Determination of the soluble form of the P2X7 receptor in aqueous humour, vitreous humour and serum under normal and pathological conditions: sP2X7R as an indicator of ocular inflammatory status.

Background:P2X7R is a nucleotide receptor, located on the cytoplasmic membrane of all inflammatory and immune cells, that binds extracellular ATP. P2X7R plays a key role in inflammation, mediating activation of the NLRP3 inflammasome and release of cytokines (IL1β), and in regulating several cell death mechanisms. Espression of P2X7R was found in both eye bulb and ocular adnexa. On the ocular surface P2X7R is activated by chemical or mechanical sources of exogenous stress, leading to inflammation, apoptosis and cell proliferation. In the retinal, abnormalities in P2X7R functions have been linked to neuronal loss in glaucoma and neuroinflammatory processes in age-related macular degeneration and diabetic retinopathy. Based on a careful analysis of the recent literature on P2X7R involvement in the pathogenesis of various ocular diseases, this observational interventional study was conducted, aiming to: asses the presence of the soluble form of the purinergic P2X7 receptor (sP2X7R) in aqueous humour and vitreous humour; compare sP2X7R levels in healthy conditions and in the presence of various ocular or systemic pathological conditions with ocular involvement, underlying an inflammatory pathogenesis, such as: glaucoma, Fuchs endothelial dystrophy, pseudoexfoliatio, age-related macular degeneration, diabetes and retinal detachment; compare sP2X7R levels in aqueous humour and vitreous humour with those in serum. Methods:The patients enrolled are represented by subjects undergoing eye surgery at the Ophthalmic Clinic Unit of the Sant'Anna Hospital of Ferrara. In the group of pathological subjects there are aqueous humour and serum samples taken from patients with different ocular pathologies who underwent planned cataract extraction surgery and vitreous humour and serum samples from patients with retinal detachment who underwent vitrectomy surgery. The control group included aqueous humour and serum samples from patients undergoing cataract extraction surgery in the absence of other eye disease and vitreous and serum samples taken from patients undergoing vitrectomy for macular hole or macular pucker. Results:The presence of sP2X7R was assessed, which was found to be detectable in aqueous humour and vitreous humour using a specific ELISA kit. The range concentrations of sP2X7R in aqueous humour control group is 3.89 to 93.87 pg/ml; in pathologic group is 3.55-277.4 pg/ml. In vitreous humour range is 4.21-76.92 pg/ml in the control group, whilst in pathological condition is 28.52 to 284.57 pg/ml. The Kruskal-Wallis Test on the equality of the medians of both control and pathology groups of the acqueous samples, estimates a p-value slightly above the test reliability limit (p-value: 0.05082), confirming that the median value of the sP2X7R concentration measured in the pathology groups is significantly different, if all the data are considered together. Despite this finding, analysing each pathology group compared to the control group, using Dunn's Test to perform the pairwise comparison with the Bonferroni's correction factor, failed to confirm a statistically significant difference. The Kruskal-Wallis Test on the equality of the median of both control and pathology groups of the vitreous samples, does not allow the initial hypothesis to be rejected with probability (p value:0.1999). The test result therefore does not allow the assertion that there is a significant difference between the sP2X7R content in the vitreous of healthy patients and patients with the studied eye diseases. Conclusion:In this study the presence of sP2X7R in aqueous humour and vitreous humour was tested for the first time, and the ELISA kit used proved to be a reliable tool. By examining comprehensively the pathologic groups it can be stated that sP2X7R is a marker of ocular inflammatory status, but the stratification analysis failed to prove a significant correlation with the single pathology. Those data could lay the groundwork for further investigationBackground: P2X7R è un recettore nucleotidico, situato sulla membrana citoplasmatica delle cellule infiammatorie e immunitarie, che lega l'ATP extracellulare. P2X7R svolge un ruolo chiave nell'infiammazione, attivando l'inflammasoma NLRP3 e il rilascio di citochine (IL1β), e nella regolazione di diversi meccanismi di morte cellulare. L'espressione di P2X7R è stata riscontrata sia nel bulbo oculare che negli annessi. Sulla superficie oculare, P2X7R è attivato da stress chimici o meccanici, determinando infiammazione, apoptosi e proliferazione cellulare. A livello retinico, anomalie nelle sue funzioni sono state collegate alla perdita neuronale nel glaucoma e ai processi neuroinfiammatori nella degenerazione maculare legata all'età (DMLE) e nella retinopatia diabetica (RD). Dopo attenta analisi della recente letteratura, è stato condotto questo studio osservazionale interventistico con l'obiettivo di: valutare la presenza della forma solubile del recettore purinergico P2X7 (sP2X7R) nell'umor acqueo e nell'umor vitreo; confrontarne i livelli in condizioni normali e in presenza di differenti patologie oculari o sistemiche con coinvolgimento oculare, con patogenesi infiammatoria: glaucoma, distrofia endoteliale di Fuchs, pseudoesfoliazione, DMLE, diabete e distacco di retina; confrontare i livelli di sP2X7R nell'umor acqueo e vitreo con quelli del siero. Metodi:I pazienti arruolati sono soggetti sottoposti a intervento chirurgico programmato presso la Clinica Oculistica dell'Ospedale Sant'Anna di Ferrara. Il gruppo patoligico comprende campioni di umore acqueo e siero prelevati da pazienti con diverse patologie oculari sottoposti a intervento di estrazione della cataratta e campioni di umore vitreo e siero prelevati da pazienti con distacco di retina sottoposti a intervento di vitrectomia. Il gruppo di controllo comprende campioni di umore acqueo e siero di pazienti sottoposti a intervento di estrazione della cataratta in assenza di altre patologie oculari e campioni di vitreo e siero prelevati da pazienti sottoposti a vitrectomia per foro o pucker maculare. Risultati:La presenza di sP2X7R è stata confermata e rilevata nell'umor acqueo e nell'umor vitreo utilizzando uno specifico kit ELISA. L'intervallo di concentrazioni di sP2X7R nel gruppo di controllo dell'umor acqueo è 3,89-93,87 pg/ml; nel gruppo patologico è 3,55-27,4 pg/ml. Nell'umor vitreo il range è 4,21-76,92 pg/ml nel gruppo di controllo, mentre nel gruppo patologico è 28,52-284,57 pg/ml. Il test di Kruskal-Wallis sull'uguaglianza delle mediane dei gruppi di controllo e patologici dell’umor acqueo stima un valore p leggermente superiore al limite di affidabilità del test (p: 0,05082), confermando che il valore mediano della concentrazione di sP2X7R misurato nei gruppi patologici è significativamente diverso, se si considerano tutti i dati insieme. Nonostante questo risultato, l'analisi di ciascun gruppo patologico rispetto al gruppo di controllo, utilizzando il test di Dunn per il confronto a coppie, applicando il fattore di correzione di Bonferroni, non ha confermato una differenza statisticamente significativa. Il test di Kruskal-Wallis sull'uguaglianza della mediana dei campioni di vitreo tra gruppi di controllo e patologici stima un valore p: 0,1999. Il risultato del test non permette quindi di affermare che esista una differenza significativa tra il contenuto di sP2X7R nel vitreo di pazienti sani e di pazienti con le patologie oculari studiate. Conclusioni: In questo studio è stata testata per la prima volta la presenza di sP2X7R nell'umor acqueo e nell'umor vitreo e il kit ELISA utilizzato si è dimostrato uno strumento affidabile. Esaminando complessivamente i gruppi patologici si può affermare che sP2X7R è un marcatore dello stato infiammatorio oculare, ma l'analisi stratificata non è riuscita a dimostrare una correlazione significativa con la singola patologia. Questi dati potrebbero gettare le basi per ulteriori indagini

