A theoretical estimate for nucleotide sugar demand towards Chinese Hamster Ovary cellular glycosylation

Glycosylation greatly influences the safety and efficacy of many of the highest-selling recombinant therapeutic proteins (rTPs). In order to define optimal cell culture feeding strategies that control rTP glycosylation, it is necessary to know how nucleotide sugars (NSs) are consumed towards host cell and rTP glycosylation. Here, we present a theoretical framework that integrates the reported glycoproteome of CHO cells, the number of N-linked and O-GalNAc glycosylation sites on individual host cell proteins (HCPs), and the carbohydrate content of CHO glycosphingolipids to estimate the demand of NSs towards CHO cell glycosylation. We have identified the most abundant N-linked and O-GalNAc CHO glycoproteins, obtained the weighted frequency of N-linked and O-GalNAc glycosites across the CHO cell proteome, and have derived stoichiometric coefficients for NS consumption towards CHO cell glycosylation. By combining the obtained stoichiometric coefficients with previously reported data for specific growth and productivity of CHO cells, we observe that the demand of NSs towards glycosylation is significant and, thus, is required to better understand the burden of glycosylation on cellular metabolism. The estimated demand of NSs towards CHO cell glycosylation can be used to rationally design feeding strategies that ensure optimal and consistent rTP glycosylation

Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Many high-value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimise the productivity of these cultures it is important to monitor cellular metabolism, for example the utilisation of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole-cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity and robustness. Importantly, our whole-cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole-cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalise on the wealth of genetic operons for metabolite sensing available in Nature for the development of other whole-cell biosensors

A Pair of Rigid Surfaces with pg= q = 2 and K2= 8 Whose Universal Cover is Not the Bidisk

We construct two complex-conjugated rigid minimal surfaces with pg = q = 2 and K2 = 8 whose universal cover is not biholomorphic to the bidisk H × H. We show that these are the unique surfaces with these invariants and Albanese map of degree 2, apart from the family of product-quotient surfaces given in [33]. This completes the classification of surfaces with pg = q = 2, K2 = 8, and Albanese map of degree 2

A NEW SYSTEM TO INCREASE THE LIFE EXPECTANCY OF OPTICAL DISCS. PERFORMANCE ASSESSMENT WITH A DEDICATED EXPERIMENTAL SETUP.

