Four decades of p53 research since the discovery of this tumor suppressor have demonstrated that p53 prevents tumorigenesis by maintaining genomic stability and eliminates cancer cells by inducing cell growth arrest, necrosis, apoptosis, and ferroptosis . p53 actKung et al. gracefully described a canonical mechanism by which oncogenic stress induces p53 activation with some new information and thoughts. Specifically, the oncogenic c-MYC or RAS signaling induces the expression of ARF, an alternate open reading frame encoded by CDKN2A, which in turn activates p53 by interacting with and inhibiting MDM2 activity. Importantly, they also summarized several potential therapeutic strategies targeting the ARF-MDM2-p53 cascade, including small molecules, peptides, and the proteolysis targeting chimera (PROTAC) tactic. In a research study, Han et al. reported that the PARP inhibitor olaparib, a targeted therapy for cancers with BRCA1/2 mutations or homologous recombination deficiencies, can induce p53 activation via RPL5/RPL11-mediated inhibition of MDM2 by triggering nucleolar stress, demonstrating an additional action mode of PARP inhibitors by targeting the nucleoli and activating the p53 pathway.Blocking the MDM2-p53 feedback loop has been considered as a promising strategy to treat cancers harboring wt p53 for decades, although it has been quite challenging as there is not an applicable drug targeting this loop as an anti-cancer therapy in clinic use. In the issue, a review article by TP53, is the most frequently mutated gene in human cancers. The cancer-derived mutations of p53 include missense, frameshift, truncation, and deletion. Most of the p53 mutants are missense mutations that often occur in the DNA-binding domain of the p53 protein. These mutants not only lose their tumor inhibitory activity, but also exert a “dominant-negative” effect on the functions of wt p53. Remarkably, several hotspot mutants, such as mt p53-R175H, G245S, R248W/Q, R249S, R273H/C, and R282W, acquire GOFs to further promote tumor growth via diverse mechanisms (Madrigal et al. in this issue, mt p53 regulation of microRNA expression involves both transcription-dependent and -independent mechanisms. Mt p53 was recently found to associate with replicating chromatin and PARP1 to facilitate aberrant DNA repair (Annor et al. in this issue demonstrated that oligomerization of mt p53-R273H is not required for its chromatin association, though oligomerization of wt p53 is indispensable for its tumor suppressive activity. In a prospective essay by von Grabowiecki et al., the authors proposed a provocative idea that mt p53 might promote endosomal trafficking of a plethora of proteins involved in tumorigenesis and cancer progression by regulating Rab11-FIP1, which is supported by some recent studies as cited in this article and will await further validation.The p53-encoding gene, chanisms . Althougchanisms . As descA repair . An inteCui et al. offered a comprehensive review on the progresses of recent studies on the coordinated regulation of p53 and mTOR pathways in response to the physiological and genotoxic conditions. This is further consolidated by another review by Nagpal and Yuan, who elegantly collected numerous previous and new findings on the role of basally expressed p53 in restraining anabolic metabolism to prevent fast cell proliferation under non-stress conditions. In accordance, the tumor suppressive function of p53 has been also attributed to its activity to regulate glucose metabolism, lipid metabolism, amino acid metabolism, and iron metabolism in cancer cells, which are nicely illustrated in a review by Yu et al. Moreover, p53 has been shown to be involved in the regulation of recycling and clearance of metabolites, nutrients, and cellular debris. This line of information on wt and mt p53’s new functions is systematically reviewed by Rahman et al. They offered another comprehensive and updated view on the roles of wt and mtp53s in the regulation of autophagy signaling and provided new insights into the therapeutic potential by modulating p53-mediated autophagy.Over the past years, growing evidence has revealed the crucial role of p53 in the maintenance of metabolic homeostasis and the prevention of cancer-associated metabolic remodeling . MammaliNagpal and Yuan described that basally expressed p53 is required for the maintenance of immune homeostasis. In addition, Shi and Jiang offered a detailed review on various mechanisms underlying wt and mt p53 regulation of inflammation and immunity. For example, p53 prevents inflammation-associated cancer development by suppressing NF-κB and STAT3 signaling pathways (Zhang et al. showed that p53 mutation is associated the increased production of chemokines, leading to infiltration of different immunocytes in breast cancer. This suggests a complex tumor microenvironment in mt p53-harboring cancers. The development of vaccines targeting p53 has been an old, yet unsolved, topic, as both wt and mtp53 epitopes can be presented on the cell surface for T cell recognition (Zhou et al. offered a thoughtful review on the recent progresses of vaccination of p53 or its peptides and discussed the possibility and application of p53-targeting vaccines to cancer treatment. Collectively, these review and research articles as published in this issue not only show recent progresses in various regulations and roles of wt and mt p53 in cancer development, progression, and immunology, but also provide more new insights into the p53 anti-cancer functions and mt p53’s oncogenic activities. Importantly, these articles also offer new thoughts and suggestions for targeting mt p53 as anti-cancer therapies, such as new potential approaches for developing p53 vaccines.The roles of wt and mt p53s in the regulation of inflammatory and immune responses have been a hot topic recently. To update this area of research, this issue has also collected several review and research articles. For instance, the review article by pathways , while mpathways . Moreoveognition . To updaAlthough the canonical functions of p53 as a key regulator of cell cycle, DNA repair, and apoptosis have been well documented, increasing studies have been continuingly unveiling novel roles of p53 in metabolic remodeling, immune surveillance, and cancer therapy, making this magic molecule as the most attractive research target as well as a promising therapeutic target for developing anti-cancer therapies in the future. We are deeply grateful to these authors who have made great efforts to our better understanding of new functions of p53 and new insights into this still mysterious molecule by contributing their comprehensive review and elegantly-designed research articles to this special issue.


Hidradenitis suppurativa/acne inversa (HS) is a chronic inflammatory disease of the pilosebaceous unit leading to formation of painful, inflammatory nodules, abscesses and tunnels in apocrine gland-bearing areas of the skin. Pain and drainage are the most important symptoms associated with reduction of quality of life in HS. On the other hand, an overlooked symptom in quality of life studies is itch, despite the fact that several studies have reported its importance. Various theories have tried to explain the pathogenesis of itch in HS, such as the presence of mast cells in the cell infiltrates and elevated Ig E levels in the lesional skin. Smoking and advanced stage of disease have been found to be associated with increased intensity of itch. A PUBMED search was conducted to perform a systematic literature review using the term “hidradenitis suppurativa” [all fields], the keywords “pruritus”, “itching”, “itch” [all fields] and with “AND” as operator. Mast cells and mTor signaling were found to be raised in both lesional and perilesional skin. Itch as a presenting symptom has been found in 35–82.6% of patients across multiple studies. It often co-presents with pain and may be misinterpreted as burning, stinging, tickling, tweaking, prickling, etc. The presence of itch is associated with reduced quality of life, depression and impairment of social life. Brodalumab, a monoclonal antibody against IL-17A receptor, produced significant improvements in itch, pain, QoL and depression in patients with moderate to severe HS. Statins have shown some reduction in itch intensity score. Further studies are required to gain a better understanding of the etiopathogenesis and optimal therapeutic modalities for itch in HS that will allow clinicians to better address issue and reduce its impact on quality of life. Hidradenitis suppurativa/acne inversa (HS) is a chronic debilitating inflammatory skin disease of the pilosebaceous unit . It is cThe clinical features reported to most reduce the QoL of HS patients are pain and malodorous purulent discharge from skin tunnels. In a study conducted on 1795 subjects, pain was found in 83.6% cases [A PUBMED search was done for systematic review of literature using the term “hidradenitis suppurativa” [all fields]; the keywords “pruritus”, “itching” and “itch” [all fields]; and “AND” as operator.The search yielded 105 articles; whereas after the removal of duplicates, 48 articles were left. A total of 28 articles were excluded due to eligibility. Finally, 20 articles were selected for the study. The selection of articles was performed independently by SBL and NSS. All articles were reviewed by the senior author, VRS, who also had the final decision in case of disagreement between the initial selectors .The presence of itch in patients with HS ranged from 62.1% to 77.5% in the selected studies ,19,25. TItch in HS has also been found to adversely affect QoL. In a study by Riis et al., a significant association was found between loss of utility and itch . A studyThe exact pathogenesis of itch in HS is not understood. In the studies by the Rotterdam group  has recently been identified as an important mediator of inflammation in HS. It has also been found to be significantly elevated in atopic dermatitis and cutaneous T-cell lymphoma, wherein it is likely to mediate the infiltration of eosinophils, basophils and T cells. Both conditions are also associated with high itch scores ,34.Itch is an important symptom in HS . In seveIn a retrospective data analysis conducted on 145 patients, itch was reported by 82.1% of the patients and correlated with Hurley stage . Vossen In a study evaluating the prodromal symptoms in HS, 20% patients experienced itching at least 24 h before the onset of any visible signs . ThroughEsmann et al.  studied Studies to assess the efficacy of various treatment modalities in HS associated itch are yet to be conducted. Few authors have associated pain with itch, and the alleviation of pain may reduce itch . Many toBrodalumab, a monoclonal antibody against IL-17A receptor, produced significant improvements in itch, pain, QoL and depression in patients with moderate to severe HS . StatinsJAK inhibitors have recently been used in treating HS successfully. INCB54707, a experimental molecule which is 52 times more selective for JAK1 than JAK2, has been tried in two multicentric phase 2 studies, wherein a single dose of 90mg oral treatment resulted in a hidradenitis suppurativa clinical response (HiSCR) (defined as at least a 50% reduction in inflammatory lesion count with no increase in abscesses or draining fistulae compared to baseline) score of 88% at week 8. A dose-dependent significant improvement in hidradenitis suppurativa quality of life scores was documented in response to the JAK1 inhibitor .Recent studies have shown that JAK inhibitors can reduce itch symptoms in patients with atopic dermatitis. It is suggested that JAK inhibitors act by blocking neuronal JAK1 signaling in non-inflammatory settings. JAK inhibition with tofacitinib were tried in five patients with severe chronic inflammatory pruritus off-label; all patients showed reduced itch scores, despite having failed to respond to multiple other treatments. These findings suggest the potential use of JAK inhibitors in alleviating itch symptoms in HS .p = 0.0001) at 24 weeks, which was statistically significant. Improvement in DLQI was significant in 64% of patients. In addition, depression, which was earlier noted to be severe in 11 patients, became nonsevere in 7 of the 11 patients [Studies have demonstrated the efficacy of metformin in treating recalcitrant HS. Metformin at a dose of 500 mg 3 times daily for 24 weeks resulted in an average reduction in Sartorius score by 12.78 (patients . Arun anpatients . The mecpatients .Despite a lack of complete understanding of itch in HS, it is an important aspect of the disease, reported in 62% to 75% of patients. Further study to gain a better understanding of the etiopathogenesis and optimal therapeutic modalities for itch in HS will allow clinicians to better address this aspect of the disease and reduce its impact on quality of life.


