Electrolyte disorders in the human ventricle. A simulation study.

Ventricular cells are immersed in a bath of electrolytes and these ions are essential for a healthy heart and a regular rhythm. Maintaining physiological concentration of them is fundamental for reducing arrhythmias and risk of sudden cardiac death, especially in haemodialysis patients and in the heart diseases treatments. Models of electrically activity of the heart based on mathematical formulation are a part of the efforts to improve the understanding and prediction of heart behaviour. Modern models incorporate the extensive and ever increasing amounts of experimental data in incorporating biophysically detailed mechanisms to allow the detailed study of molecular and subcellular mechanisms of heart disease. The goal of this project was to simulate the effects of changes in potassium and calcium concentrations in the extracellular space between experimental data and and a description incorpored into two modern biophysically detailed models (Grandi et al. Model; O’Hara Rudy Model). Moreover the task was to analyze the changes in the ventricular electrical activity, in particular by studying the modifications on the simulated electrocardiographic signal. We used the cellular information obtained by the heart models in order to build a 1D tissue description. The fibre is composed by 165 cells, it is divided in four groups to differentiate the cell types that compound human ventricular tissue. The main results are the following: Grandi et al. (GBP) model is not even able to reproduce the correct action potential profile in hyperkalemia. Data from hospitalized patients indicates that the action potential duration (APD) should be shorter than physiological state but in this model we have the opposite. From the potassium point of view the results obtained by using O’Hara model (ORD) are in agreement with experimental data for the single cell action potential in hypokalemia and hyperkalemia, most of the currents follow the data from literature. In the 1D simulations we were able to reproduce ECGs signal in most the potassium concentrations we selected for this study and we collected data that can help physician in understanding what happens in ventricular cells during electrolyte disorder. However the model fails in the conduction of the stimulus under hyperkalemic conditions. The model emphasized the ECG modifications when the K+ is slightly more than physiological value. In the calcium setting using the ORD model we found an APD shortening in hypocalcaemia and an APD lengthening in hypercalcaemia, i.e. the opposite to experimental observation. This wrong behaviour is kept in one dimensional simulations bringing a longer QT interval in the ECG under higher [Ca2+]o conditions and vice versa. In conclusion it has highlighted that the actual ventricular models present in literature, even if they are useful in the original form, they need an improvement in the sensitivity of these two important electrolytes. We suggest an use of the GBP model with modifications introduced by Carro et al. who understood that the failure of this model is related to the Shannon et al. model (a rabbit model) from which the GBP model was built. The ORD model should be modified in the Ca2+ - dependent IcaL and in the influence of the Iks in the action potential for letting it him produce a correct action potential under different calcium concentrations. In the 1D tissue maybe a heterogeneity setting of intra and extracellular conductances for the different cell types should improve a reproduction of the ECG signal.

Biologia riproduttiva di Paracentrotus lividus (Lamarck, 1816) in due zone della Sardegna

Paracentrotus lividus è un echinoderma bentonico marino appartenente alla classe Echinoidea, comune in tutto il Mar Mediterraneo e lungo le coste atlantiche dalla Scozia al Marocco. Questa specie è estremamente importante dal punto di vista ecologico in quanto specie chiave nella strutturazione della comunità bentonica macroalgale. Inoltre in tempi recenti il mercato delle sue gonadi ha subito una forte espansione rendendolo una specie di interesse economico in numerosi paesi europei. Il crescente prelievo sta gradualmente portando al depauperamento della risorsa, per questo motivo il riccio di mare è oggetto di numerosi studi e programmi a fini conservazionistici. Una maggiore conoscenza del ciclo riproduttivo della specie potrebbe risultare utile per una migliore gestione della risorsa. Pertanto, questo studio si propone di indagare sulla biologia di P. lividus in due aree distinte della Sardegna: Su Pallosu, nella Penisola del Sinis Sardegna centro-occidentale, e nell’Area Marina Protetta di Tavolara – Punta Coda Cavallo, Sardegna nordorientale, al fine di far luce sulle dinamiche del ciclo riproduttivo. Le informazioni bibliografiche a riguardo evidenziano l’influenza dei fattori ambientali quali fotoperiodo, idrodinamismo, temperatura e tipologia di pascolo sull’andamento del ciclo biologico e il momento di spawning. Diversi autori hanno osservato in Mar Mediterraneo sia un unico ciclo riproduttivo che due eventi riproduttivi annuali a conferma dell’ampia variabilità dell’andamento gonadico. Nel presente lavoro al fine di mettere in relazione le caratteristiche dell’habitat con il ciclo riproduttivo delle popolazioni in oggetto è stato analizzato l’andamento dell’Indice Gonadosomatico (IGS) su individui di diverse taglie/età, come rapporto fra il peso delle gonadi e il peso dell’individuo; inoltre a conferma dello stadio di maturazione è stata effettuata l’analisi istologica delle gonadi su campioni fissati e inclusi in paraffina, sezionati a 7μm e colorati con ematossilina-eosina prima di essere osservati al microscopio ottico. Mediante analisi d’immagine di foto subacquee utilizzando il programma Seascape è stata caratterizzata la componente macroalgale bentonica e la componente abiotica degli habitat (sabbia e roccia non vegetata) con particolare attenzione al calcolo della percentuale di ricoprimento delle specie vegetali. Inoltre i siti sono stati caratterizzati rispetto all’esposizione dei venti dominanti con elaborazione dei dati del periodo di campionamento per direzione, frequenza e intensità.

