Garbage in, garbage out: how reliable training data improved a virtual screening approach against SARS-CoV-2 MPro

Introduction: The identification of chemical compounds that interfere with SARS-CoV-2 replication continues to be a priority in several academic and pharmaceutical laboratories. Computational tools and approaches have the power to integrate, process and analyze multiple data in a short time. However, these initiatives may yield unrealistic results if the applied models are not inferred from reliable data and the resulting predictions are not confirmed by experimental evidence.Methods: We undertook a drug discovery campaign against the essential major protease (MPro) from SARS-CoV-2, which relied on an in silico search strategy –performed in a large and diverse chemolibrary– complemented by experimental validation. The computational method comprises a recently reported ligand-based approach developed upon refinement/learning cycles, and structure-based approximations. Search models were applied to both retrospective (in silico) and prospective (experimentally confirmed) screening.Results: The first generation of ligand-based models were fed by data, which to a great extent, had not been published in peer-reviewed articles. The first screening campaign performed with 188 compounds (46 in silico hits and 100 analogues, and 40 unrelated compounds: flavonols and pyrazoles) yielded three hits against MPro (IC50 ≤ 25 μM): two analogues of in silico hits (one glycoside and one benzo-thiazol) and one flavonol. A second generation of ligand-based models was developed based on this negative information and newly published peer-reviewed data for MPro inhibitors. This led to 43 new hit candidates belonging to different chemical families. From 45 compounds (28 in silico hits and 17 related analogues) tested in the second screening campaign, eight inhibited MPro with IC50 = 0.12–20 μM and five of them also impaired the proliferation of SARS-CoV-2 in Vero cells (EC50 7–45 μM).Discussion: Our study provides an example of a virtuous loop between computational and experimental approaches applied to target-focused drug discovery against a major and global pathogen, reaffirming the well-known “garbage in, garbage out” machine learning principle

Which Target Temperature for Post-Anoxic Brain Injury? A Systematic Review from “Real Life” Studies

There is a persistent debate on the optimal target temperature to use during cooling procedures in cardiac arrest survivors. A large randomized clinical trial (RCT) including more than 900 patients showed that targeted temperature management (TTM) at 33 °C had similar mortality and unfavorable neurological outcome (UO) rates as TTM at 36 °C in out-of-hospital cardiac arrest patients with any initial rhythm. Since then, several observational studies have been published on the effects of changes in target temperature (i.e., from 33 to 36 °C) on patients’ outcome. We performed a systematic literature search from 1 January 2014 to 4 December 2020 and identified ten retrospective studies (very low levels of certainty; high risk of bias), including 5509 patients, that evaluated TTM at 33 °C vs. TTM at 36 °C on the occurrence of UO (n = eight studies) and mortality (n = ten studies). TTM at 33 °C was associated with a lower risk of UO when studies assessing neurological outcome with the Cerebral Performance Categories were analyzed (OR 0.80 [95% CIs 0.72–0.98]; p = 0.03). No differences in mortality were observed within the two TTM strategies. These results suggest that an inappropriate translation of TTM protocols from large well-conducted randomized trials into clinical management may result in unexpected effects on patients’ outcome. As for all newly commercialized drugs, epidemiological studies and surveillance programs with an adequate follow-up on large databases are necessary to understand how RCTs are implemented into medical practice

Chronic Exertional Compartment Syndrome of the Forearm: Mini-invasive Technique Using an Endoscopic Soft Tissue Release System

Chronic exertional compartment syndrome of the forearm is a rare disease characterized by the recurrent onset of forearm pain and progressive weakness during exertion. Over the years many surgical techniques have been reported in literature: wide-open fasciotomy, mini-open fasciotomy, fasciotomy associated with fasciectomy, and mini-invasive endoscopic fasciotomy. Compared with traditional open techniques, mini-invasive techniques are characterized by minimal tissue trauma, less postoperative pain, faster recovery to normal activity and better esthetical result. We describe a mini-invasive endoscopic technique for the treatment of forearm chronic exertional compartment syndrome using the SmartRelease endoscopic tissue release system (MicroAire)

Efficient global optimization of reservoir geomechanical parameters based on synthetic aperture radar-derived ground displacements

none6sirestrictedComola, Francesco; Janna, Carlo; Lovison, Alberto; Minini, Marco; Tamburini, Andrea; Teatini, PietroComola, Francesco; Janna, Carlo; Lovison, Alberto; Minini, Marco; Tamburini, Andrea; Teatini, Pietr

Which target temperature for post-anoxic brain injury? A systematic review from “real life” studies

