Exploration of scalable industrial platforms for the commercial production of active molecules from microalgae cell walls

Food, nutraceutical, cosmeceutical, pharmaceutical and biomedical industries are putting significant effort into looking for new natural ingredients [1,2]. Microalgae have been recognised as potential sources of high-value chemicals, with most attention focused on antioxidants, pigments and specialty oils [3]. An under-exploited group of biochemicals produced by microalgae are extracellular polymeric substances (EPS) with hyaluronan (HA) representing one of them. Current industrial production methodologies for HA leave opportunities for the establishment of improved routes with higher molecular mass, enhanced biophysical properties, lower production costs and non-bacterial nor animal origins as key unique selling points. At present, incumbent platforms are either based upon Streptococcus spp. (pathogen) bacterial fermentation, modified (GM) Bacillus subtilis or derived from animal tissues. Furthermore, the extraction of various economically exploitable cell components from microalgal biomass is at the core of a successful microalgal biorefinery approach, and it remains a current bottleneck for the economic feasibility of microalgal biotechnological processes [4]. Cell disruption is often required to break down the hard and complicated microalgal cell walls in order to retrieve microalgal constituents such as proteins, lipids, and polysaccharides. Viral enzymes may play a beneficial role in this scenario and might be used to facilitate genetic engineering by overcoming the cell wall barrier or for biorefinery purposes. This project's hypothesis was that it was feasible for microalgae to produce HA. The objectives included investigating a stress-induced platform for the possible production of HA, learning how Chlorovirus/C. varibilis infection leads to HA formation, improving the HA production for the latter platform and looking into intriguing enzymes that can break down polysaccharides. This PhD project focused on exploring, characterizing and developing new platforms in order to achieve profitable industrial production of valuable compounds from microalgae and identify viral enzymes that could help with the processing of Chlorella cells for multiple applications. Two platforms were successfully explored for the production of valuable polysaccharides, and multiple enzymes were identified, produced, characterised and evaluated for Chlorella cell wall digestion to enable possible biorefinery approaches of a non-domesticated Chlorella vulgaris strain

Novel Capsular Polysaccharide from Lobochlamys segnis

In recent years there has been a significant effort from food, nutraceutical, cosmeceutical, pharmaceutical, and biomedical industries to discover and develop new natural ingredients. Microal�gae have been recognised as potential sources of high value chemicals, with most attention focused upon antioxidants, pigments, and specialty oils. An under-exploited group of biochemicals pro�duced by microalgae are extracellular polymeric substances (EPS). Lobochlamys segnis (formerly called Chlamydomonas segnis) was previously reported to produce a large amount of capsular polysaccharide (CPS) closely related to hyaluronan (HA) under stress conditions. In this work, the purified CPS was characterised and shown to have an average molecular mass (Mn) of about 3.7 MDa, and displayed a highly branched random coil structure unlike the simple repeating linear HA polysaccharide. Chemical analysis showed the presence of galactose, glucuronic acid, and glucose sugars confirming that the product has a different composition to that of HA. Mixotrophic growth and stress conditions were identified and improved upon with a pH control system using acetic acid solution to induce efficient production of CPS. Extraction and purification conditions were also identified exploiting the high Mn of the product. The CPS showed thickening properties and both significant antioxidant capacity and reducing power, which could have commercial applications. This is the first report on the characterization of this novel polysaccharide with non-Newtonian properties similar to HA

Effects of Extracorporeal Magnetic Stimulation in Fecal Incontinence.

Background: Fecal incontinence (FI) is a common condition that has devastating consequences for patients' QOL. In some patients, the conventional functional pelvic floor electrical stimulation has been effective but is an invasive and embarrassing treatment. The object of the study was to evaluate the feasibility of functional extracorporeal magnetic stimulation (FMS) in strengthening the pelvic floor muscles without an anal plug and the embarrassment of undressing. Materials and Methods: Thirty patients (26 female and 4 males) with FI were enrolled. All patients were assessed during a specialized coloproctology evaluation followed by endoanal ultrasonography and anorectal manometry. All patients underwent an FMS treatment once weekly for 8 weeks. Patients' outcome was assessed by the Cleveland Clinic Fecal Incontinence Score (CCFIS) and by the fecal incontinence QOL questionnaire (FIQL). Results: After 8 weeks, the number of solid and liquid stool leakage per week was significantly reduced (p<0.05) with a significant improvement of the CCFIS and of the FIQL (p<0.05). Moreover, the authors recorded a missed recruitment of the agonist and antagonists' defecation muscles. Conclusion: FMS is a safe, non-invasive and painless treatment for FI. It could be recommended for selected patients with non-surgical FI to ensure a rapid clinical improvement

Engineering The Unicellular Alga Phaeodactylum tricornutum For High-Value Plant Triterpenoid Production

