Monitoring the extracellular matrix mineralization processes in biological scaffolds using bioreactors, x-ray μct technology and 3-d modeling methods (monitoraggio dei processi di mineralizzazione della matrice extracellulare in scaffolds biologici, utilizzando bioreattore, tecnologia μct a raggi x e metodi di modellizzazione 3d)

Al fine di migliorare le tecniche di coltura cellulare in vitro, sistemi a bioreattore sono sempre maggiormente utilizzati, e.g. ingegnerizzazione del tessuto osseo. Spinner Flasks, bioreattori rotanti e sistemi a perfusione di flusso sono oggi utilizzati e ogni sistema ha vantaggi e svantaggi. Questo lavoro descrive lo sviluppo di un semplice bioreattore a perfusione ed i risultati della metodologia di valutazione impiegata, basata su analisi μCT a raggi-X e tecniche di modellizzazione 3D. Un semplice bioreattore con generatore di flusso ad elica è stato progettato e costruito con l'obiettivo di migliorare la differenziazione di cellule staminali mesenchimali, provenienti da embrioni umani (HES-MP); le cellule sono state seminate su scaffold porosi di titanio che garantiscono una migliore adesione della matrice mineralizzata. Attraverso un microcontrollore e un'interfaccia grafica, il bioreattore genera tre tipi di flusso: in avanti (senso orario), indietro (senso antiorario) e una modalità a impulsi (avanti e indietro). Un semplice modello è stato realizzato per stimare la pressione generata dal flusso negli scaffolds (3•10-2 Pa). Sono stati comparati tre scaffolds in coltura statica e tre all’interno del bioreattore. Questi sono stati incubati per 21 giorni, fissati in paraformaldehyde (4% w/v) e sono stati soggetti ad acquisizione attraverso μCT a raggi-X. Le immagini ottenute sono state poi elaborate mediante un software di imaging 3D; è stato effettuato un sezionamento “virtuale” degli scaffolds, al fine di ottenere la distribuzione del gradiente dei valori di grigio di campioni estratti dalla superficie e dall’interno di essi. Tale distribuzione serve per distinguere le varie componenti presenti nelle immagini; in questo caso gli scaffolds dall’ipotetica matrice cellulare. I risultati mostrano che sia sulla superficie che internamente agli scaffolds, mantenuti nel bioreattore, è presente una maggiore densità dei gradienti dei valori di grigio ciò suggerisce un migliore deposito della matrice mineralizzata. Gli insegnamenti provenienti dalla realizzazione di questo bioreattore saranno utilizzati per progettare una nuova versione che renderà possibile l’analisi di più di 20 scaffolds contemporaneamente, permettendo un’ulteriore analisi della qualità della differenziazione usando metodologie molecolari ed istochimiche.

Optimization of molecular and crystalline forms of drugs, agrochemicals, pesticides in relation to activity, bioavailability, patentability and to the fabrication of polymorphs, solvates, co-crystals with green chemistry methods

This doctorate was funded by the Regione Emilia Romagna, within a Spinner PhD project coordinated by the University of Parma, and involving the universities of Bologna, Ferrara and Modena. The aim of the project was: - Production of polymorphs, solvates, hydrates and co-crystals of active pharmaceutical ingredients (APIs) and agrochemicals with green chemistry methods; - Optimization of molecular and crystalline forms of APIs and pesticides in relation to activity, bioavailability and patentability. In the last decades, a growing interest in the solid-state properties of drugs in addition to their solution chemistry has blossomed. The achievement of the desired and/or the more stable polymorph during the production process can be a challenge for the industry. The study of crystalline forms could be a valuable step to produce new polymorphs and/or co-crystals with better physical-chemical properties such as solubility, permeability, thermal stability, habit, bulk density, compressibility, friability, hygroscopicity and dissolution rate in order to have potential industrial applications. Selected APIs (active pharmaceutical ingredients) were studied and their relationship between crystal structure and properties investigated, both in the solid state and in solution. Polymorph screening and synthesis of solvates and molecular/ionic co-crystals were performed according to green chemistry principles. Part of this project was developed in collaboration with chemical/pharmaceutical companies such as BASF (Germany) and UCB (Belgium). We focused on on the optimization of conditions and parameters of crystallization processes (additives, concentration, temperature), and on the synthesis and characterization of ionic co-crystals. Moreover, during a four-months research period in the laboratories of Professor Nair Rodriguez-Hormedo (University of Michigan), the stability in aqueous solution at the equilibrium of ionic co-crystals (ICCs) of the API piracetam was investigated, to understand the relationship between their solid-state and solution properties, in view of future design of new crystalline drugs with predefined solid and solution properties.

