Oxidation effects in antiaggregogenic properties of Epigallocatechingallate

Epigallocatechin-gallate (EGCG), the most abundant flavonoid in green tea, has been extensively studied for its potential in the treatment of amyloid related disorders. This molecule was found to modulate abnormal protein self-assembly, reducing resulting cellular toxicity. EGCG is known to suppress or to slow down the aggregation processes of several proteins, thus supporting the idea that general mechanisms regulate its anti-aggregogenic effects and, interestingly, in the oxidised form it demonstrated an higher efficiency in reducing protein aggregation with respect to intact molecule. We here investigate the effects of intact and oxidized EGCG the thermal aggregation pathway of Bovine Serum Albumin (BSA), a well-known model protein whose aggregation processes are known in details. By means of different spectroscopic methods, we evaluate similarities and differences of the two molecules during protein aggregation. Different solution conditions are investigated, close and away from the isoelectric point of the protein, with the aim of eliciting the role of electrostatics in the observed EGCG-Protein interaction and in the supramolecular assembly which are dramatically dependent on solution conditions

Electrostatics regulate Epigallocatechin-Gallate effects on Bovine Serum Albumin aggregation

Protein aggregation processes are complex phenomena often involved in the etiology of several pathologies. It is now assessed that all proteins, in suitable conditions, may undergo supramolecular assembly. Aggregation pathways are known to be controlled by solution conditions which regulate protein-protein and protein-solvent interactions affecting binding mechanisms, morphology and inherent toxicity of the aggregate species. In this context, the presence of small molecules was indicated as a promising method to modulate protein-protein interactions reducing pathogenic aggregation. In the light of the idea that common mechanisms regulate anti-aggregogenic properties of small molecules, we here investigate Epigallocatechin-Gallate (EGCG) effects on the thermal aggregation pathway of Bovine Serum Albumin (BSA), a well-known model protein. EGCG is a small molecule extracted from green tea, which is known to reduce aggregation of key proteins involved in neurodegenerative diseases [1]. Fundamental mechanisms which regulate EGCG effectiveness as therapeutic molecule are still not clearly elucidated. The interaction of EGCG with BSA and its effects on thermal aggregation pathway were investigated by means of spectroscopic methods and Isothermal Titration calorimetry as a function of solution conditions. Results show that electrostatic forces modulated by pH play a key role in regulating EGCG interactions with BSA. Data shows that close to the isoelectric point of the protein, EGCG is found to promote the supramolecular assembly, whilst away from the isoelectric point, EGCG is found to reduce aggregation mechanisms increasing protein conformational stability. These results reveal the large impact of electrostatics in small molecules effects on the protein aggregation phenomena requiring larger investigation aimed at rationalizing their effects on related pathogenic mechanisms

The Atopy Index Inventory: A Brief and Simple Tool to Identify Atopic Patients

Introduction: Atopy and ear, nose and throat (ENT) diseases are frequently associated; however, no clinical tool has been proposed so far to discriminate which patients could be atopic and therefore deserving of a further immunoallergological evaluation. Objective: The aim of this study was to assess and validate a set of dichotomous responses suitable for predicting the presence of atopy in adult patients. Methods: An 11-item questionnaire, i.e., the Atopy Index Inventory (AII), comprised of 4 questions regarding the clinical history for allergic disease and 7 questions evaluating the presence of the most frequent clinical signs affecting allergic patients, was developed and administered to 226 adult subjects (124 atopic subjects and 102 healthy, not atopic subjects). The atopic condition was proven by an immunoallergological evaluation according to the diagnostic criteria of the EAACI guidelines. Internal consistency and clinical validity were tested. Results: In healthy subjects, the first 4 variables of the AII returned a 100% correct response (all answered \u201cno\u201d) and were defined as \u201cdecisive\u201d responses. In the logistic regression analysis, when decisive items were negative, the atopic condition was confirmed when answering \u201cyes\u201d to at least 3 \u201cprobability\u201d items (cutoff = 2.69). The difference in AII scores between allergic and healthy group was significant using the Mann-Whitney U test (p < 0.0001). The sensitivity and specificity of the AII were 0.97 and 0.91, respectively, with a true predictive value of 0.92 and a false predictive value of 0.97. The ROC curve showed an area of 0.94, with an OR of 0.88 (95% CI 0.87\u20130.97, p = 0.0001). The internal consistency as determined by the Cronbach \u3b1 coefficient was 0.88. Conclusion: The AII has been proven to be a brief, simple and sufficiently accurate tool for screening ENT patients in search of atopic individuals and to allow their clinical management