There is a persistent debate on the optimal target temperature to use during cooling procedures in cardiac arrest survivors. A large randomized clinical trial (RCT) including more than 900 patients showed that targeted temperature management (TTM) at 33° C had similar mortality and unfavorable neurological outcome (UO) rates as TTM at 36° C in out-of-hospital cardiac arrest patients with any initial rhythm. Since then, several observational studies have been published on the effects of changes in target temperature (i.e. from 33 to 36° C) on patients’ outcome. We performed a systematic literature search from 1 January 2014 to 4 December 2020 and identified nine retrospective studies (very low levels of certainty; high risk of bias), including 3799 patients, that evaluated TTM at 33° C vs. TTM at 36° C on the occurrence of UO (n = seven studies) and mortality (n = nine studies). TTM at 33° C was associated with a lower risk of UO when studies assessing neurological outcome with the Cerebral Performance Categories were analyzed (OR 0.80 [95% CIs 0.65–0.99]; p = 0.04). No differences in mortality were observed within the two TTM strategies. These results suggest that an inappropriate translation of TTM protocols from large well-conducted randomized trials into clinical management may result in unexpected effects on patients’ outcome. As for all newly commercialized drugs, epidemiological studies and surveillance programs with an adequate follow-up on large databases are necessary to understand how RCTs are implemented into medical practice.SCOPUS: re.jinfo:eu-repo/semantics/publishe

Chronic exertional compartment syndrome of the forearm: a systematic review

The aim of this systematic review is to understand which surgical procedure provides better results in terms of pain relief and function in the treatment of chronic exertional compartment syndrome (CECS) of the forearm.We searched Medline (PubMed), Web of Science, Embase and Scopus databases on 8 July 2020. Twelve studies were included in this review.We assessed the quality of the studies using the Coleman Methodological Score.Data on demographic features, operative readings, diagnostic methods, follow-up periods, type and rates of complications, survivorship of the procedure, return to sport activity, and outcome measures were recorded.In conclusion, compared to the other techniques, endoscopic fasciotomy delivers similar success rates and lower incidence of complications. Cite this article: EFORT Open Rev 2021;6:101-106. DOI: 10.1302/2058-5241.6.200107

The activation of miR-125a-5p/IP6K1 axis in breast cancer cells upon treatment with myo-Inositol.

Several studies have been performed with the aim of identifying drugs able in inhibiting Epithelial-Mesenchymal Transition (EMT), chiefly by blocking PI3K/Akt pathway. We have already demonstrated that treatment with myo-Inositol at the pharmacological dose can block EMT in breast cancer cells by downregulating PI3K/Akt and inducing changes in cytoskeletal architecture. Herewith, we investigated the mechanism of action of myo-inositol in both highly (MDA-MB-231) and low (MCF-7) invasive human breast cancer cells. After 30’ and 24h from treatment, gene expression analysis revealed a significant downregulation of Pi3k and Psen1 after 30’ in both cell lines. Psen1 downregulation was maintained in MDA-MB-231 at 24h. Likewise, we explored the modulation of Ip6k1, Dnmt3b, Isyna1 and p53. In MDA-MB-231, a strong downregulation of Ip6k1 expression was recorded at 30’ and 24h, whilst Dnmt3b was reduced only at 30’. On the contrary, in MCF-7, Ip6k1 downregulation was unexpectedly associated to the upregulation of Dnmt3b at 30’. IP6K1 is a key enzyme of inositol metabolism, inhibits ISYNA1, probably inducing de novo DNA methylation (i.e., DNMT3B). Furthermore, IP6K1 inhibition correlates with a decrease of cancer cells motility. The upregulation of Isyna1 was observed in both cell lines at 30’, together with p53. ISYNA1 activates myo-Inositol intracellular biosynthesis starting from glucose-6-phosphate. In this activation, p53 plays a key role in binding Isyna1 promoter and eventually enabling its expression. Western-blot of MDA-MB-231 confirmed that changes in gene expression were also mirrored by concurrently modifications in IP6K1 and p53 protein levels, altogether with a decrease of both MDM2 and YAP/TAZ. It is worth noting that in MCF-7, no changes were observed in protein levels. In-silico analysis was performed using TCGA miRNA-Seq data to identify differentially expressed miRNAs between normal and tumoral tissue in breast cancer patients. To further gain mechanistic insights on myo-Inositol effects, we compared these data with main differentially expressed cancer-related miRNAs in MDA-MB-231 cells after 30’ from treatment. This analysis allowed to identify two mRNAs, downregulated in tumor tissues, that were significantly increased with myo-Inositol: miR-92a-3p and miR-125a-5p. Using DIANA tools, miR-92a-3p was predicted to interact with Notch-1 and PI3K, linking it to cytoskeletal rearrangement. Moreover, a strong interaction was predicted between miR-125a-5p and IP6K1 in 3’-UTR site. Indeed, the upregulation of miR-125a-5p is usually correlated with metastasis inhibition in breast cancer. In MDA-MB-231, miR-125a-5p upregulation was maintained at 24h, while in MCF-7 was slightly upregulated at 30’ and downregulated at 24h. Our results suggest that myo-Inositol causes early changes in gene expression, probably led by miRNAs and methylation remodeling. Elucidation of the role of miR-125a-5p/IP6K1 axis will reveal strategies for molecular targeted therapies in breast cancer