SQSTM1/p62 plays a key role in the autophagic machinery and may serve as a marker for autophagic flux in vivo. We investigated the role of p62 in neurodegeneration, analyzing its concentrations in the CSF of AD and FTD patients. We recruited 76 participants: 22 patients with AD, 28 patients with FTD, and 26 controls. CSF p62 concentrations were significantly increased in AD and FTD patients when compared to controls, which persisted after adjusting for age . In female FTD patients, p62 positively correlated with the neurodegenerative biomarkers t-Tau and p-Tau. A significant correlation between CSF p62 concentrations and several clinical features of AD was found. Our data show that p62 is increased in CSF of AD and FTD patients, suggesting a key role of autophagy in these two disorders. The levels of p62 in CSF may reflect an altered autophagic flux, and p62 could represent a potential biomarker of neurodegeneration.Several studies have revealed defects in autophagy in neurodegenerative disorders including Alzheimer’s disease (AD) and frontotemporal dementia (FTD).  Neurodegenerative disorders such as Alzheimer’s disease (AD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), and Parkinson’s disease (PD), are characterized by the progressive deposition of misfolded and aggregated proteins that lose their physiological functions and acquire neurotoxic effects . In receSQSTM1, OPTN, and VCP are directly involved in protein degradation pathways as their products contribute to recruitment of ubiquitinated proteins to the autophagosome, a key component of autophagy [Autophagy is characterized by a complex cellular process that leads to the clearance of misfolded or damaged proteins and dysfunctional organelles . This coutophagy .SQSTM1 gene and involved in several functions, including autophagy, apoptosis, inflammation, and cell survival, suggesting a role as a signaling hub that regulates cell viability in response to cytotoxic stress [SQSTM1 gene were initially described in Paget’s disease of bone [Human p62 is a scaffold protein of 440 amino acids, coded by the c stress ,11. Muta of bone , a chron of bone  and FTD  of bone ,15. In n of bone .Cerebrospinal fluid (CSF) is enriched in brain-derived substances, and CSF biomarkers might represent a valid instrument to assess neuropathology in vivo. At present, however, there are no established CSF biomarkers for monitoring brain autophagy in vivo. In 2017, Au et al. examined p62 concentrations in the CSF of 30 children with severe traumatic brain injury (TBI) and found that p62 concentrations were significantly increased, suggesting an impairment of the autophagic flux .To date, a few studies have investigated CSF p62 concentrations across different neurodegenerative disorders. Thus, the primary aim of this study was to evaluate whether p62 concentrations were detectable in the cerebrospinal fluid of patients with AD or FTD, comparing them with a control population. In addition, we investigated correlations between this autophagy biomarker and clinical or radiological characteristics of AD and FTD.Seventy-six consecutive patients referred to the “Rita Levi Montalcini” Department of Neuroscience of the University Hospital Città della Salute e della Scienza di Torino  were selected for the study. Twenty-two patients with AD  and 28 patients with FTD  were enrolled.GRN, MAPT, SQSTM1 (that encodes p62), and C9orf72 genes.Clinical diagnosis of probable AD was established according to joint Alzheimer’s Association Workgroup and National Institute of Aging guidelines  and the Twenty-six cognitive-spared patients  with neurological conditions other than neurodegenerative disorders were included as a control group, including patients with suspected acute polyneuropathy (n. 7), muscular dystrophy (n. 3), multiple sclerosis (n. 3), and normal pressure hydrocephalus (n. 13). The demographic and clinical features are summarized in g for 10 min at room temperature, subsequently transferred into aliquot tubes, and stored in a −80 °C freezer.CSF sampling and collection were performed according to previously published protocols and guidelines . After oThe CSF p62 levels were evaluated using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (Cloud-Clone Corp). The limit of detection was 0.1 ng/mL for p62, and samples below the level of detection were assigned a value of 0 ng/mL. All samples were assayed in duplicate. CSF concentrations of Aβ42, t-Tau, and p-Tau181 were also evaluated using the ELISA method with available commercially Innotest Kits (Fujirebio), according to the manufacturer’s instructions.p value of <0.05 was considered significant in biochemical and clinical comparisons.All statistical analyses were performed using SPSS version 20 . Continuous variables were presented as means ± standard deviations or medians and range, and categorical ones as count and percentage. We compared diagnostic groups  using ANOVA for normally distributed variables and Kruskal–Wallis and Mann–Whitney U tests for non-normally distributed variables, as appropriate. Correlations between p62 and CSF biomarkers were also evaluated using Pearson’s test, also adjusted for age. The statistical models were adjusted for possible confounders. A two-tailed No difference in education, comorbidities, or smoking habits was found between cases and controls. The gender distribution was comparable in the control and FTD/AD cohorts. Patients with AD and FTD did not differ in age, whereas control subjects were younger than patients with dementia .p = 0.011) (p = 0.01). The p62 concentrations were not different in AD males with respect to male controls, whereas female AD patients showed higher p62 levels when compared to female controls  (p = 0.779).The p62 concentrations were found to be significantly different between AD patients and controls  , which p= 0.044) . No signp < 0.001) (p = 0.008). When analyzing the gender effect, p62 concentrations were found to be significantly different between male FTD patients and male controls , and between female FTD patients and female controls .Crude p62 levels were significantly increased in FTD patients when compared to controls  , which pp = 0.002) and p-Tau in the female FTD subgroup alone   .p = 0.041) (p = 0.033), and depression . Furthermore, p62 concentrations were negatively correlated with temporal atrophy in the AD subgroup , mainly in the male subgroup . No correlation between p62 levels and MMSE was found  a. In AD as found b. FinallNo correlations between p62 levels and the collected clinical characteristics of FTD patients were found. In particular, no correlation between p62 CSF concentration and neuropsychological test results was found . In addition, we did not find any correlations between p62 levels and severity of the disease.SQSTM1 gene; we only found the single-nucleotide polymorphism rs186996560 in the SQSTM1 in a FTD patient. In our FTD group, p62 levels were elevated in patients both carrying and not carrying mutations in known causative genes in respect to controls. No significant correlation was found between p62 levels and the presence or absence of gene mutations (p = 0.317).In this cohort of patients, we did not find any mutations in the To our knowledge, this is the first study to investigate CSF p62 concentrations as marker of autophagy in patients with dementia. We showed that CSF p62 concentrations were significantly increased both in AD and FTD patients in comparison to controls. Furthermore, in female patients with FTD, we found that CSF p62 concentrations correlated with markers of neurodegeneration like p-Tau and t-Tau. In patients with AD, several neuropsychological characteristics of the disease showed a significant correlation with the investigated biomarker of autophagy. Taken together, our data support the hypothesis that CSF p62 concentrations may be a useful in vivo biomarker of neuronal autophagy.Increased CSF p62 concentrations in patients with neurodegenerative disorders may have several neurobiological explanations. First, p62 is involved in several cellular processes relevant to aging, including protein degradation , mitophaThe different forms of frontotemporal dementia are characterized by the aggregation of insoluble proteins within cells. The accumulation of aggregates is often due to genetic mutations in their coding genes. Most inclusions in FTD are characterized by immuno-reactivity, and the neurons and glia of FTD patients are characterized by p62 accumulations.MAPT gene mutation. In addition, we found that that CSF p62 levels correlate with biomarkers of neurodegeneration in FTD, and p62 concentrations were positively correlated with t-Tau and p-Tau. Many studies have shown that autophagy deficiency can cause tau accumulation [It has been shown that p62 represents a substrate for macroautophagy; hence, p62 levels increase when macroautophagy is inhibited, and its concentration decreases when macroautophagy is induced . In thismulation ,33. Incrmulation . In mousmulation . In a remulation . P62 mayThis is the first study to demonstrate an increase in p62 concentrations in the CSF of patients with FTD and AD; in view of the limited samples, the results obtained and their conclusions should be interpreted with caution, and further studies are needed to confirm our data. Autophagy represents a dynamic process, and our results may change according to different stages of neurodegeneration, so studies involving subjects at different stages of disease are desirable. Finally, a further investigation of other different autophagic biomarkers may be crucial for a better understanding of the autophagic machinery in neurodegenerative disorders.Our study showed a significant increase of CSF p62 concentrations in patients with Alzheimer’s disease and frontotemporal dementia, supporting an important role of autophagy alterations in these neurodegenerative conditions. In addition, a significant correlation between increased autophagy and clinical characteristics of dementia was found. However, due to the relatively small number of subjects examined, replication studies in larger series are warranted in order to confirm our data.


Sexuality is a growing field in the context of the management of chronic diseases and cancer in particular. Cancer treatments and the traumatic nature of the cancer experience frequently elicit considerable sexual difficulties.To assess the prevalence of sexual dysfunction (SD) in patients with cancer, and to determine the associated factors.This was a cross-sectional study, conducted over 1 month, involving 100 cancer patients followed in the oncology department at the Habib Bourguiba University Hospital in Sfax (Tunisia). General, clinical and therapeutic data were collected using a pre-established questionnaire. Sexual function was assessed with the “Female sexual Function Index” and the “International Index of Erectile Function”.These results showed that half of the patients were female, and 70% of them were married. Their mean age was 51.96, and 68% of them were unemployed. Unemployment in men and treatment with chemotherapy were statistically associated with erectile dysfunction (p=0.049 and p= 0.001 respectively). treatment with radiotherapy was statistically associated with decreased desire in men (p=0.048). Depression correlated with a decreased orgasm (p=0.032) and erectile dysfunction (p=0.043) in men, mean score of IIEF (p= 0.019) and with a decreased sexual arousal (p=0.006) in women.Sexual dysfunction is common in cancer patients. They can be of iatrogenic or psychological origin and can depend on the dynamics of the couple relationship. Training to raise awareness of the importance of sexuality first among cancer patients should be considered given the lack of communication between doctors and patients regarding sexuality issues.No significant relationships.


