A Novel Ras-interacting Protein Required for Chemotaxis and Cyclic Adenosine Monophosphate Signal Relay in Dictyostelium

We have identified a novel Ras-interacting protein from Dictyostelium, RIP3, whose function is required for both chemotaxis and the synthesis and relay of the cyclic AMP (cAMP) chemoattractant signal. rip3 null cells are unable to aggregate and lack receptor activation of adenylyl cyclase but are able, in response to cAMP, to induce aggregation-stage, postaggregative, and cell-type-specific gene expression in suspension culture. In addition, rip3 null cells are unable to properly polarize in a cAMP gradient and chemotaxis is highly impaired. We demonstrate that cAMP stimulation of guanylyl cyclase, which is required for chemotaxis, is reduced ∼60% in rip3 null cells. This reduced activation of guanylyl cyclase may account, in part, for the defect in chemotaxis. When cells are pulsed with cAMP for 5 h to mimic the endogenous cAMP oscillations that occur in wild-type strains, the cells will form aggregates, most of which, however, arrest at the mound stage. Unlike the response seen in wild-type strains, the rip3 null cell aggregates that form under these experimental conditions are very small, which is probably due to the rip3 null cell chemotaxis defect. Many of the phenotypes of the rip3 null cell, including the inability to activate adenylyl cyclase in response to cAMP and defects in chemotaxis, are very similar to those of strains carrying a disruption of the gene encoding the putative Ras exchange factor AleA. We demonstrate that aleA null cells also exhibit a defect in cAMP-mediated activation of guanylyl cyclase similar to that of rip3 null cells. A double-knockout mutant (rip3/aleA null cells) exhibits a further reduction in receptor activation of guanylyl cyclase, and these cells display almost no cell polarization or movement in cAMP gradients. As RIP3 preferentially interacts with an activated form of the Dictyostelium Ras protein RasG, which itself is important for cell movement, we propose that RIP3 and AleA are components of a Ras-regulated pathway involved in integrating chemotaxis and signal relay pathways that are essential for aggregation.

Immigration Law by Proxy: The Case of Colorado’s Human Smuggling Crime

Despite the federal government’s well known expansive reach in creating and enforcing immigration law, the states retain substantial authority to play an important role in migrants’ lives. Through their traditional powers to adopt criminal statutes and police their communities, states can indirectly — but intentionally — inject themselves into the incidents of ordinary life as a migrant. Colorado’s human smuggling statute, currently being challenged before the state supreme court, illustrates this type of state regulation of migration. This essay addresses the statute’s reach, its shaky constitutional footing, and places it in a broader context in which states criminalize immigration-related activity.

Modello di simulazione del processo di gestione ordini e di outbound in linguaggio Arena

La tesi ha l'obiettivo di analizzare e tradurre in un modello di simulazione il processo di evasione di ordini dal sito di stoccaggio di un’azienda di e-commerce al fine di valutarne le possibili evoluzioni. In prima analisi sono stati evidenziati i principali motivi che spingono i clienti a preferire gli store online rispetto a quelli fisici e come alcuni dei vantaggi siano il frutto dell’efficienza della gestione dell’outbound. Si è passati, quindi, all’esame del caso di studio, alla scomposizione delle singole fasi di lavorazione degli ordini e alla loro traduzione in linguaggio Arena. Il modello ottenuto è stato valutato grazie alla comparazione dei risultati di statistiche automatiche e manuali, frutto della simulazione, con i dati reali. La fase finale ha riguardato, invece, la valutazione in termini di efficienza di una possibile evoluzione del sistema che consiste nell’introduzione di un robot addetto al packing: il modello è stato modificato e le statistiche del caso as-is sono state confrontate con quelle frutto della nuova simulazione. Con i dati utilizzati è stato dimostrato come un unico robot garantisca prestazioni superiori rispetto allo stato attuale e risulti più che sufficiente per la mole di lavoro di una giornata tipo. Nei periodi dell’anno caratterizzati da un aumento esponenziale degli acquisti, tuttavia, potrebbe essere necessario affiancare alla soluzione automatica quella manuale.