Two transcriptionally and functionally distinct waves of neutrophils during mouse acute liver injury.

BACKGROUND: Neutrophils are key mediators of inflammation during acute liver injury (ALI). Emerging evidence suggests that they also contribute to injury resolution and tissue repair. However, the different neutrophil subsets involved in these processes and their kinetics are undefined. Herein, we characterized neutrophil kinetics and heterogeneity during ALI. METHODS: We used the carbon tetrachloride model of ALI and employed flow cytometry, tissue imaging, and quantitative RT-PCR to characterize intrahepatic neutrophils during the necroinflammatory early and late repair phases of the wound healing response to ALI. We FACS sorted intrahepatic neutrophils at key time points and examined their transcriptional profiles using RNA-sequencing. Finally, we evaluated neutrophil protein translation, mitochondrial function and metabolism, reactive oxygen species content, and neutrophil extracellular traps generation. RESULTS: We detected 2 temporarily distinct waves of neutrophils during (1) necroinflammation (at 24 hours after injury) and (2) late repair (at 72 hours). Early neutrophils were proinflammatory, characterized by: (1) upregulation of inflammatory cytokines, (2) activation of the noncanonical NF-κB pathway, (3) reduction of protein translation, (4) decreased oxidative phosphorylation, and (5) higher propensity to generate reactive oxygen species and neutrophil extracellular traps. In contrast, late neutrophils were prorepair and enriched in genes and pathways associated with tissue repair and angiogenesis. Finally, early proinflammatory neutrophils were characterized by the expression of a short isoform of C-X-C chemokine receptor 5, while the late prorepair neutrophils were characterized by the expression of C-X-C chemokine receptor 4. CONCLUSIONS: This study underscores the phenotypic and functional heterogeneity of neutrophils and their dual role in inflammation and tissue repair during ALI.

Trends in Fatal Opioid-Related Overdose in American Indian and Alaska Native Communities, 1999-2021.

INTRODUCTION: Opioid-related overdose mortality rates have increased sharply in the U.S. over the past two decades, and inequities across racial and ethnic groups have been documented. Opioid-related overdose trends among American Indian and Alaska Natives require further quantification and assessment. METHODS: Observational, U.S. population-based registry data on opioid-related overdose mortality between 1999 and 2021 were extracted in 2023 using ICD-10 codes from the U.S. Centers for Disease Control and Prevention's Wide-Ranging Online Data for Epidemiologic Research multiple cause of death file by race, Hispanic ethnicity, sex, and age. Segmented time series analyses were conducted to estimate opioid-related overdose mortality growth rates among the American Indian and Alaska Native population between 1999 and 2021. Analyses were performed in 2023. RESULTS: Two distinct time segments revealed significantly different opioid-related overdose mortality growth rates within the overall American Indian and Alaska Native population, from 0.36 per 100,000 (95% CI=0.32, 0.41) between 1999 and 2019 to 6.5 (95% CI=5.7, 7.31) between 2019 and 2021, with the most pronounced increase among those aged 24-44 years. Similar patterns were observed within the American Indian and Alaska Native population with Hispanic ethnicity, but the estimated growth rates were generally steeper across most age groups than across the overall American Indian and Alaska Native population. Patterns of opioid-related overdose mortality growth rates were similar between American Indian and Alaska Native females and males between 2019 and 2021. CONCLUSIONS: Sharp increases in opioid-related overdose mortality rates among American Indian and Alaska Native communities are evident by age and Hispanic ethnicity, highlighting the need for culturally sensitive fatal opioid-related overdose prevention, opioid use disorder treatment, and harm-reduction efforts. Future research should aim to understand the underlying factors contributing to these high mortality rates and employ interventions that leverage the strengths of American Indian and Alaska Native culture, including the strong sense of community.

Medication risk checklist for older adults (LOTTA) - development and validation of a self-assessment tool.

