Privileged Scaffold Decoration for the Identification of the First Trisubstituted Triazine with Anti-SARS-CoV-2 Activity

: Current therapy against severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) are based on the use of Remdesivir 1, Molnupiravir 2, and the recently identified Nirmatrelvir 3. Unfortunately, these three drugs showed some limitations regarding potency and possible drug-drug interactions. A series of derivatives coming from a decoration approach of the privileged scaffold s-triazines were synthesized and evaluated against SAR-CoV-2. One derivative emerged as the hit of the series for its micromolar antiviral activity and low cytotoxicity. Mode of action and pharmacokinetic in vitro preliminary studies further confirm the role as candidates for a future optimization campaign of the most active derivative identified with this work. © 2022 by the authors

A novel enzyme blend for efficient tissue dissociation and primary cells isolation

Tissue dissociation/primary cell isolation and cell harvesting are principal appli- cations for enzymes in tissue culture research and cell biology studies. The goal of a cell isolation procedure is to maximize the yield of functionally viable dissoci- ated cells. Among the parameters which affect the outcome of any particular dissociating procedure there are enzyme(s) used and related impurities presents in crude enzyme preparation. ABIEL srl recently produced the recombinant collagenase class I (Col G) and II (Col H) from Clostridium histolyticum (PCT WO 2011/073925 A9). The enzymes were produced in Escherichia coli and purified by affinity chromatography. The method of production adopted allows absolute control of the final composition of these enzymes, as well as their stability, purity, activity, absence of toxicity and higher reproducibility of batches. The two collagenases produced separately have been used in conjunction according to precise proportions to dissociate calvaria, liver, pancreas, retina of the BALB/c mouse; and bovine hoof. The analyses carried out on all isolated cell populations suggest that the cells maintain the structural and functional integrity of specific tissues/organs originating. Recombinant Col G and Col H enzymes produced by ABIEL are promising in the context of the tissue/cells dissociation, with the aim to make innovation in the fields of tissue engineering and transplantation medicine

Evolving trends in the management of acute appendicitis during COVID-19 waves. The ACIE appy II study

Background: In 2020, ACIE Appy study showed that COVID-19 pandemic heavily affected the management of patients with acute appendicitis (AA) worldwide, with an increased rate of non-operative management (NOM) strategies and a trend toward open surgery due to concern of virus transmission by laparoscopy and controversial recommendations on this issue. The aim of this study was to survey again the same group of surgeons to assess if any difference in management attitudes of AA had occurred in the later stages of the outbreak. Methods: From August 15 to September 30, 2021, an online questionnaire was sent to all 709 participants of the ACIE Appy study. The questionnaire included questions on personal protective equipment (PPE), local policies and screening for SARS-CoV-2 infection, NOM, surgical approach and disease presentations in 2021. The results were compared with the results from the previous study. Results: A total of 476 answers were collected (response rate 67.1%). Screening policies were significatively improved with most patients screened regardless of symptoms (89.5% vs. 37.4%) with PCR and antigenic test as the preferred test (74.1% vs. 26.3%). More patients tested positive before surgery and commercial systems were the preferred ones to filter smoke plumes during laparoscopy. Laparoscopic appendicectomy was the first option in the treatment of AA, with a declined use of NOM. Conclusion: Management of AA has improved in the last waves of pandemic. Increased evidence regarding SARS-COV-2 infection along with a timely healthcare systems response has been translated into tailored attitudes and a better care for patients with AA worldwide

RUOLO DI ENZIMI PROTEOLITICI NELLA NEOANGIOGENESI TUMORALE. CROSS-TALK TRA CELLULE ENDOTELIALI, TUMORALI E MATRICE EXTRACELLULARE