Laryngeal Squamous Cell Carcinoma (LSCC) is a squamous cancer with 2.4% new diagnoses each year, accounting for 25% of head and neck cancers, and has a high mortality rate. A deeper understanding of key mechanisms and knowledge of the putative target molecules of theranostic relevance are required. The receptor for neurotrophins p75NTR has been shown to be highly expressed in cancer stem cells (CSCs) of squamous epithelia, in LSCC as well as in other cancers. However, whether its cleavage product p75ICD expression, known to finely regulate the survival/death balance in neurons, is also a master regulator in cancer and LSCC in particular has not been directly addressed so far. To resolve this question, we performed a preliminary study using a limited number of LSCC specimens and studied p75ICD presence and expression pattern in LSCC specimens, showing that p75ICD may be a promising target in LSCC, requiring further investigation.We investigated the p75 Neurotrophin Receptor (p75NTR) expression and cleavage product p75NTR Intracellular Domain (p75ICD) as potential oncogenic and metastatic markers in human Laryngeal Squamous Cell Carcinoma (LSCC). p75NTR is highly expressed in Cancer Stem Cells (CSCs) of the laryngeal epithelia and it has been proposed as a marker for stemness, cell migration, and chemo-resistance in different squamous carcinomas. To investigate the clinical significance of p75NTR cleavage products in solid tumors, full-length and cleaved p75NTR expression was analyzed in laryngeal primary tumors from different-stage LSCC patients, diagnosed at the Policlinico Umberto I Hospital. Molecular and histological techniques were used to detect the expressions of p75NTR and p75ICD, and ATP Binding Cassette Subfamily G Member 2 (ABCG2), a CSC marker. We found regulated p75NTR cleavage during squamous epithelial tumor progression and tissue invasion. Our preliminary investigation suggests p75ICD expression and localization as possible features of tumorigenesis and metastaticity. Its co-localization with ABCG2 in squamous cells in the parenchyma invaded by the tumor formation allows us to hypothesize p75NTR and p75ICD roles in tumor invasion and CSC spreading in LSCC patients. These data might represent a starting point for a comprehensive analysis of p75NTR cleavage and of its clinical relevance as a potential molecular LSCC signature, possibly helping diagnosis, and improving prognosis and personalized therapy. Head and Neck Squamous Cell Carcinoma (HNSCC) is the sixth-most-common cancer worldwide, and Laryngeal Squamous Cell Carcinoma (LSCC) accounts for approximately 10% of all HNSCCs [Despite recent progress in the treatment of HNSCC, including surgery, radiotherapy, and medication-based therapy , a major impact on the survival rate is still lacking and it has remained substantially unchanged over the last 30 years. The general survival rate for LSCC is 61% at 5 years, but, analyzing by tumor stage, the cure rates increase up to 80–90% in case of limited disease T1 and T2). In case of local advanced diseases (T3 and T4) or regional lymph node involvement, the survival rate is <50% at 5 years [. In caseIn line with this, there is an unmet medical need for innovative treatment strategies, which are only possible through the investigation of the molecular mechanisms underlying LSCC development and progression, and the discovery of targetable signaling. In particular, characterization at the cellular and molecular levels of chemotherapy-resistance and invasion key players will be critical for the design of new drugs, as well as for the improvement of the clinical management of LSCC patients.Accumulating evidence over the last decade pinpointed the p75 Neurotrophin Receptor (p75NTR) as a potential malignant marker in cancer. For example, p75NTR has been implicated in melanoma ,4,5,6, aHowever, the described biological significance of p75NTR modulation may vary in different types of tumors, as it exerts malignant effects, such as the invasion depth of the cancerous tissue and lymph node metastasis in esophageal cancer . On the In prostatic tissues, p75NTR is absent in hyperplastic benign epithelial cells, and it is specifically overexpressed in prostatic cancer cells, where it correlates with high-risk tumors with an unfavorable prognosis (Gleason score > 7) . MoreoveOf interest for the present study, p75NTR may undergo regulated cleavage, generating intracellular fragments able to translocate to the nucleus harboring specific signaling activity, as already demonstrated for the pro-apoptotic activity of the p75NTR Intracellular Domain (p75ICD), released upon the γ- and α- secretases’ sequential cleavages of the p75NTR in neurodegenerating neurons of Alzheimer’s Disease-affected brains .p75NTR-regulated cleavage has been hypothesized to also occur in some cancer types and in tumor cellular models. For example, p75NTR enzymatic maturation has been shown to be critical for glioma invasion and spreading .However, direct proof of the regulated cleavage of p75NTR leading to the generation of p75ICD in cancer cells is still lacking in most malignant tumors, including LSCC.Here, we used the specific anti-p75ICD antibody, which does not cross react with the p75NTR Full Length (p75NTR FL) molecule, to investigate its expression and localization in primitive LSCC and its nodal metastasis in a small group of patients representative of the diverse stages of LSCC.Furthermore, since p75NTR is also present in Cancer Stem Cells (CSCs) and it is co-expressed with stemness markers, such as CD133 in LSCC , and witThe present study firstly detected the expression pattern of p75ICD and compared it to the p75NTR FL in LSCC, and revealed that p75ICD was particularly upregulated in neoplastic squamous cells expressing ABCG2 and was strongly associated with tumor progression. Moreover, p75ICD expression was cytosolic in stages II-III and became nuclear in neoplastic epithelia and in ABCG2-positive CSCs.Although these results are arising from pilot study, and therefore need to be further explored in a larger patient group, our findings indicate that p75ICD generation by p75NTR-regulated cleavage might represent a relevant marker of tumor invasion and metastasis in LSCC, and support a role for p75NTR/p75ICD in the regulation of CSC proliferation.We evaluated laryngeal specimens from LSCC surgically removed at the Otolaryngology Unit, DAI Testa-Collo, Policlinico Umberto I University Hospital, 00161 Rome, Italy, from April 2016 to April 2018. All patients included in the study were affected by LSCC and subjected to open partial horizontal laryngectomy or total laryngectomy. Specimens were processed and evaluated at the Section of Pathology of the same hospital, where a definitive histological diagnosis was provided.The inclusion criterion was confirmed laryngeal cancer treated with surgical therapy. The exclusion criteria were previous radiation therapy on the head and neck region and neoadjuvant therapy. All patients gave written informed consent for inclusion in the study. The use of these clinical materials was approved by the institutional ethics committees (RIF. 6129).The stage of LSCC was classified according to the 8th edition of the American Joint Committee on Cancer . Relapse-free survival was defined as the interval between the date of surgery and the date of recurrence or last follow-up available.The study was performed on a total of 8 LSCC patients . In 3). Different tumor characteristics were evaluated, including the location , involved side , size, pattern of growth (exophytic or endophytic), ulceration, depth of invasion, spread to extralaringeal tissues, color, and features of the mucosa near the neoplasia. Data regarding the presence/absence of neo-plastic residual on resection margins was evaluated through ink application. Different sections of each tumor were sampled and collected so as to be representative of the maximum tumor invasion and of the interface between the tumor and adjacent normal mucosa. The pathological evaluation also included the characteristics of the lining epithelium of the mucosa adjacent to the tumor . All the lymph nodes that could be found from every specimen were also sampled for histological evaluation. Four-micrometer-thick tissue sections were cut from each paraffin block and used for hematoxylin–eosin staining  . Each spn = 1), stage III (n = 1), stage IVa (n = 1), and stage IVb (n = 1) LSCC samples. Antigen retrieval was achieved by using sodium citrate buffer (pH 6.0) for 30 min at 37 °C on deparaffined sections. The slides were then cooled to Room Temperature (RT) and washed three times with phosphate-buffered saline (PBS) solution. For autofluorescence quenching, the slides were incubated with ammonium chloride  for 30 min, and then blocked with 10% normal donkey serum . The overnight incubation (4 °C) with primary antibodies  was followed by incubation with the appropriate combination of secondary antibodies. In particular, highly cross-adsorbed donkey anti-mouse-546 , donkey anti-rabbit-546 , and donkey anti-rat-488  were used for double immunofluorescence (IF) . Nuclei were counterstained with 1 µg/mL 4′,6-diamidino-2-phenylindole .Four surgical specimens of the larynx  were selSlides were coverslipped using ProLong Diamond mounting medium with anti-fading . Slide imaging was performed with the laser-scanning confocal microscope (Olympus FV1200) using a 20× air and a 40× (NA = 1.25) oil immersion lens, and visualized with FV10-ASW software . An UV diode laser operating at 405 nm, an Argon laser at 488 nm, and a HeNe laser at 543 nm were used as excitation sources. To entirely reconstruct the longitudinal tumor tissue, 14 confocal Z-stacks (1024 × 1024 pixels) were collected at 0.29 µm intervals in a 4 µm total optical depth, and then converted into max projection images. Omission of the primary antibody was routinely performed as a control for antibody specificity.Images for direct comparison were collected using the same parameters and were analyzed with the help of FIJI open-source software .In order to compare the expressions of p75ICD and p75NTR in the resected specimens at different stages, they were evaluated by means of a semi-quantitative immunohistochemical assessment based on the intensity of immunoreactivity and its distribution ,19. The For Western Blot (WB) samples, 50 µg of protein was separated on precast 4–12% gel. At the end of electrophoresis, the proteins were stained with Ponceau Solution. Non-specific protein-binding sites on the nitrocellulose blots were blocked with PBS containing 7% non-fat milk (PBS–milk) for 1 h at RT. The primary antibodies were diluted in PBS at the following dilutions: 1:1000 for p75NTR C-terminal, 1:200 for ABCG2 (BXP-53: sc-58224), and 1:1000 for GAPDH antibodies. The incubation of the nitrocellulose blots with these antibodies was performed overnight (4 °C) at RT. The blots were then washed three times with PBS and incubated for 4 h at RT with the appropriate peroxidase-conjugated antibodies diluted 1:1000 in PBS–milk. Following PBS washes, the peroxidase activity of the nitrocellulose-bound secondary antibodies was detected with the Clarity West-ern ECL chemiluminescent reagents (BioRad Laboratories), imaged by Chemidoc XRS+ Image System (BioRad Laboratories), and bands were quantified by gel densitometry using the Fiji (ImageJ 1.53c) software . All the whole Western blot figures can be found in the We analyzed the p75NTR FL level and cleavage by using a C-terminal-specific anti-p75NTR antibody able to recognize both the p75ICD and the p75NTR FL by a biochemical assay. The p75NTR