Negli ultimi decenni si \ue8 registrata una crescita esponenziale per quel che riguarda l'uso di supporti digitali per l\u2019archiviazione dei dati informativi. Tuttavia, l\u2019aspettativa di vita di questi supporti \ue8 inadeguata rispetto alle necessit\ue0 delle istituzioni che si occupano di archiviare e preservare, ad esempio, il patrimonio artistico, storico, e culturale. Sulla base di questa problematica, pi\uf9 volte sollevata dall\u2019UNESCO [1,35], proponiamo un approccio innovativo che la degradazione chimico-fisica dei dischi ottici in modo da aumentarne l\u2019aspettativa di vita. Gli obiettivi di questa tesi sono quindi la progettazione e la messa in atto di una nuova strategia intelligente, volta ad aumentare l'aspettativa di vita dei dischi ottici. Diversi apparati sperimentali sono stati sviluppati al fine di studiare i meccanismi della degradazione chimico-fisica dei dischi tramite test di invecchiamento accelerato che simulano le condizioni operative di utilizzo del disco. Nei dischi sono state identificate aree critiche, nelle quali la degradazione cresce statisticamente pi\uf9 velocemente della media, e delle aree sicure, dove la degradazione cresce invece pi\uf9 lentamente. Due apparati sperimentali sono stati costruiti. Il primo \ue8 una camera climatica in grado di indurre invecchiamento artificiale sui dischi. Il secondo \ue8 un sistema robotico in grado di rilevare la quantit\ue0 di errori in ogni blocco di dati, prima della fase di correzione degli errori effettuata dal codice "Reed Solomon" (prima e dopo la fase di invecchiamento artificiale, senza polveri, e in un ambiente con controllo di temperatura e umidit\ue0). L'analisi dei dati cos\uec raccolti ha permesso di identificare le gi\ue0 citate aree fisiche in cui i blocchi di dati hanno un numero di errori che si avvicina o addirittura supera la capacit\ue0 di correzione dei dati propria del codice Reed Solomon standard. I risultati di queste analisi hanno portato allo sviluppo di un "Codice Reed Solomon Adattivo" (codice A-RS), che consente di proteggere le informazioni memorizzate all'interno delle aree critiche. Infatti, il codice A-RS utilizza un algoritmo di redistribuzione che viene applicato ai simboli di parit\ue0. La ridistribuzione della ridondanza \ue8 stata calcolata tramite una funzione di degrado, a sua volta ottenuta fittando gli errori rilevati nella fase sperimentale. L'algoritmo di redistribuzione sposta i simboli di parit\ue0 dai blocchi di dati nelle aree sicure ai blocchi di dati nelle aree critiche. Inoltre, \ue8 interessante sottolineare come questo processo non diminuisca la capacit\ue0 di memoria dei Digital Versatile Discs (DVDs) e dei Blu-ray Discs (BDs). Questa strategia consente quindi di evitare la dismissione anticipata dei dischi ottici, dovuta alle possibili perdite di dati.The past decade has witnessed an exponential growth in the use of digital supports for big data archiving. However, the expected lifespan of these supports is inadequate with respect to the actual needs of heritage institutions. Stemming from the issues raised by UNESCO [1, 2], we address the problem of alleviating the effects of aging on optical discs. To achieve this purpose, we propose a novel logical approach that is able to conteract the physical and chemical degradation of different types of optical discs, increasing their life expectancy. In other words, the objectives of this thesis are the design and the implementation of a new intelligent strategy aimed at increasing the life expectancy of optical discs. An experimental setup has been developed in order to investigate the physical and chemical degradation processes of discs, by means of accelerated aging tests that simulated the operational disc-use conditions. Critical areas are identified where disc degradation is statistically faster than average, while in safe areas the degradation is relatively slow. To collect the needed data, two experimental devices have been built. The first is a climatic chamber, which is able to induce artificial disc aging. The second is a robotic device, which is able to detect the amount of errors in each data block prior to the \u201cReed Solomon\u201d error correction stage (before and after the accelerate aging stage, without dust and in an environment with controlled temperature and humidity). The analysis of the data allows to identify the aforementioned physical areas where data blocks have a number of errors that approaches or exceeds the data correction capability of the standard Reed Solomon code. The results of these analyses have led to develop an \u201cAdaptive Reed Solomon Code\u201d (A-RS code) that allows to protect the information stored within the critical areas. The A-RS code uses a redistribution algorithm that is applied to parity symbols. It is calculated from the fitting of the experimental errors obtained through the \u201cdegradation function\u201d. The redistribution algorithm shifts a certain number of parity symbols from Data Blocks in safe areas to Data Blocks in critical areas. Interestingly, these processes do not diminish the memory capability of Digital Versatile discs (DVDs) and of Blu-Ray discs (BDs). This strategy therefore avoids the early dismission of optical discs, due to possible losses of even minimal parts of the recorded information

SEDIMENTARY DYNAMICS OF THE MARETTIMO CONTINENTAL SHELF AND MARETTIMO CHANNEL (EGADI ISLANDS – WESTERN MEDITERRANEAN): ANALYSIS OF DEPOSITIONAL AND EROSIONAL FEATURES.

