The biological stage for this debate is set by glial cells. Although the blood-brain barrier grants the central nervous system a degree of immune privilege, the brain is actively patrolled by resident immune cells, principally microglia and astrocytes. Under healthy conditions these cells maintain tissue homeostasis, but when they detect danger signals they become the innate immune effectors of the brain, amplifying or resolving inflammatory responses. LPS engages this machinery with surgical precision: it binds Toll-like receptor 4 in cooperation with the co-receptors CD14 and MD-2, igniting a canonical signalling cascade that culminates in the production of tumour necrosis factor-alpha, interleukin-1beta, interleukin-6, nitric oxide via inducible nitric oxide synthase, and reactive oxygen species. When this activation is acute it can be adaptive, but sustained glial activation drives oxidative and nitrosative stress, and chronic microglial mediators can even push astrocytes into reactive states that erode neuronal support.

The problem, according to the review, is heterogeneity. LPS preparations differ in bacterial source, serotype, chemotype and purification grade, and studies vary wildly in dose, exposure duration, route of administration and the timing of their measurements. In vitro, most work relies on murine microglial systems, particularly the immortalised BV-2 and N9 lines and primary microglia, with LPS concentrations typically spanning 100 nanograms to 1 microgram per millilitre. Signalling switches on within minutes to hours, yet many studies sample only at 24 to 48 hours, when the biology has already moved on. The authors argue that early signalling events, such as phosphorylation of IKBalpha and nuclear translocation of the transcription factor NF-kappaB p65, are most informative within the first two hours, while later outputs like nitric oxide accumulation, prostaglandin E2 release and cytokine secretion reflect transcriptional and phenotype-level changes best captured between 12 and 48 hours. Sampling at the wrong moment conflates these distinct phases and weakens mechanistic attribution.

To build their framework, the authors conducted a structured search of PubMed, Scopus and Web of Science covering English-language publications from 2010 to 2026, ultimately synthesising evidence from 50 unique studies of isolated phytochemicals and chemically characterised natural products tested in LPS-induced neuroinflammation models. The readout landscape they map is dominated by nitric oxide biology, with nitrite quantification and iNOS induction serving as screening anchors, flanked by cytokine panels, COX-2 and prostaglandin E2 measurements, oxidative stress markers such as glutathione ratios and mitochondrial reactive oxygen species, and an essential layer of viability assays including MTT, CCK-8 and lactate dehydrogenase release to exclude non-specific cytotoxic suppression. Some studies go further, tracking microglial activation markers like Iba-1 and CD68, phagocytosis, morphology, and metabolic signatures such as extracellular acidification and oxygen consumption rates.

Mechanistically, the reviewed evidence converges on a surprisingly restricted set of signalling modules. NF-kappaB-centred signalling is the most frequently interrogated layer, assessed through p65 phosphorylation, nuclear translocation and DNA-binding activity. MAPK cascades involving ERK, JNK and p38, along with PI3K-Akt signalling, are profiled as complementary readouts. A subset of studies extends into NLRP3 inflammasome territory, measuring caspase-1 activation, gasdermin D cleavage and mature interleukin-1beta release, while others position compounds within the Nrf2-HO-1 antioxidant axis, mitophagy markers like PINK1 and Parkin, or immunometabolic nodes such as the glycolytic enzyme PKM2 and its transcriptional partner HIF-1alpha. Secondary regulatory modules, the authors stress, modulate the magnitude and resolution of the core LPS response rather than acting as primary sensing pathways, a distinction that matters when interpreting what a compound is actually doing.

The review illustrates this mechanistic positioning with concrete examples. Quercetin, tested in BV-2 microglia exposed to 100 nanograms per millilitre of LPS, reduced inflammatory cytokines and reactive oxygen species while suppressing NLRP3, cleaved caspase-1 and gasdermin D, and increasing PINK1 and LC3-II, pointing to a coupling between inflammasome control and mitochondrial quality assurance. In mice receiving repeated LPS injections, quercetin reduced hippocampal and cortical Iba-1 and CD68 and lowered immobility in behavioural tests. Sulforaphane offers a different pattern: in primary microglia it reduced TNF-alpha and interleukin-1beta within a PI3K-Akt activation framework, and in mice it alleviated depression-like behaviour in a manner abolished by pharmacological Akt inhibition. Coptisine, meanwhile, was positioned at the immunometabolic interface, reducing phosphorylated PKM2, nuclear PKM2 and HIF-1alpha, and attenuating anxiety-like behaviour in vivo.

On the animal side, the review finds that systemic intraperitoneal LPS administration predominates in rodents, with single doses ranging from 100 micrograms to 6 milligrams per kilogram and repeated schedules extending up to 14 days, while intracerebroventricular delivery and zebrafish models provide complementary spatial and kinetic insights. But the authors flag a critical interpretive caveat: systemic LPS engages peripheral immune signalling and sickness-like physiology, so it cannot be read as isolated brain inflammation. Stronger central attribution requires brain-region-specific inflammatory, glial or neuronal readouts, typically from hippocampus and cortex, alongside systemic measures. They also highlight a persistent blind spot in the literature: most rodent studies use male animals only, even though immune signalling and microglial activation are sex-dependent, potentially limiting generalisability.

To bring order to this heterogeneous evidence base, the authors propose a pragmatic four-level hierarchy of claim strength. Level 1, preliminary, covers viability-controlled changes in inflammatory markers in a single model. Level 2, mechanistic, requires concordant early-signalling and downstream mediator changes supported by replication or dose-response evidence. Level 3, cross-model, demands multi-endpoint confirmation in an independent, primary, human-relevant or multicellular platform, and is proposed as the decision point for considering progression to animal studies. Level 4, preclinical, requires coherent in vitro and in vivo findings including brain inflammatory or glial endpoints together with functional outcomes. Crucially, behaviour-only animal findings do not automatically qualify as Level 4 evidence, and studies sharing the same methodological limitations do not compound confidence.

The translational stakes are considerable. Plant-derived compounds remain a rich source of candidate anti-inflammatory agents, but the authors caution that a compound showing only nitric oxide or cytokine suppression in a single immortalised microglial line should be considered a preliminary screening hit at best. Stronger candidates link mediator suppression to pathway engagement under viability control, and the most promising ones show multi-endpoint activity across primary or human-relevant systems and animal models with brain-region and behavioural readouts. Looking forward, they call for human iPSC-derived microglia, microglia-containing organoids, microfluidic blood-brain barrier platforms and multicellular co-cultures to replace reliance on immortalised lines, and for inflammasome studies to distinguish LPS priming from secondary activation signals such as ATP, so that compound effects can be attributed to specific steps rather than the cascade as a whole.

The review also confronts an uncomfortable possibility: publication bias. Positive phytochemical findings may be preferentially published over neutral, negative or toxicity results, and testing multiple compounds, concentrations and endpoints can overstate consistency across models. Because this was a narrative review rather than a formal systematic one, the extent of such bias cannot be quantified, but the authors urge prespecified endpoints and honest reporting of null findings. Their bottom line is a call for discipline: standardised reporting of LPS source, serotype, purity, dose, timing and route; endpoints matched to inflammatory kinetics; validation in biologically distinct platforms; and cautious interpretation that treats positive results as evidence of pathway modulation under a defined challenge rather than proof of disease-modifying efficacy. In a field racing to find natural defenders of the brain, the most powerful tool may simply be a stopwatch.

Subject of Research: Lipopolysaccharide-induced neuroinflammation models and their use in phytochemical screening for anti-inflammatory drug discovery

Article Title: Kinetics and mechanisms in lipopolysaccharide-induced neuroinflammation: a framework for phytochemicals screening

Article References: Kinetics and mechanisms in lipopolysaccharide-induced neuroinflammation: a framework for phytochemicals screening. (n.d.). https://doi.org/10.1007/s11033-026-12817-4

Image Credits: AI Generated

DOI: 10.1007/s11033-026-12817-4

Keywords: neuroinflammation, lipopolysaccharide, microglia, TLR4, NF-kappaB, phytochemicals, NLRP3 inflammasome, Nrf2, BV-2 cells, cytokines, blood-brain barrier, experimental design

The research, led by Murat Çakır of Yozgat Bozok University together with colleagues at Yozgat Bozok University, Osmaniye Korkut Ata University, and Inonu University, focused on fatty acid amide hydrolase, or FAAH, the enzyme responsible for breaking down anandamide, one of the body’s principal endogenous cannabinoids. Anandamide and its molecular cousins are not merely the chemicals that give cannabis its effects; they are native signaling molecules that the body deploys to fine-tune pain, immune responses, inflammation, and cell survival. By inhibiting FAAH, URB597 prevents anandamide from being rapidly degraded, effectively amplifying the body’s own anti-inflammatory signaling without introducing any plant-derived or psychoactive compounds. This strategy of augmenting endocannabinoid tone has attracted growing interest as researchers probe its potential in inflammatory and ischemic diseases.

To test whether this approach could protect the kidneys during sepsis, the investigators used the cecal ligation and puncture model, widely regarded as the gold standard for reproducing polymicrobial sepsis in rodents. In this model, the cecum is ligated and punctured, allowing intestinal bacteria to spill into the peritoneal cavity and trigger a systemic infection that closely mirrors the clinical course of septic shock in patients. Forty male Sprague-Dawley rats were divided into four groups of ten animals each: a healthy control group, a sepsis group receiving no treatment, and two sepsis groups treated with URB597 at doses of either 0.3 or 0.6 milligrams per kilogram of body weight. Critically, the drug was administered intraperitoneally within five minutes of the sepsis induction, a timing designed to test whether early intervention could blunt the inflammatory cascade before it inflicted irreversible damage.