FL level was found to be detectable and high in all LSCC cases, while it was almost undetectable in the paratumoral tissue . MoreoveThe ABCG2 expression was also investigated in the same samples to compare the p75NTR and CSC marker expression levels in the different LSCC stages, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a control protein for normalization. ABCG2 is a family member of the ABC transporter, and its positive expression in laryngeal carcinoma has been related to tumor differentiation, age, sex, and presence of loco-regional lymph node metastasis. It has been previously suggested that the abnormal expression of ABCG2 may be related to the invasion and metastasis of laryngeal carcinoma, thus providing a good reference basis for the evaluation of LSCC invasivity .The analysis of the tumoral marker in stages III and IV, with  and without  lymph nodes metastases, confirmed that LSCC cancer showed the maximal ABCG2 expression detectable by WB at stage III and further increased at stage IVa just before metastatization, being downregulated in the more advanced stages (IVb) as compared with previous tumoral stages.Interestingly, initial p75ICD positivity could be observed at stage IVa, corresponding to the highest ABCG2 level. p75ICD was still high with an apparent slight increase at stage IVb of LSCC tumors.As mentioned in In line with the histological characteristics of LSCC tissue , p75ICD-p75ICD was weak in stage II, while a progressive increase in the number of p75ICD-positive cells was noticeable from stages III to IVb, where a further increase in the single cell expression level was found in cells with enlarged nuclei, most of them also positive for the ABCG2 cancer marker. p75ICD was co-expressed by ABCG2-positive cells from stage II onward, while p75ICD/ABCG2 co-localization could not be detected in stage II stromal cells.Indeed, the p75ICD staining could be cytosolic, as well as nuclear, while ABCG2 staining was mainly cytosolic and punctuate, in line with its expression in extracellular vesicles , aberrant epithelium (2), the carcinoma invasive front (3), and carcinoma in situ (4) .In As reported in Zones 3–4, p75ICD expression was prominent in cancer cells with a highly dysmorphic appearance and enlarged nuclei, and was more evident in Zone 4 as compared with Zone 3.Note that not all of the p75ICD positive cells with aberrant morphology also expressed ABCG2.Representative pictures of the co-localization of the p75NTR (red) and ABCG2 (green) in Zones 1–4 from a stage III LSCC patient are shown in p75NTR FL and its C-terminal fragment p75ICD showed not only differential expression levels, but also distinctive intracellular localization. In fact, while the p75NTR FL was found to be mainly cytosolic a, the p7Interestingly, the p75ICD staining was also found to be nuclear in stage II epithelial cells b.The present pilot study investigated the occurrence and, eventually, the pathological significance of the p75NTR-regulated cleavage by α- and γ-secretases, and the consequent release of the intracellular p75ICD fragment in LSCC. In particular, the study addressed the question of whether the distribution of p75ICD fragments might differ from the p75NTR FL, the expression of which has been largely described in several cancer types ,14,16, aNGF and its receptors (TrkA and p75NTR) are implicated in cancer growth, CSC proliferation, and immune system evasion . p75NTR Here, we used the specific anti-p75ICD antibody that does not cross react with the p75NTR FL molecule to investigate its expression and localization in primitive LSCC and its nodal metastasis in a small group of patients representative of the diverse stages of LSCC. Furthermore, since p75NTR is also present in CSCs and is co-expressed with stemness markers, such as CD133 in LSCC , and witTo resolve this lack of information, we first asked whether p75NTR undergoes regulated cleavage also in squamous cancers, such as LSCC, as it has been extensively reported in brain neurodegeneration and gliomas . We founIn fact, most of the findings reported by the literature do not allow discriminating between the two forms of the p75NTR, lacking the critical knowledge to target the p75NTR signaling for LSCC therapy. A deeper understanding of the opposite p75NTR and p75ICD-mediated biological functions could also help in explaining some apparently contradictory effects of p75NTR in cancers. In fact, behind the cancer-specific developmental mechanisms, p75NTR has been reported to be pro-apoptotic in gastric cancer  and prolIn order to specifically investigate p75ICD’s biological action by immunofluorescent labeling in LSCC specimens, we resorted to a validated and specific anti-p75ICD antibody , and a cThus, we performed double IF staining for p75NTR/ABCG2 and p75ICD/ABCG2, and scored immunopositivity grading by the semiquantitative assessment method based on weak (+), medium (++), and strong (+++) immunolabeling .First of all, we observed that the two molecules had distinct patterns of expression  in the fThen, we investigated p75ICD expression at different LSCC stages (II-IV) and found a progressive increase in the number and intensity of p75ICD-immunolabeled cells from stage II toward stage IV ; Table 2Additionally, we explored p75ICD expression and localization in the four different zones of the single LSCC tumor specimen ; Table 3Additionally, we studied the p75NTR level and distribution in adjacent sections and found that p75NTR expression increased in aberrant , Zone 2 These observations are partially in line with the study by Li and collaborators, in which the immunohistochemical localization of p75NTR was reported in normal, paratumoral mucosa, and LSCC without any significant difference in the p75NTR level between dysplastic epithelia and LSCC. Unlike the specific antibodies used in this study and directed against the extracellular domain (p75NTR FL) and, specifically, the ICD domain (p75ICD) of the p75NTR, Li’s study reported data obtained using an anti p75NTR C-terminal antibody, which virtually recognized p75NTR and all of its fragments, including the p75ICD . This fact might account for the difference in the trend and distribution, and also for the lack of correlation between p75NTR expression, histopathologic grading, and lymph node metastases. The same authors observed that “with the deterioration of differentiation, the ratio of p75NTR-positive cell increased partly, but the coloration intensity of each tumor cell degraded, with very faint cytoplasm coloration of p75NTR” . Since wIn accordance, we found that the p75ICD pattern of expression in the different LSCC zones correlated with ABCG2 expression better than the p75NTR profile, as reported in Furthermore, p75ICD vastly co-localized with ABCG2-positive cells, some expressing low-intensity ABCG2 immunolabeling, and others morphologically abnormal with both intracellular and extracellular ABCG2 staining . Note thBased on the critical role of p75ICD nuclear translocation for its functions in the central nervous system, the subcellular localization of p75ICD (cytosolic versus nuclear) was also assessed in the resected specimen from LSCC stages II-IV. In agreement with the augmented nuclear p75ICD staining reported in neurodegenerating neurons of Alzheimer’s Disease patients, we found prevalent p75ICD nuclear localization in aberrant epithelia from LSCC tumor stage II b, as welThe p75NTR has been demonstrated to be a central regulator of cancer invasion in highly invasive glioma , and itsTherefore, our study showing that the p75NTR and expression patterns differ in a stage-specific and zone-related manner, and that p75ICD is co-localized with the metastatic marker ABCG2 and shows a prevalent nuclear expression in aberrant and highly dysmorphic cancer cells, led us propose p75ICD as a good candidate prognostic factor in LSCC. Moreover, the presence of p75ICD in ABCG2-positive cancer cells surrounded by putative ABCG2 positive vesicles (as visible in Larger studies or meta-analyses will eventually help to reach sufficient statistical power to assess the utility of p75ICD as a prognostic factor in LSCC.Of note, p75NTR expression has been suggested by others to be prognostic when in combination with the marker CD44 in esophageal, hypopharyngeal, and oral carcinomas, as well as in HNSCC . DespiteAn in-depth study and revelation of the molecular mechanisms controlling normal epithelia and those events underlying its transformation toward metastatic SCC is of foremost importance in the quest for novel biomarkers improving early diagnosis and personalized formulation in the clinical treatment of LSCC.Although these results arose from a pilot study, and therefore need to be further explored in a larger patient group, our findings indicate that p75ICD generation by p75NTR-regulated cleavage might represent a relevant marker of tumor invasion and metastasis in LSCC, also supporting a role for p75NTR/p75ICD in the regulation of CSC proliferation.p75NTR was implicated in the control of the apoptotic machinery in neuronal cells and was indicated as a good pathological marker for the progression and spreading of both central nervous system cancers and non-neuronal carcinoma, such as LSCC. Although the exact mechanism is not clear, it is highly likely that p75NTR may predispose tumor cells to perineural invasion, conferring responsivity to NGF-producing Schwann cells and neurons, as reported in oral cancer and pancreatic cancer ,33. FurtOverall, it is advisable to foster our knowledge on p75NTR cleavage control at different stages of tumor development and specifically investigate p75ICD as a novel prognostic factor possibly improving diagnosis and instructing personalized treatment in LSCC, as well as other squamous cancers.


Literature suggests that health professionals (HPs) can play an important role in promoting physical activity (PA) in cancer patients . This study had three purposes: (1) analyzing HPs' knowledge and opinion about PA for cancer patients; (2) examining practices, barriers and roles that they identify when considering PA promotion in their patients; (3) determining factors that may enhance practices in this field.This study was designed in two phases and took place in Wallonia, Belgium. The first phase was qualitative: 12 HPs were interviewed about their experiences with PA in cancer patients. Then, in the second (and quantitative) phase, an online survey was created based upon the results of the first phase and upon the relevant literature. We sent this second-phase survey to professional organizations, hospitals and medical health centers and received 68 responses.In both phases of the study, HPs seemed to be aware of the benefits of PA for cancer patients and considered that they have to play a role in promoting PA in that specific population. However, only 25% (n = 17) of HPs were familiar with the official PA guidelines. According to our results, HPs discuss the topic of PA with more than 7 out of 10 patients (7.11 ± 2.61). Lack of time and lack of knowledge about PA were identified as barriers to PA promotion. Finally, other relationships that may help to explain the role of HPs in PA promotion were uncovered: clinicians who used more information (e.g. PA benefits) (p = >0.001), exercised referrals (p = 0.012), and prescribed PA (p = 0.007) had higher intervention rates.Our findings suggest that improving HPs' knowledge about PA guidelines and how it can be proposed to cancer patients could help them to promote PA and to encourage their patients to consult a PA specialist, and eventually adopt a physically active lifestyle.


