Natural Medicine Research Articles

Biosynthesis of unique natural product scaffolds by Fe(II)/αKG-dependent oxygenases

Abstract Fe(II)/αKG-dependent oxygenases are multifunctional oxidases responsible for the formation of unique natural product skeletons. Studies of these enzymes are important because the knowledge of their catalytic functions, enzyme structures, and reaction mechanisms can be used to create non-natural enzymes through mutation and synthesize non-natural compounds. In this review, I will introduce the research we have conducted on two fungal Fe(II)/αKG-dependent oxygenases, TlxI-J and TqaL. TlxI-J is the first Fe(II)/αKG-dependent oxygenase type enzyme heterodimer that catalyzes consecutive oxidation reactions, hydroxylation followed by retro-aldol or ketal formation, to form the complex skeletons of meroterpenoids. TqaL is the first naturally occurring aziridine synthase, and I will discuss the mechanism of its unique C–N bond formation in nonproteinogenic amino acid biosynthesis. This review will advance research on the discovery of new enzymes and the analysis of their functions by reviewing the structures and functions of these extraordinary Fe(II)/αKG-dependent oxygenases, and promote their use in the synthesis of new natural medicines. Graphical Abstract

High-Resolution LC-MS Simultaneous Quantification of Forty-Six Compounds from Jatropha podagrica Fruit Recommends Four Top Antioxidant Contributors as Q-Markers

There has been no chemical analysis of the fruit of medicinal plant Jatropha podagrica until now. The current study aimed to qualitatively and quantitatively analyze the J. podagrica fruit using a high-resolution LC-MS strategy, i.e., library-comparison ultra-high-performance liquid chromatography-Quadrupole-Orbitrap-tandem mass spectrometry. The strategy putatively identified 46 compounds from fresh fruit. During the putative identification, 10 isomers (e.g., (vitexin vs. isovitexin) were completely distinguished from each other. Thereafter, all 46 compounds were simultaneously quantified using authentic standard comparison method. Finally, they were also subjected to the 2,2′-azino bis (3-ethylbenzothiazolin-6-sulfonic acid radical (ABTS+•)-scavenging assay to characterize their relative antioxidant capacities. Their antioxidant capacities were thus multiplied by chemical contents to calculate their antioxidant contribution values, respectively. Corilagin, gallic acid, ellagic acid, and phillygenin exhibited the highest antioxidant contribution percentages and thereby were suggested as the four top antioxidant contributors. The four are recommended to build up a quality-markers (Q-markers) system of J. podagrica fruits. All these findings can help to develop J. podagrica fruits as a potential resource of natural medicine.

Exploring a new paradigm for serum-accessible component rules of natural medicines using machine learning and development and validation of a direct predictive model.

In the field of pharmaceutical research, Lipinski's Rule of Five (RO5) was once widely regarded as the prevailing standard for the development of novel drugs. Despite the fact that an increasing number of recently approved drugs no longer adhere to this rule, it continues to serve as a valuable guiding principle in the field of drug discovery. The present study aims to establish a set of rules specifically for the serum-accessible components of natural medicines. A comprehensive literature review was conducted to collect data on serum-accessible components of natural medicines, and machine learning methods were then applied to analyse and screen molecular features distinguishing serum-accessible components from non-serum-accessible ones. The most critical rules for serum-accessible components of natural medicines were identified, and these were named the "Natural Medicine's Rule of 5 (NMRO5)." We then compared the molecular property distributions and predictive performance of NMRO5 with RO5. Then, we developed a predictive model capable of directly assessing the possibility of a molecule being serum-accessible. This model was validated using in vivo experiments on multiple natural medicines. Furthermore, we performed molecular modifications on serum-accessible components to "violate" NMRO5, conducting both forward and reverse validations to confirm the reliability of NMRO5. The results obtained revealed that NMRO5 is characterised by the following: higher TPSA, MaxEState, and PEOE VSA1 values, and lower LogP and MinEState values. This indicates that natural medicine components with these properties are more likely to be serum-accessible or remain in plasma rather than being rapidly eliminated. The investigation revealed significant disparities among the five molecular properties of NMRO5, and the predictive performance of eight models based on NMRO5 consistently outperformed those based on RO5. This finding suggests that NMRO5 provides a more reliable framework for determining whether a molecule is serum-accessible compared to RO5. Finally, we developed a direct predictive model for serum-accessible components, achieving an accuracy of 0.7257, an F1 score of 0.7223, and an AUC of 0.7553.