Twenty-four hours after sepsis was induced, the researchers collected blood and kidney tissue for a battery of biochemical, histopathological, and immunohistochemical analyses. The results in the untreated septic animals were stark. Serum levels of blood urea nitrogen and creatinine, the classic clinical markers of kidney dysfunction, rose significantly, as did neutrophil gelatinase-associated lipocalin, or NGAL, an earlier and more sensitive biomarker of tubular injury that has gained traction in nephrology for flagging kidney damage before conventional blood tests change. Alongside these functional markers, the septic rats showed elevated circulating levels of the pro-inflammatory cytokines tumor necrosis factor-alpha and interleukin-1 beta, signaling molecules that drive much of the tissue destruction seen in sepsis.

Under the microscope, the kidneys of untreated septic rats displayed the characteristic hallmarks of acute injury: disrupted tubular architecture, cellular swelling, and inflammatory infiltration. But in the animals that received URB597, the picture changed dramatically. Both doses of the drug significantly reduced the sepsis-induced elevations in BUN, creatinine, NGAL, TNF-alpha, and IL-1 beta compared with untreated septic controls, and the histopathological damage to the kidney tissue was visibly attenuated. Interestingly, when the two doses were compared head to head, the researchers found no significant differences between them in the serum biochemical parameters, suggesting that even the lower dose captured most of the protective effect within the timeframe studied.

The mechanistic heart of the study lies in its immunohistochemical findings, which traced the protective effect to a specific molecular pathway. Toll-like receptor 4, or TLR4, is a pattern-recognition receptor on immune and kidney cells that acts as an alarm bell for bacterial components, launching a signaling chain that activates nuclear factor-kappa B, or NF-kappa B, a transcription factor often described as the master switch of inflammation. When NF-kappa B is activated, its inhibitory protein I-kappa B-alpha is phosphorylated and degraded, allowing the transcription factor to enter the nucleus and switch on genes encoding TNF-alpha, interleukin-1 beta, and interleukin-6. In the septic rats, the researchers detected strong renal immunoreactivity for TLR4, phosphorylated NF-kappa B, and phosphorylated I-kappa B-alpha, confirming that this inflammatory axis had been ignited in the kidney tissue itself.

URB597 treatment significantly dampened all of these molecular markers. The treated animals showed reduced renal immunoreactivity for TLR4, phosphorylated NF-kappa B, phosphorylated I-kappa B-alpha, TNF-alpha, IL-1 beta, and IL-6, indicating that the drug had effectively turned down the volume on the kidney’s local inflammatory response rather than merely masking systemic markers. This pathway-level suppression is significant because the TLR4/NF-kappa B axis has been implicated not only in septic kidney injury but also in renal damage from ischemia-reperfusion and toxic insults, meaning that a drug acting here could theoretically have broad nephroprotective applications.

Equally important was the drug’s effect on apoptosis, the programmed cell death process that contributes to the loss of functional tubular cells in septic kidneys. The researchers measured renal levels of cleaved caspase-3, the executioner enzyme that disassembles the cell from within, and caspase-8, an initiator caspase that relays death signals from inflammatory pathways. Both were elevated in the septic animals and significantly reduced by URB597 treatment. This dual action, suppressing both the inflammatory cascade and the apoptotic machinery downstream of it, suggests that boosting endocannabinoid signaling protects kidney tissue at multiple points along the injury pathway rather than targeting a single vulnerable node.

The findings dovetail with a growing body of literature on the endocannabinoid system in kidney disease. Previous work has shown that FAAH inhibition or genetic deletion protects against renal fibrogenesis after ischemia-reperfusion injury and ameliorates cisplatin-induced nephropathy in mice, while other studies have demonstrated that endocannabinoid degradation inhibitors reduce leukocyte adhesion and improve microvascular perfusion in experimental endotoxemia. Elevated levels of the endocannabinoids anandamide and 2-arachidonoylglycerol have even been proposed as prognostic markers in septic patients, hinting that the body may naturally ramp up this protective system in response to infection. The present study extends this line of inquiry directly into sepsis-associated acute kidney injury, one of the most clinically consequential settings in which the endocannabinoid system had remained comparatively underexplored.

As with all preclinical work, considerable distance separates a rat model from the intensive care unit. The cecal ligation and puncture model, while rigorous, cannot fully capture the heterogeneity of human sepsis, and the drug was given within minutes of sepsis induction, a therapeutic window that rarely exists in clinical practice where patients arrive hours or days after infection begins. Dosing, safety, and efficacy in humans remain entirely untested, and URB597 itself has had a complicated development history as a clinical candidate. Nevertheless, the study provides a clear and mechanistically grounded proof of concept: enhancing the body’s own cannabinoid signaling through FAAH inhibition can measurably reduce kidney injury, inflammation, and cell death in experimental sepsis, acting through the well-characterized TLR4/NF-kappa B pathway. For a condition in which supportive care remains the mainstay of treatment and no specific drug currently protects the kidneys from septic damage, that proof of concept is a development worth watching closely as the field moves toward translating endocannabinoid science into nephroprotective therapies.

Subject of Research: Protective effects of FAAH inhibition with URB597 in experimental sepsis-associated acute kidney injury

Article Title: URB597 attenuates experimental sepsis-associated acute kidney injury with reduced renal inflammation, apoptosis, and TLR4/NF-κB activation

Article References: Çakır, M., Aydın, A., Bircan, B., Fırat, S., & Tekin, S. (2026). URB597 attenuates experimental sepsis-associated acute kidney injury with reduced renal inflammation, apoptosis, and TLR4/NF-κB activation. Molecular Biology Reports, 53(1), Article 1631. https://doi.org/10.1007/s11033-026-12803-w

Image Credits: AI Generated

DOI: 10.1007/s11033-026-12803-w

Keywords: sepsis, acute kidney injury, URB597, FAAH, endocannabinoid system, anandamide, TLR4, NF-kappa B, inflammation, apoptosis, cytokines, rat model

The endothelium, once regarded as a passive lining of blood vessels, is now understood as an active, semi-permeable metabolic and endocrine monolayer that regulates vascular tone, leukocyte trafficking, and permeability. When endothelial cells undergo senescence—an irreversible cell cycle arrest driven by telomere attrition, oxidative stress, and DNA damage—they begin secreting a pro-inflammatory cocktail known as the senescence-associated secretory phenotype, or SASP. This mixture of cytokines, chemokines, and matrix-degrading enzymes accelerates the senescence of neighboring cells, disrupts the vascular barrier, and promotes clot formation. Mechanically, senescent endothelium diminishes the bioavailability of nitric oxide, the molecule responsible for vasorelaxation, leading to vasoconstriction, arterial stiffening, and hypertension. Deciphering the upstream triggers of this transition, the review contends, is paramount for developing effective vascular interventions.

TLR4 is a pattern recognition receptor best known as a sentinel of the innate immune system, binding both external pathogens and internally derived danger signals. Under physiological conditions, endothelial TLR4 acts as a surveillance system that coordinates tissue repair, angiogenesis, and vascular wall reconstruction. When activated by damage-associated molecular patterns, or DAMPs, it induces vascular endothelial growth factor, stimulating endothelial migration and tube formation for wound healing. In the lung, endothelial TLR4 is even essential for oxidative stress defense and tissue homeostasis. But under persistent metabolic stress—prolonged hyperglycemia or exposure to oxidized low-density lipoprotein—this protective architecture shifts into a maladaptive state, and chronic TLR4 hyperactivation begins to fuel the very senescence it evolved to prevent.

The mechanistic details are striking. When persistently stimulated by circulating DAMPs or oscillatory shear stress, endothelial TLR4 activates the NF-κB pathway through MyD88 and TRIF adaptor proteins while simultaneously assembling the NADPH oxidase 2 complex, producing a massive surge of superoxide. This oxidative flood keeps endothelial nitric oxide synthase inactive, drastically reducing local nitric oxide and generating toxic peroxynitrite. The accumulating oxidative stress induces double-stranded DNA breaks, activating the classic DNA damage response: ATM kinase phosphorylates p53, which in turn drives transcription of the cyclin-dependent kinase inhibitors p21 and p16INK4a, forcing cells into irreversible G1-phase arrest. What follows is a vicious, self-amplifying loop—senescent endothelial cells upregulate TLR4 on their own surfaces, secrete SASP factors such as IL-6, IL-1β, and TNF-α, induce bystander senescence in healthy neighbors, and further entrench the pathological state. In experimental settings, senescent endothelial cells display a ninefold upregulation of the adhesion molecule ICAM-1 upon secondary stimulation compared with healthy controls.

Perhaps the review’s most consequential contribution is its catalog of tissue-specific paradoxes. In aging mouse models, TLR4 levels rise in the heart and aorta, accelerating vascular decline, and microvascular endothelial cells—expressing far more of the TLR4 co-receptor CD14 than their macrovascular counterparts—mount an amplified IL-6 cascade that destabilizes atherosclerotic plaques. Yet in the lung, the opposite holds: endothelial TLR4 actively suppresses the senescence gene p16INK4a through HDAC2-mediated histone deacetylation, and its loss precipitates emphysema-like alveolar dilation. In diabetic retinopathy, deleting TLR4 specifically in retinal endothelial cells rescues the blood-retinal barrier, whereas deleting it in adjacent Müller cells does not. In the brain, acute activation of endothelial TLR4 triggers rapid internalization of the tight junction protein claudin-5, collapsing the blood-brain barrier—an effect entirely absent in endothelial-specific knockout mice.

