Re-Inhabiting an Industrial Ruin: Castellammare di Stabia, Italy

Site is used as form determinant in a multi-faceted exploration of parametric modeling. The Industrial Site of in the city of Castellammare di Stabia is a case study for other sites around the region. The current site sits as a relic facing three different fronts: the water to the north, the industrial shipyard to the west, and the city, east and south. Culture and contextual sustainability are other drivers of the manipulation of the site. Part of the culture in Castellammare di Stabia is to use the Bay of Naples for fresh seafood daily and natural spring water from twenty distinct springs. To integrate the site into the current culture programmatic uses that are domestic as apposed to industrial allows for the inhabitants to experience the waterfront all along the southern part of the coast, as they already do in the north. The process of reinhabitation the site has been the primary focus. This thesis deals with the various components of this industrial ruin, reconnecting the city with its amenity. The pier as a leisurely passerella extending in to the bay of Naples, the elevated silos as a housing apartment complex, the ground floor as a plaza/market place, and the roof top as a public open terrace/ theater. Much like the various components of the complex in their original use had a specific purpose, in this transformation they serve a different but equally specific purpose

Methods of NLP in arts management

The boost of digital archives and libraries in art, literature, and music; the shift of cultural marketing and cultural criticism to social media and online platforms; and the emergence of new digital art and cultural products lead to an enormous increase in digital data, creating challenges as well as opportunities for arts management practitioners and researchers. For arts practitioners, NLP can be used to improve marketing and communication for target group analysis, event evaluation, (social) media analysis, pricing, social media optimization, advertisement targeting, or search engine optimization. In the field of archives, collections, and libraries, NLP can contribute to the improvement of indexing, consistency, and quality of databases as well as the development of suitable search algorithms. In the distribution of cultural products, online platforms can be improved and the markets analyzed

Mechanism of efficient double-strand break repair by a long non-coding RNA.

Mechanistic studies in DNA repair have focused on roles of multi-protein DNA complexes, so how long non-coding RNAs (lncRNAs) regulate DNA repair is less well understood. Yet, lncRNA LINP1 is over-expressed in multiple cancers and confers resistance to ionizing radiation and chemotherapeutic drugs. Here, we unveil structural and mechanistic insights into LINP1's ability to facilitate non-homologous end joining (NHEJ). We characterized LINP1 structure and flexibility and analyzed interactions with the NHEJ factor Ku70/Ku80 (Ku) and Ku complexes that direct NHEJ. LINP1 self-assembles into phase-separated condensates via RNA-RNA interactions that reorganize to form filamentous Ku-containing aggregates. Structured motifs in LINP1 bind Ku, promoting Ku multimerization and stabilization of the initial synaptic event for NHEJ. Significantly, LINP1 acts as an effective proxy for PAXX. Collective results reveal how lncRNA effectively replaces a DNA repair protein for efficient NHEJ with implications for development of resistance to cancer therapy

Novel structural arrangement of nematode cystathionine β-synthases: characterization of Caenorhabditis elegans CBS-1

CBSs (cystathionine β-synthases) are eukaryotic PLP (pyridoxal 5 *-phosphate)-dependent proteins that maintain cellular homocysteine homoeostasis and produce cystathionine and hydrogen sulfide. In the present study, we describe a novel structural arrangement of the CBS enzyme encoded by the cbs-1 gene of the nematode Caenorhabditis elegans. The CBS-1 protein contains a unique tandem repeat of two evolutionarily conserved catalytic regions in a single polypeptide chain. These repeats include a catalytically active C-terminal module containing a PLP-binding site and a less conserved N-terminal module that is unable to bind the PLP cofactor and cannot catalyse CBS reactions, as demonstrated by analysis of truncated variants and active-site mutant proteins. In contrast with other metazoan enzymes, CBS-1 lacks the haem and regulatory Bateman domain essential for activation by AdoMet (S-adenosylmethionine) and only forms monomers. We determined the tissue and subcellular distribution of CBS-1 and showed that cbs-1 knockdown by RNA interference leads to delayed development and to an approximately 10-fold elevation of homocysteine concentrations in nematode extracts. The present study provides the first insight into the metabolism of sulfur amino acids and hydrogen sulfide in C. elegans and shows that nematode CBSs possess a structural feature that is unique among CBS proteins