Phenolic Content, Antioxidant Activity and In Vitro Anti-Inflammatory and Antitumor Potential of Selected Bulgarian Propolis Samples

Background/objectives: Propolis (bee glue) is a valuable bee product widely used as a natural remedy, a cosmetic ingredient, a nutritional value enhancer and a food biopreservative. The present research aims to investigate the phenolic content, antioxidant activity and in vitro anti-inflammatory and antitumor potential of six propolis samples from three regions of Bulgaria (Vidin, Gabrovo and Lovech). Methods: the analysis of propolis phenolic compounds was determined by high-performance liquid chromatography (HPLC); the antioxidant activity of ethanolic propolis extracts was assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and ferric-reducing antioxidant power (FRAP) assay; the in vitro anti-inflammatory activity was assessed by the inhibition of albumin denaturation method; the in vitro antitumor activity was determined in human metastatic breast cancer cell line MDA-MB-231 using 3-(4,5-Dimethyl -2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Results: The ethanolic propolis extracts exhibited the total phenolic content from 190.4 to 317.0 mg GAE/g, total flavonoid content from 53.4 to 79.3 mg QE/g and total caffeic acid derivatives content from 5.9 to 12.1 mg CAE/g. The studied propolis extracts showed significant antioxidant capacity (between 1000.3 and 1606.0 mM TE/g determined by the DPPH assay, and between 634.1 and 1134.5 mM TE/g determined by the FRAP assay). The chemical composition analysis indicated high concentrations of caffeic acid benzyl ester, chrysin, pinocembrin and pinobanksin-3-O-propionate, predominantly in three of the propolis samples originating from Gabrovo and Lovech regions. All propolis samples demonstrated promising in vitro anti-inflammatory activity, expressed as the inhibition of thermally induced albumin denaturation by 73.59% to 78.44%, which was higher than that of the conventional anti-inflammatory drugs Aspirin (58.44%) and Prednisolone Cortico (57.34%). The propolis samples exhibited high in vitro cytotoxicity against cancer cells MDA-MB-231 with IC50 values ranging between 9.24 and 13.62 µg/mL as determined by MTT assay. Conclusions: Overall, we can suggest that the high phenolic content of Bulgarian propolis from respective areas contributes to its enhanced antioxidant, anti-inflammatory and antitumor activity. Taken together, our results support the beneficial properties of Bulgarian propolis, with potential application as a promising therapeutic agent.

Investigation on bowel regulation and constipation relief based on the microbial fermentation solution of cassia seed.

As digestive health issues rise and interest in natural therapies grows, traditional herbs like Cassia Seed are gaining attention for their antioxidant, laxative, and digestive benefits. This study aimed to optimize the fermentation conditions of Cassia seed using microbial technology to enhance the content of anthraquinone compounds, thereby augmenting its pharmacological effects, particularly in promoting intestinal peristalsis and alleviating constipation. Fermentation of Cassia Seed was conducted under controlled microbial conditions. High-performance liquid chromatography (HPLC) was employed to analyze the anthraquinone profile of the fermented solution. Mouse model was utilized to evaluate biochemical markers and intestinal functions, while in the zebrafish constipation model, calcein was utilized as a marker to monitor intestinal function and excretion dynamics. Cassia Seed was fermented under controlled microbial conditions. HPLC was employed to analyze the anthraquinone profile of the fermented solution. Mouse models were utilized to evaluate biochemical markers and intestinal functions, while a zebrafish constipation model used calcein as a marker to monitor intestinal function and excretion dynamics. The optimized fermentation significantly increased the content of anthraquinone compounds, including substances like emodin, aloe-emodin, rhein, chrysophanol, and physcion, as confirmed by HPLC analysis. In a mouse constipation model, the fermented solution reduced nitric oxide (NO) levels while increasing acetylcholine (Ach) and gastrin (Gas) levels, thereby promoting intestinal peristalsis and accelerating bowel movements. Additionally, the fermented solution facilitated repair of the colonic mucosa in mice without significant pathological damage such as degeneration or fibrous tissue proliferation. In the zebrafish model, the fermented Cassia Seed solution reduced calcein fluorescence intensity, indicating enhanced intestinal peristalsis and accelerated bowel movements, thereby decreasing calcein retention time in the body. This study investigated the potential clinical applications of fermented Cassia seed ferment in digestive health, providing a theoretical basis and inspiration for further research and development. Additionally, it offers new insights and possibilities for the development of novel natural medicines or functional foods aimed at improving constipation and promoting intestinal health.