Even more paradoxical is an expression asymmetry between neighboring cell types. Senescent endothelial cells upregulate their own TLR4, driving chronic vascular decline, yet they simultaneously restrict the release of exosomal miR-326-3p, a microRNA that downregulates TLR4 in adjacent skin fibroblasts. The resulting TLR4 deficiency in fibroblasts impairs basal survival signaling and accelerates their aging. The same aging phenotype, in other words, is driven by an excess of TLR4 in one cell type and a deficit in another—a finding that demolishes any notion of the receptor as a simple pro-aging villain.

The clinical record bears out the danger of oversimplification. Epidemiological data suggest that genetically blunted TLR4 signaling protects against vascular aging: in a cohort of 2,679 patients with coronary artery disease, carriers of the loss-of-function variant rs4986790 showed a significantly attenuated rise in systolic blood pressure over time, and an Italian study of 810 subjects found that carriers of the Asp299Gly allele had lower levels of interleukin-6 and fibrinogen and reduced atherosclerosis risk. But large-scale Phase III trials of systemic TLR4 antagonists—the MD-2 blocker eritoran and the intracellular inhibitor TAK-242—failed to reduce mortality in severe sepsis and were terminated early. Complete abrogation of TLR4 signaling dismantles innate immune defense, as TLR4-deficient mice suffer fatal gram-negative pneumonia, and aged knockout mice develop spontaneous obesity driven by skewed immune profiles. Systemic blockade even disrupts dendritic cell maturation and cytotoxic T-cell priming, potentially accelerating tumor growth.

The alternative, the authors argue, is precision senotherapeutics: confining TLR4 modulation strictly to pathologically altered endothelial cells while leaving homeostatic compartments untouched. Pharmacological agents already in clinical use offer a starting point. Heparin suppresses endothelial TLR4 and MyD88 expression in a dose-dependent manner, blocking NF-κB nuclear translocation and reducing SASP components to delay senescence. The Hedgehog pathway agonist SAG rescues placental and uterine artery angiogenesis impaired by TLR4 hyperactivation, while sildenafil and nitrite supplementation bypass TLR4 injury by restoring nitric oxide signaling in neonatal necrotizing enterocolitis. Upstream strategies are equally promising: epigenetic targeting of the Ash2l gene dampens lipid uptake and stabilizes atherosclerotic plaques, RAGE silencing downregulates TLR4 expression in diabetic vasculature, and anti-eNAMPT antibodies preserve endothelial junctional integrity in acute lung injury.

The most futuristic frontier lies in targeted delivery. Nanostructured lipid carriers functionalized with anti-VCAM-1 antibodies selectively accumulate in inflamed atherosclerotic vessels while bypassing healthy ones, and have been used to deliver melatonin that silences the endothelial TLR4/NF-κB cascade and suppresses inflammatory pyroptosis. E-selectin-targeted multistep vectors ferry therapeutic microRNAs such as miR-146a and miR-181b to activated endothelium, shrinking atherosclerotic lesions, while dual-targeting lipid vehicles conjugated with both anti-VCAM-1 and anti-E-selectin antibodies achieve complete gene silencing in activated endothelial cells with no detectable systemic toxicity. Even plant-derived exosomes are entering the arena: V-Onex, engineered by grafting a VCAM-1-binding peptide onto onion-derived extracellular vesicles, homes selectively to atherosclerotic endothelial layers. Aptamer-drug conjugates that unlock only in the presence of elevated lysosomal β-galactosidase promise to release senolytic payloads exclusively inside senescent cells.

Significant hurdles remain before these platforms reach the clinic. Exosomes offer high biocompatibility but suffer from mass-production and purification challenges and risk rapid clearance in vivo; synthetic nanoparticles allow precise structural tuning but face scalability constraints and unresolved questions about long-term biodistribution and toxicity. Hybrid technologies fusing the two approaches are being proposed as a compromise. Still, the conceptual shift articulated in this review is clear: vascular aging is driven not by the mere presence of TLR4 signaling but by its localized hyperactivation and dysregulated ligand-receptor kinetics. The next generation of cardiovascular and anti-aging therapies, the authors conclude, must move past simple receptor blockade toward spatiotemporally targeted restoration—treating the endothelium not as a uniform sheet of tissue, but as a mosaic of specialized niches, each demanding its own precisely calibrated molecular rheostat.

Subject of Research: The role of endothelial Toll-like receptor 4 signaling in vascular senescence and the development of targeted senotherapeutic strategies

Article Title: Endothelial TLR4 at the Crossroads of Vascular Senescence and Targeted Senotherapeutics

Article References: Kim, H.-J., Lee, J.-H., & Hwangbo, C. (2026). Endothelial TLR4 at the Crossroads of Vascular Senescence and Targeted Senotherapeutics. Aging Cell, 25(10), Article e70757. https://doi.org/10.1111/acel.70757

Image Credits: AI Generated

DOI: 10.1111/acel.70757

Keywords: TLR4, endothelial senescence, vascular aging, SASP, senotherapeutics, senolytics, senomorphics, nanoparticle drug delivery, NF-kB, nitric oxide, atherosclerosis, targeted therapy

To understand why the finding matters, it helps to revisit how the innate immune system detects bacterial invaders. Immune cells called macrophages patrol tissues armed with pattern recognition receptors, molecular sensors that spot conserved microbial signatures. Among the most potent of these signatures is lipopolysaccharide, or LPS, a component of the outer membrane of Gram-negative bacteria. When bacteria multiply, they shed LPS-rich outer membrane vesicles, tiny blebs of membrane that can travel to distant sites in the body, sneak inside immune cells, and ignite inflammation far from the original infection. This dissemination of inflammatory triggers is a hallmark of systemic inflammatory conditions, sepsis chief among them.

Once intracellular LPS is detected, the cell deploys an inflammasome, a multiprotein machine that acts as both alarm and weapon. Inflammasomes come in two flavors. The canonical pathway relies on well-studied sensors such as NLRP3, while the non-canonical pathway is triggered directly by intracellular LPS through the inflammatory caspase-11 in mice, and caspase-4 and caspase-5 in humans. Activation of either pathway leads caspase-1 to process pro-inflammatory cytokines, most notably interleukin-1 beta, and to cleave Gasdermin-D, the executioner protein that punches holes in the cell membrane and drives inflammatory cell death. While decades of research have mapped how post-translational modifications tune the canonical pathway, the modifications governing the non-canonical route have remained largely mysterious. That gap is precisely where the McGill team, led by Michel L. Tremblay, focused its attention.

The researchers zeroed in on PTPN2, an enzyme that removes phosphate groups from tyrosine residues on target proteins, thereby switching signaling circuits on or off. To probe its role, they generated mice lacking Ptpn2 specifically in myeloid cells, the lineage that includes macrophages, and challenged these animals with LPS-rich outer membrane vesicles and with acute sepsis models. The results were striking. In macrophages deficient in Ptpn2, the transcription of interleukin-1 beta, the flagship cytokine of inflammasome biology, was markedly impaired. The defect traced back to dysregulated signaling through TLR4, the cell-surface LPS receptor, and the p38 MAPK pathway, a cascade that normally drives the priming phase of inflammasome activation by ramping up cytokine gene expression.

Intriguingly, the compromised priming did not translate into wholesale collapse of the inflammasome machinery. Levels of caspase-11, caspase-1, and Gasdermin-D remained unchanged in the knockout cells, indicating that the enzyme’s influence is exerted upstream, at the level of inflammatory gene transcription and signal tuning, rather than by altering the abundance of the core inflammasome components themselves. This distinction is biologically important: it suggests PTPN2 shapes the intensity of the inflammatory output rather than the mere presence of the detonator, a nuance that could matter greatly when designing drugs that target the pathway.

The study also uncovered a novel substrate for PTPN2, adding a fresh branch to the enzyme’s known interaction map. The team found that Lyn, a member of the Src-family of kinases, becomes hyperphosphorylated in the absence of Ptpn2. Because phosphatases and kinases operate in opposing pairs, this observation identifies Lyn as a direct or proximal target whose phosphorylation state PTPN2 normally restrains. Src-family kinases sit at hubs connecting receptor activation to downstream signaling, so their dysregulation in Ptpn2-deficient macrophages provides a mechanistic thread linking the phosphatase to the altered TLR4 and p38 MAPK signaling observed in the knockout animals.

Adding a second layer of complexity, the researchers found that loss of PTPN2 unleashes a STAT1-mediated program that boosts production of nitric oxide, a reactive molecule with a double-edged role in immunity. While nitric oxide can help kill pathogens, the study showed that in this context it undermines inflammasome activity by compromising mitochondrial fitness. Mitochondria are not merely cellular power plants; they supply signals and metabolites that feed inflammasome activation, and their dysfunction can blunt the cell’s inflammatory capacity. In Ptpn2-deficient macrophages, therefore, excessive nitric oxide erodes the very organelles the inflammasome depends upon, further dampening the response.

The in vivo consequences of these cellular defects became clear when the mice were subjected to acute sepsis. Animals lacking Ptpn2 in myeloid cells exhibited reduced numbers of small peritoneal macrophages, a key resident immune population, and lower serum levels of interleukin-1 beta following the septic challenge. Together, these findings indicate a dampened pro-inflammatory response in the absence of the phosphatase. In the context of sepsis, where excessive inflammation drives organ damage and death, a blunted response might at first glance seem protective. But the picture is nuanced: interleukin-1 beta also mobilizes host defenses, and the reduced macrophage population suggests that PTPN2 supports aspects of immune competence as well as inflammation. The enzyme, in other words, is not simply a brake or an accelerator but a fine-tuner of the response.