The nematode parasite fauna in three odontocete species: Phocoena phocoena, Lagenorhynchus albirostris, Lagenorhynchus acutus from Northern Scotland and its relationships with diet composition

Food items and nematode parasites were identified from the stomachs of 42 individuals of Phocoena phocoena, 6 of Lagenorhynchus acutus and 8 of L. albirostris stranded off the coastal waters of Northern Scotland between 2004 and 2014. Post-mortem examinations have revealed heavy parasitic worm burdens. Four nematode species complex as Anisakis spp., Contracaeucum spp., Pseudoterronova spp., and Hysterothylacium spp. were recorded. Data on presence of the anisakid species in cetaceans, reported a significative relationship between the presence of Hysterothylacium and the month of host stranding; suggesting a decrease of larval H. aduncum abundance in the period between April and August due to a seasonal effect related to prey availability. Similarly, the parasite burden of the all anisakid genera was related to the year fraction of stranding, and a relationship statistically significant was found just for L. albirostris with an increase between April and October. This finding is explained by a seasonality in occurrence of white-beaked dolphins, with a peak during August, that might be related to movements of shared prey species and competition with other species (Tursiops truncatus). Geographical differences were observed in parasites number of all anisakid species, which was the highest in cetaceans from the East area and lowest in the North coast. The parasites number also increased significantly with the length of the animal and during the year, but with a significant seasonal pattern only for P. phocoena. Regarding diet composition, through a data set consisting of 34 harbour porpoises and 1 Atlantic white-sided dolphins, we found a positive association between parasite number and the cephalopods genus Alloteuthis. This higher level of parasite infection in squid from this area, is probably due to a quantitative distribution of infective forms in squid prey, an abundance of the final host and age or size maturity of squid.

Potential effect of marine traffic on white-beaked dolphin (Lagenorhynchus albirostris) distribution and behaviour in Faxaflói Bay, southwest Iceland

Faxaflói bay is a short, wide and shallow bay situated in the southwest of Iceland. Although hosting a rather high level of marine traffic, this area is inhabited by many different species of cetaceans, among which the white-beaked dolphin (Lagenorhynchus albirostris), found here all year-round. This study aimed to evaluate the potential effect of increasing marine traffic on white-beaked dolphins distribution and behaviour, and to determine whether or not a variation in sighting frequencies have occurred throughout years (2008 – 2014). Data on sightings and on behaviour, as well as photographic one, has been collected daily taking advantage of the whale-watching company “Elding” operating in the bay. Results have confirmed the importance of this area for white-beaked dolphins, which have shown a certain level of site fidelity. Despite the high level of marine traffic, this dolphin appears to tolerate the presence of boats: no differences in encounter durations and locations over the study years have occurred, even though with increasing number of vessels, an increase in avoidance strategies has been displayed. Furthermore, seasonal differences in probabilities of sightings, with respect to the time of the day, have been found, leading to suggest the existence of a daily cycle of their movements and activities within the bay. This study has also described a major decline in sighting rates throughout years raising concern about white-beaked dolphin conservation status in Icelandic waters. It is therefore highly recommended a new dedicated survey to be conducted in order to document the current population estimate, to better investigate on the energetic costs that chronic exposure to disturbances may cause, and to plan a more suitable conservation strategy for white-beaked dolphin around Iceland.