Investigation and assessment of equilibrium solubility, Hansen solubility parameters and modeling of natural plant medicine artemisinin (Form Ⅰ) in three binary solvents at different temperatures

Investigation and assessment of equilibrium solubility, Hansen solubility parameters and modeling of natural plant medicine artemisinin (Form Ⅰ) in three binary solvents at different temperatures

Xanthone Derivatives: A Pharmacological Panorama of Versatility

Xanthones are a class of naturally occurring compounds found in various plant sources. Over the years, studies have revealed therapeutic impending xanthones in treating and managing several ailments, including cancer, inflammation, and microbial infections. Xanthones exert their therapeutic effects through various mechanisms, such as anti-inflammatory, antioxidant, and anti-proliferative activities. It has also been found that xanthones induce apoptosis, inhibit cell proliferation, and arrest cell cycles by modulating several signaling pathways. This review objective is to deliver an inclusive overview of the therapeutic essence of xanthones and their potential use as a natural remedy for managing and treating various diseases.

Screening and Verification of Blood-Activating Effective Component Group of Panax notoginseng Based on Spectrum-Effect Relationships.

Panax notoginseng (P. notoginseng) is one of the most famous natural medicines and widely used to promote blood circulation in health care. However, the active component group of P. notoginseng for activating blood is not clear. We aim to screen and validate the pharmacodynamic component group (PCG), which could exert the same blood-activating effect as P. notoginseng. To clarify the active components, the chemical components were determined by liquid chromatography-tandem mass spectrometry, and the fingerprint of P. notoginseng was established. Twenty candidate active monomers were selected through the spectrum-effect relationship analysis. Eleven active monomers, including Ginsenoside Rg1, Rb1, Rd, F1, Rh1, Rg2, Rb2, Rg3, and Rk1 and Notoginsenoside R1 and R2, were screened out as the PCG through validation by platelet aggregation test. Among them, the antiplatelet aggregation activity of Ginsenoside Rh1 was directly confirmed for the first time. The active component group could exert similar efficacy to the P. notoginseng extract invitro and invivo through the validation of invitro platelet aggregation test and the rats with cerebral ischemia. This study laid the foundation for the quality evaluation of P. notoginseng and provided a reference for the research on the material basis of the pharmacodynamics of other Chinese herbs.

A randomized phase 2 study of an individualized neoantigen-targeting immunotherapy in patients with newly diagnosed metastatic microsatellite stable colorectal cancer (MSS-CRC).