That dual role is what makes PTPN2 an attractive therapeutic candidate. Phosphatases were long considered undruggable, but recent advances in medicinal chemistry have brought PTPN2 inhibitors into clinical consideration, particularly in oncology, where the enzyme’s effects on T cells and tumor immunity have drawn intense interest. The McGill study adds a new dimension to that conversation: in sepsis and systemic inflammation, modulating PTPN2 activity could, in principle, recalibrate the non-canonical inflammasome axis and the cytokine output that fuels inflammatory injury. The authors highlight this potential, positioning the enzyme as a target for managing systemic inflammation. Any such strategy would need to account for the pathway’s complexity, since the same enzyme that supports cytokine production also, through its restraint of STAT1 and nitric oxide, protects mitochondrial fitness and inflammasome function.

The work also carries broader conceptual weight for immunology. By demonstrating that a tyrosine phosphatase governs the non-canonical inflammasome pathway, the study establishes that this arm of innate immunity, often treated as a parallel, LPS-triggered circuit, is subject to the same intricate layers of post-translational regulation that shape its canonical counterpart. It connects outer membrane vesicle biology, TLR4 signaling, Src-family kinase activity, STAT1 activation, mitochondrial health, and cytokine secretion into a single regulatory network centered on one enzyme. For clinicians confronting sepsis, a syndrome that kills millions each year and for which treatment remains largely supportive, the identification of a tunable node in the inflammatory circuitry offers a genuine reason for optimism. For basic researchers, it opens a set of new questions: how PTPN2 is itself regulated during infection, whether the Lyn and STAT1 branches operate independently, and whether human patients with altered PTPN2 activity show different sepsis trajectories. The answers could shape the next generation of anti-inflammatory therapies.

Subject of Research: Role of the phosphatase PTPN2 in regulating non-canonical inflammasome activation and inflammatory responses in sepsis

Article Title: PTPN2 sustains non-canonical inflammasome activation and shapes the inflammatory response to sepsis

Article References: Colalillo, B., Aubry, I., Aumont, P., Poirier, A. J., Hincapie, A. M., Wu, C., St-Laurent, E., Martinez Cordova, Z., & Tremblay, M. L. (2026). PTPN2 sustains non-canonical inflammasome activation and shapes the inflammatory response to sepsis. Cellular and Molecular Life Sciences. https://doi.org/10.1007/s00018-026-06469-7

Image Credits: AI Generated

DOI: 10.1007/s00018-026-06469-7

Keywords: PTPN2, sepsis, inflammasome, non-canonical pathway, interleukin-1 beta, LPS, macrophages, TLR4, p38 MAPK, Lyn kinase, STAT1, nitric oxide

Unlike vertebrates, which deploy both adaptive and innate immune responses, mollusks rely exclusively on innate immunity. Their defense toolkit includes enhanced phagocytic activity, pattern recognition receptors, and effector molecules that identify and neutralize invading microorganisms. Among these molecular sentinels is the mannose receptor, a member of the C-type lectin superfamily and a type I transmembrane protein that functions as what researchers describe as a non-standard pattern recognition receptor. Rather than serving as a conventional first-line sensor, the mannose receptor contributes to host defense primarily by recognizing and binding endogenous ligands and pathogen-associated molecules, thereby modulating immune responses and maintaining immune homeostasis under external stress. Its importance has been demonstrated previously in aquatic animals such as the red swamp crayfish Procambarus clarkii and the orange-spotted grouper Epinephelus coioides, where MR-mediated responses showed antiviral and antibacterial activity against Vibrio species.

Structurally, the mannose receptor is an intricate machine. It comprises an extracellular cysteine-rich domain, a fibronectin type II domain, and eight tandem C-type lectin-like domains, along with a transmembrane segment and a short cytoplasmic tail. Functional studies have shown that the fibronectin type II domain and the second C-type lectin-like domain act synergistically to enhance the uptake of glycosylated collagen, while the fourth domain has been implicated in modulating T cell cytotoxicity. The receptor’s C-type lectin-like domains can specifically recognize and bind carbohydrate ligands such as mannan, trehalose, and N-acetylglucosamine on both endogenous and exogenous molecules, initiating immune responses upon binding. In the bream Megalobrama amblycephala, for example, the mannose receptor binds chitosan oligosaccharide and mediates its uptake by macrophages through lectin-dependent endocytosis, modulating the expression of tumor necrosis factor receptor-associated factors, interleukins, and nitric oxide synthase.

To understand how this receptor family operates in the Manila clam, the researchers mined the clam’s genome, which was previously sequenced and made available under NCBI BioProject PRJNA479743. Using Hidden Markov Model searches based on the C-type lectin-like domain model PF00059.23, they identified a remarkably large repertoire of 13 mannose receptor genes, designated RpMR1 through RpMR13. The predicted proteins displayed considerable diversity, with molecular weights ranging from 11.27 to 320.37 kilodaltons and theoretical isoelectric points between 4.42 and 6.82. Exon numbers varied from 2 to 60, and every RpMR protein contained at least one C-type lectin domain, with copy numbers ranging from 1 to 14 per protein, hinting at substantial functional redundancy or diversification within the family. Conserved motif analysis revealed ten distinct motifs across the proteins, and multiple sequence alignment highlighted conserved cysteine residues, calcium-binding sites, acidic amino acids, and aromatic residues such as phenylalanine and tryptophan, all characteristic features of C-type lectin family proteins.

Chromosomal localization added another layer of organization to the story. Ten of the 13 RpMR genes mapped to six annotated chromosomes, with chromosome 10 harboring three genes in what appears to be a gene cluster, while the remaining three genes sat on unplaced scaffolds. Phylogenetic analysis, built from 142 mannose receptor amino acid sequences across nine representative species, revealed that MR genes cluster into two major branches, with molluscan sequences from R. philippinarum, Crassostrea virginica, C. gigas, and Biomphalaria glabrata grouping predominantly within a single large clade. This pattern suggests that mannose receptor genes in mollusks have undergone evolutionary conservation, reflecting shared ancestry and potentially conserved functional roles across the phylum. Interestingly, when the researchers examined expression across developmental stages, all RpMR genes were expressed as the clams grew, with most showing significantly increased expression at the D-larva stage, indicating that this immune machinery is active from early life onward.

The infection challenge experiments brought the gene family’s defensive role into sharp focus. Wild clams collected from Jinshitan in Dalian were immersed in V. anguillarum at a concentration of 1 × 10⁷ CFU/mL, and hepatopancreas tissues were sampled at intervals from 0 to 96 hours post-challenge. The results showed that RpMR expression surged following infection, with RpMR1, RpMR2, RpMR3, RpMR4, and RpMR6 all peaking at 72 hours post-infection, reaching 3.2-, 6.8-, 8.3-, 1.68-, and 1.1-fold increases respectively relative to baseline. Tissue-specific analysis revealed that the genes were expressed throughout the clam body, including the adductor muscle, mantle, foot, gill, siphon, and digestive gland, but expression was most pronounced in the hepatopancreas, where RpMR2 reached twelvefold higher levels than in the adductor muscle. This organ, the researchers conclude, likely serves as a key site for mannose receptor-mediated immune responses.

The team then zeroed in on RpMR1 as a functional candidate. They cloned the gene’s coding region into a PET-28A(+) vector, expressed it in E. coli Rosetta (DE3) cells, and purified the recombinant protein, which appeared at its theoretical 123 kilodalton position on SDS-PAGE gels and was confirmed by Western blot. In vitro antibacterial assays tested the protein against eight bacterial strains, and the results were strikingly specific. RpMR1 significantly inhibited growth of three Gram-negative Vibrio pathogens: V. splendidus, with significant suppression observed at 4 hours, and V. anguillarum, with sustained inhibition at 6, 8, and 10 hours, along with V. alginolyticus. No inhibitory activity was detected against Bacillus subtilis, Staphylococcus aureus, Vibrio parahaemolyticus, V. harveyi, or Escherichia coli. The researchers attribute this bactericidal specificity to RpMR1’s affinity for lipopolysaccharides, the dominant outer membrane component of Gram-negative bacteria and a conserved pathogen-associated molecular pattern recognized by Toll-like receptor 4.

The most dramatic evidence came from in vivo experiments. Clams were divided into four groups receiving different injections: phosphate-buffered saline plus recombinant protein, V. anguillarum plus recombinant protein, V. anguillarum plus buffer, and buffer alone. By 96 hours, the cumulative mortality rate in the infected group that received only buffer reached 78.6 percent, while the infected group that also received RpMR1 protein saw mortality drop to 52.7 percent, a 26 percent reduction in deaths. Clams receiving protein without infection showed the same survival as untreated controls, confirming the protein’s safety. Complementary molecular measurements showed that injection of RpMR1 rapidly activated the Toll-like receptor signaling pathway, with the genes TLR, MyD88, TRAF, NF-κB, IKK, and AP-1 all peaking at 6 hours post-infection in the infected, protein-treated group, while most pathway genes in the infected, untreated group did not peak until 96 hours. Nitric oxide synthase activity was also significantly elevated at 12 and 72 hours in the protein-treated infected group, consistent with nitric oxide’s established role in promoting phagolysosome maturation and microbicidal activity.