BACKGROUND: Patient safety strategies highlight patients' own active involvement in ensuring medication safety. A prerequisite for involving patients in their medication therapy is having tools that can assist them in ensuring safe medicine use. Older home-dwelling adults with multiple medications are at high risk for medication-related problems, yet only a few age-specific patient self-administered medication risk screening tools exist. This study aimed to develop, validate, and assess the feasibility of a self-administered medication risk checklist for home-dwelling older adults ≥65 years. MATERIALS AND METHODS: The draft checklist was formed based on a validated practical nurse-administered Drug Related Problem Risk Assessment Tool supplemented with findings from two systematic literature reviews. The content validity of the draft checklist was determined by a three-round Delphi survey with a panel of 19 experts in geriatric care and pharmacotherapy. An agreement of ≥80% was required. A feasibility assessment (i.e. understandability of the items, fill-out time of the checklist) of the content-validated checklist was conducted among older adults ≥65 years (n = 87) visiting community pharmacies (n = 4). Data were analysed using qualitative content analysis. RESULTS: The final validated and feasibility-tested Medication Risk Checklist (LOTTA) for home-dwelling older adults consists of eight items screening the highest priority systemic risks (three items), potentially drug-induced symptoms (one item), adherence, and self-management problems (four items). The checklist proved feasible for self-administration, the mean fill-out time being 6.1 min. CONCLUSIONS: A wide range of potential medication risks related to the medication use process can be identified by patient self-assessment. Screening tools such as LOTTA can enhance early detection of potential medication risks and risk communication between older adults and their healthcare providers. A wider and more integrated use of the checklist could be facilitated by making it electronically available as part of the patient information systems.

Patient safety strategies highlight patients' own active involvement in ensuring medication safety, which in turn, requires easy-to-use tools to self-assess potential medication risks and communicate them with healthcare providers.This study produced a short, age-specific eight item Medication Risk Checklist (LOTTA) to be self-administered by home-dwelling older adults to identify major systemic risks, potential drug-induced symptoms, adherence, and self-management problems related to medication taking.To facilitate the use of the checklist in early detection of potential medication risks, future studies should focus on converting the LOTTA list into electronic form and pilot its use as an integrated part of the electronic patient information system.

The multifaceted role of macrophages during acute liver injury.

The liver is situated at the interface of the gut and circulation where it acts as a filter for blood-borne and gut-derived microbes and biological molecules, promoting tolerance of non-invasive antigens while driving immune responses against pathogenic ones. Liver resident immune cells such as Kupffer cells (KCs), a subset of macrophages, maintain homeostasis under physiological conditions. However, upon liver injury, these cells and others recruited from circulation participate in the response to injury and the repair of tissue damage. Such response is thus spatially and temporally regulated and implicates interconnected cells of immune and non-immune nature. This review will describe the hepatic immune environment during acute liver injury and the subsequent wound healing process. In its early stages, the wound healing immune response involves a necroinflammatory process characterized by partial depletion of resident KCs and lymphocytes and a significant infiltration of myeloid cells including monocyte-derived macrophages (MoMFs) complemented by a wave of pro-inflammatory mediators. The subsequent repair stage includes restoring KCs, initiating angiogenesis, renewing extracellular matrix and enhancing proliferation/activation of resident parenchymal and mesenchymal cells. This review will focus on the multifaceted role of hepatic macrophages, including KCs and MoMFs, and their spatial distribution and roles during acute liver injury.

Memantine/Aripiprazole Combination Alleviates Cognitive Dysfunction in Valproic Acid Rat Model of Autism: Hippocampal CREB/BDNF Signaling and Glutamate Homeostasis.