LBA13 Background: Neoantigen-targeting immunotherapy aims to provide therapeutic benefit to patients by generating potent neoantigen-specific T cells. GRANITE is an immunotherapy regimen that uses chimpanzee adenovirus and self-amplifying mRNA vaccines encoding patient-specific neoantigens in combination with immune checkpoint inhibitors. Preliminary efficacy was noted in MSS-CRC (Palmer et al Nature Medicine 2022) and this regimen is being assessed in a randomized Phase 2 study in 1L metastatic MSS-CRC (NCT05141721). Methods: Patients (pts) with 20 neoantigens based on the EDGEÔ prediction model were randomized 1:1 to receive the GRANITE regimen in addition to 1L standard of care (SOC) (GRANITE vaccine arm), or 1L SOC alone (control arm). Results: Baseline demographics and disease characteristics were balanced between arms. As of 15 Oct 2024, GRANITE pts had an improvement in PFS relative to control pts (HR=0.73, 90% CI [0.44, 1.21]). In this study, high and low disease burden groups were identified using baseline ctDNA assessed by Gritstone’s validated, highly sensitive, tumor-informed assay. Patients with low disease burden benefited more than those with high disease burden, consistent with emerging data suggesting neoantigen-targeting immunotherapy having greater activity in minimal disease settings (e.g., adjuvant setting). Furthermore, in the low disease burden group at the most recent study visit, more patients were free of progression with very low ctDNA levels (ie, below the limit of quantification of the assay) in the GRANITE arm, 41% (7/17), versus the control arm, 21% (3/14). The greater proportion of GRANITE patients both free of progression and with very low ctDNA, suggests potential further separation of PFS cuves with additional follow up. Neoantigen-specific T cell responses were observed in 100% of patients in the GRANITE arm with evaluable PBMC samples (17/17 assessed by ex vivo and in vitro stimulation ELISpots). Molecular responses based on a reduction in ctDNA at a single timepoint was similar between arms (39% in GRANITE and 40% in control arms). Most common (≥20%) treatment-related adverse events (TRAE) included pyrexia and influenza like illness and no pts discontinued due to a TRAE. Conclusion: This randomized study shows GRANITE may benefit patients, especially those with low burden disease, in this front line setting of metastatic MSS-CRC that has not benefitted from immunotherapy. Updated analysis will be presented. Clinical trial information: NCT05141721 .

Safety and efficacy of oral administrated cepharanthine in non-hospitalized, asymptomatic or mild COVID-19 patients: a Double-blind, randomized, placebo-controlled trial

Cepharanthine (CEP) is a natural remedy that potently inhibits SARS-CoV-2 activity both in vitro and in vivo. To evaluate the efficacy and safety of CEP compared with placebo in adults with asymptomatic or mild coronavirus disease 2019 (COVID-19), we conducted a proof-of-concept, double-blind, randomized, placebo-controlled trial. Patients were randomized to receive 120 mg/day of CEP, 60 mg/day CEP or placebo for 5 days. Main outcome was the time from randomization to negative nasopharyngeal swab and safety. Among 262 randomized participants, 188 completed the trial among group of 120 mg/day CEP (n = 65), 60 mg/day CEP (n = 68) and placebo (n = 55). Neither 120 mg/day or 60 mg/day CEP shortened the time to negative significantly compared with placebo. However, 60 mg/day CEP showed a slight trend (difference=-0.77 days, hazard ratio (HR) = 1.40, 95% CI 0.97–2.01, p = 0.072). In analysis of participants with good medication compliance, 60 mg/day CEP significantly shortened the time to negative (difference=-0.87 days, HR = 1.56, 95% CI 1.03–2.37, p = 0.035). Adverse events were not different among the three groups, and no serious adverse events occurred. In conclusion, treatment of asymptomatic or mild Covid-19 with 120 mg/day or 60 mg/day did not shorten the time to negative significantly. However, 60 mg/day CEP showed a slight trend which needs future confirmatory trials to validate. (NCT05398705).