To establish causality rather than mere correlation, the researchers turned to RNA interference. Injecting synthetic double-stranded RNA targeting RpMR1 successfully silenced the gene, reducing its expression significantly. The knockdown had cascading effects: TRAF6 expression dropped significantly, while TLR4 and AP-1 expression levels were also significantly reduced. This demonstrates that RpMR1 positively regulates components of the TLR signaling pathway during the immune response, confirming a functional interaction between the mannose receptor and TLR4. The finding fits with a broader literature on pattern recognition receptor crosstalk: mannose-binding lectin, another C-type lectin family member, has been shown to potentiate TLR4 signaling through direct interaction with its leucine-rich repeat domain, and cooperative interactions between mannose receptors and TLR4 have been documented in orchestrating pro-inflammatory mediator release, including interleukin-1β, tumor necrosis factor-alpha, and interleukin-6. Because the mannose receptor itself lacks intrinsic signaling capacity, such cooperative interactions with other receptors may be essential for transducing immune activation signals while maintaining immune homeostasis.

The implications for aquaculture are considerable. The authors suggest that the immunological functions of recombinant RpMR1 protein could eventually be applied in Manila clam farming, potentially incorporated into feed as an antimicrobial agent, though they caution that further research is needed to determine optimal dosage and whether sustained antibacterial activity can be maintained at scale. More broadly, the study marks the first functional demonstration of mannose receptor-mediated immunity in mollusks, bridging receptor-mediated pathogen recognition with downstream effector mechanisms such as nitric oxide production and TLR signaling in bivalve host defense. The researchers note that the precise molecular interplay by which RpMR1 confers anti-Vibrio immunity through nitric oxide synthase-dependent mechanisms remains to be fully delineated, and targeted investigations into the tripartite relationship between receptor activation, effector enzyme regulation, and pathogen clearance are still needed. Nevertheless, by identifying a concrete molecular target linked to survival during infection, the work opens a promising avenue for breeding or engineering disease-resistant clam strains, offering a potential lifeline for an aquaculture industry under relentless bacterial pressure.

Subject of Research: Mannose receptor-mediated innate immunity in the Manila clam against Vibrio anguillarum infection

Article Title: Mannose receptor RpMR1 of Manila clam (Ruditapes philippinarum) defense against Vibrio anguillarum infection

Article References: Yin, Z., & Nie, H. (2025). Mannose receptor RpMR1 of Manila clam (Ruditapes philippinarum) defense against Vibrio anguillarum infection. Advanced Biotechnology, 3(3), Article 23. https://doi.org/10.1007/s44307-025-00075-7

Image Credits: AI Generated

DOI: 10.1007/s44307-025-00075-7

Keywords: Manila clam, Ruditapes philippinarum, mannose receptor, RpMR1, Vibrio anguillarum, pattern recognition receptor, innate immunity, TLR4, nitric oxide synthase, RNA interference, aquaculture, bivalve defense

Glucagon is the hormonal mirror image of insulin. While insulin ushers glucose out of the bloodstream and into tissues, glucagon does the opposite, instructing the liver to release stored glucose when fasting or hypoglycemia threatens the brain’s energy supply. It is produced by alpha cells, a small but critical population within the pancreatic islets that has historically received far less attention than the insulin-secreting beta cells. In type 2 diabetes, the glucagon axis is often deranged: alpha cells fail to suppress glucagon after meals, contributing to the hyperglycemia that defines the disease, while paradoxically overreacting during hypoglycemic episodes. Understanding what drives alpha cell dysfunction has therefore become a pressing question, and the new study suggests that a circulating liver-derived protein may be part of the answer.

Fetuin-A, encoded in humans by the AHSG gene, is produced almost exclusively by hepatocytes and circulates at high concentrations in the blood. Previous research had established associations between elevated Fetuin-A levels, insulin resistance, fatty liver disease, and low-grade inflammation, but its influence on alpha cell function had never been systematically explored. To probe that gap, the investigators turned to Alpha TC1 clone 6 cells, a mouse pancreatic alpha cell line widely used to study glucagon biology. They exposed the cells to high concentrations of Fet-A under hyperglycemic conditions and then switched them to low glucose to trigger glucagon synthesis, a protocol designed to mimic the metabolic swings that alpha cells experience in vivo.

The results were striking. Fetuin-A treatment increased the expression of preproglucagon mRNA, the genetic template from which glucagon is made, indicating that the protein acts at the level of gene transcription rather than merely prompting the release of preformed hormone. Mechanistically, the team found that Fet-A activated inflammatory signaling through Toll-like receptor 4, or TLR4, the same innate immune receptor that recognizes bacterial lipopolysaccharide. When the researchers silenced the TLR4 gene using small interfering RNA, the stimulatory effect of Fetuin-A on glucagon synthesis vanished, demonstrating that the receptor is a necessary intermediary. Pharmacological inhibitors corroborated the finding, and Western blotting confirmed that the alpha cells express TLR4 basally, giving the pathway a plausible structural foothold.

But the study did not stop at inflammation. A second, and arguably more novel, mechanism emerged from the interaction between Fetuin-A and insulin-like growth factor 1, or IGF-1. Under normal circumstances, IGF-1 suppresses glucagon synthesis during hypoglycemia through the PI3K/Akt/FoxO1 signaling cascade, a well-characterized intracellular pathway that transmits growth factor signals from the cell surface to the nucleus. The researchers found that Fetuin-A disrupted this braking mechanism, impairing IGF-1’s ability to dampen glucagon production precisely when suppression matters most. In other words, when blood sugar drops and the body needs to restrain glucagon’s counter-regulatory surge, elevated Fetuin-A appears to loosen the leash.

To understand how a circulating glycoprotein could interfere with a receptor-driven pathway, the team turned to cell-surface confocal microscopy and molecular docking. The imaging experiments revealed a dose-dependent co-localization of Fetuin-A with the IGF-1 receptor on the alpha cell membrane, suggesting that the two molecules physically occupy overlapping territory. Docking analysis predicted potential structural overlap within the receptor’s extracellular domain, and sequence alignment studies showed that the relevant beta-subunit domain is spatially conserved across human and mouse insulin and IGF-1 receptors. Taken together, the data support a model of steric interference: Fetuin-A binds at or near the receptor’s ligand-binding region, physically obstructing IGF-1 from engaging its target and thereby blunting the downstream signal that normally curtails glucagon synthesis.

The cell culture findings were then tested in living organisms. The researchers administered Fetuin-A to CD-1 mice for three consecutive days, with a saline-treated control group for comparison. The treated animals showed increased circulating glucagon concentrations, along with elevated levels of inflammatory cytokines, mirroring the dual inflammatory and hormonal signature observed in the cell experiments. All animal protocols were approved by the local Animal Care Committee and conducted in accordance with European directive 2010/63/EU, the ARRIVE guidelines, and the 3R principle, lending regulatory rigor to the in vivo component of the work.

The final and most clinically consequential piece of evidence came from humans. The team analyzed data from 93 non-diabetic adults enrolled in the CATAMERI study, the CAtanzaro MEtabolic RIsk cohort, a long-running observational project examining cardiometabolic risk factors. Fasting plasma Fetuin-A concentrations were positively associated with fasting glucagon levels, and the association held up after statistical adjustment for age, sex, body mass index, insulin, and IGF-1. That independence matters: it argues that the relationship is not simply a byproduct of obesity, insulin status, or growth factor levels, but reflects a direct physiological link between the liver-derived protein and alpha cell output. Glucagon was measured using a chemiluminescence immunoassay, a sensitive and standardized method that strengthens confidence in the clinical measurement.

What emerges from the combined evidence is a coherent mechanistic story with potential implications for metabolic disease. If Fetuin-A, elevated in obesity and fatty liver, both inflames alpha cells through TLR4 and disables the IGF-1-mediated brake on glucagon synthesis, then the protein could help explain why hyperglucagonemia persists in insulin-resistant states even when glucose is abundant. The liver, in this framing, is not just a victim of hormonal miscommunication but an active participant, secreting a signal that reshapes the behavior of the very cells responsible for glucose counter-regulation. The authors also verified that Fet-A was not cytotoxic to the alpha cells across the concentration range tested, using MTT viability assays over 48 hours, which rules out the trivial explanation that the hormone changes simply reflect cell damage or death.

Cautions remain, as they always do in translational research. The cell line is murine, the mouse experiments involved exogenous protein administration rather than chronic endogenous elevation, and the human data are cross-sectional, showing association rather than causation. Whether lowering Fetuin-A would normalize glucagon levels in people with diabetes is a question for future interventional studies. Still, the convergence of evidence across three biological levels, from transcriptional regulation in cultured cells to hormone measurements in mice and statistical associations in a well-characterized human cohort, gives the hypothesis unusual solidity. The work was supported by Italian national research funding programs, including PNRR and PRIN grants, and the authors report no competing interests. As the field increasingly recognizes alpha cells as active drivers of dysglycemia rather than passive bystanders, Fetuin-A now sits squarely on the list of molecular suspects worth pursuing, and therapies aimed at interrupting its interaction with TLR4 or the IGF-1 receptor may one day earn a place in the metabolic medicine arsenal.