Significant efforts are increasingly directed towards identifying novel therapeutic targets for autism spectrum disorder (ASD) with a rising role of aberrant glutamatergic transmission in the pathogenesis of ASD-associated cellular and behavioral deficits. This study aimed at investigating the role of chronic memantine (20 mg/kg/day) and aripiprazole (3 mg/kg/day) combination therapy in the management of prenatal sodium valproate (VPA)-induced autistic-like/cognitive deficits in male Wistar rats. Pregnant female rats received a single intraperitoneal injection of VPA (600 mg/kg) to induce autistic-like behaviors in their offspring. Prenatal VPA induced autistic-like symptoms (decreased social interaction and the appearance of stereotyped behavior) with deficits in spatial learning (in Morris water maze) and cognitive flexibility (in the attentional set-shifting task) in addition to decreased hippocampal protein levels of phosphorylated cAMP response element-binding protein (p-CREB), brain-derived neurotrophic factor (BDNF), and gene expression of glutamate transporter-1 (Glt-1) with a decline in GABA/glutamate ratio (both measured by HPLC). These were accompanied by the appearance of numerous neurofibrillary tangles (NFTs) with enhanced apoptosis in hippocampal sections. Memantine/aripiprazole combination increased the protein levels of p-CREB, BDNF, and Glt-1 gene expression with restoration of GABA/glutamate balance, attenuation of VPA-induced neurodegenerative changes and autistic-like symptoms, and improvement of cognitive performance. This study draws attention to the favorable cognitive effects of memantine/aripiprazole combination in autistic subjects which could be mediated via enhancing CREB/BDNF signaling with increased expression of astrocytic Glt-1 and restoration of GABA/glutamate balance, leading to inhibition of hippocampal NFTs formation and neuronal apoptosis.

Cascaded Hough Transform-Based Hair Mask Generation and Harmonic Inpainting for Automated Hair Removal from Dermoscopy Images.

Restoring information obstructed by hair is one of the main issues for the accurate analysis and segmentation of skin images. For retrieving pixels obstructed by hair, the proposed system converts dermoscopy images into the L*a*b* color space, then principal component analysis (PCA) is applied to produce grayscale images. Afterward, the contrast-limited adaptive histogram equalization (CLAHE) and the average filter are implemented to enhance the grayscale image. Subsequently, the binary image is generated using the iterative thresholding method. After that, the Hough transform (HT) is applied to each image block to generate the hair mask. Finally, the hair pixels are removed by harmonic inpainting. The performance of the proposed automated hair removal was evaluated by applying the proposed system to the International Skin Imaging Collaboration (ISIC) dermoscopy dataset as well as to clinical images. Six performance evaluation metrics were measured, namely the mean squared error (MSE), the peak signal-to-noise ratio (PSNR), the signal-to-noise ratio (SNR), the structural similarity index (SSIM), the universal quality image index (UQI), and the correlation (C). Using the clinical dataset, the system achieved MSE, PSNR, SNR, SSIM, UQI, and C values of 34.7957, 66.98, 42.39, 0.9813, 0.9801, and 0.9985, respectively. The results demonstrated that the proposed system could satisfy the medical diagnostic requirements and achieve the best performance compared to the state-of-art.

Distinct spatial distribution and roles of Kupffer cells and monocyte-derived macrophages in mouse acute liver injury.

Macrophages are key regulators of inflammation and repair, but their heterogeneity and multiple roles in the liver are not fully understood. We aimed herein to map the intrahepatic macrophage populations and their function(s) during acute liver injury. We used flow cytometry, gene expression analysis, multiplex-immunofluorescence, 3D-reconstruction, and spatial image analysis to characterize the intrahepatic immune landscape in mice post-CCl4-induced acute liver injury during three distinct phases: necroinflammation, and early and late repair. We observed hepatocellular necrosis and a reduction in liver resident lymphocytes during necroinflammation accompanied by the infiltration of circulating myeloid cells and upregulation of inflammatory cytokines. These parameters returned to baseline levels during the repair phase while pro-repair chemokines were upregulated. We identified resident CLEC4F+ Kupffer cells (KCs) and infiltrating IBA1+CLEC4F- monocyte-derived macrophages (MoMFs) as the main hepatic macrophage populations during this response to injury. While occupying most of the necrotic area, KCs and MoMFs exhibited distinctive kinetics, distribution and morphology at the site of injury. The necroinflammation phase was characterized by low levels of KCs and a remarkable invasion of MoMFs suggesting their potential role in phagoctosing necrotic hepatocytes, while opposite kinetics/distribution were observed during repair. During the early repair phase, yolksac - derived KCs were restored, whereas MoMFs diminished gradually then dissipated during late repair. MoMFs interacted with hepatic stellate cells during the necroinflammatory and early repair phases, potentially modulating their activation state and influencing their fibrogenic and pro-repair functions that are critical for wound healing. Altogether, our study reveals novel and distinct spatial and temporal distribution of KCs and MoMFs and provides insights into their complementary roles during acute liver injury.