Myo-Inositol treatment inhibits motility in triple negative breast cancer via miR-125a-5p/IP6K1 axis

Background: Several researches have been performed with the aim of identifying drugs able in blocking PI3K/Akt pathway. We have already demonstrated that myo-Inositol (myo-Ins) treatment can block EMT in breast cancer cells by downregulating PI3K/Akt and inducing changes in cytoskeletal architecture. Aim: Herein, we set our experiments to investigate migration/invasiveness inhibition though in vitro and in vivo models upon myo-Ins administration. Methods: In vitro experiments were performed using both mesenchymal-like (MDA-MB-231) and epithelial-like (MCF-7) invasive human breast cancer cells. We used transwell assays for in vitro and Zebrafish embryos as in vivo models to evaluate migration and invasiveness. The expression of key genes involved in the mechanism was evaluated by qPCR, while gain- and loss-of-function approaches allowed identifying the specific dynamical relationships. Results: Myo-Ins inhibits motility and invasiveness only in MDA-MB-231 cells both in vitro and in vivo. In MDA cells, miR-125a-5p upregulation was linked to IP6K1 downregulation triggered by myo-Ins treatment. Silencing and overexpression experiments confirmed the key role of miR-125a-5p/IP6K1 axis in blocking cell motility. This effect was demonstrated to be myo-Ins-dependent MDM2 inhibition. Given that MDM2 in MCF-7 cells was unaffected by treatment, in these cells myo-Ins was unable in antagonizing motility. Conversely, both miR-125a-5p and IP6K1 were not modulated. However, MDM2 silencing restore sensitivity to myo-Ins, thus leading to a significant inhibition of the MCF-7 cells motility capability. Conclusions: Our results suggest that myo-Ins can inhibit motility in triple negative breast cancer. Such an effect is likely mediated by MDM2 inhibition, which, in turn, triggers a complex tumor reversion promoted by the miR-125a-5p/IP6K1 axis modulation. Elucidation of the role of miR-125a- 5p/IP6K1 axis will reveal strategies for molecular targeted therapies in breast cancer

Pain pupillary index to prognosticate unfavorable outcome in comatose cardiac arrest patients

Background: The prognostic role of the Pupillary Pain Index (PPI), derived from automated pupillometry, remains unknown in post-anoxic brain injury. Methods: Single-center retrospective study in adult comatose cardiac arrest (CA) patients. Quantitative PPI and Neurologic Pupil Index (NPi) were concomitantly recorded on day 1 and day 2 after CA. The primary outcome was to assess the prognostic value of PPI to predict 3-month unfavourable outcome (UO, defined as Cerebral Performance Category of 3 & ndash;5). Secondary outcome was the agreement between PPI and NPi to predict unfavourable outcome. Results: A total of 102 patients were included; patients with UO (n = 69, 68%) showed a lower NPi (4.2 [3.5 & ndash;4.5] vs. 4.6 [4.3 & ndash;4.7]; p < 0.01 on day 1 & ndash;4.3 [3.8 & ndash;4.7] vs 4.6 [4.3 & ndash;4.8] on day 2), and PPI (3 [1 & ndash;6] vs. 6 [3 & ndash;7]; p < 0.01 on day 1 & ndash;3 [1 & ndash;6] vs 6 [4 & ndash;8]; p < 0.01 on day 2) than others. A PPI =1 on day 2 showed a sensitivity of 26 [95% CI 16 & ndash;38]% and a specificity of 100 [95% CI 89 & ndash;100]% to predict UO (p = 0.003 vs. NPi < 2). On day 2, a total of 6 patients had concomitant PPI = 1 and NPi < 2, while 12 showed NPi > 2 and PPI = 1; the coefficient of agreement was 0.42. Moreover, NPi and PPI values showed a moderate correlation both on day 1 and day 2. Conclusions: In this study, PPI = 1 on day 2 could predict UO in comatose CA patients with 100% specificity, but with a low sensitivity (yet higher than NPi). The agreement between PPI and NPi values was moderate