Cystathionine beta-synthase mutants exhibit changes in protein unfolding: conformational analysis of misfolded variants in crude cell extracts

Protein misfolding has been proposed to be a common pathogenic mechanism in many inborn errors of metabolism including cystathionine β-synthase (CBS) deficiency. In this work, we describe the structural properties of nine CBS mutants that represent a common molecular pathology in the CBS gene. Using thermolysin in two proteolytic techniques, we examined conformation of these mutants directly in crude cell extracts after expression in E. coli. Proteolysis with thermolysin under native conditions appeared to be a useful technique even for very unstable mutant proteins, whereas pulse proteolysis in a urea gradient had limited values for the study of the majority of CBS mutants due to their instability. Mutants in the active core had either slightly increased unfolding (p.A114V, p.E302K and p.G307S) or extensive unfolding with decreased stability (p.H65R, p.T191M, p.I278T and p.R369C). The extent of the unfolding inversely correlated with the previously determined degree of tetrameric assembly and with the catalytic activity. In contrast, mutants bearing aminoacid substitutions in the C-terminal regulatory domain (p.R439Q and p.D444N) had increased global stability with decreased flexibility. This study shows that proteolytic techniques can reveal conformational abnormalities even for CBS mutants that have activity and/or a degree of assembly similar to the wild-type enzyme. We present here a methodological strategy that may be used in cell lysates to evaluate properties of proteins that tend to misfold and aggregate and that may be important for conformational studies of disease-causing mutations in the field of inborn errors of metabolism

Systems-Thinking: Formalization of parametric process

This paper details a design process focused on explicit digital parametric modelling as an integral system-outcome design. This investigation isolates and alters a simple geometric form (cylinder) in a constructed architectural design method. Systems are defined as logical, sequential operations inherent to the resultant effects. These operations within each system are composed of various parameters, singular entities containing or referencing data. Given specific data, operations are preformed culminating with corresponding outcomes. The two main components of this research pertain to object extraction and transformation. A single grain silo (cylinder), as the architectural/geometric object under examination, is tested using a system of varied parameters inputted into the program Grasshopper, an “explicit historyi graphic plug-in for Rhinoceros. This application is utilized to digitally manipulate parameters as objects in a nodal arrangement. Throughout the operations execution, this isolated silo will be transformed into a multitude of versions, then regrouped into the original cluster of silos to expose the implications from patterning, adjacency, and repetition given the proximity of the each silo and its new parametric characteristics. As the various parameters in specific operations affect the system as a whole, so is each adjacent silo in proximity given the same or similar operation? This then is translated and reflected in the outcome. This research seeks to explore design process by applying constant digital 3-D reductive geometric, modular forms inviting systems thinking in parametric environments that can lead to architectural design implications. By focusing on the technical aspect of the parameterization and valuing functionality rather then style, the process becomes focused on formal qualities as the system-outcome relationships. This research tests the “aesthetic implicationsi of a varied mode of digital design, namely the investigation of an architectural process utilizing parametric design

Stages, scaffolds and strings in the spatial organisation of non-homologous end joining: Insights from X-ray diffraction and Cryo-EM.

Non-homologous end joining (NHEJ) is the preferred pathway for the repair of DNA double-strand breaks in humans. Here we describe three structural aspects of the repair pathway: stages, scaffolds and strings. We discuss the orchestration of DNA repair to guarantee robust and efficient NHEJ. We focus on structural studies over the past two decades, not only using X-ray diffraction, but also increasingly exploiting cryo-EM to investigate the macromolecular assemblies