Facial rejuvenation by microneedling with irradiated amniotic collagen matrix compared to platelet rich plasma.

Many modalities are used for treatment of facial wrinkles, such as microneedling that enhances collagen production, and platelet-rich plasma (PRP) which contains concentrated levels of growth factors. The human amniotic membrane isolated from the placentae of donors (during elective cesarean sections) has high levels of growth factors that help in rejuvenation by improving the migration and proliferation of keratinocytes, fibroblasts and increased collagen synthesis. Was to confirm the efficacy of irradiated amniotic collagen matrix (IACM) versus platelet rich plasma (PRP) delivered via microneedling in facial rejuvenation. The present study included 20 patients with facial wrinkles divided into two groups using split face technique: Group A subjected to microneedling with topical IACM on the right side of the face. Group B subjected to microneedling with topical PRP on the left side of the face. Patients received six sessions 2 weeks apart. Photos by Antera camera and skin biopsies were taken to assess the clinical results. There were a statistically significant improvement in both sides after than before treatment; with better improvement in patients treated with IACM more than patients treated with PRP using microneedling in both sides as proved clinically (assessed by WSRS and GAIS scale), pathologically (Orcein and Masson trichrome stain) and by Antera camera (texture and pigmentation). Microneedling using IACM is a new, safe and effective method for facial rejuvenation, more effective when compared to microneedling using PRP; in need for further studies to evaluate the correct dose and number of sessions to get the best outcome.

Novel Functions of Integrins as Receptors of CD154: Their Role in Inflammation and Apoptosis.

CD154, an inflammatory mediator also known as CD40 ligand, has been identified as a novel binding partner for some members of the integrin family. The αIIbß3, specifically expressed on platelets, was the first integrin to be described as a receptor for CD154 after CD40. Its interaction with soluble CD154 (sCD154) highly contributes to thrombus formation and stability. Identifying αIIbß3 opened the door for investigating other integrins as partners of CD154. The αMß2 expressed on myeloid cells was shown capable of binding CD154 and contributing as such to cell activation, adhesion, and release of proinflammatory mediators. In parallel, α5ß1 communicates with sCD154, inducing pro-inflammatory responses. Additional pathogenic effects involving apoptosis-preventing functions were exhibited by the CD154-α5ß1 dyad in T cells, conferring a role for such interaction in the survival of malignant cells, as well as the persistence of autoreactive T cells. More recently, CD154 receptors integrated two new integrin members, αvß3 and α4ß1, with little known as to their biological significance in this context. This article provides an overview of the novel role of integrins as receptors of CD154 and as critical players in pro-inflammatory and apoptotic responses.