Tp53 gene. These mutations cause a disturbance in cellular response to stress, and eventually, cancer development. Apart from the full-length p53, at least twelve isoforms of p53 have been characterized. They are able to modulate p53 activity under stress conditions. In 2020, almost a half of million people around the world were diagnosed with renal cancer. One genetic disturbance which is linked to the most common type of kidney cancer, renal cell carcinoma, RCC, occurs from mutations in the VHL gene. Recent data has revealed that the VHL protein is needed to fully activate p53. Disturbance of the interplay between p53 and VHL seems to explain the lack of efficient response to chemotherapy in RCC. Moreover, it has been observed that changes in the expression of p53 isoforms are associated with different stages of RCC and overall survival. Thus, herein, an attempt was made to answer the question whether p53 and its isoforms are important factors in the development of RCC on the one hand, and in positive response to anti-RCC therapy on the other hand.p53 is a transcription al factor responsible for the maintenance of cellular homeostasis. It has been shown that more than 50% of tumors are connected with mutations in the  Consequently, kidney cancer is the 16th most common cancer globally . Kidney cancer  is one of the most common tumors, aside from bladder cancer of the urinary system . In 2020globally A. The stglobally B. In 202There are several types of kidney cancer. The most common type, however, is renal cell carcinoma (RCC) which arises from renal tubular epithelial cells, and it accounts for approximately 90–95% of all kidney cancer cases ,2. The mVHL gene result in the deregulation of expression of the hypoxia-inducible factor (HIF), which plays a crucial role as a transcription factor in oxygen regulation . M. MTp53 gTp53 gene/p53 protein (Tp53/p53) observed in renal cancer? Surprisingly, the level of mutations in Tp53/p53 seems to be quite low in kidney cancer compared to other cancers. According to the Catalogue Of Somatic Mutations In Cancer, COSMIC, those mutations are observed in 8.75% of total mutated samples from kidney  treatment has been varied [Are p53 isoform expression patterns associated with renal cancer development? Earlier studies using tissue samples of 45 RCC patients at different tumor stages and 25 non-neoplastic tissues have shown that expression of almost all p53 isoforms changed during cancer development and progression . An incrn varied . Thus, tn varied .More recently, it has been shown that p53β is associated with better recurrence-free survival, RFS (the length of time after primary treatment for a cancer ends that the patient survives without any signs or symptoms of that cancer), and overall survival . AnalysiIt emerged that changes in p53 isoforms are somehow associated with different stages of RCC; in particular, a high level of p53β seems to be an important molecular indicator of better prognosis for patients. Analyses of p53 isoform patterns in RCC patients could help predict how cancer will progress and what treatment is potentially effective.Tp53 gene is infrequently mutated in RCC, the presence of a mutated version of p53 influences clinical prognosis and overall survival [It has been shown that the crosstalk between p53 and VHL is a pivotal element in the DNA-damage response mediated by p53 in RCC . VHL-delsurvival ,39,40,41survival . It seemsurvival ,65. The survival , and by survival , which csurvival ,63,65. A


The larvae of green lacewings and brown lacewings are called ‘aphidlions’, as they consume aphids. They play also an economic role as biological pest control. Aphidlions have, mostly, elongated spindle-shaped bodies, and similarly to most lacewing larvae, they possess a pair of venom-injecting compound jaws, also called stylets. Fossils that have been interpreted as aphidlions are known from amber of different ages . In this study, new aphidlion-like larvae are reported from about 100 million-year-old amber from Myanmar and about 35 million-year-old Baltic amber. The shapes of head and stylets were compared between the different time slices. With the newly described fossils and specimens from the literature, a total of 361 specimens could be included in the analysis: 78 fossil larvae, 188 extant larvae of brown lacewings, and 95 extant larvae of green lacewings. The results indicate that the diversity of head shapes stays about the same over time besides a certain increase in diversity of the head shapes in brown lacewing larvae. In certain other lacewings, a distinct decrease in the diversity of head shapes was observed in the larvae.Aphidlions are larvae of certain lacewings (Neuroptera), and more precisely larvae of the groups Chrysopidae, green lacewings, and Hemerobiidae, brown lacewings. The name ‘aphidlion’ originates from their ecological function as specialised predators of aphids. Accordingly, they also play an economic role as biological pest control. Aphidlions have, mostly, elongated spindle-shaped bodies, and similarly to most lacewing larvae they are equipped with a pair of venom-injecting stylets. Fossils interpreted as aphidlions are known to be preserved in amber from the Cretaceous (130 and 100 million years ago), the Eocene (about 35 million years ago) and the Miocene (about 15 million years ago) ages. In this study, new aphidlion-like larvae are reported from Cretaceous amber from Myanmar (about 100 million years old) and Eocene Baltic amber. The shapes of head and stylets were compared between the different time slices. With the newly described fossils and specimens from the literature, a total of 361 specimens could be included in the analysis: 70 specimens from the Cretaceous, 5 from the Eocene, 3 from the Miocene, 188 extant larvae of Chrysopidae, and 95 extant larvae of Hemerobiidae. The results indicate that the diversity of head shapes remains largely unchanged over time, yet there is a certain increase in the diversity of head shapes in the larvae of Hemerobiidae. In certain other groups of Neuroptera, a distinct decrease in the diversity of head shapes in larval stages was observed. The group Holometabola, with well-known representatives, such as beetles, bees, butterflies, or flies, represents a large share of animal biodiversity, especially in terrestrial ecosystems. Moreover, some smaller lineages are part of this diversity, such as Neuroptera, the group of lacewings. Neuroptera is generally understood as having been part of the early radiation of Holometabola, and as more diverse and species-rich in the past ,2,3. ForMany holometabolans spend most of their lifetime in their larval stages; this is also true for many lacewings. Hence, a supposed loss of biodiversity within a holometabolan ingroup should also be expected to manifest in a loss of the morphological and ecological diversity of larvae. Indeed, the Cretaceous has provided numerous lacewing larvae with peculiar morphologies, clearly not represented in the extant fauna ,18,19,20Furthermore, a loss of diversity was demonstrated with quantitative measures of morphology for the larvae of silky lacewings  and of A type of lacewing larva that is quite common in Cretaceous amber is that of the aphidlion. The term is used for extant larvae of the two groups Chrysopidae (green lacewings) and Hemerobiidae (brown lacewings). The name ‘aphidlion’ refers to the habit of larvae to consume aphids ,27. HencOlder studies still indicated a sister-group relationship between Chrysopidae and Hemerobiidae, mostly based on morphological characters e.g., ,31), but, but31])The mouthparts of aphidlions differ from those of the more commonly known lacewing larvae, such as those of antlions, in being simpler. Similarly to other lacewings, each upper jaw (mandible) and lower jaw (maxilla) form a stylet . These sChrysopa slossonae can sneak across ants guarding their prey, as they use the wax of their pre as part of the camouflaging cloak [Many larvae of Chrysopidae camouflage themselves by attaching different objects to their back. This camouflaging cloak or package can be composed of rather different objects, including the remains of consumed prey, snail shells , or planng cloak . Fossils of aphidlions with a very modern appearance are known from ambers from the Miocene ,49,50 anWe here summarise all known occurrences of aphidlions and aphidlion-like lacewing larvae in the fossil record and report numerous new specimens from different ambers. Similar to earlier studies, we compare the morphological diversity of the heads of these larvae over time in order to recognise possible decreases of diversity.Data for the analysis originated from different sources. Numerous specimens were based on images from the literature or databases (image repositories). Other specimens were directly inspected and documented. Extant specimens of aphidlions came from two zoological collections: the Zoologische Staatsammlung München (ZSM) and the Centrum für Naturkunde Hamburg (CeNak), Leibniz-Institut zur Analyse des Biodiversitätswandels (LIB). Fossil specimens, all preserved in amber, came from various collections: the Palaeo-Evo-Devo Research Group Collection of Arthropods, Ludwig-Maximilians-Universität München (PED), the Senckenberg Forschungsinstitut und Naturmuseum, Frankfurt/Main (SMF Be), and collections of some of the authors, namely C.G. (CCGG), Ch.H. + H.-W.H. (CCHH), P.M. (BUB), T.W. (Weiterschan BuB), and J.W. (F xxx BU CJW) . Polarisers were placed on flashes and in front of the lens to provide cross-polarised light ,58.All images are composite images ; a stackOutlines of all head capsules and stylets were redrawn in Adobe Illustrator CS2 or Inkscape. The more accessible half of the head capsule and the more accessible stylet were outlined. The stylet was artificially rotated forward so that the tip was in a straight line with the proximal insertion on the head capsule. Afterwards, the drawn half of the head was mirrored.In cases in which the head was partly concealed by the thorax (the head was retracted) we only considered the visible or free part. This area reflects the functional part of the head, but also considers that for cases of fossils only the outer visible part can be considered. For many  extant specimens, often the entire head is additionally available, either in the literature or on cleared mounted specimens. Yet, to have an even dataset we did not consider this type of information, as it was not available for some of the other specimens.The analysis was performed in SHAPE. The procedure was outlined in . The app(1)Specimen 4819 (BUB 3060) is preserved in Myanmar amber. It is accessible in a dorsal B,C and v(2)Specimen 4821 (BUB 3066) is preserved in Myanmar amber. It is accessible in a ventral view A,B. No c(3)Specimen 4822 (BUB 3347) is preserved in Myanmar amber. It is accessible in a dorsal B,C and v(4)Specimen 4823 (BUB 3358) is preserved in Myanmar amber. It is accessible in a dorsal A and ven(5)Specimen 4824 (BUB 3359) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(6)Specimen 4820 (BUB 3361) is preserved in Myanmar amber. It is accessible in a dorsal C and ven(7)Specimen 4825 (BUB 3379) is preserved in Myanmar amber. It is accessible in a dorsal C and ven(8)Specimen 4826 (BUB 3393) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(9)Specimen 4827 (F 3196 BU CJW) is preserved in Myanmar amber. It is accessible in a dorsal A,B and l(10)Specimen 4829 (PED 0038) is preserved in Myanmar amber. It is accessible in a dorsal E,F and v(11)Specimen 4828 (PED 0034) is preserved in Myanmar amber. It is accessible in a dorsal C and ven(12)Specimen 4830 (PED 0065) is preserved in Myanmar amber. It is only accessible in a lateral view A–C, henc(13)Specimen 4832 (PED 0248) is preserved in Myanmar amber. It is accessible in a dorsal E,F and v(14)Specimen 4831 (PED 0149) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(15)Specimen 4833 (PED 0251) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(16)Specimen 4834 (PED 0252) is preserved in Myanmar amber. It is accessible in a dorsal to dorso-lateral A,B and v(17)Specimen 4835 (PED 0253) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(18)Specimen 4836 (PED 0315) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(19)Specimen 4837 (PED 0323) is preserved in Myanmar amber. It is accessible in a dorsal E,F and v(20)Specimen 4838 (PED 0330) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(21)Specimen 4839 (PED 0375) is preserved in Myanmar amber. It is accessible in a dorsal D and ven(22)Specimen 4840 (PED 0427) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(23)Specimen 4841 (PED 0433) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(24)Specimen 4842 (PED 0441) is preserved in Myanmar amber. It is accessible in a dorsal C and ven(25)Specimen 4843 (PED 0455) is preserved in Myanmar amber. It is accessible in a dorsal B,C and v(26)Specimen 4844 (PED 0518) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(27)Specimen 4845 (PED 0541) is preserved in Myanmar amber. It is accessible in dorsal C and ven(28)Specimen 4846 (PED 0580) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(29)Specimen 4848 (PED 0642) is preserved in Myanmar amber. It is accessible in a dorsal A,B, vent(30)Specimen 4849 (PED 0666) is preserved in Myanmar amber. It is accessible in a dorsal B,C and v(31)Specimen 4850 (PED 0667) is preserved in Myanmar amber. It is only accessible in dorsal view A,B. No c(32)Specimen 4851 (PED 0696) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(33)Specimen 4852 (PED 0715) is preserved in Myanmar amber. It is accessible in a dorsal F and ven(34)Specimen 4854 (PED 0782) is preserved in Myanmar amber. It is only accessible in dorsal view A,B. No c(35)Specimen 4855 (PED 0793) is preserved in Myanmar amber. It is accessible in a dorsal C,D and v(36)Specimen 4856 (PED 0807) is preserved in Myanmar amber. It is accessible in a dorsal H and ven(37)Specimen 4853 (PED 0754) is preserved in Myanmar amber. It is only accessible in dorsal view F,G. No c(38)Specimen 4857 (PED 0837) is preserved in Myanmar amber. It is accessible in a ventral