Nuovi derivati chinossalinici inibitori della Diidrofolato Reduttasi (DHFR) e della Timidilato Sintetasi (TS) della <i>Leishmania major</i>: risultati preliminari

L’incidenza delle malattie protozoarie imputate al genere Leishmania è aumentata enormemente con la diffusione dell’AIDS. In particolare la forma viscerale della Leishmaniosi sta emergendo come malattia nuova e sempre più frequente. Oltre gli antimoniali altri farmaci importanti sono le pentammidine e l’amfotericina B. Un approccio abbondantemente esplorato in passato è stato lo sviluppo di farmaci antifolici antiprotozoari che bloccano il ciclo dell’acido folico, che, previa attivazione metabolica di enzimi folato dipendenti, attraverso il trasporto di unità carboniose, è responsabile della sintesi de novo degli acidi nucleici e degli amminoacidi. In questo senso sono stati usati il trimetoprim e la pirimetamina che sono attivi contro i plasmodi e i toxoplasmi, ma non contro le Leishmanie e i tripanosomi. Questi due protozoi appartenenti alla classe dei Kinetoplastidae sono sensibili agli antifolici antitumorali come il metotrexato (MTX), ma sviluppano immediatamente una chemioresistenza in quanto riattivano l’autoproduzione di acido folico mediante una via di salvataggio che impiega l’enzima Pteridina riduttasi, recentemente scoperto, appunto, nelle Leishmanie major e nel tripanosoma cruzi. Sulla base di queste premesse ci è sembrato che le nostre molecole a struttura chinossalinica, analoghe al metotrexato, potessero essere impiegate come inibitori enzimatici di questa famiglia di folato

Haplotype affinities resolve a major component of goat (<i>Capra hircus</i>) MtDNA D-loop diversity and reveal specific features of the Sardinian stock

Goat mtDNA haplogroup A is a poorly resolved lineage absorbing most of the overall diversity and is found in locations as distant as Eastern Asia and Southern Africa. Its phylogenetic dissection would cast light on an important portion of the spread of goat breeding. The aims of this work were 1) to provide an operational definition of meaningful mtDNA units within haplogroup A, 2) to investigate the mechanisms underlying the maintenance of diversity by considering the modes of selection operated by breeders and 3) to identify the peculiarities of Sardinian mtDNA types. We sequenced the mtDNA D-loop in a large sample of animals (1,591) which represents a non-trivial quota of the entire goat population of Sardinia. We found that Sardinia mirrors a large quota of mtDNA diversity of Western Eurasia in the number of variable sites, their mutational pattern and allele frequency. By using Bayesian analysis, a distance-based tree and a network analysis, we recognized demographically coherent groups of sequences identified by particular subsets of the variable positions. The results showed that this assignment system could be reproduced in other studies, capturing the greatest part of haplotype diversity. We identified haplotype groups overrepresented in Sardinian goats as a result of founder effects. We found that breeders maintain diversity of matrilines most likely through equalization of the reproductive potential. Moreover, the relevant amount of inter-farm mtDNA diversity found does not increase proportionally with distance. Our results illustrate the effects of breeding practices on the composition of maternal gene pool and identify mtDNA types that may be considered in projects aimed at retrieving the maternal component of the oldest breeds of Sardinia.</br

Differentiation impairs Bach1 dependent HO-1 activation and increases sensitivity to oxidative stress in SH-SY5Y neuroblastoma cells

Neuronal adaptation to oxidative stress is crucially important in order to prevent degenerative diseases. The role played by the Nrf2/HO-1 system in favoring cell survival of neuroblastoma (NB) cells exposed to hydrogen peroxide (H2O2) has been investigated using undifferentiated or all-trans retinoic acid (ATRA) differentiated SH-SY5Y cells. While undifferentiated cells were basically resistant to the oxidative stimulus, ATRA treatment progressively decreased cell viability in response to H2O2. HO-1 silencing decreased undifferentiated cell viability when exposed to H2O2, proving the role of HO-1 in cell survival. Conversely, ATRA differentiated cells exposed to H2O2 showed a significantly lower induction of HO-1, and only the supplementation with low doses of bilirubin (0,5-1 \uce\ubcM) restored viability. Moreover, the nuclear level of Bach1, repressor of HO-1 transcription, strongly decreased in undifferentiated cells exposed to oxidative stress, while did not change in ATRA differentiated cells. Furthermore, Bach1 was displaced from HO-1 promoter in undifferentiated cells exposed to H2O2, enabling the binding of Nrf2. On the contrary, in ATRA differentiated cells treated with H2O2, Bach1 displacement was impaired, preventing Nrf2 binding and limiting HO-1 transcription. In conclusion, our findings highlight the central role of Bach1 in HO-1-dependent neuronal response to oxidative stress

Sintesi di una nuova serie di n-[4-(1<i>H</i>(2<i>H</i>)-benzotriazol-1(2)- il)fenil]alchilcarbossiammidi e n,n'-bis[4-(1<i>H</i>(2<i>H</i>)-benzotriazol- 1(2)-il)fenil]alchildicarbossiammidi a potenziale attività antivirale