Subject of Research: The role of the liver-derived glycoprotein Fetuin-A in regulating pancreatic alpha cell glucagon synthesis and secretion

Article Title: Alpha 2-HS glycoprotein increases glucagon synthesis and secretion in cells, mice, and humans

Article References: Mancuso, E., Averta, C., Rubino, M., Citraro, R., Palummo, A., Servello, A., Belviso, S., Massimino, M., Mannino, G. C., De Sarro, G., Sesti, G., & Andreozzi, F. (2026). Alpha 2-HS glycoprotein increases glucagon synthesis and secretion in cells, mice, and humans. Journal of Translational Medicine. https://doi.org/10.1186/s12967-026-08965-7

Image Credits: AI Generated

DOI: 10.1186/s12967-026-08965-7

Keywords: Fetuin-A, glucagon, pancreatic alpha cells, TLR4, IGF-1 receptor, inflammation, insulin resistance, type 2 diabetes, liver, metabolic disease, PI3K/Akt/FoxO1, CATAMERI study

TLR4 sits at the front line of the body’s defense against bacteria. When the portion of the receptor exposed outside the cell detects a bacterial molecule, the receptor undergoes changes that prompt two TLR4 proteins to come together as dimers, and those dimers can then trigger cascades inside the cell, including the NF-κB pathway, one of the principal drivers of inflammation. The receptor can also respond to certain non-bacterial molecules associated with tissue damage, which helps explain why its misfiring is tied to such a broad range of inflammatory conditions. Despite decades of interest, no FDA-approved drug specifically blocks TLR4, in part because the receptor’s most druggable-looking surfaces are also the hardest to engage with precision. The Scripps team’s strategy was to aim not at the well-studied extracellular region but at the stretch of the protein buried inside the membrane, a site long considered beyond the reach of rational design.

The biological rationale for this approach emerged from a simple question posed by first author Colleen Maillie, a research project analyst at Scripps Research. Scientists had generally assumed that the regions of TLR4 exposed outside and inside the cell were the main signaling drivers, with the membrane-spanning segment serving a purely structural role. To test that assumption, the researchers introduced a fragment of TLR4, comprising the membrane-spanning region plus a small neighboring section, into human cells grown in the laboratory. The fragments readily associated with full-length TLR4 within the membrane, an interaction the team detected using a screening method recently developed in the Mravic lab that emits light when tagged proteins come into close proximity. Crucially, the presence of these fragments reduced overall NF-κB signaling responses, providing the first concrete evidence that the transmembrane region is not a passive tether but an active determinant of the receptor’s inflammatory output.

That screening assay matters as much as the molecules it helped find. Measuring whether two proteins interact inside a lipid bilayer is far harder than doing so in water, because most conventional biochemical tools are optimized for aqueous environments. The Mravic lab’s light-up approach was built specifically to identify which synthetic proteins target membrane proteins, and the TLR4 fragments provided an early demonstration that the method works in practice. With that tool in hand, the team could move from observation to engineering: if simply presenting the receptor with its own transmembrane sequence could dampen signaling, then a purpose-built synthetic protein designed to bind that region more tightly might dial the response down even further.

Designing such a molecule required confronting a stubborn gap in computational biology. Protein design software has matured enormously for soluble proteins, whose folding rules in water are increasingly well captured by physics-based equations and machine learning models. Membranes are a different story. The cell membrane consists of two compact layers of oily molecules, an environment with biochemical properties radically different from the water-based settings where most proteins reside, and many of the rules governing how proteins fold and function within those greasy layers remain poorly understood. As Mravic explained, models for protein interactions in water have become increasingly accurate, but for membrane proteins they are not, because there are unique atomic details underlying molecular biophysics in lipid bilayers that current equations and AI models do not accurately capture.

Unable to rely on the software’s raw predictions, the team treated the computer-generated three-dimensional blueprints as starting points rather than finished designs. They then applied custom design criteria developed in the Mravic lab to optimize what the researchers call apolar packing, the tightness with which the chemical structures of the designed proteins fit together with their TLR4 target. The underlying theory, which the team encoded into software, holds that maximizing apolar packing produces more stable protein interactions within the hydrophobic membrane environment. From the many structural options generated, the researchers narrowed the field to the nine candidates with the best predicted biophysical and chemical features and took those forward into tests in living cells.

The results validated the strategy with striking efficiency. Using the same light-up screening approach, the team found that eight of the nine synthetic proteins showed signs of associating with TLR4. Three emerged as the strongest candidates, and one, designated Design-6, showed the most robust evidence of interaction along with several other appealing qualities. Unlike the naturally occurring transmembrane fragments tested earlier, Design-6 did not aggregate as much, a critical property for any molecule that might eventually be developed as a biologic. Most importantly, Design-6 substantially reduced NF-κB inflammatory signaling, demonstrating that a rationally designed transmembrane protein could not merely bind to its target but measurably alter the receptor’s biological function.

The implications extend well beyond TLR4 itself. The study establishes that the membrane-embedded regions of immune receptors can be treated as legitimate, programmable drug sites, opening what the researchers describe as a potential launchpad for a new class of biologics that operate from within the membrane. It also delivers a set of computational tools, built around the apolar packing design criteria, that other laboratories can apply to proteins embedded in membranes, a category that includes many of the most sought-after targets in modern pharmacology. Because roughly a quarter of known drug targets are membrane proteins, design methods that work in lipid bilayers rather than despite them could reshape how researchers approach targets that conventional antibodies and small molecules have failed to reach.

Considerable work remains before any of this reaches patients. The experiments were conducted in human embryonic kidney cells, which are convenient for laboratory study but not especially relevant to inflammation-driven disease, so the team must confirm that the approach holds in more disease-relevant cell types such as liver cells and immune cells. Delivery poses another obstacle: these are oily synthetic proteins that must reach and insert into cell membranes, and no established clinical method yet exists for getting them there. Maillie acknowledged the challenge candidly, noting that this is an innovative space that carries a lot of risk and a long roadmap to the clinic, but emphasizing that the team has shown that with clever design and biophysics, they are getting closer. Maillie, Ward and Mravic are inventors on a provisional patent application titled Compositions and Methods for Inhibiting Toll-Like Receptor 4 Mediated Inflammation, signaling that the group intends to pursue the therapeutic potential of the platform.

What makes the study resonate beyond its immediate findings is the way it reframes a familiar target. TLR4 has long been described as a key sensor of bacteria that activates and mobilizes immune cells to fight infection, and, in Ward’s framing, as both a sensor and a dial to tune innate immunity, one that vaccine adjuvants can turn up and inhibitors can turn down. The Scripps work shows that the dial’s most sensitive setting may lie in the part of the receptor nobody thought to touch. By proving that a designed transmembrane protein can find its target, bind it, and quiet the inflammatory signal it produces, the study turns a long-standing blind spot in protein design into a working blueprint, and hands the field both a molecule and a method for building the next ones.

Subject of Research: De novo computational design of transmembrane proteins that inhibit the innate immune receptor TLR4

Article Title: Computer-designed protein targets immune receptor linked to inflammation at a new “undruggable” site

Article References: Computer-designed protein targets immune receptor linked to inflammation at a new “undruggable” site. (n.d.). Original publication

Image Credits: AI Generated

DOI: Not provided

Keywords: TLR4, protein design, transmembrane proteins, innate immunity, inflammation, NF-κB signaling, Scripps Research, membrane biophysics, apolar packing, drug design, computational biology, PNAS

The biological logic behind this approach begins with a phenomenon the authors call dysbiosis—a collapse of the gut’s normally diverse microbial community. Chemotherapy agents such as 5-fluorouracil, oxaliplatin, and cisplatin, the workhorses of gastric cancer regimens, do not discriminate neatly between tumor cells and the bacteria lining the intestine. The resulting microbial disruption damages the intestinal barrier, allowing bacterial components to leak into the bloodstream. Among these, lipopolysaccharide (LPS), a molecule embedded in the outer membrane of certain bacteria, is particularly consequential. Once LPS crosses a compromised gut wall, it binds to Toll-like receptor 4 (TLR4) on immune cells, triggering a signaling cascade through the transcription factor NF-κB and culminating in the activation of the NLRP3 inflammasome—a multiprotein molecular machine that ignites some of the body’s most potent inflammatory responses. The review identifies this LPS/TLR4/NF-κB/NLRP3 axis as a central driver of the systemic inflammation that underlies many chemotherapy side effects, from mucositis to immune dysfunction.

At the same time, chemotherapy starves the gut of one of its most valuable protective resources: short-chain fatty acids, or SCFAs. These small molecules—chiefly acetate, propionate, and butyrate—are produced by beneficial bacteria as they ferment dietary fiber. SCFAs are far more than metabolic waste; they act as signaling molecules and, remarkably, as inhibitors of histone deacetylases (HDACs), enzymes that control gene expression by chemically modifying chromatin. When SCFA levels fall, HDAC activity rises unchecked, anti-inflammatory gene programs are silenced, and the intestinal mucosa loses a critical shield against injury. Experimental work cited in the review shows that butyrate supplementation can prevent chemotherapy-induced gastrointestinal toxicity and microbial dysbiosis, while SCFA receptors such as GPR41, GPR43, and GPR109A on immune and epithelial cells mediate much of this protection. In other words, the difference between a tolerable chemotherapy course and a debilitating one may hinge, in part, on the metabolic output of a patient’s gut bacteria.

Against this mechanistic backdrop, the authors propose that TCM operates through what they term a structure–metabolite–immune framework. The idea is elegant in its simplicity: herbal compounds reshape the structure of the microbial community, the restructured community alters its production of metabolites, and those metabolites recalibrate the immune system. Restored microbial diversity means fewer LPS-producing pathogens and more SCFA-generating commensals. Lower LPS means less TLR4 activation; higher SCFAs means restored HDAC inhibition and calmer inflammatory signaling. The framework offers a unifying explanation for how wildly different herbal preparations—from complex multi-herb decoctions to single purified polysaccharides—can converge on similar protective outcomes. It also provides testable predictions: if the framework is correct, microbial sequencing and metabolite measurements should reveal consistent signatures in patients who respond well to TCM adjunct therapy.