Uncovering The Pharmacological Mechanism of Ficus elastica as Anti-hyperlipidemia Candidate: LC-HRMS, Network Pharmacology, In vitro and In vivo Studies

Hyperlipidemia is a major risk factor for cardiovascular diseases. While conventional treatments exist, there is a growing interest in natural remedies with fewer side effects. Ficus elastica has promising medicinal properties, yet its potential as an anti-hyperlipidemic agent remains unexplored. This study aimed to investigate the anti-hyperlipidemic effects of F. elastica using an integrated approach of LC-HRMS-based chemical bioinformatics and in vitro/in vivo experimental validation. The anti-hyperlipidemic potential of F. elastica and its mechanism of action were screened using integrative computational network pharmacology followed by in vitro HMG-CoA reductase inhibition and in vivo lipid-lowering activity in a hyperlipidemia rat model. Network pharmacology analysis identified STAT3, HSP90AA1, and TLR4 as potential core targets involved in lipid and atherosclerosis-related KEGG pathways. Molecular docking simulations revealed high-affinity interactions between F. elastica compounds and the identified targets, notably compound 41 and compound 61. In vitro assay demonstrated that ethanolic extract of F. elastica inhibited HMG-CoA reductase with an IC50 of 297.73 µg/mL. In vivo experiment using a hyperlipidemic rat model showed significant reductions in total cholesterol, triglycerides, and increased HDL levels. The reduction of triglycerides and elevation of HDL level after F. elastica ethanolic extract supplementation is similar to the effect from supplementation of simvastatin. These findings suggest that F. elastica ethanolic extract possesses notable anti-hyperlipidemic properties, likely mediated through multiple molecular targets and pathways. The study highlights the potential of F. elastica ethanolic extract as a promising candidate for anti-hyperlipidemic therapy and underscores the efficacy of integrating computational and experimental approaches in natural product research.

Smart Drug Recommendation with AI Chatbot for Personalized Healthcare

This AI-driven web application combines machine learning with an advanced chatbot to deliver personalized healthcare solutions The chatbot communicates with users in a conversational manner to collect information about their symptoms, which is subsequently processed by a machine learning system to suggest possible health conditions. Following this assessment, the application suggests suitable medications for the identified issues and offers natural home remedies for temporary symptom relief. A unique health reminder system checks in with users periodically, tracking symptom progression and offering follow- up guidance. Based on user feedback, the system may recommend consulting a healthcare professional, supporting a proactive, user-centered approach to self-care and health management.

Pomegranate as a natural remedy for gastric ulcers prevention: a review of its gastroprotective mechanisms and pharmacological benefits.

Gastric ulcers (GUs) represent a common gastrointestinal disorder characterized by mucosal damage and inflammation, often precipitated by factors such as Helicobacter pylori infection and the consumption of COX inhibitors. This comprehensive review investigates the role of oxidative stress and inflammation in the pathogenesis of GUs and assesses the potential therapeutic effects of Punica granatum (pomegranate, Pg) supplementation. Utilizing a series of experimental models, including indomethacin, aspirin, and alcohol-induced ulcers, we demonstrate that Pg extracts possess significant gastroprotective properties. The antioxidant activity of Pg is ascribed to its capacity to neutralize reactive oxygen species (ROS), enhance the activity of endogenous antioxidants such as superoxide dismutase (SOD) and catalase (CAT), and diminish lipid peroxidation. Furthermore, the anti-inflammatory effects of Pg are mediated through the suppression of pro-inflammatory cytokines such as TNF-α and IL-1β, in conjunction with the promotion of gastric mucosal protective agents. Histological analyses indicate that Pg extract preserves the architecture of gastric tissue and alleviates ulcer severity. These findings highlight the potential of Pg as a natural remedy for GUs, thereby necessitating further investigation into its mechanisms of action and optimal therapeutic formulations.

Chromatographic Analysis of Polyherbal Extract and Formulation by HPTLC and GC-MS Methods