Immunotherapy for patients with advanced urothelial cancer: Current evidence and future perspectives

In recent years, immunotherapy has produced encouraging results in a rapidly increasing number of solid tumors. The responsiveness of bladder cancer to immunotherapy was first established in nonmuscle invasive disease in 1976 with intravesical instillations of bacillus Calmette-Guérin (BCG). Very recently immune checkpoint inhibitors demonstrated good activity and significant efficacy in metastatic disease. In particular the best results were obtained with programmed death-ligand-1 (PD-L1) and programmed death-1 (PD-1) inhibitors, but many other immune checkpoint inhibitors, including anti-cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) antibodies, are currently under investigation in several trials. Simultaneously other therapeutic strategies which recruit an adaptive immune response against tumoral antigens or employ externally manipulated tumor infiltrating lymphocytes might change the natural history of bladder cancer in the near future. This review describes the rationale for the use of immunotherapy in bladder cancer and discusses recent and ongoing clinical trials with checkpoint inhibitors and other novel immunotherapy agents

Ischemic Preconditioning Modulates the Peripheral Innate Immune System to Promote Anti-Inflammatory and Protective Responses in Mice Subjected to Focal Cerebral Ischemia

The development of tolerance triggered by a sublethal ischemic episode (preconditioning, PC) involves a complex crosstalk between neurons, astrocytes and microglia, although the role of the peripheral immune system in this context is largely unexplored. Here, we report that severe cerebral ischemia caused by transient middle cerebral artery occlusion (MCAo) in adult male mice elevates blood counts of inflammatory neutrophils and monocytes, and plasma levels of miRNA-329-5p. These inflammatory responses are prevented by ischemic PC induced by 15 min MCAo, 72h before the severe insult (1h MCAo). As compared with sham-operated animals, mice subjected to either ischemic PC, MCAo or a combination of both (PC+MCAo) display spleen contraction. However, protein levels of Ym1 (a marker of polarization of myeloid cells towards M2/N2 protective phenotypes) are elevated only in spleen from the experimental groups PC and PC+MCAo, but not MCAo. Conversely, Ym1 protein levels only increase in circulating leukocytes from mice subjected to 1h MCAo, but not in preconditioned animals, which is coincident with a dramatic elevation of Ym1 expression in the ipsilateral cortex. By immunofluorescence analysis, we observe that expression of Ym1 occurs in amoeboid-shaped myeloid cells, mainly representing inflammatory monocytes/macrophages and neutrophils. As a result of its immune-regulatory functions, ischemic PC prevents elevation of mRNA levels of the pro-inflammatory cytokine interleukin (IL)-1β in the ipsilateral cortex, while not affecting IL-10 mRNA increase induced by MCAo. Overall, the elevated anti-inflammatory/pro-inflammatory ratio observed in the brain of mice pre-exposed to PC is associated with reduced brain infarct volume and ischemic edema, and with amelioration of functional outcome. These findings reaffirm the crucial and dualistic role of the innate immune system in ischemic stroke pathobiology, extending these concepts to the context of ischemic tolerance and underscoring their relevance for the identification of novel therapeutic targets for effective stroke treatment