The review catalogs a remarkable range of TCM components that fit this model. Polysaccharides from Astragalus membranaceus, a staple of Chinese herbal oncology, have been shown through multi-omics studies to protect against chemotherapeutic intestinal mucositis in animal models, and their structure–immunomodulation relationships are increasingly well mapped. Poria cocos polysaccharides exert a genuine prebiotic function, attenuating the adverse effects of 5-fluorouracil while improving its therapeutic outcome in mouse models of intestinal cancer. Polysaccharides from Atractylodes macrocephala protect against LPS-induced intestinal injury and appear to work partly by promoting gut bacterial production of tryptophan metabolites that activate the aryl hydrocarbon receptor, another key immunoregulatory pathway. On the flavonoid side, curcumin has been shown to suppress colorectal tumorigenesis partly by restoring gut microbiota and metabolites, while baicalin, derived from Scutellaria baicalensis, rebalances the regulatory T cell–Th17 axis and modulates SCFA profiles in models of colonic inflammation. Classical formulations such as Banxia Xiexin decoction and Shenling Baizhu powder have likewise been shown to reshape intestinal microbiota and SCFA metabolism in preclinical studies.

What elevates this review beyond a catalog of laboratory curiosities is its assessment of clinical evidence. The authors report that oral and injectable TCM formulations have accumulated moderate- to high-quality evidence for specific benefits in gastric cancer patients receiving chemotherapy. Shenqi Fuzheng injection (SFI), one of the most widely studied Chinese herbal injectables, has been the subject of multiple systematic reviews and meta-analyses showing improved immune function when combined with chemotherapy. Chinese herbal injections combined with SOX chemotherapy regimens—a common oxaliplatin plus S-1 protocol for advanced gastric cancer—have shown favorable effectiveness and safety profiles in Bayesian network meta-analyses. Oral preparations such as Shenqi Xiangyi granules have demonstrated benefits in advanced gastric cancer chemotherapy, and decoctions including Yipi Huayu and Zhipu Liujunzi have been associated in retrospective and propensity-matched studies with improved immune markers, favorable tumor marker trends, and reduced adverse reactions. The review also notes that these interventions appear to alleviate myelosuppression—the dangerous depletion of blood cell production in the bone marrow—and reduce gastrointestinal toxicities, two of the most clinically burdensome chemotherapy side effects.

The evidence is not uniformly strong, however, and the authors are candid about the gaps. External TCM therapies, including acupuncture and moxibustion, have generated some supportive data—a meta-analysis found benefits for gastrointestinal function and adverse events in gastric cancer patients after surgery and chemotherapy—but the overall quality of evidence for external approaches remains limited compared with oral and injectable formulations. Many of the most compelling mechanistic studies rely on animal models whose microbial and immune systems do not perfectly recapitulate human biology. Clinical trials are frequently small, single-center, and heterogeneous in their TCM preparations, dosing, and outcome measures, making it difficult to generalize. The review’s authors also point out that standard supportive care for chemotherapy toxicity—growth factors for neutropenia, antiemetic regimens for nausea—addresses symptoms rather than the underlying microbial and inflammatory mechanisms, leaving a genuine therapeutic vacuum that microbiota-targeted strategies could fill if properly validated.

Looking forward, the review charts several ambitious research frontiers. The authors highlight the STING signaling pathway—a DNA-sensing immune circuit with intimate connections to both inflammation and cell death—as an underexplored mechanistic link between microbial dysbiosis and chemotherapy toxicity. They similarly call for deeper investigation of the gut–brain axis, noting emerging evidence that chemotherapy-induced microbial disruption can exacerbate cancer-related fatigue through neuroimmune-endocrine signaling, and that interventions such as acupuncture and Lycium barbarum–probiotic combinations have shown promise in preclinical fatigue models by acting on this axis. Perhaps most consequentially, the authors advocate for large-scale randomized controlled trials and for the development of microbiota-based personalized TCM interventions—strategies in which a patient’s individual microbial profile would guide the selection of herbal therapy, much as tumor genomics guides targeted cancer drugs today. Such an approach would represent a genuine fusion of ancient pharmacopeia and modern precision medicine.

The implications extend well beyond gastric cancer. Dysbiosis has been implicated in chemotherapy-induced peripheral neuropathy, cardiotoxicity, and neurocognitive disorders, and microbial signatures of dysbiosis appear across gastrointestinal carcinogenesis. If the structure–metabolite–immune framework holds up under rigorous testing, microbiota modulation could become a standard pillar of supportive oncology care, alongside antiemetics and growth factors. For now, the review by Li and Ji stands as a careful synthesis of a rapidly maturing field—one that reframes chemotherapy toxicity not as an inevitable consequence of cancer treatment, but as a modifiable disorder of the gut ecosystem. The trillions of microbes that share our bodies, long overlooked in oncology, are emerging as both culprits and allies, and traditional Chinese medicine, whatever its ultimate clinical standing, has provided a rich and testable pharmacological toolkit for engaging them. The next decade of trials will determine whether this ancient-meets-modern strategy earns a permanent place in the cancer clinic.

Subject of Research: Gut microbiota modulation by traditional Chinese medicine to reduce chemotherapy-induced toxicity in gastric cancer

Article Title: Modulation of the gut microbiota by traditional Chinese medicine to attenuate chemotherapy-induced toxicity in gastric cancer: a review

Article References: Li, J., & Ji, F. (2026). Modulation of the gut microbiota by traditional Chinese medicine to attenuate chemotherapy-induced toxicity in gastric cancer: a review. Supportive Care in Cancer, 34(10), Article 1041. https://doi.org/10.1007/s00520-026-11290-w

Image Credits: AI Generated

DOI: 10.1007/s00520-026-11290-w

Keywords: gastric cancer, chemotherapy toxicity, gut microbiota, traditional Chinese medicine, short-chain fatty acids, LPS, TLR4, NF-κB, NLRP3 inflammasome, dysbiosis, polysaccharides, integrative oncology

The central finding of the study is that heat stress reshapes the gut microbiome of mice in two complementary ways. First, the microbial disruption is accompanied by increased levels of lipopolysaccharide, or LPS, a potent inflammatory molecule embedded in the outer membrane of many gut bacteria. When the intestinal barrier weakens, LPS can slip into the bloodstream and activate immune cells throughout the body. Second, the team found that heat stress enhances microbial catabolism of arginine, an amino acid that the new work identifies as a natural brake on inflammatory signaling. In other words, heat does not just add fuel to the inflammatory fire; it removes one of the fire extinguishers at the same time.

To establish that these microbial changes are not merely a byproduct of heat injury but an active driver of it, the researchers performed fecal microbiota transplantation experiments. When they transferred gut microbes from heat-stressed mice into healthy recipient animals, the recipients went on to mount exaggerated inflammatory responses after experiencing heat stress themselves. This result provides some of the strongest causal evidence yet that the gut microbial alterations induced by heat are functionally responsible for worsening systemic inflammation, rather than simply correlating with it. It suggests that the microbiome acts as an intermediary that can be reprogrammed by environmental temperature and then transmit that reprogramming into immune consequences.

The mechanistic heart of the paper concerns how arginine protects tissues at the molecular level. Using cell culture experiments in macrophage-like RAW264.7 cells and microglial BV2 cells, the team showed that arginine reduces the abundance of MyD88, a key adaptor protein that sits at the crossroads of multiple innate immune signaling pathways. MyD88 physically interacts with Toll-like receptor 4, the receptor that recognizes LPS, and passes the alarm signal onward to the transcription factor NF-kB, whose p65 subunit must move into the nucleus to switch on genes encoding pro-inflammatory cytokines. The researchers found that arginine promotes ubiquitination of MyD88, the molecular tag that marks proteins for degradation, thereby lowering MyD88 protein levels. With less MyD88 available, the interaction between MyD88 and TLR4 weakens, p65 nuclear translocation is curtailed, and the expression of pro-inflammatory genes falls.

Experiments in living mice reinforced this mechanism. Pretreating animals with exogenous arginine significantly suppressed inflammation in the liver and in the cortex of the brain after heat stress, the two organs highlighted in the study as major sites of heat-related damage. When the researchers knocked down TLR4 or MyD88 in their cellular models, the protective effect of arginine was lost or blunted, indicating that the amino acid’s benefit depends on the very pathway it modulates. Taken together, the data sketch a coherent circuit: heat raises LPS and lowers arginine, LPS engages TLR4, and the resulting MyD88-dependent signaling cascade runs unchecked when arginine is scarce.

Importantly, the team did not stop at animal models. They examined serum samples from patients suffering from heatstroke and found that arginine levels were lower in these patients than expected. Moreover, the degree of arginine depletion was positively related to markers of liver injury and inflammation, meaning that patients with the least circulating arginine tended to show the worst clinical indicators. While the human component of the study is observational and cannot by itself prove causation, it aligns closely with the mouse and cellular data and raises the possibility that arginine status could serve as a biomarker for heatstroke severity or as a guide for nutritional intervention.

The translational ambition of the work culminates in a delivery system the authors developed to counteract the microbial imbalance: an arginine-enriched oral inulin hydrogel, abbreviated Arg_OIH. Inulin is a dietary fiber with prebiotic properties, meaning it can nourish beneficial gut microbes and help maintain microbial homeostasis. By embedding arginine within an inulin-based hydrogel, the researchers aimed to achieve two goals at once. The inulin component stabilizes the gut microbiota, which in turn reduces LPS levels, while the hydrogel matrix provides a sustained, slow release of arginine to keep the amino acid available to tissues over time rather than delivering a single spike that the body would quickly clear.