Background: The World Health Organization (WHO) acknowledges that natural plants and their bioactive components have been recognized for their potential therapeutic benefits. The main issue facing the herbal industry is the lack of effective quality control for herbal extracts and formulations Establishment standards are essential to ensuring the efficacy and integrity of natural medicine development. Determining the precise compound responsible for therapeutic action poses an equally challenging task, as these compounds frequently work synergistically to produce medicinal benefits. Aim: In this exploration, GC-MS and high-performance thin-layer chromatography were used to analyze the formulation and polyherb extract. The HPTLC technique is often employed to analyze the sample and compare its fingerprint to reference standards to verify plant extracts’ quality and formulations’ quality. Using GC-MS, the phytochemical ingredients in the formulation and the extract have been determined. Methods: This research assessed a mixture of herb extract and formulation using analytical approaches. The combined hydroalcoholic extracts of the herbs were used to produce the polyherb extract of seeds of Azadirachta indica, Carica papaya, Trigonella foenum-graecum, Gossipium herbaceum, fruit of Piper nigrum. The required excipients were added before compressing the herbal mixture extract into tablets. Results: The extract and formulation’s HPTLC chromatograms displayed Rf values of 0.14, 0.34, 0.48, 0.63, and 0.76.The results of the GC-MS, N-Hexadecanoic, 1,19-Eicosadiene, 9,12-Octadecadienoic Acid, Z, Z-4,16-Octadecadien-1-Ol Acetate, Piperine that were analyzed provide different peaks to determining the presence of phytochemicals found in the extract and formulation were confirmed. Conclusion: HPTLC and GC-MS aid in the identification of individual chemical constituents within herbal formulations by coupling chromatographic and mass spectral data of the sample with reference standards or databases, the identity of key compounds can be confirmed, ensuring authenticity, and has shown an intense correlation between the mixture of herb extract and formulation. Major Findings: In this Research, GC-MS and high-performance thin-layer chromatography were used to analyze the formulation and polyherbal extract of key compounds can be confirmed, ensuring authenticity, and has shown an intense correlation between the mixture of herb extract and formulation.

Fluorescence paper sensor meets magnetic affinity chromatography: discovering potent neuraminidase inhibitors in herbal medicines.

Given the inherent complexity of natural medicines, finding a straightforward and efficient method for identifying active ingredients remains a significant challenge, yet it is of paramount importance. Influenza virus neuraminidase (NA), a primary target for anti-influenza drug development, plays a crucial role in the infection process, making it essential to develop rapid and facile methods for screening NA inhibitors. Herein, we developed a novel and efficient analytical technique for the identification of NA inhibitors from complex herbal medicines by integrating dual sensing with affinity chromatography. This approach simplifies the experimental process and highlights the benefits of being quicker, more sensitive, and cost-effective. Regarding the biosensing section, the innovative concept of a 4-methylumbelliferyl-N-acetylneuraminic acid-NA-based fluorescence paper sensor strategy enables the rapid detection of NA inhibitors in complex herbal samples. In affinity chromatography, bioactive compounds were precisely captured, separated, and identified. The efficacy and reliability of the developed method were confirmed using both negative and positive controls. Then, the method was applied to screen for NA inhibitors in 20 different herbal medicines. The results revealed that Bupleurum chinense DC. exhibited the most pronounced inhibitory effect on NA. Subsequent analysis utilizing affinity chromatography identified three bioactive compounds, namely saikosaponin a, saikosaponin d, and baicalin, as the active agents responsible for this inhibitory effect, with IC50 values of 177.3μM, 262.9μM, and 241.4μM, respectively. Molecular docking studies further indicated that these three bioactive compounds exhibit a strong binding affinity with NA. This research provides novel insights into the screening of enzyme inhibitors within herbal medicines.

Affinity Ultrafiltration Mass Spectrometry for Screening Active Ingredients in Traditional Chinese Medicine: A Review of the Past Decade (2014–2024)

Discovering targets in natural products is a critical and challenging task in new drug development. Rapid and efficient screening of active ingredients from complex systems like traditional Chinese medicine (TCM) is now crucial in drug research. Affinity ultrafiltration (AUF) technology is widely used to screen active ingredients in natural medicines. AUF-liquid chromatography–mass spectrometry (AUF-LC-MS) leverages the affinity between natural medicine extracts and targets to isolate active ingredients from complex matrices, employing LC-MS for detection and activity assessment. This review discusses the developments in employing AUF-LC-MS to analyze TCM and TCM compound preparations over the last decade. This review succinctly presents the advantages and limitations of AUF-LC-MS, illustrating its benefits through the example of screening for active ingredients in natural pharmaceuticals.

