Natural compounds Camptothecin and Triptolide: highly specific enzyme inhibitors and tools to dissect transcriptional functions

With this work I elucidated new and unexpected mechanisms of two strong and highly specific transcription inhibitors: Triptolide and Campthotecin. Triptolide (TPL) is a diterpene epoxide derived from the Chinese plant Trypterigium Wilfoordii Hook F. TPL inhibits the ATPase activity of XPB, a subunit of the general transcription factor TFIIH. In this thesis I found that degradation of Rbp1 (the largest subunit of RNA Polymerase II) caused by TPL treatments, is preceded by an hyperphosphorylation event at serine 5 of the carboxy-terminal domain (CTD) of Rbp1. This event is concomitant with a block of RNA Polymerase II at promoters of active genes. The enzyme responsible for Ser5 hyperphosphorylation event is CDK7. Notably, CDK7 downregulation rescued both Ser5 hyperphosphorylation and Rbp1 degradation triggered by TPL. Camptothecin (CPT), derived from the plant Camptotheca acuminata, specifically inhibits topoisomerase 1 (Top1). We first found that CPT induced antisense transcription at divergent CpG islands promoter. Interestingly, by immunofluorescence experiments, CPT was found to induce a burst of R loop structures (DNA/RNA hybrids) at nucleoli and mitochondria. We then decided to investigate the role of Top1 in R loop homeostasis through a short interfering RNA approach (RNAi). Using DNA/RNA immunoprecipitation techniques coupled to NGS I found that Top1 depletion induces an increase of R loops at a genome-wide level. We found that such increase occurs on the entire gene body. At a subset of loci R loops resulted particularly stressed after Top1 depletion: some of these genes showed the formation of new R loops structures, whereas other loci showed a reduction of R loops. Interestingly we found that new peaks usually appear at tandem or divergent genes in the entire gene body, while losses of R loop peaks seems to be a feature specific of 3’ end regions of convergent genes

Enduring Fluoride Health Hazard for the Vesuvius Area Population: The Case of AD 79 Herculaneum

Background: The study of ancient skeletal pathologies can be adopted as a key tool in assessing and tracing several diseases from past to present times. Skeletal fluorosis, a chronic metabolic bone and joint disease causing excessive ossification and joint ankylosis, has been only rarely considered in differential diagnoses of palaeopathological lesions. Even today its early stages are misdiagnosed in endemic areas. Methodology/ Principal Findings: Endemic fluorosis induced by high concentrations of fluoride in water and soils is a major health problem in several countries, particularly in volcanic areas. Here we describe for the first time the features of endemic fluorosis in the Herculaneum victims of the 79 AD eruption, resulting from long-term exposure to high levels of environmental fluoride which still occur today. Conclusions/ Significance: Our observations on morphological, radiological, histological and chemical skeletal and dental features of this ancient population now suggest that in this area fluorosis was already endemic in Roman times. This evidence merged with currently available epidemiologic data reveal for the Vesuvius area population a permanent fluoride health hazard, whose public health and socio-economic impact is currently underestimated. The present guidelines for fluoridated tap water might be reconsidered accordingly, particularly around Mt Vesuvius and in other fluoride hazard areas with high natural fluoride levels

Malignant Glomus Tumour: A Case Report and Review of the Literature

Purpose: Glomus tumours are characteristically benign solitary tumours. At our knowledge, about 23 reports are present in literature regarding the malignant counterpart, but only a minority developed metastases. We describe a locally aggressive glomus tumour with lymphnode metastasis

La morfologia carsica della provincia di Lecce e la sua influenza sulla idrografia superficiale e profonda (studio preliminare)

ItIl censimento delle cavità carsiche epigee presenti nell’ambito del territorio della provincia di Lecce e la loro tipicizzazione in relazione alla costituzione del substrato, hanno permesso di valutare l’incidenza della locale morfologia carsica nel deflusso delle acque in superficie e nella alimentazione delle falde nel sottosuolo. Le differenti situazioni riscontrate sono state comparate con una mappa della piovosità ottenuta tramite l’analisi statistica dei dati esistenti, nonché con un quadro sinoptico delle opere di canalizzazione e di recapito delle acque superficiali, con lo scopo di portare un preliminare contributo alla soluzione dei problemi connessi sia con lo smaltimento delle acque meteoriche e dei reflui urbani sia con i possibili processi di desertificazione.EnInvestigation concerned in karst holes which are present in the territory of Salento and their relationship with the characteristics of the substratum, has allowed to appreciate the incidence of the local karstic morphology in the outflow of the waters on surface and in the feeding in the groundwater. The different situations have been comparative with a map of the rains got through the statistic analysis of the existing data, as well as with a picture of the works of canalization of superficial waters. The purpose is to bring a preliminary contribution to the problem connected with the disposal of the meteoric waters and the urban refluis and with the possible processes of desertification. The integration of all the geological, stratigraphical, hydrogeological, geomorfological and climatic data in elaboration will allow us to effect a verify of the actual state of the territory. A first analysis of all the elaborate data has put in evidence that: - in the oriental zone, among Otranto, Minervino and Presicce, in the EST and Manduria in the WEST, characterized by the maximum precipitations, the meteoric waters quickly are absorbed in the subsoil for the nature of the permeable calcareous substratum for fractures and particularly for karst; here the outflows are also very scarce because of the planar morphology. - in the western zone, that is among Taviano, Nardò and Copertino, in the EST and Gallipoli and Manduria in the WEST, where are weak precipitations, the geological and particularly morphological character condition the outflow of the waters determining the formation of hydrographic networks which are more diffused in comparison to the east zone and mainly developed along the borders of the watershed (Fig. 1); on the contrary, where coverages of clayey grounds are present and morphologies are planar, superficial stagnations are verified