Plant triterpenoids constitute a diverse class of organic compounds that play a major role in development, plant defense and environmental interaction. Several triterpenes have demonstrated potential as pharmaceuticals. One example is betulin, which has shown promise as a pharmaceutical precursor for the treatment of certain cancers and HIV. Major challenges for triterpenoid commercialization include their low production levels and their cost‐effective purification from the complex mixtures present in their natural hosts. Therefore, attempts to produce these compounds in industrially relevant microbial systems such as bacteria and yeasts have attracted great interest. Here we report the production of the triterpenes betulin and its precursor lupeol in the photosynthetic diatom Phaeodactylum tricornutum, a unicellular eukaryotic alga. This was achieved by introducing three plant enzymes in the microalga: a Lotus japonicus oxidosqualene cyclase and a Medicago truncatula cytochrome P450 along with its native reductase. The introduction of the L. japonicus oxidosqualene cyclase perturbed the mRNA expression levels of the native mevalonate and sterol biosynthesis pathway. The best performing strains were selected and grown in a 550L pilot scale photobioreactor facility. To our knowledge, this is the most extensive pathway engineering undertaken in a diatom and the first time that a sapogenin has been artificially produced in a microalga, demonstrating the feasibility of the photo‐bio‐production of more complex high‐value, metabolites in microalgae

The Italian Unitary Society of Colon-proctology (SIUCP: Società Italiana Unitaria di Colonproctologia) guidelines for the management of anal fissure

Introduction: The aim of these evidence-based guidelines is to present a consensus position from members of the Italian Unitary Society of Colon-Proctology (SIUCP: Società Italiana Unitaria di Colon-Proctologia) on the diagnosis and management of anal fissure, with the purpose to guide every physician in the choice of the best treatment option, according with the available literature. Methods: A panel of experts was designed and charged by the Board of the SIUCP to develop key-questions on the main topics covering the management of anal fissure and to performe an accurate search on each topic in different databanks, in order to provide evidence-based answers to the questions and to summarize them in statements. All the clinical questions were discussed by the expert panel in different rounds through the Delphi approach and, for each statement, a consensus among the experts was reached. The questions were created according to the PICO criteria, and the statements developed adopting the GRADE methodology. Conclusions: In patients with acute anal fissure the medical therapy with dietary and behavioral norms is indicated. In the chronic phase of disease, the conservative treatment with topical 0.3% nifedipine plus 1.5% lidocaine or nitrates may represent the first-line therapy, eventually associated with ointments with film-forming, anti-inflammatory and healing properties such as Propionibacterium extract gel. In case of first-line treatment failure, the surgical strategy (internal sphincterotomy or fissurectomy with flap), may be guided by the clinical findings, eventually supported by endoanal ultrasound and anal manometry

Sacral nerve stimulation in slow-transit constipation: effectiveness at 5-year follow-up

Aim: The aim of this study is to evaluate the short- and long-term efficacy of sacral nerve stimulation (SNS) for treating slow-transit constipation (STC). Method: This is a retrospective cohort analysis of the efficacy of SNS in treating patients affected by STC, who previously failed to respond to conservative therapies. Only patients free of concomitant diseases were enrolled in our study. A temporary stimulation lead was initially implanted; patients with a &gt; 50% symptom reduction were eventually deemed eligible for a permanent implant. Results: This study enrolled 25 patients who underwent a SNS test stimulation; 21 patients (13 women; median age 32&nbsp;years) eventually got a permanent implant. The median preoperative Cleveland Clinic Constipation Score (CCCS) was 21 (16–25). Preoperative colorectal transit time recorded a median of 10 markers (7–19) retained in the colorectal tract. At 6-month postoperative follow-up, the total number of markers retained in the colorectal tract decreased to 3 (0–4). The CCCS score improved during the first postoperative year (P &lt; 0.001), but progressively worsened over the longer term. The SF-36 questionnaire showed an improvement in all 8 scales measuring physical and psycho-emotional states; all parameters recorded into the bowel diary also improved. Overall, at 60-month follow up, the overall neuromodulator removal rate was 48%. Conclusions: The SNS is a minimally invasive surgical procedure that we tested for treating STC. The short-term outcome was promisingly after 6&nbsp;months; however, there was a declining trend beyond this interval. Thus, the long-term efficacy of SNS needs to be further assessed

D-shape asymmetric excision in recurrent pilonidalis disease: an analytic longitudinal long-term evaluation

Sacrococcigeal pilonidalis disease (SPD) recurrence is a major factor influencing surgical outcomes. Several different surgical treatments have been reported, however, there is a lack of long-term data on reoperation. Aim of this study was to analyze outcomes of a single center adopting a standardized off-midline asymmetric procedure (D-shape). Analytic longitudinal assessment of 83 patients (median age 35&nbsp;years, range 23–59&nbsp;years) with recurrent SPD that completed the 5-year study design following D-shape reoperation. Among a cohort of 607 patients, we enrolled 83 recurrent SPD. After D-shape reoperation, second recurrence rate was 9.6% (8/83). Second recurrence rate was not statistically significantly different among patients undergone D-shape as first surgery compared to patients of symmetric excision group (11.8% vs. 7.4%, p = 0.57). Similarly, there was no statistical difference among patients who underwent D-shape as first surgery compared to patients who underwent symmetric excision elsewhere (11.8% vs. 9.1%, p =.75). D-shape is a safe and effective when adopted as revisional surgery at a long-term follow-up. Comparative evaluation is warranted to establish the potential superiority over different surgical surgery in case of recurrence