Determination of the soluble form of the P2X7 receptor in aqueous humour, vitreous humour and serum under normal and pathological conditions: sP2X7R as an indicator of ocular inflammatory status.

Background:P2X7R is a nucleotide receptor, located on the cytoplasmic membrane of all inflammatory and immune cells, that binds extracellular ATP. P2X7R plays a key role in inflammation, mediating activation of the NLRP3 inflammasome and release of cytokines (IL1β), and in regulating several cell death mechanisms. Espression of P2X7R was found in both eye bulb and ocular adnexa. On the ocular surface P2X7R is activated by chemical or mechanical sources of exogenous stress, leading to inflammation, apoptosis and cell proliferation. In the retinal, abnormalities in P2X7R functions have been linked to neuronal loss in glaucoma and neuroinflammatory processes in age-related macular degeneration and diabetic retinopathy. Based on a careful analysis of the recent literature on P2X7R involvement in the pathogenesis of various ocular diseases, this observational interventional study was conducted, aiming to: asses the presence of the soluble form of the purinergic P2X7 receptor (sP2X7R) in aqueous humour and vitreous humour; compare sP2X7R levels in healthy conditions and in the presence of various ocular or systemic pathological conditions with ocular involvement, underlying an inflammatory pathogenesis, such as: glaucoma, Fuchs endothelial dystrophy, pseudoexfoliatio, age-related macular degeneration, diabetes and retinal detachment; compare sP2X7R levels in aqueous humour and vitreous humour with those in serum. Methods:The patients enrolled are represented by subjects undergoing eye surgery at the Ophthalmic Clinic Unit of the Sant'Anna Hospital of Ferrara. In the group of pathological subjects there are aqueous humour and serum samples taken from patients with different ocular pathologies who underwent planned cataract extraction surgery and vitreous humour and serum samples from patients with retinal detachment who underwent vitrectomy surgery. The control group included aqueous humour and serum samples from patients undergoing cataract extraction surgery in the absence of other eye disease and vitreous and serum samples taken from patients undergoing vitrectomy for macular hole or macular pucker. Results:The presence of sP2X7R was assessed, which was found to be detectable in aqueous humour and vitreous humour using a specific ELISA kit. The range concentrations of sP2X7R in aqueous humour control group is 3.89 to 93.87 pg/ml; in pathologic group is 3.55-277.4 pg/ml. In vitreous humour range is 4.21-76.92 pg/ml in the control group, whilst in pathological condition is 28.52 to 284.57 pg/ml. The Kruskal-Wallis Test on the equality of the medians of both control and pathology groups of the acqueous samples, estimates a p-value slightly above the test reliability limit (p-value: 0.05082), confirming that the median value of the sP2X7R concentration measured in the pathology groups is significantly different, if all the data are considered together. Despite this finding, analysing each pathology group compared to the control group, using Dunn's Test to perform the pairwise comparison with the Bonferroni's correction factor, failed to confirm a statistically significant difference. The Kruskal-Wallis Test on the equality of the median of both control and pathology groups of the vitreous samples, does not allow the initial hypothesis to be rejected with probability (p value:0.1999). The test result therefore does not allow the assertion that there is a significant difference between the sP2X7R content in the vitreous of healthy patients and patients with the studied eye diseases. Conclusion:In this study the presence of sP2X7R in aqueous humour and vitreous humour was tested for the first time, and the ELISA kit used proved to be a reliable tool. By examining comprehensively the pathologic groups it can be stated that sP2X7R is a marker of ocular inflammatory status, but the stratification analysis failed to prove