The clinically relevant CHK1 inhibitor MK-8776 induces the degradation of the oncogenic protein PML-RARα and overcomes ATRA resistance in acute promyelocytic leukemia cells.

Acute promyelocytic leukemia (APL) is a hematological disease characterized by the expression of the oncogenic fusion protein PML-RARα. The current treatment approach for APL involves differentiation therapy using all-trans retinoic acid (ATRA) and arsenic trioxide (ATO). However, the development of resistance to therapy, occurrence of differentiation syndrome, and relapses necessitate the exploration of new treatment options that induce differentiation of leukemic blasts with low toxicity. In this study, we investigated the cellular and molecular effects of MK-8776, a specific inhibitor of CHK1, in ATRA-resistant APL cells. Treatment of APL cells with MK-8776 resulted in a decrease in PML-RARα levels, increased expression of CD11b, and increased granulocytic activity consistent with differentiation. Interestingly, we showed that the MK-8776-induced differentiating effect resulted synergic with ATO. We found that the reduction of PML-RARα by MK-8776 was dependent on both proteasome and caspases. Specifically, both caspase-1 and caspase-3 were activated by CHK1 inhibition, with caspase-3 acting upstream of caspase-1. Activation of caspase-3 was necessary to activate caspase-1 and promote PML-RARα degradation. Transcriptomic analysis revealed significant modulation of pathways and upstream regulators involved in the inflammatory response and cell cycle control upon MK-8776 treatment. Overall, the ability of MK-8776 to induce PML-RARα degradation and stimulate differentiation of immature APL cancer cells into more mature forms recapitulates the concept of differentiation therapy. Considering the in vivo tolerability of MK-8776, it will be relevant to evaluate its potential clinical benefit in APL patients resistant to standard ATRA/ATO therapy, as well as in patients with other forms of acute leukemias.

Thalassemias: from gene to therapy.

Thalassemias (α, ß, γ, δ, δß, and εγδß) are the most common genetic disorders worldwide and constitute a heterogeneous group of hereditary diseases characterized by the deficient synthesis of one or more hemoglobin (Hb) chain(s). This leads to the accumulation of unstable non-thalassemic Hb chains, which precipitate and cause intramedullary destruction of erythroid precursors and premature lysis of red blood cells (RBC) in the peripheral blood. Non-thalassemic Hbs display high oxygen affinity and no cooperativity. Thalassemias result from many different genetic and molecular defects leading to either severe or clinically silent hematologic phenotypes. Thalassemias α and ß are particularly diffused in the regions spanning from the Mediterranean basin through the Middle East, Indian subcontinent, Burma, Southeast Asia, Melanesia, and the Pacific Islands, whereas δß-thalassemia is prevalent in some Mediterranean regions including Italy, Greece, and Turkey. Although in the world thalassemia and malaria areas overlap apparently, the RBC protection against malaria parasites is openly debated. Here, we provide an overview of the historical, geographic, genetic, structural, and molecular pathophysiological aspects of thalassemias. Moreover, attention has been paid to molecular and epigenetic pathways regulating globin gene expression and globin switching. Challenges of conventional standard treatments, including RBC transfusions and iron chelation therapy, splenectomy and hematopoietic stem cell transplantation from normal donors are reported. Finally, the progress made by rapidly evolving fields of gene therapy and gene editing strategies, already in pre-clinical and clinical evaluation, and future challenges as novel curative treatments for thalassemia are discussed.

Effect of earthworms and plants on the efficiency of vertical flow systems treating university wastewater.

One of the possible ways to improve the operation efficiency of constructed wetlands and to prevent their clogging is the application of earthworms. They have already been successfully applied for vermicomposting and for sludge dewatering and treatment. A few studies have already examined the effect of earthworms on the treatment of wastewater by vertical flow constructed wetlands (VFCWs), but none of them have provided a yearlong research result from an open-air system or compared the effect that different seasons in a temperate climate area can have on these invertebrates. The goal of this research was to estimate the effect that earthworms and plants have on VFCW's operation. Four mesocosms (a filter, a filter with earthworms, a VFCW and a VFCW with earthworms) were built and their influent and effluent water quality was monitored for a period of 1 year. They were fed with wastewater coming from a building of the University of Bologna (Italy). The results have shown that the presence of earthworms in this specific system did not reduce the organic matter content of the substrate, but it has positively influenced plants' growth. However, since neither earthworms nor plants had a statistically significant effect on the effluent quality, it can be concluded that the integration of these invertebrates cannot improve wastewater treatment of vertical flow filters or constructed wetlands.