Mechanical modeling of Intraneural Ganglion Cyst

Intraneural Ganglion Cyst is a 200 year old mystery related to nerve injury which is yet to be solved. Current treatments for the above problem are relatively simple procedures related to removal of cystic contents from the nerve. However, these treatments may result into neuropathic pain and recurrence of the cyst. The articular theory proposed by Spinner et al., (Spinner et al. 2003) takes into consideration the neurological deficit in Common Peroneal Nerve (CPN) branch of the sciatic nerve and affirms that in addition to the above treatments, ligation of articular branch results into foolproof eradication of the deficit. Mechanical Modeling of the Affected Nerve Cross Section will reinforce the articular theory (Spinner et al. 2003). As the cyst propagates, it compresses the neighboring fascicles and the nerve cross section appears like a signet ring. Hence, in order to mechanically model the affected nerve cross section; computational methods capable of modeling excessively large deformations are required. Traditional FEM produces distorted elements while modeling such deformations, resulting into inaccuracies and premature termination of the analysis. The methods described in this Master’s Thesis are effective enough to be able to simulate such deformations. The results obtained from the model adequately resemble the MRI image obtained at the same location and shows an appearance of a signet ring. This Master’s Thesis describes the neurological deficit in brief followed by detail explanation of the advanced computational methods used to simulate this problem. Finally, qualitative results show the resemblance of mechanical model to MRI images of the Nerve Cross Section at the same location validating the capability of these methods to study this neurological deficit.

Langfristig sinnvoll: Digitale Nachhaltigkeit fuer die vierte industrielle Revolution

Ende des 18. Jahrhunderts veraenderte die Dampfmaschine den Transportsektor, die Produktion und viele andere Bereiche der Wirtschaft grundlegend. Aufgrund der Erfindung der Spinnmaschinen konnte die Garnproduktion um ein Vielfaches erhoeht werden, mit Wasser und Dampf konnten Webmaschinen betrieben werden, der Wohlstand der Bevoelkerung wuchs. Gleichzeitig verloren viele Arbeiter ihre Jobs als Spinner, Weber oder Faerber, es gab Proteste und Aufstaende. Nach dieser ersten industriellen Revolution folgte mit der Elektrifizierung Ende 19. Jahrhundert die zweite Revolution. Wieder gewann die Gesellschaft als Ganzes an Wohlstand, aber viele Menschen verloren ihr Einkommen. Die dritte industrielle Revolution folgte Ende des 20. Jahrhunderts mit den Informationstechnologien. Und nur wenig spaeter befinden wir uns bereits an der Schwelle zur vierten industriellen Revolution, der Vernetzung physischer und digitaler Systeme.

Etwas über den Stand der Cultur bei den Juden in Polen im XVI. Jahrhundert

von S. Spinner

(Table T1) Magnetic properties of MORB from ODP Leg 187 sites

The core samples of mid-ocean-ridge basalts (including Indian and Pacific type) recovered from the Southeast Indian Ridge (SEIR) area near the Australian Antarctic Discordance during Ocean Drilling Program Leg 187 were studied using rock magnetism, mineralogy, and petrography methods. On the basis of thermomagnetic analyses and low-temperature magnetometry, the dominant magnetic carrier in most of the basalt samples (pillow basalts) is characterized as titanomaghemite, which presumably formed by low-temperature oxidation of primary titanomagnetite. Some samples from unaltered massive basalts contain nearly unoxidized titanomagnetite as the main magnetic mineral. A metadiabase sample showing greenschist facies metamorphism contains magnetic minerals dominated by magnetite. The pillow basalts contain titanomaghemite ranging from stable single-domain to pseudosingle-domain (PSD) grains, and the majority are characterized by a single stable component of remanence. The massive basalts show hysteresis features of larger PSD grains and contain a very low coercivity remanence. The values of natural remanent magnetization (NRM) of the samples in this SEIR area are on the same order as those of other oceanic ridge basalts. They show a general decreasing trend of NRM with increasing crust age. However, the values of NRM show no correlation either with the tectonic zonations (Zone A vs. Zone B) or with the mantle provinces (Pacific vs. Indian types).

