Consecutive day dosing of high-dose cytarabine consolidation over 3 days is resource-efficient and safe in older adult patients with acute myeloid leukemia.

High-dose cytarabine (HDAC) is conventionally delivered on days 1, 3 and 5 (HDAC-135) as acute myeloid leukemia (AML) post-remission therapy. Limited data is available on alternative HDAC schedules such as HDAC-123 (given consecutively for 3 days). We retrospectively compared the tolerability and efficacy of HDAC-135 and HDAC-123 delivered in sequential cohorts of adult AML patients. Seventy-three patients were included with 33% aged ≥60 years. HDAC-123 was associated with faster hematological recovery, reduced bacteremia and shorter hospitalization. No differences in safety profile or hematological recovery were seen between patients ≥60 years and <60 years receiving HDAC-123 except a shorter median time to neutrophil count recovery after cycle 1 in the latter group. Three patients (8%) receiving HDAC-123, all aged <60 years, required a change in schedule to HDAC-135 due to transient cytarabine-related side effects. HDAC-123 consolidation was well-tolerated by AML patients, including those ≥60 years, and associated with tangible reductions in resource utilization.

Improved Triplex-Forming Isoorotamide PNA Nucleobases for A-U Recognition of RNA Duplexes.

Four new isoorotamide (Io)-containing PNA nucleobases have been designed for A-U recognition of double helical RNA. New PNA monomers were prepared efficiently and incorporated into PNA nonamers for binding A-U in a PNA:RNA2 triplex. Isothermal titration calorimetry and UV thermal melting experiments revealed slightly improved binding affinity for singly modified PNA compared to known A-binding nucleobases. Molecular dynamics simulations provided further insights into binding of Io bases in the triple helix. Together, the data revealed interesting insights into binding modes including the notion that three Hoogsteen hydrogen bonds are unnecessary for strong selective binding of an extended nucleobase. Cationic monomer Io8 additionally gave the highest affinity observed for an A-binding nucleobase to date. These results will help inform future nucleobase design toward the goal of recognizing any sequence of double helical RNA.

In Vivo Genome Editing in Type I and II Methanotrophs Using a CRISPR/Cas9 System.

Methanotrophic bacteria are Gram-negative, aerobic organisms that use methane as their sole source of carbon and energy. In this study, we constructed and exemplified a CRISPR/Cas9 genome editing system and used it to successfully make gene deletions and insertions in the type I methanotroph Methylococcus capsulatus Bath and the type II methanotroph Methylocystis parvus OBBP. High frequencies of gene deletions and insertions were achieved in combination with homology-directed repair. In M. parvus OBBP, we also investigated the impact of several parameters on the CRISPR/Cas9 genome editing, where the ligD gene was targeted with various PAM sequences and guide RNA spacer sequences, homology arms of variable length, differences in the duration of mating during conjugation, and exploiting promoters of different strengths to control the expression of cas9 and sgRNA. Although not the first attempt to develop a CRISPR/Cas system in methanotrophs, this work demonstrated for the first time an efficient CRISPR/Cas9 system generating scarless clean gene deletions and insertions in methanotroph genomes.

The effects of responsible drinking messages on attentional allocation and drinking behaviour.

AIMS: Four experiments were conducted to assess the acute impact of context and exposure to responsible drinking messages (RDMs) on attentional allocation and drinking behaviour of younger drinkers and to explore the utility of lab-based methods for the evaluation of such materials. METHODS: A simulated bar environment was used to examine the impact of context, RDM posters, and brief online responsible drinking advice on actual drinking behaviour. Experiments one (n = 50) and two (n = 35) comprised female non-problem drinkers, whilst Experiments three (n = 80) and 4 (n = 60) included a mixed-gender sample of non-problem drinkers, recruited from an undergraduate student cohort. The Alcohol Use Disorders Identification Test (AUDIT) was used to assess drinking patterns. Alcohol intake was assessed through the use of a taste preference task. RESULTS: Drinking in a simulated bar was significantly greater than in a laboratory setting in the first two studies, but not in the third. There was a significant increase in alcohol consumption as a result of being exposed to RDM posters. Provision of brief online RDM reduced the negative impact of these posters somewhat; however the lowest drinking rates were associated with being exposed to neither posters nor brief advice. Data from the final experiment demonstrated a low level of visual engagement with RDMs, and that exposure to posters was associated with increased drinking. CONCLUSIONS: Poster materials promoting responsible drinking were associated with increased consumption amongst undergraduate students, suggesting that poster campaigns to reduce alcohol harms may be having the opposite effect to that intended. Findings suggest that further research is required to refine appropriate methodologies for assessing drinking behaviour in simulated drinking environments, to ensure that future public health campaigns of this kind are having their intended effect.

Utilization of communication technology by patients enrolled in substance abuse treatment.

BACKGROUND: Technology-based applications represent a promising method for providing efficacious, widely available interventions to substance abuse treatment patients. However, limited access to communication technology (i.e., mobile phones, computers, internet, and e-mail) could significantly impact the feasibility of these efforts, and little is known regarding technology utilization in substance abusing populations. METHODS: A survey was conducted to characterize utilization of communication technology in 266 urban, substance abuse treatment patients enrolled at eight drug-free, psychosocial or opioid-replacement therapy clinics. RESULTS: Survey participants averaged 41 years of age and 57% had a yearly household income of less than $15,000. The vast majority reported access to a mobile phone (91%), and to SMS text messaging (79%). Keeping a consistent mobile phone number and yearly mobile contract was higher for White participants, and also for those with higher education, and enrolled in drug-free, psychosocial treatment. Internet, e-mail, and computer use was much lower (39-45%), with younger age, higher education and income predicting greater use. No such differences existed for the use of mobile phones however. CONCLUSIONS: Concern regarding the digital divide for marginalized populations appears to be disappearing with respect to mobile phones, but still exists for computer, internet, and e-mail access and use. Results suggest that mobile phone and texting applications may be feasibly applied for use in program-client interactions in substance abuse treatment. Careful consideration should be given to frequent phone number changes, access to technology, and motivation to engage with communication technology for treatment purposes.

The l2 minor capsid protein of human papillomavirus type 16 interacts with a network of nuclear import receptors.

The L2 minor capsid proteins enter the nucleus twice during viral infection: in the initial phase after virion disassembly and in the productive phase when, together with the L1 major capsid proteins, they assemble the replicated viral DNA into virions. In this study we investigated the interactions between the L2 protein of high-risk human papillomavirus type 16 (HPV16) and nuclear import receptors. We discovered that HPV16 L2 interacts directly with both Kapbeta(2) and Kapbeta(3). Moreover, binding of Ran-GTP to either Kapbeta(2) or Kapbeta(3) inhibits its interaction with L2, suggesting that the Kapbeta/L2 complex is import competent. In addition, we found that L2 forms a complex with the Kapalpha(2)beta(1) heterodimer via interaction with the Kapalpha(2) adapter. In agreement with the binding data, nuclear import of L2 in digitonin-permeabilized cells could be mediated by either Kapalpha(2)beta(1) heterodimers, Kapbeta(2), or Kapbeta(3). Mapping studies revealed that HPV16 L2 contains two nuclear localization signals (NLSs), in the N terminus (nNLS) and C terminus (cNLS), that could mediate its nuclear import. Together the data suggest that HPV16 L2 interacts via its NLSs with a network of karyopherins and can enter the nucleus via several import pathways mediated by Kapalpha(2)beta(1) heterodimers, Kapbeta(2), and Kapbeta(3).