Sedimentary dynamics of the Marettimo continental shelf and Marettimo Channel (Egadi Island – Western Mediterranean), have been unveiled based on the erosive and depositional features observed at the seafloor. Different morphologic features were recognized through side scan sonar, multibeam and high-resolution seismics, acquired in the framework of national Research Projects (CARG, GebecSud, MaGIC) carried out by the Department of Earth and Marine Science (former Department of Geology and Geodesy) of the University of Palermo. Sediment samples have been also collected and analyzed. The Egadi Islands are part of the NW Sicilian offshore, and belong to the mountain chain that connects the Sicilian chain to the Maghrebides one. The Marettimo Island represents an isolated portion of the Egadi continental shelf and is separated from the rest of the archipelago by the Marettimo Channel. Previous studies allowed to recognize different kind of sedimentary structures along the seafloor. Lo Iacono and Guillén (2008) described extensive field of subaqueous dunes (Ashley, 1990) along the Marettimo inner shelf, forming on gravelly and pebbly sediments and related to strong storm events (H:5.5 m, T: 12 s). Larger dunes have been observed in the north-western sector of the inner shelf with wavelengths ranging from 1 to 2.5 m, heights between 15 cm and 30 cm and extending laterally for hundreds of meters. Moreover in the north-eastern sector of the inner shelf, sorted bedforms have been detected at a depth of 40 m: they are characterized by a width varying between 15 m and 50 m and a length that reaches hundreds of meters. In the south-eastern sector of the Marettimo outer shelf, Colantoni et al. (1993) map two groups of sedimentary structures. In the same area, Lo Iacono (2004) recognized two-dimensional and three-dimensional subaqueous dunes. 2D dunes are found at depths between 60 and 90 m and show a wavelength variable between 15 and 50 m. 3D dunes, which are found at a depth of 80 m, are characterized by a wavelength of 30 m and a lateral extent of 50 m. Morphological evidences of a strong hydrodynamic regime come also from the morphobathymetric analysis of the Marettimo Channel, that is a 24 km long submarine valley oriented NNW-SSE, with a depth ranging from a minimum of 180 m to a maximum of 370 m and a width from 2 km to 14 km. Inside the channel, south of its narrowest point, a 180 m deep bathymetric threshold separates two areas, which deepens towards opposite directions: north-west and south. The flanks of the sector that deepens towards the NW are affected by mass-wasting features (Lo Iacono et al., 2007). The western flank of the channel, which delimits the Marettimo continental shelf, is carved by gullies, making the shelf-edge very uneven. Gullies develop to a maximum depth of 280 m and have a length of 250 m and a width of 50 m. At the bottom of the channel a linear incision 20 m deep, 400 m wide and almost 3 km long, is probably generated by erosive bottom currents. On the whole, the morpho-sedimentary structures here described reveal a strong hydrodynamic regime. The morphological and textural features of the bedforms observed on the Marettimo inner continental shelf suggest the occurrence of sporadic medium to strong storm events coming from western sectors. The morphological characteristics of the Marettimo Channel reveal the occurrence of strong bottom currents probably related to the severe interchange of water masses between the Tyrrhenian Sea and the Sicily Channel. References: Ashley, G., 1990. Journal of Sedim. Petrology 60, 160–172. Colantoni, P., Ligi, M., Morsiani, M.P., Penitenti, D., 1993. UNESCO Reports in Mar. Science 58, 93–98. Lo Iacono, C., 2004. Università di Napoli ‘‘Federico II,’’ Naples, Italy. Ph.D. Thesis, 150pp... Lo Iacono, C., Catalano, R., Agate, M., 2007. VI Conv. FIST GeoItalia, Rimini (Italy), 12-14 Sept. Lo Iacono C., Guillén J., 2008. Continental Shelf Research 28, 245–256

Functioning of family system in pediatric oncology during treatment phase

The study focuses on parents\u2019 psychological implications caused by the treatment of their children su\ufb00ering from tumor. It investigates some speci\ufb01c mothers\u2019 resource factors such as their strategies of coping and the perception of their own family functioning in terms of cohesion and adaptability. The study was performed with 34 mothers of children su\ufb00ering from acute lymphoblastic leukemia(ALL),duringthetreatmentphase.TheusedtoolsweretheCopingOrientationtoProblem Experienced\u2014New Italian Version, to investigate coping strategies, and the Family Adaptability and Cohesion Evaluation Scale-III, to analyze both real and ideal perception of family functioning. The data related to coping, show how the involved mothers tend to mainly use the strategies of positive aptitude, orientation toward problem and social support ( F = 99.88, d f= 4, P< .01). The family functioning, in terms of adaptabilit y, is described as chaotic relating to both the real (\u3c72 = 13.29,d f =3, P=.004)andideal(\u3c72=11.52,d f=2, P=.003)family,whereasintermsof cohesion, itisperceivedaschie\ufb02ydisengagedintherealfamily(\u3c72 =12.3,d f=3, P=.006)andasenmeshed in the ideal one (\u3c72 = 12.58, d f= 3, P= .006). Statistically positive correlations were only detected between adaptability and avoidance ( r = 0.49, P< .01); adaptability and orientation toward problem (r = 0.36, P< .05); and adaptability and transcendent orientation ( r = 120.04, P< .05). Despitethecriticalsituation,themothershaveshownoptimisticview,careforproblemmanagementandcapabilitytoaskforhelp.Thesecopingstrategiesallowthetherapeuticalliancebetween families and health care workers, so useful for the quality of childcarThe study focuses on parents\u2019 psychological implications caused by the treatment of their children su\ufb00ering from tumor. It investigates some speci\ufb01c mothers\u2019 resource factors such as their strategies of coping and the perception of their own family functioning in terms of cohesion and adaptability. The study was performed with 34 mothers of children su\ufb00ering from acute lymphoblastic leukemia(ALL),duringthetreatmentphase.TheusedtoolsweretheCopingOrientationtoProblem Experienced\u2014New Italian Version, to investigate coping strategies, and the Family Adaptability and Cohesion Evaluation Scale-III, to analyze both real and ideal perception of family functioning. The data related to coping, show how the involved mothers tend to mainly use the strategies of positive aptitude, orientation toward problem and social support ( F = 99.88, d f= 4, P< .01). The family functioning, in terms of adaptabilit y, is described as chaotic relating to both the real (\u3c72 = 13.29,d f =3, P=.004)andideal(\u3c72=11.52,d f=2, P=.003)family,whereasintermsof cohesion, itisperceivedaschie\ufb02ydisengagedintherealfamily(\u3c72 =12.3,d f=3, P=.006)andasenmeshed in the ideal one (\u3c72 = 12.58, d f= 3, P= .006). Statistically positive correlations were only detected between adaptability and avoidance ( r = 0.49, P< .01); adaptability and orientation toward problem (r = 0.36, P< .05); and adaptability and transcendent orientation ( r = 120.04, P< .05). Despitethecriticalsituation,themothershaveshownoptimisticview,careforproblemmanagementandcapabilitytoaskforhelp.Thesecopingstrategiesallowthetherapeuticalliancebetween families and health care workers, so useful for the quality of childcar