Modelling growth dynamics and toxin production in Ostreopsis cf. ovata

The benthic dinoflagellate O. ovata represents a serious threat for human health and for the ecology of its blooming areas: thanks to its toxicity this microalga has been responsible for several cases of human intoxication and mass mortalities of benthic invertebrates. Although the large number of studies on this dinoflagellate, the mechanisms underpinning O. ovata growth and toxin production are still far to be fully understood. In this work we have enriched the dataset on this species by carrying out a new experiment on an Adriatic O. cf. ovata strain. Data from this experiment (named Beta) and from another comparable experiment previously conducted on the same strain (named Alpha), revealed some interesting aspects of this dinoflagellate: it is able to grow also in a condition of strong intracellular nutrient deficiency (C:P molar ratio > 400; C:N > 25), reaching extremely low values of chlorophyll-a to carbon ratio (0.0004). Was also found a significant inverse relationships (r > -0.7) between cellular toxin to carbon and cellular nutrient to carbon ratios of experiment Alpha. In the light of these result, we hypothesized that in O. cf. ovata nutrient-stress conditions (intended as intracellular nutrient deficiency) can cause: i) an increase in toxin production; ii) a strong decrease in chlorophyll-a synthesis; iii) a lowering of metabolism associated with the formation of a sort of resting stage. We then used a modelling approach to test and critically evaluate these hypotheses in a mechanistic way: newly developed formulation describing toxin production and fate, and ad hoc changes in the already existent formulations describing chlorophyll synthesis, rest respiration, and mortality, have been incorporated in a simplified version of the European Regional Seas Ecosystem Model (ERSEM), together with a new ad hoc parameterization. The adapted model was able to accurately reproduce many of the trends observed in the Alpha experiment, allowing us to support our hypotheses. Instead the simulations of the experiment Beta were not fully satisfying in quantitative terms. We explained this gap with the presumed different physiological behaviors between the algae of the two experiments, due to the different pre-experimental periods of acclimation: the model was not able to reproduce acclimation processes in its simulations of the experiment Beta. Thus we attempt to simulate the acclimation of the algae to nutrient-stress conditions by manual intervention on some parameters of nutrient-stress thresholds, but we received conflicting results. Further studies are required to shed light on this interesting aspect. In this work we also improve the range of applicability of a state of the art marine biogeochemical model (ERSEM) by implementing in it an ecological relevant process such as the production of toxic compounds.

Biologia ed ecologia dello squalo di profondità Dalatias licha (Bonnaterre, 1788)

La presente tesi vuole colmare le conoscenze biologiche ed ecologiche dello squalo di profondità Dalatias licha. Attualmente le conoscenze riguardanti le popolazioni di questo squalo nel Mediterraneo sono scarse dato il numero limitato di studi affrontati. La biologia e l’ecologia di questo squalo è stata studiata attraverso la stima di alcune caratteristiche importanti come la sex ratio, la maturità sessuale degli individui, gli indici di condizione (epatosomatico e gonadosomatico) e le analisi dei contenuti stomacali. I 78 esemplari (16 femmine e 62 maschi) osservati sono stati campionati tra il 2001 e il 2003 dal bycatch di motopescherecci a strascico della marineria di Genova porto, operanti sul piano batiale del Golfo di Genova (Mar Ligure). Le catture sono state eseguite ad una batimetria compresa tra -450 e -800 m, tra Genova e Sestri Levante. I risultati, sebbene basati su un numero limitato di individui, sono di particolare interesse visto il deficit conoscitivo inerente a questa specie in ambito nazionale e mediterraneo

Complessi tetrazolici di Re(I): design e preparazione di una nuova classe di probes luminescenti

In this experimental work we report the design, the synthesis and characterization of a new class of Re(I) complexes of the general formula fac-[Re(CO)3(N^N)(2-QTZ)], where N^N = 2,2’ bipyridine or 1,10 phenantroline, whereas 2-QTZ is the anion 2-quinolyl-tetrazolate. The complexes and, in particular, the tetrazolate ligand 2-QTZ were designed in order to investigate their specific interaction with biologically and toxicologically relevant metal ions, as Zn(II), Cd(II) e Cu(II). The addition of such ions led to substantial variations of the photophysical properties of these complexes, suggesting their application as luminescent sensors. The photophysical performance of the complexes proved to remain unchanged inside cellular substrates, as Yarrowia Lipolytica cultures. Within these yeasts, the complexes show unchanged ability to perform luminescent sensing towards Zn(II) and Cd(II) ions.