Organic Solvent Nano ltration in the peptide industry

In recent years the application of membrane technology to molecular separation processes has stimulated interest and showed great potential in a number of industrial fields. Ultrafiltration membranes have been successfully applied to downstream separation of therapeutically active peptides, to overcome some of the limitations of the conventional techniques in terms of costs, scale-up, selectivity and solvent recovery. In this research project, Organic Solvent Nanofiltration of peptide solutions is studied, and this understanding is applied to the development of innovative membrane-based purification strategies for industrial case studies. Basic understanding of transport mechanisms was approached by investigating solvent transport through ceramic nano- and ultrafiltration membranes, and developing a predictive phenomenological model for the transport of solvents and solvent mixtures. Effects of solvent-membrane interactions strongly affected the solvent permeation through nanofiltration membranes, while they were found to be negligible in the ultrafiltration range. The effect of the organic solvent on the permeation of neutral and charged solutes (monovalent salts, a small molecule and peptides) in organic/water mixtures was studied, with particular attention to the role of preferential solvation in the solvent mixture. It was found that the solvent composition and the complex association of counter-ions and buffers highly affect membrane permeation and rejection of organic molecules. It is proposed that all these components change the relative solute-membrane affinity. Since permeation of peptides in organic/water mixtures is affected by complicated matrices of input parameters, a Design of Experiment approach was proposed to efficiently investigate the nanofiltration of model peptides in acetonitrile/water solutions. Statistical models for solvent flux, peptide and ion rejection were obtained by Analysis of Variance and interpreted from a phenomenological point of view. The statistical models were used to asisst process development for two industrial case studies: (1) concentration and salt/solvent exchange of a first therapeutic peptide were optimised, based on the integration of the statistical DoE models with the process simulation for concentration and diafiltration; (2) the nanofiltration-assisted synthesis of a second therapeutic peptide, based on the coupling between nanofiltration and reaction in one unique process, was developed and compared to the established process by techno-economical analysis. The so-called "Reactive Peptide Nanofiltration" was found to be advantageous in terms of economics, efficacy, impact on the market, and on the environment. In conclusion, nano ltration was found to be a solid and competitive technique for application to peptide processes. On the basis of the results of this research, Lonza decided to invest in a new nano ltration plant for the downstream of peptides with ceramic membranes. The advantages of nanofiltration technology, in terms of development of more efficient materials (stable in critical solvents and harsh acid/basic conditions), improvement of membrane performances (selectivity, lifetime) and integration of nanofiltration with other techniques in hybrid processes seem therefore promising in overcoming the hesitancy of industries to modify the established processes and invest in new nano ltration plants, by making the payback period for the return of investment more attractive. It is plausible to think that this technology will shortly become a primary choice for new separation and purification processes

A Systematization of Cybersecurity Regulations, Standards and Guidelines for the Healthcare Sector

The growing adoption of IT solutions in the healthcare sector is leading to a steady increase in the number of cybersecurity incidents. As a result, organizations worldwide have introduced regulations, standards, and best practices to address cybersecurity and data protection issues in this sector. However, the application of this large corpus of documents presents operational difficulties, and operators continue to lag behind in resilience to cyber attacks. This paper contributes a systematization of the significant cybersecurity documents relevant to the healthcare sector. We collected the 49 most significant documents and used the NIST cybersecurity framework to categorize key information and support the implementation of cybersecurity measures.Comment: 14 page

Tecnica e prospettive di utilizzo della citometria a flusso per la tipizzazione linfocitaria del liquido sinoviale di cane