Urtica dioica Aqueous Leaf Extract: Chemical Composition and In Vitro Evaluation of Biological Activities

Urtica dioica L. has been used as a natural remedy due to its healing properties for over 2000 years. The aim of this study is to investigate the chemical composition, antimicrobial, antioxidant, and antitumor properties in vitro of the aqueous extract of Urtica dioica leaves (AEUD). The chemical composition was assessed by an ultra-high-performance liquid chromatography system coupled to a benchtop QExactive high-resolution accurate mass spectrometry operating in a data-dependent acquisition mode as a non-target approach. Minimal inhibitory concentration (MIC) and disc diffusion were used to assess the antibacterial efficacy against nine bacterial strains. The antioxidant impact was assessed using DPPH, ABTS, FRAP, and ferrous ion-chelating ability assays. By using the MTT method, the cytotoxicity effect of AEUD on colon cancer cell HCT-116 was evaluated. Flow cytometry was used to analyze the cell cycle. Finally, the anti-migration and anti-invasion properties of AEUD on HCT-116 cells were estimated using the wound healing test and Transwell assays. AEUD is a rich source of phenolic compounds. The results of disc diffusion and MIC showed that the AEUD is more active against Gram-positive bacteria than against Gram-negative bacteria. MTT assay confirmed that the AEUD inhibited HCT-116 colon cancer cell proliferation. Findings of flow cytometry confirmed that cell cycle arrest occurred at the G2 phase. Additionally, AEUD had anti-migration and anti-invasion effects. This study shows that Urtica dioica aqueous leaf extract exhibits potential antibacterial, antioxidant, and antitumoral activities on HCT-116 colon cancer cells.

High Rosmarinic Acid Content Melissa officinalis L. Phytocomplex Modulates Microglia Neuroinflammation Induced by High Glucose

Diabetic patients experience hyperglycemia, which can affect multiple organs, including brain function, leading to disabling neurological complications. Hyperglycemia plays a key role in promoting neuroinflammation, the most common complication in diabetic individuals, through the activation of microglia. Attenuating hyperglycemia-related neuroinflammation in microglia may reduce diabetes-associated neurological comorbidities. Natural remedies containing phenolic compounds have shown efficacy in mitigating microglia-mediated neuroinflammation. The aim of this study was to investigate the potential of a Melissa officinalis L. (MO) phytocomplex, obtained from plant cell cultures and enriched in its main polyphenolic constituent, rosmarinic acid (RA), in attenuating hyperglycemia-induced neuroinflammation in microglia. A time-course morphological analysis of BV2 microglial cells exposed to high glucose (HG) levels showed a shift towards a proinflammatory phenotype, peaking after 48 h, which was reversed by pretreatment with MO. Biochemical assays revealed increased expression of the microglial marker CD11b (187%), activation of the NF-κB pathway (179%), expression of iNOS (225%), enhanced phosphorylation of ERK1/2 (180%), and increased expression of the proinflammatory cytokine IL-6 (173%). Pretreatment with MO prevented the aberrant expression of these proinflammatory mediators and restored SIRT1 levels. Exposure of neuronal SH-SY5Y cells to the conditioned medium from HG-exposed microglia significantly reduced cell viability. MO counteracted this effect, exhibiting neuroprotective activity. RA showed efficacy comparable to that of MO. In conclusion, MO and RA attenuated microglia-mediated oxidative imbalance and neuroinflammation under HG exposure by inhibiting the morphological shift toward a proinflammatory phenotype induced by HG and abrogating the subsequent activation of the downstream ERK1/2–NF-κB–iNOS pathway.

α-Phellandrene: A Promising Natural Remedy for Rotenone-Induced Parkinson's Disease.