(Table 3) Concentrations of polybrominated diphenyl ether and congeners in blubber samples from whales, seals and porpoises between 1986-2009

A selection of PBDE congeners was analyzed in pooled blubber samples of pilot whale (Globicephala melas), ringed seal (Phoca hispida), minke whale (Balaenoptera acutorostrata), fin whale (Balaenoptera physalus), harbor porpoise (Phocoena phocoena), hooded seal (Cystophora cristata) and Atlantic white-sided dolphin (Lagenorhynchus acutus), covering a time period of more than 20 years (1986-2009). The analytes were extracted and cleaned-up using open column extraction and multi-layer silica gel column chromatography, and the analysis was performed on a GC-MS system operating in the NCI mode. The highest PBDE levels were found in the toothed whale species pilot whale and white-sided dolphin, and the lowest levels in fin whales and ringed seals. One-sided analyses of variance (ANOVA) followed by Tukey comparisons of means were applied to test for differences between years and sampling areas. Due to inter-year sampling variability, only general comparisons of PBDE concentrations between different sampling areas could be made. Differences in PBDE concentrations between three sampling periods, from 1986 to 2007, were evaluated in samples of pilot whales, ringed seals, white-sided dolphins and hooded seals. The highest PBDE levels were found in samples from the late 1990s or beginning of 2000, possibly reflecting the increase in the global production of technical PBDE mixtures in the 1990s. The levels of BDE #153 and #154 increased relative to the total PBDE concentration in some of the species in recent years, which may indicate an increased relative exposure to higher brominated congeners. In order to assess the effect of measures taken in legally binding international agreements, it is important to continuously monitor POPs such as PBDEs in sub-Arctic and Arctic environments.

Magnetic properties of basalts from DSDP Hole 69-504B

The NRM intensity, AF demagnetization characteristics, hysteresis parameters, initial susceptibility, and thermomagnetic characteristics of 18 basalt specimens from Deep Sea Drilling Project Hole 504B were determined. In six samples, the grain size was large enough to allow microprobe analysis. We conclude (1) that the dominant magnetic mineral is titanomagnetite/titanomaghemite; (2) that, except for the upper few meters of the core where the grains are in the stable monodomain state, the grain size of the magnetic mineral lies in the pseudo-single-domain range (< 10 µm); (3) that maghemitization (i.e., low-temperature (< 350°C) oxidation) has taken place. We discuss possible geological histories.

Paleomagnetic of igneous rocks of ODP Leg 114

A paleomagnetic study was made of 12 samples of trachytic basalt from the base of ODP Hole 698A on the Northeast Georgia Rise (southwest Atlantic) and four samples of andesitic basalt and nine samples of volcanic breccia from the base of ODP Hole 703A on the Meteor Rise (southeast Atlantic). The magnetic intensities of the Hole 703A samples are anomalously low, possibly reflecting alteration effects. The mean magnetic intensity of the Hole 698A samples is high, and compatible with the model of Bleil and Petersen (1983) for the variation of magnetic intensity with age in oceanic basalts, involving progressive low-temperature oxidation of titanomagnetite to titanomaghemite for some 20 m.y. followed by inversion to intergrowths of magnetite and other Fe-Ti oxides during the subsequent 100 m.y. These results support the interpretation of the Hole 698A basalts as true oceanic basement of Late Cretaceous age rather than a younger intrusion. Well-defined stable components of magnetization were identified from AF and thermal demagnetization of the Hole 698A basalts, and less well-defined components were identified for the Hole 703A samples. Studies of the magnetic homogeneity of the Hole 698A basalts, involving harmonic analysis of the spinner magnetometer output, indicate the presence of an unevenly distributed low-coercivity component superimposed on the more homogeneous high-coercivity characteristic magnetization. The former component is believed to reside in irregularly distributed multidomain magnetite grains formed along cracks within the basalt, whilst the latter resides in more uniformly distributed finer magnetic grains. The inclination values for the high-coercivity magnetization of five Hole 698A basalt samples form an internally consistent set with a mean value of 59° ± 5°. The corresponding Late Cretaceous paleolatitude of 40° ± 5° is shallower than expected for this site but is broadly compatible with models for the opening of the South Atlantic involving pivoting of South America away from Africa since the Early Cretaceous. The polarity of the stable characteristic magnetization of the Site 698 basalts is normal. This is consistent with their emplacement during the long Campanian to Maestrichtian normal polarity Chron C33N.