Industrial Cooling Tower Disinfection Treatment to Prevent Legionella spp.

The contamination of industrial cooling towers has been identified as one cause of legionellosis, but the real risk has been underestimated. Two different disinfection treatments were tested on Legionella colonization in an industrial Cooling Tower System (CTS). Environmental monitoring of Legionella, P. aeruginosa, and a heterotrophic plate count (HPC) at 36 °C was performed from June to October 2016. The disinfection procedures adopted were based on hydrogen peroxide (H2O2) and silver salts (Ag⁺), in addition to an anti-algal treatment, then using hyperclorination as a shock, and then continuous treatment by sodium hypochlorite (NaClO). L. pneumophila serogroup 8 was found at a concentration of 5.06 Log cfu/L after the CTS filling; a shock treatment performed by H2O2/Ag⁺ produced a rapid increase in contamination up to 6.14 Log cfu/L. The CTS activity was stopped and two subsequent shock treatments were performed using NaClO, followed by continuous hyperclorination. These procedures showed a significant decrease (p < 0.05) in Legionella concentration (1.77 Log cfu/L). The same trend was observed for P. aeruginosa (0.55 Log cfu/100 mL) and HPC (1.95 Log cfu/mL) at 36 °C. Environmental monitoring and the adoption of maintenance procedures, including anti-scale treatment, and physical, chemical, and microbiological control, ensure the good performance of a CTS, reducing the Legionella risk for public health.

Effect of monochloramine treatment on colonization of a hospital water distribution system by Legionella spp.: a 1 year experience study.

Contamination of hot water distribution systems by Legionella represents a great challenge due to difficulties associated with inactivating microorganisms, preserving the water characteristics. The aim of this study was to examine over the course of 1 year in 11 fixed sites, the impact of monochloramine disinfection on Legionella, heterotrophic bacteria (36 °C), Pseudomonas aeruginosa contamination, and chemical parameters of a plumbing system in an Italian hospital. Three days after installation (T0), in the presence of monochloramine concentration between 1.5 and 2 mg/L, 10/11 sites (91%) were contaminated by L. pneumophila serogroups 3 and 10. After these results, the disinfectant dosage was increased to between 6 and 10 mg/L, reducing the level of Legionella by three logarithmic unit by 2 months postinstallation (T2) until 6 months later (T3). One year later (T4), there was a significant reduction (p = 0.0002) at 8/11 (73%) sites. Our data showed also a significant reduction of heterotrophic bacteria (36 °C) in 6/11 (55%) sites at T4 (p = 0.0004), by contrast the contamination of P. aeruginosa found at T0 in two sites persisted up until T4. The results of the present study show that monochloramine is a promising disinfectant that can prevent Legionella contamination of hospital water supplies.

Self and other in the human motor system.

Observation of another's action can selectively facilitate the brain's motor circuits for making the same action . A "mirror-matching mechanism" might map observed actions onto the observer's own motor representations . Crucially, this view suggests that the brain represents others' actions like one's own. However, this hypothesis has been difficult to test because the experience of one's own body differs from that of others' bodies with respect to viewpoint, morphological features, familiarity, and the hallmark feature of kinaesthetic experience. We used an established method for manipulating the sense of body ownership ("rubber-hand illusion") to compare effects of observing actions that either were or were not illusorily attributed to the subject's own body. We show that observing another's actions facilitated the motor system, whereas observing identical actions, which were illusorily attributed to the subject's own body, showed the opposite pattern. Thus, motor facilitation strongly depends on the agent to whom the observed action is attributed. This result contradicts previous concepts of equivalence between one's own actions and actions of others and suggests that social differentiation, not equivalence, is characteristic of the human action system.