Recentemente abbiamo riportato la sintesi e l'attività antivirale di un'ampia serie di N-[4- (1H(2H)-benzotriazol-1(2)-il)fenil]alchilcarbossiammidi e N,N’-bis[4-(1H(2H)-benzotriazol- 1(2)-il)fenil]alchildicarbossiammidi. La valutazione biologica nei confronti di vari RNA e DNA virus, ha messo in evidenza una generale interessante attività su alcune specie rappresentative dei Picornavirus, quali Polio Sabin 1 e Coxsackie B1. I derivati più attivi hanno mostrato una EC50 lt; 10 μM, associata con una bassa o nulla citotossicità. Come ulteriore sviluppo di questa ricerca è stata ora progettata la nuova serie di derivati benzotriazolici

Rheumatoid arthritis patient antibodies highly recognize IL-2 in the immune response pathway involving IRF5 and EBV antigens

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by a progressive joint damage due to largely unknown environmental factors acting in concert with risk alleles conferring genetic susceptibility. A major role has been attributed to viral infections that include past contacts with Epstein-Barr virus (EBV) and, more recently, to non-protein coding sequences of human endogenous retrovirus K (HERV-K) integrated in the human genome. Molecular mimicry between viral and self proteins is supposed to cause the loss of immune tolerance in predisposed hosts. There are evidences that anti-IL-2 antibodies (Abs) are present in subjects affected by autoimmune diseases and may be responsible for alterations in regulatory T cell responses. In this study, we evaluated the levels of Abs against IL-2, viral epitopes and interferon regulatory factor 5 (IRF5) in 140 RA patients and 137 healthy controls (HCs). Ab reactivity reached the highest levels for IRF5, EBV and IL-2 (56%, 44% and 39%, respectively) in RA with significantly lower values among HCs (7–9%, p &lt;  0.0001), which suggests a possible cross-reaction between IRF5/EBV homologous antigens and shifts in T cell balance disrupted by anti-IL-2 Abs

Survival as a clinical outcome and its spiritual significance in a cohort of patients with advanced central pelvic neoplastic disease undergoing total pelvic evisceration: a poorly debated issue

Background Patients with either treatment-resistant or relapsing advanced central pelvic neoplastic disease present with a condition responsible for debilitating symptoms and consequently poor quality of life (QoL). For these patients, therapeutic strategies are very limited and total pelvic evisceration is the only option for relieving the symptoms and increasing survival. Of note, taking charge of these patients cannot be limited to increasing their lifespan but must also be aimed at improving the clinical, psychological, and spiritual conditions. This study aimed to prospectively evaluate the improvement in survival and QoL, focusing on spiritual wellbeing (SWB), in patients with poor life expectancy who underwent total pelvic evisceration for advanced gynecological cancers at our center. Patients and methodsThe QoL and SWB were assessed using the European Organisation for Research and Treatment of Cancer QoL questionnaire (EORTC QLQ-C30), EORTC QLQ-SWB32, and SWB scale, which were repeatedly administered: 30 days before surgery, 7 days after the procedure, 1 and 3 months after surgery, and then every 3 months until death or the last follow-up assessment. Operative outcomes (blood loss, operative time, hospitalization, and incidence of complications) were evaluated as secondary endpoints. The patients and their families were included in a dedicated psycho-oncological and spiritual support protocol, which was managed by specifically trained and specialized personnel who accompanied them during all phases of the study. ResultsA total of 20 consecutive patients from 2017 to 2022 were included in this study. Of these patients, 7 underwent total pelvic evisceration by laparotomy and 13 underwent laparoscopy. The median survival was 24 months (range: 1-61 months). After a median follow-up of 24 months, 16 (80%) and 10 patients (50%) were alive at 1 year and 2 years after surgery, respectively. The EORTC-QLQ-C30 scores significantly improved yet at 7 days and at 1, 3, 6, and 12 months, as compared with the preoperative values. In particular, an early improvement in pain, overall QoL, and physical and emotional functions was observed. With respect to the SWB, the global SWB item score of the EORTC QLQ-SWB32 questionnaire significantly increased after 1 month and 3 months, as compared with preoperative values (p = 0.0153 and p = 0.0018, respectively), and remained stable thereafter. The mean SWB scale score was 53.3, with a sense of low overall SWB in 10 patients, a sense of moderate SWB in eight patients, and a sense of high SWB in two patients. The SWB scale score significantly increased after 7 days, 1 month, and 3 months, as compared with the preoperative value (p = 0202, p = 0.0171, and p = 0.0255, respectively), and remained stable thereafter. ConclusionTotal pelvic evisceration is a valid approach for improving both survival and QoL in selected patients with advanced pelvic neoplasms and poor life expectancy. Our results particularly underline the importance of accompanying the patients and their families during the journey with dedicated psychological and spiritual support protocols

HO-1 up-regulation: a key point in high-risk neuroblastoma resistance to bortezomib.