According to the study, this hydrogel formulation was able to prevent the inflammatory responses exacerbated by heat-induced gut microbial alterations, and the supplementary data document its characterization, its effects on the gut microbiota of heat-stressed mice, and histopathological evidence of tissue protection. The authors also compared the hydrogel against a simple oral arginine solution, suggesting that the sustained-release design offers advantages over free amino acid supplementation. If the approach can be translated safely to humans, it could offer a practical prophylactic strategy for vulnerable populations, such as outdoor workers, athletes, the elderly, and people living in regions where extreme heat events are becoming routine.

The study also carries broader conceptual weight for microbiome science. It adds to a growing body of evidence that the gut microbiome functions as a metabolic buffer between the environment and the immune system, converting external stressors into shifts in microbial chemistry that either calm or inflame the host. Here, the balance between LPS and arginine emerges as a previously underappreciated axis: one side pushes TLR4-MyD88 signaling toward inflammation, while the other restrains it through post-translational modification of a central adaptor protein. Disrupting that balance, as heat stress appears to do, converts a manageable physiological response into a self-amplifying inflammatory loop capable of damaging multiple organs.

Caveats remain, as they do in any preclinical study. The mouse models of heat stress, however carefully controlled, cannot fully reproduce the complex physiology of human heatstroke, and the clinical samples, while suggestive, came from a patient cohort whose characteristics are detailed in the study’s supplementary tables. The authors note that the published version was shared early as accepted peer-reviewed research, subject to final editorial processing. Nevertheless, the convergence of evidence across fecal transplantation, metabolomics, cellular signaling assays, genetic knockdown experiments, and human serum measurements gives the arginine-MyD88 axis a solid evidentiary foundation. As global temperatures continue their upward climb, interventions that restore gut microbial balance and replenish protective metabolites may become an increasingly important part of the public health arsenal against heat, and this study offers a concrete, mechanistically grounded starting point for developing them.

Subject of Research: Gut microbial arginine catabolism and LPS balance in heat stress-induced inflammatory responses

Article Title: Heat stress enhances gut microbial arginine catabolism to amplify MyD88-dependent inflammatory responses

Article References: Ye, X., Cai, Q., Pan, Y., Xu, M., Li, Y., You, M., Yang, J., Chen, S., He, H., Hong, G., & Zheng, H. (2026). Heat stress enhances gut microbial arginine catabolism to amplify MyD88-dependent inflammatory responses. Microbiome, 14(1), Article 215. https://doi.org/10.1186/s40168-026-02514-6

Image Credits: AI Generated

DOI: 10.1186/s40168-026-02514-6

Keywords: heat stress, gut microbiota, arginine, lipopolysaccharide, MyD88, TLR4, inflammation, heatstroke, inulin hydrogel, NF-kB, liver injury, metabolomics

Obesity has long been recognized as a state of chronic low-grade inflammation, a smoldering immune activation that contributes to insulin resistance, fatty liver disease, and cardiovascular risk. At the same time, obesity is marked by circadian disruption: the molecular clocks that orchestrate daily cycles of metabolism, hormone secretion, and immune activity fall out of sync. What has remained unclear is whether these two phenomena are merely parallel consequences of excess weight or whether they are mechanistically intertwined, and, crucially, whether a dietary intervention can pull both back into alignment at the same time.

The new study, led by Claudia Pivonello and senior author Annamaria Colao, set out to address exactly that question. Thirty-eight adults with obesity completed a twelve-week hypocaloric Mediterranean diet program. Before and after the intervention, the researchers collected detailed anthropometric and metabolic measurements, including body weight, body mass index, waist circumference, fat mass percentage, fat-free mass, muscle mass, and HDL cholesterol. They also assessed each participant’s chronotype using the Morningness–Eveningness Questionnaire, a validated instrument that captures whether a person’s natural tendencies lean toward morning or evening activity.

The most technically ambitious part of the study involved the participants’ peripheral blood mononuclear cells, or PBMCs, a mixed population of immune cells that includes lymphocytes and monocytes. Blood samples were drawn at two fixed time points, eight in the morning and four in the afternoon, allowing the researchers to track how gene expression shifted across the day. Using quantitative reverse-transcription PCR, they measured the expression of five core clock genes: BMAL1, PER1, PER2, CRY1, and CRY2. These genes form the interlocking feedback loops that constitute the molecular circadian oscillator, with BMAL1 acting as the positive driving element and the PER and CRY proteins as the negative limbs that restrain its activity on a roughly twenty-four-hour cycle. Alongside the clock genes, the team quantified expression of two innate immune receptors, TLR4 and TLR8, members of the Toll-like receptor family that detect microbial components and trigger inflammatory signaling.

The metabolic results were consistent with what clinicians have come to expect from a well-executed Mediterranean diet intervention. Participants showed significant reductions in body weight, body mass index, waist circumference, and fat mass percentage, together with increases in fat-free mass, muscle mass, and HDL cholesterol. Their MEQ scores also rose significantly, indicating a shift toward greater morningness, although the distribution of chronotype categories did not change in a statistically significant way. In other words, participants drifted somewhat toward morning preference without crossing into a new chronotype classification.

The molecular findings were more striking. At baseline, before any dietary change, expression of TLR4 and TLR8 in PBMCs was significantly higher in the afternoon than in the morning, a diurnal pattern in innate immune receptor expression that the researchers documented for the first time in this cohort. After twelve weeks of the Mediterranean diet, the picture had changed in a coordinated fashion. BMAL1 expression increased significantly in the morning samples, while PER1, TLR4, and TLR8 expression decreased significantly in the afternoon samples. Moreover, all five clock genes examined, BMAL1, PER1, PER2, CRY1, and CRY2, displayed a more pronounced morning-to-afternoon expression pattern following the intervention, which the authors interpret as evidence of improved diurnal organization of the peripheral clock.

Perhaps the most intriguing observation was the correlation structure linking the two systems. The researchers found significant positive correlations between BMAL1 expression and both TLR4 and TLR8 expression at both sampling time points. This statistical relationship supports the hypothesis of a bidirectional interaction between the molecular clock and innate immune pathways, a connection that has been suggested by animal and cellular studies but rarely examined in the context of a human dietary intervention. If BMAL1 activity helps govern the daily rhythm of Toll-like receptor expression, then dietary signals that strengthen the clock could, in principle, dampen the inappropriate immune activation that characterizes obesity.

The authors frame their results as evidence of partial restoration of diurnal–immune homeostasis. Increased morning BMAL1 together with reduced afternoon TLR4 and TLR8 suggests that the intervention did not simply suppress inflammation indiscriminately but instead rebalanced the temporal architecture of immune readiness. They propose that dietary modulation of what they call the BMAL1–TLR axis may represent a mechanism linking metabolic improvement to diurnal regulation, and they identify peripheral clock gene expression as a potential translational biomarker of response to lifestyle interventions. If validated, such a biomarker could allow clinicians to monitor, at the level of gene expression in a simple blood draw, whether a patient’s body clock and immune system are responding to dietary therapy.

Important caveats temper these conclusions. The study lacked a comparator arm, meaning there was no control group of patients with obesity who did not receive the intervention or who received an alternative diet. Without such a group, the findings must be interpreted as associations rather than proof of causation, and the authors are explicit on this point, calling for confirmation in randomized controlled studies. The sample size of thirty-eight participants, while adequate for an exploratory interventional design, is modest, and the two-time-point sampling strategy captures only a coarse snapshot of the full twenty-four-hour rhythm. The study was registered at ClinicalTrials.gov as NCT06236932 and was approved by the Ethical Committee of Federico II University of Naples, and it was funded by the Italian Ministry of University and Research through a Projects of Relevant National Interest grant.

Even with those limitations, the study adds a compelling piece to a rapidly growing body of work on the circadian–immune axis. It suggests that the Mediterranean diet, already celebrated for its cardiovascular and metabolic benefits, may exert part of its influence by entraining the peripheral clocks embedded in immune cells and by smoothing out the aberrant daily oscillations of innate immune receptors. For patients with obesity, the practical message is familiar but now carries deeper mechanistic weight: what you eat, and presumably when you eat it, may help decide not only how much you weigh but how well your body’s internal timekeeping and immune surveillance work together. For researchers, the message is that the blood-borne clock may be a readable, and potentially modifiable, target of nutritional therapy.

Subject of Research: Effects of a hypocaloric Mediterranean diet on circadian clock gene and innate immune receptor expression in patients with obesity

Article Title: A 12-week hypocaloric Mediterranean diet is associated with metabolic improvement and modulation of diurnal clock gene and innate immune receptor expression in peripheral blood mononuclear cells of patients with obesity

Article References: Pivonello, C., Magnacca, N., Graziadio, C., Negri, M., Vetrani, C., Amatrudo, F., Vozza, E., Vecchio, G. D., Tini, S., Celano, S., Prodam, F., Auriemma, R. S., Pivonello, R., & Colao, A. (2026). A 12-week hypocaloric Mediterranean diet is associated with metabolic improvement and modulation of diurnal clock gene and innate immune receptor expression in peripheral blood mononuclear cells of patients with obesity. Journal of Translational Medicine. https://doi.org/10.1186/s12967-026-09013-0

Image Credits: AI Generated

DOI: 10.1186/s12967-026-09013-0

Keywords: Mediterranean diet, obesity, circadian rhythm, clock genes, BMAL1, Toll-like receptors, TLR4, TLR8, peripheral blood mononuclear cells, inflammation, chronotype, nutritional intervention