Exploring Outcome Priorities and Real-Life Management of Chemotherapy-Induced Peripheral Neurotoxicity: A Survey of the Italian Association for the Study of Pain members

Introduction: Chemotherapy-induced peripheral neurotoxicity (CIPN) affects nearly 70% of cancer patients after chemotherapy, causing sensory, motor, autonomic dysfunction, and neuropathic pain. The Desirability of Outcome Ranking (DOOR) framework is proposed as a better way to assess preventive or therapeutic interventions for CIPN. Methods: A survey was conducted among Italian healthcare professionals and researchers affiliated to the Italian Chapter of the International Association for the Study of Pain (AISD) to identify the most important outcomes in clinical management and research. Results: Among the 73 respondents, 61 qualified for the survey, with an overall response rate of 1.2%. The vast majority were physicians (77%), most of whom were anesthesiologists (47.5%). The results showed that pain, survival, sensory impairment, motor impairment, and quality of life were consistently ranked as the most important outcomes, but there was significant disagreement in the outcomes relative ranking, making it difficult to develop a DOOR algorithm. The study also revealed that clinicians commonly use structured interviews to evaluate patients with CIPN, and the most prescribed drugs or supplements were palmitoylethanolamide, pregabalin, gabapentin and alpha lipoic acid as preventive agents and pregabalin, palmitoylethanolamide, duloxetine, gabapentin, and amitriptyline as therapeutic agents. However, many of these drugs have not been clinically proven to be effective for CIPN. Discussion: This study suggests that the implementation of a DOOR framework for CIPN using healthcare professionals is more difficult than expected, given the significant disagreement in our respondents' ranking of outcomes. Our work provides interesting topics for future research in CIPN, but its limitations include a small sample size, a low response rate, and a possible selection bias

Tramadol/dexketoprofen (TRAM/DKP) compared with tramadol/paracetamol in moderate to severe acute pain: results of a randomised, double-blind, placebo and active-controlled, parallel group trial in the impacted third molar extraction pain model (DAVID study)

Objectives: To compare efficacy/safety of oral tramadol 75 mg/dexketoprofen 25 mg (TRAM/DKP) and TRAM 75 mg/paracetamol 650 mg (TRAM/paracetamol) in moderate to severe pain following surgical removal of impacted lower third molar. Design: Multicentre, randomised, double-blind, placebo-controlled, phase IIIb study. Participants: Healthy adult patients scheduled for surgical extraction of at least one fully/partially impacted lower third molar requiring bone manipulation. 654 patients were randomised and 653 were eligible for analysis. Interventions: Surgery was performed under local anaesthetic. No sedation was permitted. Patients rated pain intensity (PI) using an 11-Numerical Rating Scale (NRS) (0 no pain; 10 worst pain). Participants experiencing moderate/severe pain (≥4) within 4 hours of surgery were randomised (2:2:1 ratio) to a single oral dose of TRAM/DKP 75/25 mg, TRAM/paracetamol 75/650 mg or placebo. Main outcome measures: Efficacy was based patients' electronic diaries. Analgesia and pain were recorded as follows: pain relief (PAR) on a 5-point Verbal Rating Scale (0='no relief', 1='a little (perceptible) relief', 2='some (meaningful) relief', 3='lot of relief', 4='complete relief') at the predefined postdose time points t15 min, t30 min, t1 hour, t1.5 hour, t2 hour, t4 hour, t6 hour and t8 hour and PI on the 11-point NRS at t0 and at the same predefined postdose time points. Onset of analgesia documented using double stopwatch method over a 2-hour period. Primary endpoint was total pain relief over 6 hours (TOTPAR6). Rescue medication was available during the treatment period. Results: TRAM/DKP was superior to TRAM/paracetamol and placebo at the primary endpoint TOTPAR6 (p<0.0001). Mean (SD) TOTPAR6 in the TRAM/DKP group was 13 (6.97), while those in the active control and placebo groups were 9.2 (7.65) and 1.9 (3.89), respectively. Superiority of TRAM/DKP over active comparator and placebo was observed at all secondary endpoints. Incidence of adverse events was comparable between active groups. Conclusions: TRAM/DKP (75/25 mg) is effective and superior to TRAM/paracetamol (75/650 mg) in relieving moderate to severe acute pain following surgical removal of impacted lower third molar, with a faster onset of action, greater and durable analgesia, together with a favourable safety profile