AbstractHigh-risk neuroblastoma (NB) is characterized by the development of chemoresistance, and bortezomib (BTZ), a selective inhibitor of proteasome, has been proposed in order to overcome drug resistance. Considering the involvement of the nuclear factor-erythroid-derived 2-like 2 (Nrf2) and heme oxygenase-1 (HO-1) in the antioxidant and detoxifying ability of cancer cells, in this study we have investigated their role in differently aggressive NB cell lines treated with BTZ, focusing on the modulation of HO-1 to improve sensitivity to therapy. We have shown that MYCN amplified HTLA-230 cells were slightly sensitive to BTZ treatment, due to the activation of Nrf2 that led to an impressive up-regulation of HO-1. BTZ-treated HTLA-230 cells down-regulated p53 and up-regulated p21, favoring cell survival. The inhibition of HO-1 activity obtained by Zinc (II) protoprophyrin IX (ZnPPIX) was able to significantly increase the pro-apoptotic effect of BTZ in a p53- and p21-independent way. However, MYCN non-amplified SH-SY5Y cells showed a greater sensitivity to BTZ in relation to their inability to up-regulate HO-1. Therefore, we have shown that HO-1 inhibition improves the sensitivity of aggressive NB to proteasome inhibition-based therapy, suggesting that HO-1 up-regulation can be used as a marker of chemoresistance in NB. These results open up a new scenario in developing a combined therapy to overcome chemoresistance in high-risk neuroblastoma

Clot characterization by multidisciplinary approach: biochemical and imaging parameters in a hypocoagulative setting. A pilot study.

Background: Clot characterization is, to the present days, a multimodal approach: scanning the clot by electron microscopy (SEM) is helpful for the visualization of fibrin structure along with laboratory parameters such as the clot waveform analysis (CWA) and thrombin generation in different settings of clot abnormalities. This study aimed to assess whether the coagulative parameters were consistent with the clot images texture acquired by SEM, and therefore to propose a more generalist and integrative approach to clots classification.Design and Methods: In this pilot study, the examined population consists of eight healthy subjects, seven patients affected by Acquired Hemophilia A (AHA) and seven patients treated with Vitamin K Antagonists (VKAs), similar for age and gender. We studied the velocity and acceleration (1st and 2nd derivative of the aPTT) of clot formation (CWA), the thrombin generation, and the clots' scanning by SEM. Images acquired with SEM were then analyzed with the MATLAB software with the "Texture Analysis" methods to perform classification. Among the various texture parameters, we reported Contrast and Energy.Results: Significant differences among healthy subjects, patients with AHA and those treated with VKAs were detected for the coagulative parameters. We found no differences between VKAs and AHA patients. Contrast and energy highlighted a significant difference among the three groups in agreement with the laboratory's parameters. We found no significant differences between VKAs and AHA patients.Conclusions: The use of SEM, CWA and thrombin generation parameters may be a starting point for studies aimed to demonstrate the general characteristics of clot formation in different clinical conditions with a multiparametric approach

<i>Thymus catharinae Camarda</i>: comparazione fitochimica della composizione dell'olio essenziale ottenuto da popolazioni puntuali vegetanti spontanee in Sardegna e valutazione della attività biologica

Sono stati intrapresi studi fitochimici al fine di descrivere compiutamente diverse popolazioni spontanee in Sardegna di Thymus catharinae. In diverse zone dell’interno della Sardegna, sono state effettuate raccolte puntuali della specie, al fine di verificare eventuali differenze fitochimiche legate al substrato, all’altitudine ed all’esposizione delle piante e infine valutarne l’attività antiradicalica ed antimicrobica

Low back pain among healthcare workers in a surgical environment: a research protocol

BACKGORUND: Nurses and surgeons are among the healthcare categories with the highest risk of developing low back pain (LBP). The aim of this study is to present the research protocol of a survey for assessing the presence of low back pain among healthcare workers of a surgical ward and its associated factors. METHODS: The survey will be conducted in the Orthopaedics ward of the teaching hospital “Policlinico Umberto I” in Rome, during the year 2018. The low-back pain will be measured using the Italian version of the Nordic Questionnaire musculoskeletal disorders. Socio-demografic and clinical data will be collected, such as: aghe, gender, heaight, weight, body mass index (BMI), type of work, years of work, working timetable. LBP intensity will be assessed with the Numerical Rating Scale. This is a one-dimension numerical scale with possible score from 0 (no pain) to 10 (the worst possible pain). The general health status will be assessed with the SF-12 questionnaire, for calculating the Mental and Physical component scales, MCS and PCS respectively. The physical activity will be assessed with the Italian version of the International Physical Activity Questionnaire. Moreover, work-related stress will be assessed using the Karasec-15 questionnaire. CONCLUSION: This research protocol will be useful for assessing the prevalence of LBP and its associated factors. These informations will be used for planning primary and tertiary prevention