Chemical inhibitor targeting the replication protein A-DNA interaction increases the efficacy of Pt-based chemotherapy in lung and ovarian cancer

Platinum-based chemotherapeutics exert their therapeutic efficacy via the formation of DNA adducts which interfere with DNA replication, transcription and cell division and ultimately induce cell death. Repair and tolerance of these Pt-DNA lesions by nucleotide excision repair (NER) and homologous recombination (HR) can substantially reduce the effectiveness of therapy. Inhibition of these repair pathways, therefore, holds the potential to sensitize cancer cells to Pt treatment and increase clinical efficacy. Replication Protein A (RPA) plays essential roles in both NER and HR, along with its role in DNA replication and DNA damage checkpoint activation. Each of these functions is, in part, mediated by RPA binding to single-stranded DNA (ssDNA). Here we report the synthesis and characterization of novel derivatives of RPA small molecule inhibitors and their activity in models of epithelial ovarian cancer (EOC) and non-small cell lung cancer (NSCLC). We have synthesized analogs of our previously reported RPA inhibitor TDRL-505 and determined the structure-activity relationships. These data led us to the identification of TDRL-551, which exhibited a greater than 2-fold increase in in vitro activity. TDRL-551 showed synergy with Pt in tissue culture models of EOC and in vivo efficacy, as a single agent and in combination with platinum, in a NSCLC xenograft model. These data demonstrate the utility of RPA inhibition in EOC and NSCLC and the potential in developing novel anticancer therapeutics that target RPA-DNA interactions

Reflecting on Algorithmic Bias with Design Fiction:the MiniCoDe Workshops

In an increasingly complex everyday life, algorithms-often learned from data, i.e., machine learning (ML)-are used to make or assist with operational decisions. However, developers and designers usually are not entirely aware of how to reflect on social justice while designing ML algorithms and applications. Algorithmic social justice-i.e., designing algorithms including fairness, transparency, and accountability-aims at helping expose, counterbalance, and remedy bias and exclusion in future ML-based decision-making applications. How might we entice people to engage in more reflective practices that examine the ethical consequences of ML algorithmic bias in society? We developed and tested a design-fiction-driven methodology to enable multidisciplinary teams to perform intense, workshop-like gatherings to let potential ethical issues emerge and mitigate bias through a series of guided steps. With this contribution, we present an original and innovative use of design fiction as a method to reduce algorithmic bias in co-design activities.</p

Testing the metal of ERCC2 in predicting the response to platinum-based therapy

DNA repair has been shown to affect the cellular response to platinum-based therapy in a variety of cancers; however, translating this knowledge to the clinic has proven difficult and yielded mixed results. In this issue of Cancer Discovery, Van Allen and colleagues have analyzed responders and nonresponders to neoadjuvant platinum-based therapy with locally advanced urothelial cancer and identified a series of mutations in the nucleotide excision repair (NER) gene ERCC2 that correlate with the response to platinum-based therapy. This work provides evidence that defects in NER can be exploited to maximize the efficacy of conventional platinum-based chemotherapy

Recognition of DNA Termini by the C-Terminal Region of the Ku80 and the DNA-Dependent Protein Kinase Catalytic Subunit

DNA double strand breaks (DSBs) can be generated by endogenous cellular processes or exogenous agents in mammalian cells. These breaks are highly variable with respect to DNA sequence and structure and all are recognized in some context by the DNA-dependent protein kinase (DNA-PK). DNA-PK is a critical component necessary for the recognition and repair of DSBs via non-homologous end joining (NHEJ). Previously studies have shown that DNA-PK responds differentially to variations in DSB structure, but how DNA-PK senses differences in DNA substrate sequence and structure is unknown. Here we explore the enzymatic mechanisms by which DNA-PK is activated by various DNA substrates and provide evidence that the DNA-PK is differentially activated by DNA structural variations as a function of the C-terminal region of Ku80. Discrimination based on terminal DNA sequence variations, on the other hand, is independent of the Ku80 C-terminal interactions and likely results exclusively from DNA-dependent protein kinase catalytic subunit interactions with the DNA. We also show that sequence differences in DNA termini can drastically influence DNA repair through altered DNA-PK activation. These results indicate that even subtle differences in DNA substrates influence DNA-PK activation and ultimately the efficiency of DSB repair

NOVEL PROCESS FOR REMOVAL AND RECOVERY OF VAPOR-PHASE MERCURY

The goal of this project is to investigate the use of a regenerable sorbent for removing and recovering mercury from the flue gas of coal-fired power plants. The process is based on the sorption of mercury by noble metals and the thermal regeneration of the sorbent, recovering the desorbed mercury in a small volume for recycling or disposal. The project was carried out in two phases, covering five years. Phase I ran from September 1995 through September 1997 and involved development and testing of sorbent materials and field tests at a pilot coal-combustor. Phase II began in January 1998 and ended September 2000. Phase II culminated with pilot-scale testing at a coal-fired power plant. The use of regenerable sorbents holds the promise of capturing mercury in a small volume, suitable for either stable disposal or recycling. Unlike single-use injected sorbents such as activated carbon, there is no impact on the quality of the fly ash. During Phase II, tests were run with a 20-acfm pilot unit on coal-combustion flue gas at a 100 lb/hr pilot combustor and a utility boiler for four months and six months respectively. These studies, and subsequent laboratory comparisons, indicated that the sorbent capacity and life were detrimentally affected by the flue gas constituents. Sorbent capacity dropped by a factor of 20 to 35 during operations in flue gas versus air. Thus, a sorbent designed to last 24 hours between recycling lasted less than one hour. The effect resulted from an interaction between SO{sub 2} and either NO{sub 2} or HCl. When SO{sub 2} was combined with either of these two gases, total breakthrough was seen within one hour in flue gas. This behavior is similar to that reported by others with carbon adsorbents (Miller et al., 1998)