Wetland selection, niche breadth and overlap in 10 duck species wintering in northern Campania (Italy)

Analysis of the selection index, the niche breadth and the niche overlap of 10 species of wintering ducks in 11 wetland areas of the Campania region between January 2003 and 2015 revealed that the distribution of the species in the examined sites was not casual. In fact, each species actively chose their wintering area. Using selection index, we were able to measure the species’ preferences and non-preferences for the 11 examined sites. By calculating Shannon-Wiener entropy and its normalized value (Evenness), we determined niche breadth; the Morisita index allowed us to check niche overlap

Design, synthesis and preliminary pharmacological evaluation of new imidazolinonesas L-DOPA prodrugs

L-DOPA, the immediate biological precursor of dopamine, is still considered the drug of choice in the treatment of Parkinson's disease. However, therapy with L-DOPA is associated with a number of acute problems. With the aim to increase the bioavailability after oral administration, we designed a multi-protected L-DOPA prodrugs able to release the drug by both spontaneous chemical or enzyme catalyzed hydrolysis. The new compounds have been synthesized and preliminarily evaluated for their water solubility, log P, chemical stability, and enzymatic stability. The results indicate that the incorporation of the amino acidic moiety of L-DOPA into an imidazoline-4-one ring provides prodrugs sufficiently stable to potentially cross unchanged the acidic environment of the stomach, and to be absorbed from the intestine. They also might be able to release L-DOPA in human plasma after enzymatic hydrolysis. The ability of prodrugs 6a-b to increase basal levels of striatal DA, and influence brain neurochemistry associated with dopaminergic activity following oral administration, as well as the radical-scavenging activity against DPPH for compounds 6a-b and 15a are also reported

Malignant glomus tumour: a case report and review of the literature. Sarcoma

Abstract Purpose: Glomus tumours are characteristically benign solitary tumours. At our knowledge, about 23 reports are present in literature regarding the malignant counterpart, but only a minority developed metastases. We describe a locally aggressive glomus tumour with lymphnode metastasis. Patient: The patient was a 40 year-old man presenting a 1.5-cm lesion on the right wrist incompletely excised and a recurrent tumour, 4 Â 2 cm in size, removed after 9 months, for which he received radiotherapy. After 2 years he developed an axillary lymphnode metastasis. Results: Histologically, both tumours (primary and metastasis) were similar. There were sheets and nests of uniform small cells with scant eosinophilic cytoplasm and round to polygonal nuclei; there was some degree of pleomorphism and the mitotic index was high (up to 18 m/10 HPF). The tumour cells were positive for vimentin and smooth muscle actin, but negative for desmin, NSE, Factor VIII, chromogranin, cytokeratin. Remarkably, in the primary, the cells strongly expressed p53 (70%) and MIB-1 (35%). Discussions: In many reported malignant cases, the histology of the tumour cells suggested that they were malignant, yet the clinical course has been benign. Carefully reviewing the literature, it seems that actually we have enough histological criteria to identify the cases with biological adverse outcome. Those unfortunate cases behave as high grade sarcomas and therefore may deserve an aggressive therapeutic treatment

Physical Activity in Patients with Neuromuscular Disease Three Years after COVID-19, a Longitudinal Survey: The After-Effects of the Quarantine and the Benefits of a Return to a Healthier Life-Style

Background: Quarantine was one of the strategies adopted by governments against the spread of COVID-19. This restriction has caused an increase in sedentary behaviors and a decrease in the practice of physical activity (PA), with a consequent negative impact on lifestyle both in healthy people and in those who need constant practice of PA to combat diseases, such as patients suffering from neuromuscular diseases (NMDs). Hence, this study aimed to compare PA levels among patients with NMD during and after quarantine. Methods: An adapted version of the International Physical Activity Questionnaire Short-Form and the Short-Form Health Survey were administered during COVID-19 quarantine (T0) and after 3 years (T1) to 91 Italian patients with NMDs. Results: We found a significant increase in the total PA level at T1, with no significant changes in vigorous-intensity PA. Moreover, a significant decrease in the PA level was found among the patients with different NMDs. No significant changes in physical component scores and mental component scores were detected. Conclusions: Our results suggest that it would be necessary to provide alternative indoor exercise settings to prevent the adoption of sedentary behaviors