβ3 expression decreased early (day 6) and stabilized thereafter, whereas CD9 remained unchanged. TO staining indicated a gradual loss of RNA-rich platelets, signifying aging. ELISA revealed increased sGPVI levels from day 6 to day 14, inversely correlating with surface GPVI loss. Conclusions: Prolonged storage leads to receptor degradation and platelet senescence, notably affecting GPIbα, GPVI, and αIIbβ3. Elevated sGPVI levels and reduced RNA content reflect progressive PSL. Flow cytometry and ELISA offer reliable monitoring tools, and sGPVI may serve as a biomarker for platelet quality during storage.

: Plasma kallikrein (PKa), a serine proteinase in the kallikrein–kinin system, is involved in vascular inflammation and edema. In this work, three natural flavonoids, Fisetin, Morin, and Rutin, were screened as prospective human plasma kallikrein inhibitors and compared to the standard drug Sebetralstat by means of integrated in vitro and in silico assays. Chromogenic assay revealed concentration-dependent inhibition of plasma Kallikrein (IC50 = 300.13 µM for Fisetin, 377.36 µM for Morin, and 566.04 µM for Rutin). Satisfactory binding energies (–9.1, –8.6, and –7.5 kcal mol⁻¹ respectively) were also predicted, along with similar interaction patterns by molecular docking analyses. MD simulations demonstrated that Morin and Fisetin formed a stable complex, whereas Rutin induced local dynamic properties. The ADMET predictions revealed acceptable pharmacokinetic profiles for Fisetin and Morin, while Rutin showed low permeability. These results point to Fisetin and Morin as candidate natural scaffolds for selective inhibition of plasma kallikrein, with moderate in vitro potency but good in silico stability and pharmacokinetics. Additional confirmation in cellular models and structure-guided optimization of these flavonoid scaffolds are required to develop safer anti-inflammatory drugs that act on the kallikrein–kinin system.

Background: Alzheimer’s disease remains without an effective disease-modifying therapy, largely due to its multifactorial pathology involving tau hyperphosphorylation, amyloid dysregulation, neuroinflammation and neuronal atrophy. Dual inhibition of glycogen synthase kinase-3[ ] (GSK-3[ ] and histone deacetylase 6 (HDAC6) — two enzymes central to AD progression — has recently emerged as a promising strategy for simultaneous modulation of these pathogenic pathways. This study aims to identify natural molecules capable of concurrently targeting GSK-3[ ] and HDAC6 using an integrated computational pipeline. Methods:Receptor-guided pharmacophore models were generated from crystallographic complexes of GSK-3[ ] and HDAC6 to screen large natural product libraries. Common hits were subjected to molecular docking, MM-GBSA binding free energy estimation, molecular dynamics (MD) simulations and ADMET profiling to assess binding strength, stability and drug-likeness. Results: Eleven phytochemicals satisfied both pharmacophore hypothesis, among which four — Cardamonin, Isorhapontigenin, Neobavaisoflavone (NBI) and Pinostilbene — demonstrated the strongest dual-target complementarity. These compounds achieved docking scores ranging from –7.96 kcal/mol to –9.32 kcal/mol for GSK-3[ ] and –6.85 kcal/mol to –7.97 kcal/mol for HDAC6. MM-GBSA [ ] values ranged from –61.04 kcal/mol to –67.37 kcal/mol for GSK-3[ ] and –31.04 kcal/mol to –38.96 kcal/mol for HDAC6, surpassing co-crystal controls. MD simulations confirmed sustained structural integrity with average RMSD of 1.67–2.01 Å for GSK-3[ ] and 1.26–2.03 Å for HDAC6. ADMET analysis revealed acceptable CNS permeability, metabolic stability and safety profiles. Conclusion: This study identified four natural-product chemotypes with favorable dual-binding profiles for GSK-3[ ] and HDAC6. These findings will provide a promising starting point for future biochemical, enzymatic and cellular validation studies, which are essential to confirm their therapeutic potential.