RIASSUNTO L’utilizzo della citometria a flusso, o fluorimetria, per l’immunofenotipizzazione linfocitaria è una metodica in uso in medicina umana soprattutto nel settore reumatologico, dove riveste un’importanza notevole nella comprensione dei meccanismi degenerativi cartilaginei in corso di artrite reumatoide, mentre è poco utilizzata nella più comune artrosi. In medicina veterinaria la tecnica non è a tutt’oggi utilizzata nella diagnostica collaterale dei problemi ortopedici e reumatologici. Sono disponibili solo due segnalazioni di lavori sperimentali in argomento. Sono state fatte numerose prove su campioni di liquido sinoviale di cane, prelevato sia durante la visita clinica che in sede intraoperatoria, in corso di patologie articolari diverse. Il liquido è stato sottoposto a conta cellulare con contaglobuli elettronico; per essere processato a tal fine e per l’analisi citofluorimetrica, è stato sottoposto a pretrattamento con jaluronidasi. La componente cellulare mononucleata è stata quindi separata e concentrata mediante centrifugazione in gradiente di densità su LymphoprepTM (Ficoll-Isopaque) e studiata con citofluorimetro. Contemporaneamente l’immunofenotipizzazione linfocitaria è stata condotta anche su un campione di sangue periferico. Prove successive finalizzate a rendere il campione un fluido acquoso hanno permesso di evidenziare l’assoluta necessità di pretrattare il campione con 2 gocce di jaluronidasi alla concentrazione di 150UI/ml per 0,25 cc di liquido sinoviale. La quantità di liquido prelevabile è in generale scarsa nei fatti degenerativi cronici, più abbondante nei fenomeni infiammatori acuti, e questo può rappresentare un limite nell’applicazione di questa metodica nella pratica clinica. Per ottenere una buona lettura è necessario avere infatti almeno 1cc di liquido articolare e/o almeno 1000 cellule/μl. La distribuzione delle sottopopolazioni linfocitarie CD3+ e CD21+, ed in particolare del rapporto fra i linfociti CD4+ e CD8+ nel liquido sinoviale e nel sangue, si presenta indicativamente diversa nei fenomeni prevalentemente degenerativi rispetto a quelli squisitamente infiammatori. La tipizzazione linfocitaria nel liquido sinoviale del cane può aprire la strada ad ulteriori indagini per studiare più a fondo i meccanismi eziopatogenetici alla base della condrodegenerazione e della risposta locale e sistemica nei fenomeni osteoartritici e immunitari, oltreché fornire una ausilio nell’individuazione e nel monitoraggio delle terapie. SUMMARY The use of flow cytometry, also called cytofluorimetry, for lymphocyte immunophenotyping is a technique adopted in uman medicine, especially in the rheumatoid field, where is of paramount importance to understand the cartilage degenerative mechanisms during rheumatoid arthritis, while is less used in degenerative joint disease. Till now in veterinary medicine this technique is not included in the ancillary diagnostic methods for orthopaedic and rheumatologic problems. To date only two papers are available about its experimental use. Several tests were done on canine synovial fluid samples, collected during the clinical examination or intraoperatively, in subjects with different joint diseases. The synovial fluid was submit for total cell count with electronic cell-counter; for this purpose and for cytofluorimetric examination too was pre-treated with jaluronidasis. The mononuclear cell phase was then separated and concentrated by centrifugation on LymphoprepTM (Ficoll-Isopaque) and studied with cytofluorimetry. In the mean time the lymphocyte immunophenotyping was performed also on a peripheral blood sample. Further tests aimed to transform the sample in an aqueous fluid has shown the absolute necessity to pre-treat the sample with two drops of jaluronidasis 150 UI/ml in 0,25 cc of synovial fluid. In chronic degenerative joint disease only few drops of synovial fluid can be collected from joints, while more abundant quantities could came from acute inflammatory cases, and this could represent a limit for the applications of the technique in the clinical practice. To perform a good examination we need at least 1cc of synovial fluid and/or at least 1000 cells/μl. Lymphocyte CD3+ and CD21+ subpopulation distribution, and especially CD4+/CD8+ ratio in blood and synovial fluid, shows differences in prevalently degenerative and mainly inflammatory phenomena. The lymphocyte immunophenotyping of canine synovial fluid can represent an alternative instrument to deeply analyze the aetiopathogenetic mechanisms settled at the basis of chondrodegeneration and of the local and systemic response to osteoarthritic and immunological phenomena, and to help in therapeutic choice and monitoring

Clinically Mild Encephalopathy with a Reversible Splenial Lesion Caused by Influenza B Virus in an Unvaccinated Child

: Reversible lesions involved in the splenium of corpus callosum (RESLES) are a rare clinic-radiological condition, whose pathogenesis could be related to infectious events (such as in mild encephalopathy with reversible splenial lesion-MERS), epilepsy or metabolic/electrolyte disorders. MERS is characterized by an acute mild encephalopathy associated with lesions in the splenium of corpus callosum on brain magnetic resonance imaging. Viral infections are commonly associated with this condition and type A influenza is the most common cause. The prognosis is generally favorable with spontaneous resolution of clinical and radiological abnormalities. We report a case report of type B influenza MERS in an 8-year-old unvaccinated girl with complete clinical and radiological recovery

Occurrence of mislabelling in prepared fishery products in Southern Italy

Fish authentication is a major concern not only for the prevention of commercial fraud, but also for the assessment of safety risks deriving from the undeclared introduction of potentially dangerous toxic or allergenic substances or environmentally damaging fish where endangered species are involved. Moreover, food authentication might affect the diet of certain groups of consumers, such as followers of religious practices. Considering the authentication of fish products is one of the key issues in food safety, quality and sustainability, the aim of this work was to investigate the prevalence of mislabelling in sole (Solea solea), plaice (Pleuronectes platessa), Atlantic salmon (Salmo salar), and hake (Merluccius merluccius) fillets from markets and supermarkets located in Apulia (Southern Italy) using DNA barcoding. The results of the molecular investigations reveal that 42/98 (42.8%) fillet samples were not correctly labelled. In particular, 12/27 (44.4%) fillets of sole (Solea solea) were identified as belonging to Solea senegalensis. In addition, 13/28 (46.4%) plaice (Pleuronectes platessa) samples were identified as Pangasius hypophtalmus. All Atlantic salmon (Salmo salar) samples were correctly labelled. Post-sequencing data analysis revealed that 17/30 (56.6%) hake fillets (Merluccius merluccius) were not correctly labelled, of which 8/30 samples identified as Merluccius hubbsi, 5/30 samples as Merluccius products and 4/30 as Merluccius capensis. The study reveals a high occurrence of species mislabelling in the prepared fish fillet products, further evidence of the need for increased traceability and assessment of the authenticity of food products