Probing diffuse radio emission in bridges between galaxy clusters with uGMRT

Aims. Recent X-ray and Sunyaev-Zeldovich (SZ) observations have detected thermal emission between early-stage merging galaxy clusters. The main purpose of this work is to investigate the properties of the non-thermal emission in the interacting cluster pairs Abell 0399-Abell 0401 and Abell 21-PSZ2 G114.9. Methods. These two unique cluster pairs have been found in an interacting state. In both cases, their connection along a filament is supported by an SZ effect detected by the Planck satellite and, in the special case of Abell 0399-Abell 0401, the presence of a radio bridge has been already confirmed by LOFAR observations at 140 MHz. Here, we analyse new high-sensitivity, wideband (250-500 MHz) uGMRT data of these two systems and describe an injection procedure to place limits on the spectrum of Abell 0399-Abell 0401 and on the radio emission between Abell 21-PSZ2 G114.9. Results. In both cases, the low-surface-brightness diffuse emission is not detected in Band 3 (250-500 MHz). For the A399-A401 pair, we are able to constrain the steep spectral index of the bridge emission to be α > 2:2 with a 95% confidence level between 140MHz and 400 MHz. We also detect a small patch of the bridge with a flatter spectral index, which may suggest a variable spectral index distribution across the bridge area. For the A21-PSZ2 G114.9 pair, we are able to place an upper limit on the flux density of the bridge emission with two different methods, finding at the central frequency of 383MHz a conservative value of f1u < 260 mJy at a 95% confidence level, and a lower value of f2u < 125 mJy at an 80% confidence level, based on visual inspection and a morphological criterion. Conclusions. Our work provides a constraint on the spectrum in the bridge A399-A401 that disfavours shock acceleration as the main mechanism for the radio emission. The methods that we propose for the limits on the radio emission in the A21-PSZ2 G114.9 system represent a first step towards a systematic study of these sources

Pre-amyloid oligomers budding:a metastatic mechanism of proteotoxicity

The pathological hallmark of misfolded protein diseases and aging is the accumulation of proteotoxic aggregates. However, the mechanisms of proteotoxicity and the dynamic changes in fiber formation and dissemination remain unclear, preventing a cure. Here we adopted a reductionist approach and used atomic force microscopy to define the temporal and spatial changes of amyloid aggregates, their modes of dissemination and the biochemical changes that may influence their growth. We show that pre-amyloid oligomers (PAO) mature to form linear and circular protofibrils, and amyloid fibers, and those can break reforming PAO that can migrate invading neighbor structures. Simulating the effect of immunotherapy modifies the dynamics of PAO formation. Anti-fibers as well as anti-PAO antibodies fragment the amyloid fibers, however the fragmentation using anti-fibers antibodies favored the migration of PAO. In conclusion, we provide evidence for the mechanisms of misfolded protein maturation and propagation and the effects of interventions on the resolution and dissemination of amyloid pathology

High-grade serous carcinoma of unknown primary origin associated with STIC clinically presented as isolated inguinal lymphadenopathy: a case report

Serous tubal intraepithelial carcinoma (STIC) is a precancerous lesion of high-grade serous ovarian carcinoma (HGSOC). Usually, it arises from the fimbrial end of the tube, and it is associated with metastatic potential. On average, the time to progress from STIC to HGSOC is 6.5 years. Therefore, whenever a STIC lesion is found, surgical staging and prophylactic salpingectomy are recommended in order to prevent ovarian cancer. We report a rare case of a 45-year-old female patient who clinically presented an isolated right inguinal lymphadenopathy. The remaining clinical examination was normal. Therefore, an excisional biopsy of the lymph node was performed. Pathological analysis revealed a high-grade serous carcinoma, most likely of gynecological origin. Due to histological evidence, a computed tomography (CT) scan was carried out. There was no CT evidence of ovarian disease, pelvic involvement, intra-abdominal lymphadenopathies, metastatic disease, or ascites. All tumor markers were negative. The patient underwent laparoscopic hysterectomy and bilateral salpingo-oophorectomy followed by surgical staging. Surprisingly, pathological examination showed a STIC lesion in the fimbria of the left fallopian tube. We aim to report the potential capability of STIC to spread particularly through lymphatic pathways rather than peritoneal dissemination