Novel role of peroxisome proliferator activated receptor-α in valproic acid rat model of autism: Mechanistic study of risperidone and metformin monotherapy versus combination.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder of heterogenous etiology exhibiting a challenge in understanding its exact neuro-pathophysiology. Recently, peroxisome proliferator activated receptor (PPAR)-α activation was found to play a fundamental role in neuroprotection and improving autistic-like-behaviors in experimental animal models of ASD through alleviating neuroinflammation, oxidative-stress, astrocyte reactivity, tauopathy in addition to its favorable role in metabolic regulation, thus attracting attention as a possible target in treatment of ASD. This study aimed to investigate the role of PPAR-α, astrocytic dysfunction and tauopathy in ASD and detect the possible neuroprotective effects of metformin (MET), through PPAR-α activation, and risperidone (RIS) either monotherapy or in combination in alleviating autistic-like-changes at behavioral and neurobiological levels in male Wistar rats. Pregnant female Wistar rats received valproic-acid (VPA) to induce autistic-like-behavioral and neurobiological alterations in their offspring. Chronic intra-peritoneal MET (100 mg/kg/day) and RIS (1 mg/kg/day) either monotherapy or in combination started from postnatal day (PND) 24 till PND61 (38 days). Prenatal VPA exposure simulated the autistic core behaviors associated with neurochemical and histopathological neurodevelopmental degenerative changes. Both MET and RIS either monotherapy or in combination were able to reverse these changes. The effect of MET was comparable to RIS. Moreover, MET was able to alleviate the RIS induced weight gain and improve cognitive functions highlighting its promising adjunctive role in alleviating ASD pathophysiology. Our study highlighted the favorable effects of MET and RIS both in monotherapy and in combination in alleviating the autistic-like-changes and proposed PPAR-α activation along with restoring astrocytes homeostasis as promising targets in novel therapeutic strategies in ASD.

New tilomisole-based benzimidazothiazole derivatives as anti-inflammatory agents: Synthesis, in vivo, in vitro evaluation, and in silico studies.

New tilomisole-based benzimidazothiazole derivatives were designed and synthesized in this work. Their anti-inflammatory activity was assessed through the in vivo carrageenan rat paw edema model, and the in vitro COX inhibition assay. Compounds 13, 20, 30, 40, 43, and 46 demonstrated values of inhibition of induced edema in the in vivo assay comparable to celecoxib. All the synthesized compounds expressed their activity on COX-2 enzyme more than COX-1, proving their advantageous selectivity. In addition, compounds 13, 16, 20, 25, and 46 displayed lower IC50 values than celecoxib as a reference drug against COX-2 enzyme; having values of 0.09, 13.87, 32.28, 33.01, and 5.18 nM respectively vs 40.00 nM for celecoxib. Particularly, the most active compound (13) with its extreme potency (400 folds more potent than celecoxib) exhibited a notable high selectivity index (SI = 159.5). In silico studies, including ADMET prediction, compliance to Lipinski's rule of five, and molecular docking into the active site of both COX isozymes were conducted for the synthesized compounds. The results suggested that these compounds are good candidates for orally active drugs, and docking revealed higher number of interactions with COX-2 for 13 as the most active compound compared with COX-1 reflecting its advantageous selectivity and explaining its extreme potency.

Treatment with oral vitamin D alone, topical minoxidil, or combination of both in patients with female pattern hair loss: A comparative clinical and dermoscopic study.

BACKGROUND: One of the most common dermatological complaints among female is female pattern hair loss (FPHL). Serum vitamin D is a factor lately taken into consideration in approaching patients complaining of hair loss. AIM: To evaluate the serum level of 25-hydroxy vitamin D in patients with FPHL and to evaluate the efficacy of vitamin D therapy alone or combined with minoxidil in the treatment of this disease. METHODS: 45 patients with FPHL and 15 controls to measure serum level of vitamin D were enrolled in the study. Patients then were subdivided into 3 groups: group I received topical minoxidil and oral vitamin D, group II received topical minoxidil, and group III received oral vitamin D for 6 months. Clinical and dermoscopic evaluation was done for the three groups before and after treatment. RESULTS: Vitamin D level was significantly decreased in patients compared to controls. After treatment, as regard Ludwig scale, there was statistically significant improvement in group I than II while no significant improvement was found in group III. Dermoscopy revealed that thin hair and single-hair unit were significantly improved in groups I and II, while it was not significantly improved in group III. CONCLUSION: Oral vitamin D combination to topical minoxidil is recommended to treat patients with FPHL; they had better results than vitamin D or topical minoxidil alone.