Background: Platelet concentrates (PCs) are vital for treating hematologic disorders and thrombocytopenia, yet their short shelf life (3–5 days) is limited by platelet storage lesion (PSL)-a process involving biochemical and structural deterioration that reduces post-transfusion efficacy. This study aimed to characterize alterations in platelet surface receptors and RNA content during storage to better understand PSL mechanisms. Methods: Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from healthy donors and stored PCs. Flow cytometry was used to assess the expression of GPIbα, GPVI, Integrin αIIbβ3, and CD9. Thiazole orange (TO) staining evaluated RNA content to distinguish young from aged platelets, while soluble GPVI (sGPVI) levels were quantified by ELISA. Data were analyzed using one-way ANOVA and Student’s t-test (P < 0.05). Results: Baseline receptor profiles were established from fresh donor platelets. Stored PCs showed a progressive decline in GPIbα and GPVI expression from day 6, with significant reductions by day 11 (P < 0.05). αIIbβ3 expression decreased early (day 6) and stabilized thereafter, whereas CD9 remained unchanged. TO staining indicated a gradual loss of RNA-rich platelets, signifying aging. ELISA revealed increased sGPVI levels from day 6 to day 14, inversely correlating with surface GPVI loss. Conclusion: Prolonged storage leads to receptor degradation and platelet senescence, notably affecting GPIbα, GPVI, and αIIbβ3. Elevated sGPVI levels and reduced RNA content reflect progressive PSL. Flow cytometry and ELISA offer reliable monitoring tools, and sGPVI may serve as a biomarker for platelet quality during storage .

Gastric ulcers are among the most widespread gastrointestinal diseases, often caused by NSAIDs. Natural compounds with less toxicity and more safety are being explored to minimize their impact. The main goal is to examine the gastroprotective effects of Myricetin in indomethacin-induced gastric ulcers in rats, and the mechanism involved. The rats were allocated into control, negative, standard, Myricetin 10 mg/kg, and Myricetin 20 mg/kg groups. The effect of Myricetin on indomethacin–induced ulcers was examined for 14 days. The Ulcer index, gastric acidity, and PGE₂ level were evaluated. The inflammatory markers (IL-1β, IL-6, and TNF-α), oxidative stress levels (MDA, GSH, CAT, and SOD), histological study, and apoptotic markers (Bax, NF-κB, and caspase-3) were also assessed. Pre-treatment with Myricetin showed a protective effect towards indomethacin-induced gastric ulceration via reducing Ulcer index, gastric juice volume, and increasing gastric pH, and protective PGE₂ level. It also alleviated the level of TNF-α, IL-1β, and IL-6 and modulated MDA, GSH, CAT, and SOD activity and the apoptotic markers (BAX, Caspase-3, and NF-κB) in the gastric mucosa. It was also examined by histopathological study that Myricetin reduced mucosal ulcers, inflammatory infiltration, and degeneration of cell membranes. The results showed that Myricetin has a gastroprotective effect on indomethacin-induced gastric ulcers through anti-inflammatory, antioxidant, and antiapoptotic mechanisms that should be further investigated.

Thioacetamide (TAA), an established liver toxic chemical, is used to develop experimental models of liver injury by inducing inflammation, oxidative stress, and apoptosis. The current study investigates the hepatoprotective effect of diosmetin (DSM), a bioflavonoid molecule, with mechanistic investigations using an in vivo TAA-induced liver injury model because there is no precedence. In the current investigation, 30 Wistar rats were randomly assigned to five groups. The rats were given TAA (200 mg/kg) and DSM (50 and 100 mg/kg body weight) for 8 weeks. Liver function biomarkers (ALT, ASP, AST, bilirubin, GGT, CRP, albumin, globulin, and total protein) and inflammatory markers were analyzed on serum, whereas antioxidant levels, histological evaluation, and apoptotic markers were evaluated on liver tissue. The finding of this study indicated that DSM ameliorates TAA-induced increases in ALT, ASP, AST, bilirubin, GGT, CRP, MDA, albumin, globulin, total protein, and antioxidant enzyme activity such as SOD, CAT, and GSH levels. DSM reduced alterations in inflammatory biomarkers (IL-6, IL-1β, TNF-α) and apoptotic markers (Bax, NF-κB, caspase-3, Nrf-2, PPAR_ϒ) in liver tissues. Furthermore, DSM therapy resulted in significant major changes in liver histology, fibrosis, and cell death. The current study’s findings demonstrated that DSM significantly decreases hepatotoxicity by enhancing liver structure and function while suppressing oxidative stress, inflammation, and apoptosis.

