Sara Ferraiuolo

Experimental thesis in Chemistry and Pharmaceutical Technology (CTF), at the University of Camerino (Unicam);

In particular, the experimental thesis carried out involved the synthesis of Inhibitors (and possible Activators) of SARM-1, an enzyme implicated in the pathways of multiple Neurodegenerative diseases (ALS, Wallerian degeneration, Alzheimer's, Parkinson's...), and subsequent analysis of the same by NMR spectroscopy instrumentation, of the 1H and 13C type, as well as by mass spectrometry (MS).

• Organizing and carrying out the workload independently and efficiently.
• Support and collaboration with colleagues in handling assigned tasks.
• Carrying out assigned tasks quickly and efficiently.
• Handling unforeseen events and work problems with speed and efficiency.
• Organizing activities according to urgency and assigned workload.
• Consistent compliance with laboratory safety regulations and procedures.
• Performing laboratory analysis and testing for data acquisition and determination of quantitative and qualitative composition of substances.

Presented the first Scientific Poster, among whose authors is my name, by Doctoral student Matteucci Federica (at Unicam, University of Camerino), on the occasion of the transition from the first to the second year of her doctoral program, at the ChIP, Chemistry Interdisciplinary Project (Unicam, Camerino);

Poster title: NOVEL STERILE ALPHA AND TOLL INTERLEUKIN RECEPTOR MOTIF CONTAINING-1 (SARM-1) AND NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE (NAPRT) MODULATORS AS POTENTIAL NEUROPROTECTIVE AND ANTITUMOR AGENTS .

• Licensed to practice as a pharmacist
• Achieved grade: 100/100

• Grade achieved: 110/110 cum laude with honors awarded by the Commission.

Presented scientific poster, whose authors include my name, by Professor Cappellacci Loredana, Professor at Unicam (Camerino).
The Poster was presented at the NMMC in Chieti (2023), a national meeting convened by the Division of Medicinal Chemistry of the Italian Chemical Society, in conjunction with which advances in medicinal chemistry and drug discovery are discussed, within the main therapeutic areas, including infectious, neurodegenerative, rare and cancer diseases.
Poster title: NOVEL STERILE ALPHA AND TOLL INTERLEUKIN RECEPTOR MOTIF CONTAINING-1 (SARM-1) INHIBITORS AS POTENTIAL NEUROPROTECTIVE AGENTS

• PF24 Training Pathway and subsequent acquisition of 24 CFUs in anthropo-psycho-pedagogical disciplines and educational methodologies and technologies.

43rd Advanced Course of Medicinal Chemistry and Seminar for PhD students

• Presented the third Scientific Poster, whose authors include my name, by, once again, PhD student Matteucci Federica, at the European School of Medicinal Chemistry ESMEC in Urbino 2024, whose mission is to update PhD students and young researchers in academia and industry on the latest advances in medicinal chemistry and organic chemistry, pharmacology, molecular biology and analytical/structural chemistry.

Poster title: NEW INHIBITORS OF STERILE ALPHA AND TOLL INTERLEUKIN RECEPTOR MOTIF CONTAINING-1 (SARM1) AS POTENTIAL NEUROPROTECTIVE AGENTS .

• The above Poster was also brought and presented at the 1st PhD Summer Day (2024) "Chemical and Pharmaceutical Sciences and Biotechnology," Congress held at ChIP, Unicam Research Center (Camerino).

• Research activity (with Fellowship), at the Department of Medical Chemistry and Biophysics, within the laboratory of Professor Anders Hofer (Umeå University, Sweden), taking advantage of a Fellowship for advanced training abroad.
  
  Activities performed:
  1. In vitro assays of compounds (natural extracts and essential oils) tested against Trypanosoma Brucei, a parasite responsible for African Human Trypanosomiasis (also known as "sleeping sickness");
  2. In vitro assays of compounds (natural extracts and essential oils) whose cytotoxicity was tested on mammalian cells;
  3. HPLC analysis of nucleotide pools.

• I have, in addition, had the occasion to investigate the inherent aspect of performing (under the supervision of Professor Anders Hofer) protein-type assays:
  
  1. Investigation of the allosteric regulation of Ribonucleotide Reductase (RNR) protein, having one of the most complex allosteric regulations known in nature, knowledge of which would be of extreme utility for medicinal purposes, such as the treatment of bacterial diseases;
  2. Studied reductive chelation procedures, and carried out studies on the protein activity of the VcR1 and VcR2 subunits, the modulation of which was analyzed by addition of co-factors, such as Iron (III) and Manganese (IV).
  3. The whole analysis was correlated by subsequent HPLC runs, and subsequent analysis of the obtained chromatograms.

Skills Acquired:

• GraphPad Prism (data processing program)
• Microsoft Excel
• PowerPoint
• Cell counting
• Cell culture
• In vitro assays on Mammalian cells
• In vitro assays on Trypanosoma brucei parasites
• Use of Fluorescence microplate reader
• HPLC
• Use of laboratory instrumentation
• Drafting of research reports and final presentations of results.

Opportunity provided to me by Dr. Orsomando Giuseppe (moreover, one of the supervisors of my thesis), and, on the occasion of which, I had the chance to carry out Research inherent to potential activators/inhibitors (in general, regulators) of the enzyme SARM-1, representing an NAD+ hydrolase and cyclase involved in the process of axonal degeneration.

Recent results have made it possible to attribute to the NAD+ coenzyme the task of counteracting the processes underlying several neurodegenerative diseases, such as those occurring in Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis.

• Conducting laboratory experiments using specific instruments and equipment. In particular:

1. Use and management of preparative FPLC (particularly, I employed columns of type C18 tsk DEAE (anion exchange column) );
2. Subsequent Spectrophotometer analysis;
3. Analysis of the chromatograms obtained, following the performance of the runs, in HPLC and/or FPLC;
4. Management of the lyophilization process (use of the lyophilizer);
5. Use and management of ANALYTIC FPLC (columns used: C18 column TEKNOKROMA (KROMASIL 100 5μm 25x0.46), C18 column VARIAN (5μm 90A 25x0.46) ), and CATIONIC EXCHANGE COLUMN (SourceS);

• Injection (by HPLC) of samples, such as Standards: NAR (Nicotinamide Riboside), Nicotinate, Isoquinoline, various Nucleotide-analogues, etc.

I was, in addition, given the opportunity to delve into the aspect of performing enzymatic tests on the aforementioned SARM-1 enzyme; specifically:

1. Performing tests on the activity of potential activators/inhibitors of SARM-1
(personally synthesized);
2. Purification of the various Nucleosides and Nucleoside analogs (of the FM series), whose activity was tested on the above enzyme;
3. Cloning of the recombinant human NRK1 protein.

• Analysis and interpretation of data collected during research activities;
• Drafting of research reports and final presentation of results.

Professional internship, held at Olivieri Pharmacy (Address: Via Velino, 57, 63074 San Benedetto del Tronto AP), totaling 7 months (> 1000 hours).

• Management and terminal data entry of products in the National Health Service registry.
• Application in the profession of university notions and ethical practices.
• Greeting clients, measuring pressure and other parameters when required.
• Checking inventory and sending replenishment orders in a timely manner to avoid shortages.
• Unpacking of orders, checking their integrity and arranging them on the counter, shelf or in chests and refrigerated spaces, according to how they are stored.
• Setting up and restocking the sales area and any displays.