Parkinson's Disease (PD) is a neurodegenerative disorder characterized by the pro-gressive loss of dopaminergic neurons in the substantia nigra, leading to motor dysfunction and non-motor symptoms. Current treatments primarily offer symptomatic relief without halt-ing disease progression. This has driven the exploration of natural compounds with neuropro-tective properties. In previous studies, α-phellandrene, a monoterpene present in essential oils of various aromatic plants, has shown promise in mitigating neurodegenerative processes. This study focuses on alpha-phellandrene's therapeutic potential in a rotenone-induced Parkinson's Disease model. Rotenone, a mitochondrial complex I inhibitor, is commonly used to induce PD-like symptoms in experimental models due to its ability to mimic the neurodegenerative processes observed in human PD. Our review explores the neuroprotective effects of alpha-phellandrene, focusing on its antioxidant, anti-inflammatory, and anti-apoptotic properties. Experimental groups of rodents received rotenone to induce PD-like symptoms, followed by alpha-phellandrene treatment. Biochemical analyses were performed to measure oxidative stress markers, inflammatory cytokines, and apoptotic signals in brain tissues. Results indi-cated that alpha-phellandrene administration significantly improved motor function and re-duced rotenone-induced oxidative stress, inflammation, and apoptosis in dopaminergic neu-rons. Histopathological examinations revealed a notable preservation of neuronal integrity in alpha-phellandrene-treated groups compared to controls. In conclusion, alpha-phellandrene demonstrates considerable neuroprotective effects in a rotenone-induced Parkinson's dmodel. These findings suggest that alpha-phellandrene could be a promising natural therapeutic agent for PD, warranting further investigation into its mechanisms of action and potential clinical applications. Specifically, our review indicates that alpha-phellandrene may exert neuropro-tective effects by various mechanisms, such as reducing oxidative stress, modulating neuro-transmitter levels, or inhibiting neuroinflammation. These mechanisms highlight its potential to alleviate PD symptoms and slow disease progression, underscoring the need for in-depth studies to validate these therapeutic effects in clinical settings.

380. Implementation of Pooled Saliva Tests for Universal Screening of Congenital Cytomegalovirus (cCMV) Infection: Lessons Derived Over a 25-Months Period

Abstract Background cCMV infection is the most common intrauterine infection, leading to infant neurodevelopmental disabilities and hearing loss, which can manifest at birth or develop later during childhood. Currently, targeted screening of cCMV by PCR testing of saliva samples is employed in many centers. However, targeted screening of high-risk neonates misses the majority of infants with cCMV who are asymptomatic at birth, yet are at risk for late-onset sequelae. Thus, universal newborn screening of cCMV has been increasingly advocated. In the absence of a high-throughput screening test that can identify all infected neonates, the development of an accurate and efficient testing strategy for universal cCMV screening has remained an ongoing challenge. We have recently reported on the development of a new saliva pooling setup for universal screening of cCMV (Nature Medicine 2024). Here we report on lessons derived over a 25-months period of its routine implementation. Methods This prospective study took place in 2 hospitals of Jerusalem from April 2022 through April 2024. Saliva samples of all newborns with parental consent were tested for cCMV in 8-sample pools. Positive saliva results were confirmed by urine testing. Results During the study period, 32,413 infants (94.4% of all live newborns) were screened for cCMV using the pooled approach. The pooling strategy proved highly efficient, with each PCR test covering an average of 6 newborns during the first 13 months of the study, thus reducing the number of required saliva tests by 83%. PCR loss of sensitivity was minor, in accordance with the theoretical prediction for an 8-sample pool. cCMV was identified in 123 newborns, with a birth prevalence of 3.8 per 1000. Over half (57.7%) of infants with cCMV identified by universal screening were asymptomatic at birth and would have been missed by targeted screening. Conclusion The study demonstrates the wide feasibility and benefits of pooled-saliva cCMV testing as an accurate and cost-sparing approach, which could revolutionise the implementation of universal cCMV screening. Beyond the clinical implications toward early diagnosis, monitoring and treatment of cCMV, data derived from the implemented universal screening will serve to define the burden, risk factors, and long-term clinical outcomes of cCMV. Disclosures Dana Wolf, MD, GSK: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Grant/Research Support|MSD: Advisor/Consultant|Takeda: Advisor/Consultant