Magnetic properties, grain sizes and minerals at DSDP Site 83-504B

Twenty-seven samples from the Leg 83 section of Hole 504B have been investigated using magnetic, optical, and electron optical methods. The primary magnetic mineral to crystallize was titanomagnetite of approximate composition Fe2.4Ti0.6O4 (TM60), but none survives, nor is there evidence of titanomaghemite produced by oxidation of TM60. The average measured magnetic properties can be interpreted in terms of magnetite, Fe3O4, having average grain size of <1 µm and present in average volume concentration of - 0.5%. The intensity of the natural remanent magnetization (NRM) of the rocks could also be accounted for as being a thermoremanence carried by this mineral. Although the heterogeneity of the titanomagnetite grains could be detected optically, the texture of the intergrown phases is poorly developed. In some samples from the massive units of the lower part of the section, trellis patterns were visible. The Fe3O4 present in the intergrowths is too intimately mixed with the other intergrown phases to be revealed by electron microprobe analysis that simply returns the bulk composition of the intergrowth (oxidized TM60). The path by which the mineral assemblage evolved from TM60 to an Fe304-containing intergrowth, under the temperature and pressure conditions obtaining in the Leg 83 section, makes interesting speculation. Deuteric oxidation, maghemitization/inversion, or some hypothetical low-temperature/high-pressure oxidation by a leaching-of-iron process may all play roles.

Susceptibility and time-dependent magnetization of basalts at DSDP Hole 76-534A

Basalts from Hole 534A are among the oldest recovered from the ocean bottom, dating from the opening of the Atlantic 155 Ma. Upon exposure to a 1-Oe field for one week, these basalts acquire a viscous remanent magnetization (VRM), which ranges from 4 to 223% of their natural remanent magnetization (NRM). A magnetic field of similar magnitude is observed in the paleomagnetic lab of the Glomar Challenger, and it is therefore doubtful if accurate measurements of magnetic moment in such rocks can be made on board unless the paleomagnetic area is magnetically shielded. No correlation is observed between the Konigsberger ratio (beta), which is usually less than 3, and the ability to acquire a VRM. The VRM shows both a log t dependence and a Richter aftereffect. Both of these, but especially the log t dependence, will cause the susceptibility measurements (made by applying a magnetic field for a very short time) to be minimum values. The susceptibility and derived Q should therefore be used cautiously for magnetic anomaly interpretation, because they can cause the importance of the induced magnetization to be underestimated.

Paleomagnetism at DSDP Leg 85 Holes

On Leg 85, 16 holes were cored at five sites. Thirteen of the holes were cored with the hydraulic piston corer (HPC) or the variable-length hydraulic piston corer (VLHPC) or both; the remainder were rotary drilled. Partially duplicating stratigraphic sections were successfully recovered by hydraulic piston coring at Sites 572 to 575. Sub-bottom penetration was deepest (about 210 m) at HPC Hole 575A, which bottomed in lower Miocene sediments. Penetration by hydraulic piston coring was limited at all sites not by the failure of the corer to stroke out but by the excessive force (overpull) necessary to retrieve the core barrel from the hole. The sediments recovered are relatively uniform siliceous-calcareous oozes to calcareous ooze-chalks. Paleomagnetic measurements were made at all stratigraphic levels, but magnetostratigraphic sequences could be resolved only for the Pleistocene-Pliocene and for brief upper, middle, and lower Miocene sections. In the younger and less consolidated sediments, the declination often shows large-scale azimuthal rotations downcore. These smooth trends vary from core to core, indicating either rotation between the sediment and the core liner or the rotation of the core barrel during the coring or retrieval process. Thus, azimuthal orientation of the samples was impossible even though a Kuster azimuthal orientation tool was used during the hydraulic piston coring. At all sites, the downhole shift from mainly siliceous to mainly calcareous ooze-chalk coincided with a decrease in NRM intensity of at least one order of magnitude, to 1.0*10**-8 G. Diagenesis is the probable reason for this behavior, although the dilution of magnetic carriers as the result of higher accumulation rates may also be a factor. A tectonic analysis using data from samples with stable remanence indicates a northward plate motion of about 0.3 deg/m.y. during the last 18 m.y., a rate that agrees with most previous reconstructions of Pacific Plate motion.