Orthorexia nervosa, eating patterns and personality traits: a cross-cultural comparison of Italian, Polish and Spanish university students

Background: The amount of research about orthorexic attitudes and behaviours has increased in the last five years, but is still mainly based on descriptive and anecdotal data, yielding a variety of prevalence data and inconsistent results. The interplay between socio-cultural context and orthorexia has been poorly investigated and is still far from being understood. Method: Multicentre, cross-sectional study involving Italian (N = 216), Polish (N = 206) and Spanish (N = 242) university students, assessed through a protocol including informed consent, socio-demographic and anamnestic data sheet and self-administered questionnaires (ORTO-15, Eating Attitudes Test- 26 [EAT-26], Temperament and Character Inventory [TCI]). Results: Higher prevalence of orthorexia (as described by the ORTO-15 cutoff) was found in Poland. Female gender, Body Mass Index (BMI), current Eating Disorder, dieting, EAT-26 score ≥ 20 and low/medium Persistence were associated with orthorexia in the whole sample. The cross-cultural comparison showed several differences among the three subgroups of students. Conclusions: The associations found between orthorexic attitudes, self-reported current eating disorder, BMI and adherence to a dieting need to be supported by further research. The differences among students from the three countries seem to suggest a possible rolve for cultural elements in the construct of orthorexia

Up-Cycling of Olea europaea L. Ancient Cultivars Side Products: Study of a Combined Cosmetic-Food Supplement Treatment Based on Leaves and Olive Mill Wastewater Extracts

In recent years, a reversal of the global economic framework has been taking place: from the linear model, there has been a gradual transition to a circular model where by-products from the agri-food industry are taken and transformed into value products (upcycling) rather than being disposed of. Olive tree pruning represents an important biomass currently used for combustion; however, the leaf part of the olive tree is rich in phenolic substances, including hydroxytyrosol. Mill wastewater is also discarded, but it still contains high amounts of hydroxytyrosol. In this study, cosmetic and food supplement formulations were prepared using biophenols extracted from leaves and wastewater and were tested in a placebo-controlled study on healthy volunteers using a combined cosmetic and food supplement treatment. A significant improvement in skin health indicators (collagen density, elasticity, etc.) and a 17% improvement against Photo-induced Irritative Stimulus was observed

Fasting glucose and body mass index as predictors of activity in breast cancer patients treated with everolimus-exemestane: the EverExt study

Evidence on everolimus in breast cancer has placed hyperglycemia among the most common high grade adverse events. Anthropometrics and biomarkers of glucose metabolism were investigated in a observational study of 102 postmenopausal, HR + HER2- metastatic breast cancer patients treated with everolimus-exemestane in first and subsequent lines. Best overall response (BR) and clinical benefit rate (CBR) were assessed across subgroups defined upon fasting glucose (FG) and body mass index (BMI). Survival was estimated by Kaplan-Meier method and log-rank test. Survival predictors were tested in Cox models. Median follow up was 12.4 months (1.0-41.0). The overall cohort showed increasing levels of FG and decreasing BMI (p < 0.001). Lower FG fasting glucose at BR was more commonly associated with C/PR or SD compared with PD (p < 0.001). We also observed a somewhat higher BMI associated with better response (p = 0.052). More patients in the lowest FG category achieved clinical benefit compared to the highest (p < 0.001), while no relevant differences emerged for BMI. Fasting glucose at re-assessment was also predictive of PFS (p = 0.037), as confirmed in models including BMI and line of therapy (p = 0.049). Treatment discontinuation was significantly associated with changes in FG (p = 0.014). Further research is warranted to corroborate these findings and clarify the underlying mechanisms

Multidisciplinary management of neuroendocrine neoplasia: a real-world experience from a referral center

Purpose: Multidisciplinary approach is widely advised for an effective care of patients with neuroendocrine neoplasia (NEN). Since data on efficacy of multidisciplinary management of NENs patients in referral centers are scanty, this study aimed at analyzing the modality of presentation and clinical outcome of patients with NENs managed by a dedicated multidisciplinary team. Methods. In this prospective observational study, we included all consecutive new patients visiting the Sant'Andrea Hospital in Rome (ENETS-Center of Excellence) between January 2014 and June 2018. Results. A total of 195 patients were evaluated. The most frequent sites were pancreas (38.5%), small bowel (22%), and lung (9.7%). Median Ki67 was 3%. After the first visit at the center, additional radiological and/or nuclear medicine procedures were requested in 163 patients (83.6%), whereas histological data revision was advised in 84 patients (43.1%) (revision of histological slides: 27.7%, new bioptic sampling: 15.4%). After that, disease imaging staging and grading was modified in 30.7% and 17.9% of patients, respectively. Overall, a change in therapeutic management was proposed in 98 patients (50.3%). Conclusions. Multidisciplinary approach in a dedicated team may lead to change of disease imaging staging and grading in a significant proportion of patients. Enhancing referral routes to dedicated-NEN center should be promoted, since it may improve patients' clinical outcome