Background: This study aimed to evaluate the effects of selected phosphodiesterase-4 inhibitors (PDE-4 inhibitors)—roflumilast, ibudilast, and crisaborole—on the activity of blood coagulation factor XII (FXII). In the intrinsic coagulation pathway, FXII is known to initiate the kallikrein–kinin system (KKS), causing an increase in the system expression, which ultimately leads to inflammation and coagulation states. Additionally, the activation of KKS downstream effectors leads to inflammation. Inflammation signaling was found to be initiated when the bradykinin (BK) protein binds to its B2 receptor because of the FXII-dependent pathway activation. BK abnormalities can cause a critical condition, hereditary angioedema (HAE), which is characterized by recurring serious swelling. While it is considered unnecessary for hemostasis, FXII is an important enzyme for pathogenic thrombosis. Because of this special characteristic, FXII is a desirable therapeutic target. Our hypothesis is to identify the inhibitory effects of roflumilast, ibudilast, and crisaborole on the activated FXII and to reveal their beneficial impacts in the reduction of the pathogenesis of FXII-related conditions, HAE, and thrombosis. In a current study, we presented the inhibitory effect of tested drugs on the main target activated factor XII (FXIIa) as well as two other plasma protease enzymes included in the target pathway, plasma kallikrein and FXIa. Methods: To achieve our aim, in vitro chromogenic enzymatic assays were utilized to assess the inhibitory effects of these drugs by monitoring the amount of para-nitroaniline (pNA) chromophore released from the substrate of FXIIa, FXIa, or plasma kallikrein. Results: Our study findings exhibited that among assessed PDE-4 inhibitor drugs, roflumilast at micromolar concentrations significantly inhibited FXIIa in a dose-dependent manner. The FXIIa was clearly suppressed by roflumilast, but not the other related KKS members, plasma kallikrein, or the activated factor XI. On the other hand, ibudilast and crisaborole showed no inhibitory effects on the activities of all enzymes. Conclusions: Overall, roflumilast could be used as a lead compound for developing a novel multifunctional therapeutic drug used for the prevention of HAE or thrombotic disorders.

Cervical Cancer remains a women’s health concern worldwide and ranks among the most prevalent cancers, particularly in developing countries. Many women are diagnosed with cervical cancer, with a substantial number succumbing to the disease even after the availability of vaccines and drugs. The tumour microenvironment often exhibits immune evasion, including suppression of T-cell activity and altered cytokine, impacting the efficacy of therapeutic interventions and highlighting the need for treatments to modulate the immune response. Despite efforts to promote HPV vaccination and regular screenings, it causes many deaths, underscoring the urgent need for continued research, healthcare access, and rapid drug development or repurposing. In this study, we identified various proteins involved in cervical cancer cell cycle regulation and DNA replication proteins, performed the multitargeted docking with an FDA-approved library, and identified Oxidopamine HBr as a multitargeted drug. Studies extended with pharmacokinetics and compared with the standard values followed by DFT, which supported the compound as a multitargeted inhibitor. Further, the docked complexes were taken for the interaction fingerprints, and it was identified that there are many 9 polar, 5 hydrophobic, 2 aromatic, and 2 basic residues. We extended our studies for 100ns MD Simulation in water, and the computations explored the deviation and fluctuations under 2Å and many intermolecular interactions; the same trajectory files were used for the MM\GBSA studies. All the studies have supported the Oxidopamine HBr as a cervical cancer multitargeted inhibitor—however, experimental studies are needed before human use.

Recent studies have indicated that microRNA and VEGF are considered to be genetic modifiers and are associated with elevated levels of fetal haemoglobin HbF, and thus they reduce the clinical impact of sickle haemoglobin (HbS) patients. This cross-sectional study was performed on clinical confirmed subjects of SCD cases. miR-423-rs6505162 C>T and VEGF-2578 C>A genotyping was conducted by ARMS-PCR in SCD and healthy controls. A strong clinical significance was reported while comparing the association of miR-423 C>T genotypes between SCD patients and controls (p = 0.031). The microRNA-423 AA genotype was associated with an increased severity of SCD in codominant model with odd ratio (OR = 2.36, 95% CI, (1.15–4.84), p = 0.018) and similarly a significant association was observed in recessive inheritance model for microRNA-423 AA vs (CC+CA) genotypes (OR = 2.19, 95% CI, (1.32–3.62), p < 0.002). The A allele was associated with SCD severity (OR = 1.57, 95% CI, (1.13–2.19), p < 0.007). The distribution of VEGF-2578 C>A genotypes between SCD patients and healthy controls was significant (p < 0.013). Our results indicated that in the codominant model, the VEGF-2578-CA genotype was strongly associated with increased SCD severity with OR = 2.56, 95% CI, (1.36–4.82), p < 0.003. The higher expression of HbA1 (65.9%), HbA2 (4.40%), was reported in SCD patients carrying miR-423-AA genotype than miR-423 CA genotype in SCD patients carrying miR-423 CA genotype HbA1 (59.98%), HbA2 (3.74%) whereas SCD patients carrying miR-423 CA genotype has higher expression of HbF (0.98%) and HbS (38.1%) than in the patients carrying AA genotype HbF (0.60%), HbS (36.1%). ARMS-PCR has been proven to be rapid, inexpensive and is highly applicable to gene mutation screening in laboratories and clinical practices. This research highlights the significance of elucidating genetic determinants that play roles in the amelioration of the HbF levels that is used as an indicator of severity of clinical complications of the monogenic disease. Further well-designed studies with larger sample sizes are necessary to confirm our findings.