A study of maternal competence in preterm birth condition, during the transition from hospital to home: An early intervention program’s proposal

The study was conducted with 104 mothers (average age 32.5 years, SD 6.1) of preterm infants (very and moderately preterm but still healthy) to monitor the perceived maternal role competence from the time of hospitalisation to post-discharge, in order to define an intervention program to support mothers during this transition. A targeted Q-Sort tool (Maternal Competence Q-Sort in preterm birth) was applied at two different times as a self-observation tool for parenting competence in neonatology. A tendency towards dysregulation of the maternal role competence was detected, mainly in terms of low self-assessment and was found to worsen during post-discharge, particularly with regard to caregiving ability. This study suggests the importance of accompanying parenting competence in preterm birth conditions, not only during hospitalisation in the Neonatal Intensive Care Unit (NICU) but also following discharge in order to promote the development of premature infants. This paper reports in the last part a specific integrated psychoeducational intervention program (psychologist and nurses), which we defined precisely in light of the suggestions offered by the study data on perceived maternal competence created with the Q-sort

Parental Resources in Parents of Children with Special Needs (SNs) at the Time of COVID-19

Background. The limitations imposed by governments for containing the spread of COVID-19 have affected familial relationships, especially those of families dealing with children with special needs or chronic illness conditions. The current study aims to better understand what pathological/disability condition has impacted parental resources, sense of competence, and perception of children's executive functioning the most. Methods. A sample of 648 parents was asked to answer a survey assessing children's condition (typical development, specific learning disorder, autism spectrum syndrome, chronic illness), parental resources, parenting sense of competence (distinguished into parental satisfaction and self-efficacy), and parents' perception of their children's executive functioning. A MANOVA model was performed to assess differences in parental resources, sense of competence, and perception of the children's executive functions according to their condition. A path analysis model was performed to examine the impact of sense of competence and children's condition on parental resources and children's executive functions. Results. Parents with children with specific learning disorder reported lower scores of parental resources in terms of total scores, common antecedents, and specific antecedents compared to parents with typically developed children (total scores: MD = 56.45, p &lt; 0.001; common antecedents: MD = 22.28, p &lt; 0.00; specific antecedents: MD = 34.17, p &lt; 0.001), parents with autistic children (total scores: MD = 62.79, p = 0.01; common antecedents: MD = 24.03, p = 0.01; specific antecedents: MD = 38.76, p = 0.02) and parents of children with chronic illness (total scores: MD = 37.42, p = 0.04; common antecedents: MD = 16.59, p = 0.01). The path analysis model shows a direct effect of parental satisfaction (beta = 0.26) and pathology/disability (beta = -0.10) on parental resources that, in turn, influence parents' perception of their children's executive functioning (beta = 0.24). Conclusion. Although no data about the prepandemic situation is available, the current study highlights that parental stress influence children's cognition. Moreover, parents of children with special needs seemed to be challenged during COVID-19, especially parents of children with specific learning disorders, that are already stressed out by their children's condition. Therefore, academic services should undertake preventive measures to preserve parental well-being and to provide a supportive environment for children, especially for those with atypical development