a significant correlation with the single pathology. Those data could lay the groundwork for further investigationBackground: P2X7R è un recettore nucleotidico, situato sulla membrana citoplasmatica delle cellule infiammatorie e immunitarie, che lega l'ATP extracellulare. P2X7R svolge un ruolo chiave nell'infiammazione, attivando l'inflammasoma NLRP3 e il rilascio di citochine (IL1β), e nella regolazione di diversi meccanismi di morte cellulare. L'espressione di P2X7R è stata riscontrata sia nel bulbo oculare che negli annessi. Sulla superficie oculare, P2X7R è attivato da stress chimici o meccanici, determinando infiammazione, apoptosi e proliferazione cellulare. A livello retinico, anomalie nelle sue funzioni sono state collegate alla perdita neuronale nel glaucoma e ai processi neuroinfiammatori nella degenerazione maculare legata all'età (DMLE) e nella retinopatia diabetica (RD). Dopo attenta analisi della recente letteratura, è stato condotto questo studio osservazionale interventistico con l'obiettivo di: valutare la presenza della forma solubile del recettore purinergico P2X7 (sP2X7R) nell'umor acqueo e nell'umor vitreo; confrontarne i livelli in condizioni normali e in presenza di differenti patologie oculari o sistemiche con coinvolgimento oculare, con patogenesi infiammatoria: glaucoma, distrofia endoteliale di Fuchs, pseudoesfoliazione, DMLE, diabete e distacco di retina; confrontare i livelli di sP2X7R nell'umor acqueo e vitreo con quelli del siero. Metodi:I pazienti arruolati sono soggetti sottoposti a intervento chirurgico programmato presso la Clinica Oculistica dell'Ospedale Sant'Anna di Ferrara. Il gruppo patoligico comprende campioni di umore acqueo e siero prelevati da pazienti con diverse patologie oculari sottoposti a intervento di estrazione della cataratta e campioni di umore vitreo e siero prelevati da pazienti con distacco di retina sottoposti a intervento di vitrectomia. Il gruppo di controllo comprende campioni di umore acqueo e siero di pazienti sottoposti a intervento di estrazione della cataratta in assenza di altre patologie oculari e campioni di vitreo e siero prelevati da pazienti sottoposti a vitrectomia per foro o pucker maculare. Risultati:La presenza di sP2X7R è stata confermata e rilevata nell'umor acqueo e nell'umor vitreo utilizzando uno specifico kit ELISA. L'intervallo di concentrazioni di sP2X7R nel gruppo di controllo dell'umor acqueo è 3,89-93,87 pg/ml; nel gruppo patologico è 3,55-27,4 pg/ml. Nell'umor vitreo il range è 4,21-76,92 pg/ml nel gruppo di controllo, mentre nel gruppo patologico è 28,52-284,57 pg/ml. Il test di Kruskal-Wallis sull'uguaglianza delle mediane dei gruppi di controllo e patologici dell’umor acqueo stima un valore p leggermente superiore al limite di affidabilità del test (p: 0,05082), confermando che il valore mediano della concentrazione di sP2X7R misurato nei gruppi patologici è significativamente diverso, se si considerano tutti i dati insieme. Nonostante questo risultato, l'analisi di ciascun gruppo patologico rispetto al gruppo di controllo, utilizzando il test di Dunn per il confronto a coppie, applicando il fattore di correzione di Bonferroni, non ha confermato una differenza statisticamente significativa. Il test di Kruskal-Wallis sull'uguaglianza della mediana dei campioni di vitreo tra gruppi di controllo e patologici stima un valore p: 0,1999. Il risultato del test non permette quindi di affermare che esista una differenza significativa tra il contenuto di sP2X7R nel vitreo di pazienti sani e di pazienti con le patologie oculari studiate. Conclusioni: In questo studio è stata testata per la prima volta la presenza di sP2X7R nell'umor acqueo e nell'umor vitreo e il kit ELISA utilizzato si è dimostrato uno strumento affidabile. Esaminando complessivamente i gruppi patologici si può affermare che sP2X7R è un marcatore dello stato infiammatorio oculare, ma l'analisi stratificata non è riuscita a dimostrare una correlazione significativa con la singola patologia. Questi dati potrebbero gettare le basi per ulteriori indagini