Platelet rich plasma injection versus topical erythromycin 2% in treatment of acne vulgaris.

BACKGROUND: Acne vulgaris is a common inflammatory skin disease that affects the pilosebaceous glands. There are different modalities of treatment of acne but there is no standard treatment free of side effects. Platelet rich plasma (PRP) is an autologous concentration of platelets in a small volume of plasma. When platelets are activated, multiple growth factors are released. They play an important role in angiogenesis, inflammatory process and wound healing. AIM: was to evaluate and compare the therapeutic efficacy of platelet rich plasma versus topical erythromycin 2% in treatment of acne vulgaris. METHODS: 40 patients with inflammatory acne lesions were included. All patients received PRP injection sessions in one side of the face (group A) every 2 weeks for 6 sessions and topical erythromycin 2% in the other side (group B). RESULTS: There was significant difference between both groups in which better improvement was reported in group A (55% of patients showed good to excellent improvement and 35% showed moderate improvement, especially the inflammatory lesions). Group A showed better patients' satisfaction and lower rate of recurrence than group B. CONCLUSION: PRP is effective and safe treatment option for inflammatory acne and alternative to other systemic modalities especially if they are contraindicated.

Synthesis and Biological Evaluation of Thiazole-Based Derivatives as Potential Acetylcholinesterase Inhibitors.

Nineteen new thiazole-based derivatives were synthesized and their structures characterized with analytical and spectral data. The in vitro assessment of their acetylcholinesterase (AChE) inhibitory activity revealed that compounds 10 and 16 produced potent AChE inhibitory activities with IC50 values of 103.24 and 108.94 nM, respectively. Compounds 13, 17, 18, 21, 23, 31, and 33 displayed moderate activity with 25-50% relative potency compared to the known potent AChE inhibitor donepezil. Molecular docking studies of the active compounds docked within the active site cavity of AChE showed a binding orientation similar to that of donepezil, with good predicted binding affinities. These compounds could therefore be considered as potential lead compounds for the development of new and potentially improved AChE inhibitors.

Evaluation of the efficacy and safety of platelet rich plasma injection in treatment of rosacea.

Rosacea is a chronic relapsing inflammatory skin disease, with a high prevalence among adults. Treatment of rosacea is difficult, with high rate of recurrence. Due to the strong anti- inflammatory and antibacterial effects of platelet rich plasma (PRP), it was used in the medicine for treating many inflammatory diseases. To evaluate the role of PRP injection in treatment of rosacea. The study was carried on 40 patients with rosacea. They were treated by PRP injection in right side of the face (group A) and platelet poor plasma injection in left side (group B). They underwent one session every 2 weeks for 3 months (6 sessions). The patients were assessed clinically before and after treatment by the rosacea grading scale. Skin biopsies were taken to evaluate the clinical results. There was a statistically significant decrease in rosacea grading scale after treatment with PRP injection, 50% of the patients showed excellent improvement and 50% showed good improvement. The improvement was significantly better in group A than B. There was marked decrease in inflammatory cells by hematoxylin and eosin stain, and decrease in expression of nuclear factor kappa ßeta after treatment with PRP. PRP was effective and safe technique in treatment of rosacea and alternative to other systemic modalities, especially if they are contraindicated.

Monoclonal Antibody Targeting the CD154 Cleavage Site Inhibits CD40-Dependent and -Independent Cleavage of CD154 from the Cell Surface.