A,B and a(39)Specimen 4858 (PED 0901) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(40)Specimen 4859 (PED 0952) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(41)Specimen 4860 (PED 0983) is preserved in Myanmar amber. It is accessible in a dorsal A,B and l(42)Specimen 4861 (PED 0989a) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(43)Specimen 4862 (PED 0989b) is preserved in Myanmar amber, in the same amber piece as PED 0989a. It is accessible only in a dorso-lateral view F,G. No c(44)Specimen 4863 (PED 1000) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(45)Specimen 4864 (PED 1223) is preserved in Myanmar amber. It is accessible in a dorsal K and ven(46)Specimen 4865 (PED 1229a) is preserved in Myanmar amber. It is accessible only in a ventral view A,B. No c(47)Specimen 4866 (PED 1229b) is preserved in Myanmar amber, in the same amber piece as PED 1229a. It is only accessible in ventral view E,F. The (48)Specimen 4867 (PED 1258a) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(49)Specimen 4868 (PED 1258b) is preserved in Myanmar amber, in the same amber piece as PED 1258a. It is accessible in a dorsal G and ven(50)Specimen 4869 (PED 1287) is preserved in Myanmar amber. It is only accessible in a dorsal view A,B. Many(51)Specimen 4870 (PED 1301) is preserved in Myanmar amber. It is accessible in a dorso-lateral C and ven(52)Specimen 4871 (PED 1311) is preserved in Myanmar amber. It is accessible in a dorsal E,F and v(53)Specimen 4872 (PED 1322) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(54)Specimen 4873 (PED 1323) is preserved in Myanmar amber. It is accessible in a dorsal C,D and v(55)Specimen 4874 (PED 1333) is preserved in Myanmar amber. It is accessible in a ventral A,B,D and(56)Specimen 4875 (PED 1335) is preserved in Myanmar amber. It is accessible in a lateral A–C and p(57)Specimen 4876 (Weiterschan BuB 11) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(58)Specimen 4877 (Weiterschan BuB 31) is preserved in Myanmar amber. It is accessible in a dorsal A,B and v(59)Specimen 4703 (SMF Be 2021) is preserved in Baltic amber. It is accessible in a dorsal C, ventra(60)Specimen 4702 (CCGG 7615) is preserved in Baltic amber. It is accessible in a dorsal view A,B. A pr(61)Specimen 4704 (SMF Be 1861) is preserved in Baltic amber. It is accessible in a dorsal view C,D. No c(62)Specimen 4753 (CCHH 1786-3) is preserved in Baltic amber. It is accessible in dorsal A,B and vAs an expanded basis for the shape analysis, numerous new aphidlions and aphidlion-like larvae are presented here. The terminology follows earlier studies ,23,24,25The analysis of the head shape resulted in four effective principal components . They toPC1 explains 79.69% of the overall variation. It is strongly influenced by the relative length of the stylets. A low value indicates shorter stylets, a high value indicates longer stylets .PC2 explains 9.24% of the overall variation. It is strongly influenced by the posterior rim of the head capsule. A low value indicates a concave posterior rim, a high value indicates a convex posterior rim .PC3 explains 3.90% of the overall variation. It is strongly influenced by the anterior rim of the head capsule. A low value indicates a convex anterior rim, a high value indicates a concave posterior rim .PC4 explains 2.55% of the overall variation. It is strongly influenced by the proximal region of the stylets at the transition to the head capsule. A low value indicates a concave shape, a high value indicates a convex shape .2-value of 0.40. For type 2 larvae, only two specimens were available, providing no coefficient of determination. For type 3 larvae, the correlation is quite strong with an R2-value of 0.88. For type 4 larvae, the correlation is weaker with an R2-value of 0.63. Finally, for Hemerobiidae-type larvae there is a strong correlation with an R2-value of 0.95.As PC1 was so strongly dominating , we looked for a possible correlation of it to size. We did this for different types of larvae (explained in more detail in the discussion section). For type 1 larvae , such a correlation is weak with an RAs outlined, aphidlions are larvae of Chrysopidae and Hemerobiidae, and these two groups seem not to be closely related to each other. What does that mean for the term aphidlion? What does it in fact refer to?We already know that, from the ecological functional side, the term refers to a larva with specific feeding habits. Yet, apparently it also refers to a certain combination of morphological characters, otherwise we could not recognise a fossil as “aphidlion-like”. These characters include a simple head with curved, rather slender, but toothless stylets, well-developed antennae and labial palps, and a simple spindle-shaped body.Most of these characters very likely characterise a rather basal node within Neuroptera, which seems so far unnamed, and is namely the group of lacewings excluding Coniopterygidae, Nevrorthidae, Sisyridae, and Osmylidae (following the phylogeny of ). When aSimple-curved toothless stylets would become derived (“lost”) in the lineages of Dilaridae, Mantispidae + Berothidae, and Myrmeleontiformia. A spindle-shaped body is retained in most lineages; within Myrmeleontiformia, a broader shape has evolved . Thi“Aphidlion” is therefore likely a kind of ecological category, but it also refers to a rather unspecialised terrestrial predator that likely represents the ancestral condition of larval morphology and ecology for a large group within Neuroptera. Within the group, the larvae evolved more specialised morphologies and, coupled to this, strategies, such as pit traps Larvae of type 1 have relatively long protrusions on the anterior and posterior trunk, which are all about the same length. Many of the not-that-well-preserved specimens may represent type 1 larvae, yet they cannot be reliably interpreted.(2)Larvae of type 2 have relatively long protrusions on the anterior trunk, but even longer ones on the posterior trunk.(3)Larvae of type 3 have relatively long protrusions on the anterior trunk, but rather short ones on the posterior trunk.Many of the Cretaceous fossils bear protrusions on the dorsal side, similar to many fossil larvae that have been generally interpreted as relatives of Chrysopidae ,9,10,11.Another type of larvae, type 4, lacks dorsal protrusions. These larvae have shorter lateral protrusions and relatively long stylets. Their trunk region is often much better preserved than those of the first three morphotypes. None of these larvae seem to bear a camouflaging cloak.The last distinct type has a slender body, no protrusions, and shorter stylets. This feature is reminiscent of the larvae of Hemerobiidae. Therefore, we will use “Hemerobiidae-type” as a reference. Details further supporting or contradicting this interpretation will be given in the following.Ankyloleon, [The stylets of all new specimens are gently curved and rather slender. In many ingroups of Neuroptera, the stylets are more or less straight , or curved, but shorter and stouter . In some neuropteran ingroups, the stylets are proximally straight, but curved distally . Simpleyloleon, ,16 unlikyloleon, ; some layloleon, ). FurtheThe head capsule shape of most of the new fossils clearly differs from that of long-nosed antlions (larvae of Psychopsidae) and long-necked antlions (larvae of Crocinae). Long-nosed antlions have a prominent labrum region projecting forward from the head capsule , which iOther prominent head structures are the antennae and the labial palps. The antenna of many fossils, especially of types 1–3, bears a prominent distal seta e.g., A. Such aThe labial palps of larvae of Chrysopidae consist of three elements, with the second one being the longest. The labium of larvae of Hemerobiidae often has a proximally conjoined part projecting forward, which seems rather unique within Neuroptera. Moreover, in Hemerobiidae, the palps have three elements, with the third one being the longest; in some modern larvae this element appears swollen .Yet, the case is in fact a bit more complicated. Extant larvae of Chrysopidae have empodia only in larval stages 2 and 3 ,38, but Some extant larvae of Chrysopidae have dorsal processes and use these to attach a camouflaging cloak . Larvae The structure of the processes in the new fossils differs from those of other groups, but also from those in the modern-day larvae of Chrysopidae. Still, it seems most parsimonious to assume that all Cretaceous specimens discussed here with such processes are closely related to Chrysopidae, basically following earlier interpretations of comparable fossils e.g., ,10,11).,11.10,11As to be expected in such a larger sample, there is quite some variation within the material. It seems likely that the sample contains different larval stages (see further below for this aspect), but probably also different species. As the state of preservation in many of these larvae is at best challenging, we refrain from erecting formal species based on these specimens. We hope that expanding the data set further in the future will allow for a stricter phylogenetic approach, as performed in Badano et al. ,15.Most specimens reported here seem to be relatives of modern green lacewings (Chrysopidae). Types 1–3 are characterised by the prominent processes of the back and hence are quite similar to the modern aphidlions of Chrysopidae. Moreover, type 4 specimens strongly resemble modern green lacewing aphidlions, but are clearly of a non-camouflaging type. The differences within these specimens (as outlined above) may indicate that this type in fact represents at least two different species. Indeed, the fossils of the Hemerobiidae-type possess characters that further support an interpretation of these larvae as at least closer relatives of these lineages.In earlier quantitative studies of lacewing larvae, no real effects of shape and size could be recognised , but alComparing the head shapes of the different groups included here reveals that all have large overlapping areas when plotting them in two dimensions. No group separates strongly from the others, yet certain differences can be well recognised . Extant The fossil larvae plot largely within the range of green lacewing aphidlions . Only a The comparison of the different time slices is partly complicated by the fact that the sample sizes for the Miocene and Eocene are much smaller than those of the Cretaceous and especially the modern fauna . The difWe can therefore mostly recognise that the fossils from the Miocene and Eocene fall within the area of the morphospace occupied by modern larvae. It is, however, noteworthy that the range of the green lacewing aphidlions in the Miocence and Eocene spans more or less the entire space occupied by modern green lacewing aphidlions in the PC1. This large range indicates that although the overall number of specimens is rather low, the overall morphological diversity of green lacewing aphidlions in the Eocene/Miocene was already about as large as today. Moreover, for the Cretaceous, the area occupied by modern green lacewing aphidlions was already largely occupied, at least along the PC1 .The larvae in the Eocene and Miocene already have a very modern appearance and are indeed likely representatives of modern lineages. The Cretaceous larvae are not only similar in head shapes, but also partly in possessing structures on the back to carry a camouflaging cloak. Yet, in the details these Cretaceous larvae are quite different from the modern ones. For example, the head is much more distinctly set off in the fossils; moreover, the exact morphology of the camouflage-cloak-carrying processes is quite different .These morphologies indicate that the fossils performed a very similar ecological role, yet are not closely related to the modern forms. We therefore assume that the Cretaceous species, probably entire lineages, went extinct, possibly at the end of the Cretaceous, and became ecologically replaced by the modern green lacewings before the Eocene. This combination of a severe extinction of some lineages in combination with a diversification of another lineage, basically substituting the extinct forms, is generally addressed as “faunal turnover”. Yet, this expression is usually used to address taxonomic changes of a community.For brown lacewing aphidlions (Hemerobiidae), the case is different. The area exclusively occupied by modern brown lacewing aphidlions (not overlapping with green lacewing aphidlions) is basically unoccupied by fossils, besides a single specimen from the Eocene . The basIt is unlikely that this observation is due to a preservation bias. Brown lacewing larvae nowadays have a quite similar lifestyle to green lacewing larvae in the aspects of living on plants and hunting their preferred prey there. With this, they should not have had a lower preservation potential. One could speculate that larvae with such a morphology back in the Cretaceous had a different lifestyle that is making a preservation in amber less likely (see discussion in ). This wIt is well established that lacewings were more diverse in the past ,2, whichAs apparent in the here presented data set, the most common types of aphidlion larvae in the Cretaceous are those with protrusions on the back (types 1–3) that allow them to carry a camouflaging cloak . It is sLarvae of type 4 have only short protrusions on the abdomen segments. These may still have acted as a support for a camouflaging cloak, as observed in other lacewing larvae (cf. ). Hence,(1)A certain loss of very extreme forms, most likely coupled to a loss of specific life habits A loss of many further forms, especially of camouflaging larvae; yet this loss is factually compensated by:(3)A diversification of the modern lineages of green lacewings and their larval forms.In summary, we observe in the case of Chrysopidae and their closer relatives almost no change in head shape over the last 100 million years. Nevertheless, in detail we observe:In Hemerobiidae, the case is less clear due to a significantly smaller sample size. Still, the results indicate an increase in larval brown lacewings after the Cretaceous.