Inflammaging: should this term be suitable for age related macular degeneration too?

INTRODUCTION: Inflammaging is a phenomenon triggered by the conjunction of chronic repetitive and subclinical inflammation from external aggressors and internal inflammatory mechanisms due to the progressive degradation of systems such as the mitochondrial function. Age-related macular degeneration is the leading cause of blindness and visual impairment in patients older than 60 years in developed countries. DISCUSSION: Remarkable correlations have been documented between common or rare immunological/inflammatory gene polymorphisms and AMD, unequivocally indicating the involvement of inflammation and immune-mediated processes (complement activation) in the pathogenesis of this disease. CONCLUSION: Altogether these factors also drive this pathologic condition under the general heading of "Inflammaging"

Who is Afraid of a Bug Mantled Like Zorro?

Obligate intracellular microorganism in the form of Elementary bodies, Chlamydia trachomatis, in the extracellular environment surrounds itself with a mantle, like Zorro and can adapt in a balanced activity as in a niche, letting the cells survive but determin- ing multisite and long-lasting subclinical forms, causing inflammatory pathologies that go beyond the trachoma itself to one way of Inflammaging. This means alerting the medical doctor to verify signs or consequences typical of C. trachomatis infection such as tri- chiasis, corneal ulcer and trachomatous scar, spontaneous abortion, inflammatory pelvic disease, infertility and cervical cancer, also paying attention to the correlation with ocular lymphomas, the rare oropharyngeal disease also supported by Neisseria gonorrheae and introducing the need to control patients also for sexual attitude. This suggests the opportunity to add an S. (Sexual Behaviour) to the S.A.F.E. strategy prescribed by The WHO to combat Trachom

About Chlamydia trachomatis. Reply to Garcia-Teillard et al. Trachoma and the Importance of Sexual Infective Route in Developed Countries. Comment on “Gallenga et al. Why the SAFE—S Strategy for Trachoma? Are Musca sorbens or Scatophaga stercoraria Really the Culprit?—A Brief Historical Review from an Italian Point of View. Pathogens 2023, 12, 1419”