In addition to the membrane-bound molecule, soluble CD154 (sCD154) is also detected at high levels in the medium of activated T cells and platelets and in the serum of patients suffering from different inflammatory diseases. This sCD154 is the result of cleavage of the full-length molecule between the glutamic acid residue at position 112 (E112) and methionine at position 113 (M113) and can be derived from the intracellular milieu and from cleavage of cell surface molecules. We have recently reported that substitution of both E112 and M113 by alanine inhibits intracellular and CD40-induced membrane cleavage of CD154 and procures to CD154 an increased biological function as compared with cleavable CD154. Thus, in this study, and in the aim of developing tools inhibiting cleavage of CD154 from the cell surface, we generated a panel of anti-human CD154 mAbs. One of the derived mAbs that did not alter the binding of sCD154 to CD40, named in this study Clone 8 mAb, totally lost its binding activity against cells expressing CD154 mutated at its E112 and M113 residues. Treatment with Clone 8 mAb was shown to completely abolish CD40-dependent and -independent cleavage of CD154 from the cell surface. Our study is highlighting the development and characterization of an innovative therapeutic tool capable of inhibiting the release/cleavage of CD154 from cells and thus maintaining its availability on the cell surface and the high probably of increasing its potency as an activator of CD40-induced responses.

Design, synthesis, and antitumor activity of PLGA nanoparticles incorporating a discovered benzimidazole derivative as EZH2 inhibitor.

PURPOSE: Targeting enhancer of zeste homolog 2 (EZH2) can represent a hopeful strategy for oncotherapy. Also, the use of PLGA-based nanoparticles as a novel and rate-controlling carrier system was of our concern. METHODS: Benzimidazole derivatives were synthesized, and their structures were clarified. In vitro antitumor activity was evaluated. Then, a modeling study was performed to investigate the ability of the most active compounds to recognize EZH2 active sites. Compound 30 (Drug) was selected to conduct pre-formulation studies and then it was incorporated into polymeric PLGA nanoparticles (NPs). NPs were then fully characterized to select an optimized formula (NP4) that subjected to further evaluation regarding antitumor activity and protein expression levels of EZH2 and EpCAM. RESULTS: The results showed the antitumor activity of some synthesized derivatives. Docking outcomes demonstrated that Compound 30 was able to identify EZH2 active sites. NP4 exhibited promising findings and proved to keep the antitumor activity of Compound 30. HEPG-2 was the most sensitive for both Drug and NP4. Protein analysis indicated that Drug and NP4 had targeted EZH2 and the downstream signaling pathway leading to the decline of EpCAM expression. CONCLUSIONS: Targeting EZH2 by Compound 30 has potential use in the treatment of cancer especially hepatocellular carcinoma.

Multisubstituted pyrimidines effectively inhibit bacterial growth and biofilm formation of Staphylococcus aureus.

Biofilms are multicellular communities of microorganisms that generally attach to surfaces in a self-produced matrix. Unlike planktonic cells, biofilms can withstand conventional antibiotics, causing significant challenges in the healthcare system. Currently, new chemical entities are urgently needed to develop novel anti-biofilm agents. In this study, we designed and synthesized a set of 2,4,5,6-tetrasubstituted pyrimidines and assessed their antibacterial activity against planktonic cells and biofilms formed by Staphylococcus aureus. Compounds 9e, 10d, and 10e displayed potent activity for inhibiting the onset of biofilm formation as well as for killing pre-formed biofilms of S. aureus ATCC 25923 and Newman strains, with half-maximal inhibitory concentration (IC50) values ranging from 11.6 to 62.0 µM. These pyrimidines, at 100 µM, not only decreased the number of viable bacteria within the pre-formed biofilm by 2-3 log10 but also reduced the amount of total biomass by 30-50%. Furthermore, these compounds were effective against planktonic cells with minimum inhibitory concentration (MIC) values lower than 60 µM for both staphylococcal strains. Compound 10d inhibited the growth of S. aureus ATCC 25923 in a concentration-dependent manner and displayed a bactericidal anti-staphylococcal activity. Taken together, our study highlights the value of multisubstituted pyrimidines to develop novel anti-biofilm agents.