Ruolo di enzimi proteolitici nella neoangiogenesi tumorale. “Cross-talk” tra cellule endoteliali, tumorali e matrice extracellulare. S. Saladino e G. Ghersi Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università di Palermo, Viale delle Scienze, edificio 16, Palermo-Italia. [email protected] L’angiogenesi è un fenomeno complesso che consiste nella creazione di nuovi vasi sanguigni a partire da quelli pre-esistenti (Folkman J, 1992). Tappa critica di questo processo multifasico è il rimodellamento dell’ECM, ad opera di enzimi proteolitici quali metalloproteasi, sistema attivatore del plasminogeno (PA)-plasmina, serino-proteasi transmembranarie di tipo II (Yoon SO et al, 2003; Pepper MS et al, 2001). Il ruolo degli enzimi litici e le modalità di attivazione finora non sono stati pienamente elucidati. La comprensione di questi meccanismi potrebbe sicuramente rivelarsi utile per fini terapeutici anti-tumorali, probabilmente medinte l’inibizione di bersagli specifici o l’inibizione di pathway molecolari. Sono stati realizzati sistemi di co-coltura tra cellule endoteliali (ECV304) e cellule di carcinoma mammario con fenotipo invasivo (MDA-MB-231 e 8701-BC), mediante l’utilizzo di transwells. Le cellule endoteliali sono state seminate all’interno di transwell sia stratificati con collagene di tipo I (coltura 2D), sia ricoperti con un sottile gel tridimensionale dello stesso collagene (coltura 3D). Sulle co-colture, sono stati effettuati saggi di migrazione ed invasività cellulare, mediante microscopia ottica e confocale. Inoltre, ECV304 seminate su gel di collagene sono state incubate con i mezzi condizionati delle linee tumorali. I lisati cellulari e i surnatanti ottenuti da queste colture sono state sottoposte a zimografia, per studiare l'espressione specifici enzimi litici. I saggi di invasività in transwell suggeriscono che le cellule endoteliali migrano in misura maggiore quando coltivate con le linee tumorali mammarie, piuttosto che da sole (controllo). In presenza del tumore, le ECV304 acquisiscono un fenotipo più mesenchimale rispetto al controllo, e tendono ad invadere la matrice extracellulare, come si evince dalle misurazioni in fluorescenza. Gli zimogrammi mostrano in particolare un'espressione della metalloprotesi MMP9 maggiore nei campioni che derivano dalle colture con i mezzi condizionati tumorali. Nessun incremento di espressione significativo è stato osservato per MMP2. Il modello di co-coltura tridimensionale adottato rivela, dunque, un'attivazione delle cellule ECV304 da parte delle linee di tumore mammario invasivo MDA-MB-231 e 8701-BC. Le cellule cancerose potrebbero indurre un potenziale migratorio ed invasivo in quelle endoteliali; stimolando l'espressione di MMP9. Analisi Western-Blot e saggi enzimatici specifici potrebbero fornire dati più utili alla comprensione del fenomeno. In futuro, lo stesso sistema sperimentale verrà allestito con cellule endoteliali derivanti da microvasi del derma, più adatta per lo studio dell'angiogenesi tumorale

3D collagen matrix improves islets of Langerhans differentiation and function

The importance of preserve cell-cell contact and cell-matrix interaction are both fundamental for maintain the differentiation state. We use both standardized method of isolation and culture in 3D matrix which improves islet cell survival and function for more than one week in colture

Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II

Collagenases class I (Col G) and class II (Col H) currently available for tissue dissociation are produced from Clostridium histolyticum (human pathogen) strains. In the processes of extraction of the cells from the tissue, combined activity of both classes of enzymes is required. CI and CII are complementary in degrading collagen. ABIEL recently produced the collagenase class I and II using the recombinant DNA technologies (PCT WO 2011/073925 A9). The enzymes were produced in E. coli and purified by affinity chromatography. The method of production adopted allows absolute control of the final composition of these enzymes, as well as their stability, purity, activity, absence of toxicity and higher reproducibility of batches of collagenase. The two collagenases produced separately have been used in conjunction according to precise proportions to dissociate calvaria and liver of the BALB/c mouse and bovine hoof. The analysis carried out on all isolated cell populations suggest that the cells maintain the structural and functional integrity of specific tissues/organs originating. Recombinant Col G and Col H enzymes represent a promising tool for tissue dissociation