Non-alcoholic fatty liver disease (NAFLD) presents the most common chronic liver disease globally; it is estimated that 25.24% of the world’s population has NAFLD. NAFLD is a multi-factorial disease whose development involves various processes, such as insulin resistance, lipotoxicity, inflammation, cytokine imbalance, the activation of innate immunity, microbiota and environmental and genetic factors. Numerous clinical studies have shown that the Mediterranean diet produces beneficial effects in NAFLD patients. The aim of this review is to summarize the beneficial effects of lycopene, a soluble pigment found in fruit and vegetables, in NAFLD. Non-alcoholic fatty liver disease (NAFLD) presents the most common chronic liver disease globally. It is estimated that 25.24% of the world’s population has NAFLD with the highest prevalence rates in the Middle East and South America . NAFLD iIn NAFLD pathogenesis, mitochondrial dysfunction is characterized by structural alterations, such as the depletion of mitochondrial DNA and morphological and ultra-structural changes and functional alterations, involving the respiratory chain and mitochondrial β-oxidations . In factp < 0.01) between the baseline and the end of treatment. In a study of non-diabetic patients with NAFLD, the subjects underwent 16 weeks of MD, 16 weeks of washout and 16 weeks of low-fat diet . LY. LY58]. Several studies have shown that the white adipose tissue is an endocrine organ capable of releasing numerous adipokines and pro-inflammatory factors. It has been shown that high levels of adipokines, including IL-6 and TNF-α, can contribute to the reduction in lipid oxidation leading to lipotoxicity and insulin resistance. TNF-α is a key modulator of adipocyte metabolism owing to its direct role in glucose homeostasis and lipid metabolism. In addition, both IL-6 and TNF-α promote the production of leptin by adipose tissue, with an increase in inflammatory cytokines and the induction of the absorption of cholesterol by macrophages. In a study conducted by Luvizzotto et al., supplementation with LYC was observed to reduce leptin levels and restore plasma IL-6 concentrations significantly in obese rats .Moreover, some LYC metabolites are bioactive, particularly apo-10′-lycopenoic acid (ALA), the major metabolite of LYC known to suppress hepatic steatosis and inflammation by stimulating sirtuin 1 (SIRT1). A study conducted by Chung et al. showed that ALA supplementation increased hepatic SIRT1 protein with a concomitant increase in the deacetylation of NF-κB p65, which in turn decreased the hepatic protein levels of IL-6 and TNFα by a mechanism not yet known . The prop < 0.05). In a study conducted by Jiang et al., 65 rats with NAFLD were divided into a control group and three groups treated with 5, 10 and 20 mg with LYC, respectively [p < 0.01). Furthermore, the serum levels of AST and ALT were significantly decreased in a dose-dependent manner after the LYC treatment. Regarding the serum lipid levels, a remarkable reduction in the concentrations of LDL-C and TG in dose-dependent manners was observed, showing the lipid-lowering effects of LYC. Additionally, this study reported a significant increase in SOD and glutathione (GSH) levels in rats treated with LYC. Finally, the results of this study showed the decreased expression of TNF-α and CYP2E1, involving a reduction in inflammatory response [p = 0.002), and serum LDL levels (p = 0.008) and TC concentrations in the liver were normalized (p = 0.001). Regarding oxidative stress enzymes, it was observed that SOD and CAT activities were increased in the group on a normal diet supplemented with LYC. The macroscopic analysis found a complete recovery of injured livers. In a study conducted by Negri et al., 61 obese children with fatty liver were enrolled; all participants underwent a washout low carotenoid diet for two weeks, and then they were randomly assigned to a restricted calorie regimen (RCR) (Group 1) or RCR supplemented with tomato juice and basil and oregano extracts in extra virgin olive oil (Group 2) for 60 days [p < 0.01) accompanied with the loss of subcutaneous and visceral fat and decreased liver size. Additionally, the serum levels of insulin decreased by 34% in group 2. Total cholesterol showed a decrease in group 2 of 6.7%. Regarding the LYC antioxidant activity, it was detected that GSH levels increased in both groups after being exposed to LYC. Instead, the anti-inflammatory activity was ascertained by the decrease in IL-4 and increase in adiponectin.Several studies have been carried out to verify the effects of LYC on NAFLD . In a stectively . After tresponse . In a stresponse . After i 60 days . Subsequp < 0.01 and p < 0.05, respectively). Another study by Wang et al. showed that LYC can inhibit hepatocarcinogenesis induced by NASH [p < 0.01), suggesting that both supplementations have anticarcinogenic actions. In addition, a reduction in hepatocytes, which exposed the proliferating cell nuclear antigen (PCNA), was observed in all groups where LYC or tomato extract was administered (p < 0.05), indicating that LYC could inhibit cell proliferation induced by NASH. Since oxidative stress contributes to the onset of liver cancer, the levels of certain cellular components involved in oxidative stress were analyzed. In particular, it was noted that the HFD and LYC group showed an increased expression of transcription factor 2 related to nuclear factor E2 (Nrf2), a key regulator of cellular response to oxidative stress (p < 0.05). Nrf2 is the main regulator of the enzyme heme oxygenase-1 (HO-1) that acts as an antioxidant and eliminates free radicals [NASH is closely associated with hepatic fibrosis and, if not adequately treated, can evolve into HCC ,81. The  by NASH . In partradicals . The abiradicals . The purradicals . ALA seeCurrently, NAFLD presents a new challenge for scholars and physicians. A balanced and healthy diet with reduced amounts of saturated fat is the strategy that gives beneficial effects both in the prevention and in the treatment of NAFLD. Several studies are available in the literature that confirm the effectiveness of the MD in improving NAFLD owing to a great variety of nutraceutical components, among which LYC is one of the most abundant components. Several LYC biological functions in the body can be particularly effective against a multifactorial pathology, such as NAFLD. Particular attention is paid to the antioxidant activity, which is already known to act efficiently in NAFLD. In fact, many studies have reported an increase in the activity of SOD and CAT as a final result of the administration of LYC. The anti-inflammatory activity of LYC has also been confirmed in several studies performed for the reduction in TNF-α and CYP1E2 expressions. Further, the reduction in serum levels of lipids and hepatic levels of cholesterol by LYC was confirmed, indicating a possible action on the lipid metabolism. The mechanism for these results has not yet been understood fully. Many preclinical studies are available in the literature with the aim to clarify the mechanism of LYC action. However, there is a great lack of clinical studies. In conclusion, we can state that LYC currently presents a very promising natural compound in the prevention and treatment of NAFLD, but more studies by the scientific communities are certainly indispensable to investigate some of its effects in a more useful way. A large multicenter clinical study is needed to confirm the efficacy of LYC in the treatment of NAFLD.