Differential activation of DNA-PK based on DNA strand orientation and sequence bias

DNA-PKcs and Ku are essential components of the complex that catalyzes non-homologous end joining (NHEJ) of DNA double-strand breaks (DSBs). Ku, a heterodimeric protein, binds to DNA ends and facilitates recruitment of the catalytic subunit, DNA-PKcs. We have investigated the effect of DNA strand orientation and sequence bias on the activation of DNA-PK. In addition, we assessed the effect of the position and strand orientation of cisplatin adducts on kinase activation. A series of duplex DNA substrates with site-specific cisplatin–DNA adducts placed in three different orientations on the duplex DNA were prepared. Terminal biotin modification and streptavidin (SA) blocking was employed to direct DNA-PK binding to the unblocked termini with a specific DNA strand orientation and cisplatin–DNA adduct position. DNA-PK kinase activity was measured and the results reveal that DNA strand orientation and sequence bias dramatically influence kinase activation, only a portion of which could be attributed to Ku-DNA binding activity. In addition, cisplatin–DNA adduct position resulted in differing degrees of inhibition depending on distance from the terminus as well as strand orientation. These results highlight the importance of how local variations in DNA structure, chemistry and sequence influence DNA-PK activation and potentially NHEJ

New onset status epilepticus in influenza associated encephalopathy: The presenting manifestation of genetic generalized epilepsy

We hereby present a case of a young woman with no history of seizures or epilepsy who experienced a de novo generalized Non Convulsive Status Epilepticus (NCSE) followed by encephalopathy lasting for several days during influenza B infection. Influenza can have a broad spectrum of presentation ranging from an uncomplicated illness to many serious conditions as is the case of influenza associated encephalitis/encephalopathy (IAE). In this context however, it is possible to observe seizures and/or status epilepticus as the presenting manifestation of a genetic generalized epilepsy

Effect of a Lactobacillus sakei and Staphylococcus xylosus protective culture on Listeria monocytogenes growth and quality traits of Italian fresh sausage (salsiccia) stored at abusive temperature

Fresh sausages are not always thoroughly cooked before consumption and can support the growth of pathogenic microorganisms, especially when stored at incorrect temperatures. The aim of this study was to verify the growth of Listeria monocytogenes in Italian salsiccia stored at 2 °C, 7 °C and 12 °C for 9 days (t9) with (PC+) and without (PC–) a commercial protective culture (Lactobacillus sakei and Staphylococcus xylosus). L. monocytogenes PC + counts were statistically different from PC–, after 7 days (t7) at 7 °C and at 12 °C. At 2 °C, they increased in PC + by 0.03 and 0.36 log CFU/g vs. 0.25 and 0.91 in PC–, at t7 and t9. Moreover, quality characteristics (total aerobic counts, colour parameters, TBARS values, pH, sensory attributes) were assessed in fresh sausages stored at 7 °C. Significant differences were obtained in PC + samples at t7 for Pseudomonadaceae and Enterobacteriaceae (about 2 log CFU/g). Yeasts and moulds and Brochothrix thermosphacta were also significantly lower in PC + samples. PC + samples were more acidic than PC–, with statistically different colour parameters values particularly at the external surface; raw sausages resulted sensorially discernible at t7, whereas PC + and PC– cooked samples did not show any significant sensory difference. The studied protective culture proved to be a useful tool to increase safety and microbiological quality of salsiccia at abusive storage temperature, effectively limiting the growth of L. monocytogenes and Gram negative spoilage microorganisms, with some sensory drawbacks, especially at the end of the shelf life.HIGHLIGHTS The tested culture improves the microbial safety of salsiccia. It reduces the growth of spoilage microorganisms, especially Pseudomonadaceae and Enterobacteriaceae. It mainly influences colour and acidity evolution of the raw product with no sensory drawbacks in the cooked product

Effects of magnetic field and disorder on electronic properties of Carbon Nanotubes

Electronic properties of metallic and semiconducting carbon nanotubes are investigated in presence of magnetic field perpendicular to the CN-axis, and disorder introduced through energy site randomness. The magnetic field field is shown to induce a metal-insulator transition (MIT) in absence of disorder, and surprisingly disorder does not affect significantly the MIT. These results may find confirmation through tunneling experimentsComment: 4 pages, 6 figures. Phys. Rev. B (in press

Listeria monocytogenes contamination of Tenebrio molitor larvae rearing substrate: Preliminary evaluations

Today, edible insects represent a hot topic as an emerging and eco-friendly source of protein. The mealworm (Tenebrio molitor L.) is among the most employed insects for human consumption and feed purposes. So far Listeria monocytogenes, have never been detected either in products sold on the market or during the rearing process. In this study, the substrate employed for mealworm rearing was deliberately contaminated with L. monocytogenes and the bacterium was enumerated during the rearing period and after technological treatments of the larvae. L. monocytogenes persisted during the rearing period. Washing the larvae did not produce any significant effect, while fasting the larvae for 24 or 48 h reduced the L. monocytogenes load (P &lt; 0.001). Oven cooking eliminated L. monocytogenes cells from the product, reducing the risk associated to this foodborne pathogen to zero