The confirmatory comment of Garcia-Teillard et al. [1] has led us to formulate an answer based on three points: - The treatment of infection with targeted and prolonged antibiotic therapy that can in- terfere with the Chlamydia trachomatis life cycle, reducing antibiotic inappropriateness and, thus, efficacy; - Reducing indiscriminate pesticide pollution that interferes with the food chain and reduces biodiversity; - Evaluating the active role of dipterans in the transmission of infection in urban areas by raising awareness of sexually transmitted infection among Health Authorities through information and prevention, differentiating between protection and contraception, with a current increase in the spread of infection in the competent population

New Insights into Molecular Oncogenesis and Therapy of Uveal Melanoma

Uveal melanoma (UM), which is the most common cancer of the eye, was investigated in recent years by many teams in the field of biomedical sciences and eye clinicians. New knowledge was acquired on molecular pathways found to be dysregulated during the multistep process of oncogenesis, whereas novel therapeutic approaches gave significant results in the clinical applications. Uveal melanoma-affected patients greatly benefited from recent advances of the research in this eye cancer. Tumour biology, genetics, epigenetics and immunology contributed significantly in elucidating the role of different genes and related pathways during uveal melanoma onset/progression and UM treatments. Indeed, these investigations allowed identification of new target genes and to develop new therapeutic strategies/compounds to cure this aggressive melanoma of the eye. Unfortunately, the advances reported in the treatment of cutaneous melanoma have not produced analogous benefits in metastatic uveal melanoma. Nowadays, no systemic adjuvant therapy has been shown to improve overall survival or reduce the risk of metastasis. However, the increasing knowledge of this disease, and the encouraging results seen in clinical trials, offer promise for future effective therapies. Herein, different pathways/genes involved in uveal melanoma onset/progression were taken into consideration, together with novel therapeutic approaches

Macrophage Activation in Follicular Conjunctivitis during the COVID-19 Pandemic

Among the symptoms of SARS-CoV-2, follicular conjunctivitis has become relevant. The conjunctiva acts as an open lymph node, reacting to the viral antigen that binds the epithelial cells, forming follicles of B cells with activated T cells and NK cells on its surface, which, in turn, talk to monocyte-derived inflammatory infected macrophages. Here, the NLRP3 inflammasome is a major driver in releasing pro-inflammatory factors such as IL-6 and caspase-1, leading to follicular conjunctivitis and bulbar congestion, even as isolated signs in the ‘asymptomatic’ patient

Progression in migraine: Role of mast cells and pro-inflammatory and anti-inflammatory cytokines

Migraine is a common painful neurovascular disorder usually associated with several symptoms, such as photophobia, phonophobia, nausea, vomiting and inflammation, and involves immune cells. Mast cells (MCs) are immune cells derived from hematopoietic pluripotent stem cells which migrate and mature close to epithelial, blood vessels, and nerves. In almost all vascularized tissues there are MCs that produce, contain and release biologically active products including cytokines, arachidonic acid compounds, and proteases. In addition, MCs participate in innate and adaptive immune responses. Innate responses in the central nervous system (CNS) occur during neuroinflammatory phenomena, including migraine. Antigens found in the environment have a crucial role in inflammatory response, causing a broad range of diseases including migraine. They can be recognized by several innate immune cells, such as macrophages, microglia, dendritic cells and MCs, which can be activated trough Toll-like receptor (TLR) signaling. MCs reside close to primary nociceptive neurons, associate with nerves, and are capable of triggering local inflammation. MCs are involved in the pathophysiology of various tissues and organs, especially where there is an increase of angiogenesis. Activated MCs release preformed mediators include histamine, heparin, proteases (tryptase, chimase), hydrolases, cathepsin, carboxypeptidases, and peroxidase, and they also generate pro-inflammatory cytokines/chemokines. In addition, activated macrophages, microglia and MCs in the CNS release pro-inflammatory cytokines which provoke an increase of arachidonic acid product levels and lead to migraine and other neurological manifestations including fatigue, nausea, headaches and brain fog. Innate immunity and pro-inflammatory interleukin (IL)-1 cytokine family members can be inhibited by IL-37, a relatively new member of the IL-1 family. In this article, we report that some pro-inflammatory cytokines inducing migraine may be inhibited by IL-37, a natural suppressor of inflammation, and innate and acquired immunit