FLS in each point were calculated. A level of p < 0.05 was accepted as statistically significant. As a whole, FLS was significantly higher in FEs vs. ERMEs (p < 0.001); FS in both SCP and DCP was not significantly different between ERMEs and FEs . Correlation in focal retinal areas between FLS and FS in ERMEs was significant in SCP (p = 0.002) and not significant in DCP (p = 0.205); in FEs was significant in both SCP (p < 0.001) and DCP (p = 0.022). As previously described, these defective areas were located mainly in sites of distortion of retinal layers; therefore, it can be hypothesized that a focal change in FS, occurring mostly in SCP, could be involved in the onset of the functional defect.Inner macular layers are the most involved in the retinal distortion caused by idiopathic epiretinal membrane (iERM). They represent the anatomical structures in which the superficial (SCP) and deep (DCP) capillary plexus are embedded. We quantified flow signal (FS) in these capillary plexuses using Swept Source OCT angiography to identify possible markers for postoperative outcome. The software ImageJ was used to quantify the FS in a 150 µm radius area around each point analyzed by MAIA microperimeter. In 16 patients with unilateral iERM, focal light sensitivity (FLS) in the para- and perimacular areas was measured to evaluate macular function in 24 points overlapping macular plexuses and compared with normal fellow eyes (FEs).  Idiopathic epiretinal membranes (iERM) are a frequent macular pathology with a prevalence of 7–11.8% of the population. Epiretinal membranes are made of glial cells, retinal pigment epithelial (RPE) cells, macrophages, fibrocytes and collagen in varying proportions. IERM can stretch the underlying retina, depending on the cell type present in the ERM , causingPatients with ERMs usually show visual symptoms as the conditions worsen, including reduced visual acuity and metamorphopsia. A thickening of inner nuclear layer (INL) has been reported more in patients with metamorphopsia than in those without . A furthPrevious works identified preoperative markers correlated with postoperative visual function in order to predict visual recovery after surgical treatment . Recent Inner retinal layers are the anatomical structures in which the superficial (SCP) and deep (DCP) capillary plexus of the macula are located; therefore, we quantified flow signal (FS) in these retinal capillary plexuses using both Swept Source-OCT (SS-OCT) and Swept Source SS-OCT angiography (OCTA) to assess the preoperative anatomical status and identify possible markers for postoperative outcome in patients affected by iERM.The aim of this study is, therefore, to quantify by means of OCTA the FS in the SCP and DCP in eyes affected by iERM and evaluate a possible correlation with FLS.The study was conducted in a consecutive series of patients with unilateral idiopathic ERMs; 31 eyes of 16 patients, 16 eyes with ERM (ERMEs) and 15 normative fellow eyes (FEs), 9 women and 7 men, mean age 63.21 ± 5.34 years, were studied.All procedures in this study adhered to the tenets of the Declaration of Helsinki and were approved by the investigational review board of Central Ethic Committee IRCCS Lazio. All subjects gave their informed consent after the aim of the study had been fully explained.Before imaging, all patients underwent ophthalmic examination, including best-corrected visual acuity and fundus examination, performed by a retina specialist using a +90 diopters (D) lens. Inclusion criteria were the presence of iERM in one eye and normal morphology without any retinal changes in the fellow eye. Exclusion criteria were the presence of any retinal or choroidal disease such as retinal detachment, retino-vascular disease, AMD, diabetic retinopathy, glaucoma or ocular hypertension, a history of previous ocular laser or surgery, eyes with refractive errors > 3D, media opacities that prevented good visualization of the fundus, any associated systematic disorders  or vascular diseases without retinopathy, which would affect the FS.® Elite 9000 . It is a swept-source OCT (SS-OCT) and OCT angiography (SS-OCTA) that provides automated segmented enface OCT with analysis of different plexus: Superficial Capillary Plexus (SCP), Deep Capillary Plexus (DCP), Avascular Retina, Choriocapillaris (CC) and Choroid (Ch). OCTA on the PLEX® Elite 9000 is generated with the OMAG® algorithm  [The macula was assessed using the split-spectrum amplitude-decorrelation angiography with PLEXography) ,9, whichography) ,11. The ® Elite 9000 analyzes retinal layers using infrared (IR) light produced by a swept source laser. This IR laser ray penetrates in deeper retinal layers with less bias than light used in Spectral Domain OCT. The instrument further reduces the segmentation errors related to the projection artifacts on DCP using a software, that subtracts the artifacts related to the SCP flow signal from the enface image of the DCP.The PLEXThe software ImageJ , was used to quantify the FS in the focal area around each point analyzed by MAIA microperimeter . The MAIA microperimeter checks the fixation stability and gives an output of 3 possible fixation stability: stable, relatively stable and unstable; we repeated the microperimetry if necessary to analyze only the examinations with stable fixation and to have valid sensitivity values in each patient. To evaluate macular functional status, we measured the focal light sensitivity (FLS) in the para- and perimacular areas, as reported in previous papers ,13,14, bWe decided to assess the visual function outside the central fovea, assessed in previous studies ,16, as tAn ImageJ circle with a radius of 150 micron was used to select the focal area around each one of the 24 points analyzed by microperimeter MAIA. Thereafter, in each binarized focal area, we calculated the density of white pixels, representing the focal FS, and density was then correlated with the FLS at each point. As the aim of the study is to evaluate the correlation between morphological and functional parameters in each focal areas of the retina, the N of focal areas examined in every one of the 16 patients are 24; therefore, 24 × 16 = 384 for each capillary plexus were analyzed.The eccentricity of the 24 points analyzed in each eye was between 2.5° and 5°, the 1° grid was not included for its location on the border of the FAZ. The 12 points with eccentricity 2.5° represent inner grid, 12 points at 5° outer grid .Images with visible eye motion or blinking artefacts and with poor image quality were excluded . OCTs were visually assessed by the same retina specialist (A.M.C.) to ensure proper segmentation of SCP and DCP.By measuring macular capillary network with OCTA, an association was found between the presence of epiretinal membranes in pediatric age and the reduction in FS at the level SCP and DCP . In ordet-Test for independent samples iERMEs vs. FEs and the Pearson correlation coefficient (R2) of FS (density) vs. FSL (dB) in each point was calculated, comparing the same focal areas of the retina according to the grid between iERMEs and FEs. A level of p < 0.05 was accepted as statistically significant. The statistical analyses were performed using SPSS software version 15.0 .The p < 0.001), with a statistically significant difference, as displayed in FS and FLS values of each 24 focal areas from anyone of the 16 patients were evaluated and compared as a whole between affected and fellow eyes; in iERMEs, mean FLS was 24.39 ± 4.19 dB, in FEs 25.57 ± 4.18 dB   .A significant mean reduction in light sensitivity was present in affected eyes .p = 0.002) and not significant in DCP (p = 0.205); in FEs, was significant in both SCP (p < 0.001) and DCP (p = 0.022). The correlation in iERMEs is negative in both SCP (R2 = −0.164) and DCP (R2 = −0.69), whereas in FEs, it is positive in SCP (R2 = 0.279) and negative in DCP (R2 = −0.121), as reported in Interesting findings were standard deviations (SD) differences, for both SCP and DCP, between ERMEs and FEs, probably associated with focal changes in the retinal structure related to ERM contraction. In iERMEs, the correlation between FLS and FS was significant in SCP  and fellow eyes (FEs).Differences are present for FLS and SCP FS between inner and outer grids: inner vs. outer grid in FEs FLS Comparison between affected and fellow eyes:p < 0.004). Mean FS in iERMEs was 106.57 ± 21.91 for SCP and 71.85 ± 16.34 for DCP; in FEs, 107.41 ± 15.03 for SCP and 70.55 ± 11.01 for DCP without reaching a statistically significant difference between the two groups . Correlation between FLS and FS is significant in SCP in both iERMEs (p = 0.047) and FEs (p < 0.001), and not significant in DCP, iERMEs (p = 0.182) and FEs (p = 0.195). Correlation is positive only in SCP of FEs (R2 = 0.371) and negative in DCP of FEs (R2 = −0.097), SCP (R2 = −0.153) and DCP (R2 = −0.103) of iERMEs.Inner grid: in iERMEs, mean FLS was 24.94 ± 3.71 dB; in FEs, 26.12 ± 3.95 dB, statistically significant between fellow and affected eyes (p = 0.006), with a statistically significant difference between fellow and affected eyes. Mean FS in iERMEs were 115.43 ± 21.21 for SCP and 71.21 ± 16.74 for DCP, in FEs, 115.24 ± 18.12 for SCP and 73.05 ± 10.58 for DCP without a statistically significant difference . Correlation between FLS and FS is statistically significant in FEs only in SCP (p = 0.0001) and not significant in DCP (p = 0.115), whereas it is not significant in both capillary plexuses of iERMEs: SCP (p = 0.081) and DCP (p = 0.535). Correlation is positive only in SCP of FEs (R2 = 0.283) and negative in DCP of FEs (R2 = −0.118), SCP (R2 = −0.135) and DCP (R2 = −0.048) of iERMEs. Based on these data, results showed that, except for the change in the means values of FLS and FS, there is no significant difference between fellow and affected eyes related to the eccentricity of focal retinal areas  of affected eyes were analyzed to evaluate if a higher correlation between FLS and FS was present.p = 0.003 R2 = −0.370 and in DCP (N = 45) p = 0.006 R2 = −0.402 were statistically significant. In these focal areas, therefore, negative correlation is significant in both capillary plexuses  plexuses .The data of this study are consistent with the hypothesis that FS could be a marker to assess the anatomical status and predict the functional status of focal retinal areas in eyes affected by iERM. ERM contraction causes focal functional defects, as shown by the mean FLS which is statistically lower in affected vs. fellow eyes. On the contrary, the mean FS were not significantly different between the two groups in both SCP and DCP; however, this apparent discrepancy can be explained by the fact that in eyes with ERM, the irregular contraction of the ERM causes focal displacement of blood flow, thus not affecting the mean values, mainly in SCP, and in DCP. Additionally, retinal blood flow in SCP and light sensitivity are significantly different in relation to eccentricity of the focal retinal areas.Analysis of the correlation between morphological and functional parameters shows that focal functional alterations in the inner retina can occur as a consequence of focal structural change. The correlation between FLS and FS is significant in SCP in both affected and fellow eyes. These data validate a relation between the blood flow density and the functional status in focal retinal areas. Focal blood flow could be related to the conformation of the retinal structure, which can be modified by inner retinal layer shrinkage due to the entity of ERM contraction. An increase in blood flow, mainly in the SCP, is present in focal areas in which the structure of inner retina, such as in eyes affected by ERM, is more altered. We found a significant and negative correlation between FS and FLS in SCP of iERMs, whereas in FEs, the correlation between these parameters is significant and positive, showing, therefore, a different relation between FS and FLS in healthy and affected areas of the retina, as shown in 2 is higher when the correlation is calculated in groups of focal areas in which the values of FS differ more (±2SD) from the mean  layer, and subsequently affecting bipolar and amacrine cells and both inner and outer plexiform layers, fed by DCP. The GC layer is more injured by the mechanical damage and a previous study of ours reported that the GC layer can also be affected by an alteration occurring before the mechanical damage related to the contraction on the iERM . MoreoveThe correlation between FS and FLS is more significant in patients with a higher difference from mean value of FS; therefore, it can be hypothesized that the severity of the functional retinal cell damage is correlated to the amount of FS change .Furthermore, the FS change and the statistically significant correlation found between SCP and FLS in affected eyes, support the hypothesis that FS focal alterations could be helpful to assess the anatomical integrity and the corresponding functional status of inner retinal layers. Consequently, FS and FLS parameters could be considered predictive markers to evaluate when patients should undergo surgical treatment and to predict post-operative outcome.® Elite 9000, infrared light SS-OCT, is more accurate with respect to SD-OCT. Additionally, the more significant capillary plexus in the affected eyes is the superficial, in which lower projection artifacts are involved, related just to the presence of epiretinal (clear) tissue. Even if the presence of projection artefacts can bias the evaluation of the FS in the DCP, in the PLEX® Elite 9000, the new software added to the infrared light of the SS-OCT significantly reduces the interference in the evaluation of the DCP. A limitation of the study is related to the series with N = 16; however, it should be considered that the number of focal retinal areas is 24 in each patient; therefore, as previously reported, 384 is the total number of focal areas examined in the study. Each focal area is an example of what happens in the retinal tissues as a consequence of ERM contraction. Following this model, we considered the N of 384 focal areas could be statistically significant to support a hypothesis reported in this pilot study. On the basis of these data, a further study in larger series is recommended to confirm that SCP could represent an interesting biomarker to assess focal retinal defects in eyes affected by iERM.It has been previously reported that the segmentation of SCP and DCP  in patieThe evaluation of FS with new generation SS-OCTA, could represent a reliable technique to analyze the macular function in eyes affected by iERM, as demonstrated by the findings of correlation between FLS and FS in macular capillary network. These results allowed us to suggest that FS, mainly in SCP, can be a biomarker to detect morpho-functional worsening in patients affected by iERM and to decide when it is the optimal time, with best benefit-risk ratio, to perform pars plana vitrectomy and peeling of the macular epiretinal membrane.