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Drugs are designed to bind their target proteins in physiologically relevant tissues and organs to modulate biological functions and elicit desirable clinical outcomes. Information about target engagement at cellular and subcellular resolution is therefore critical for guiding compound optimization in drug discovery, and for probing resistance mechanisms to targeted therapies in clinical samples. We describe a target engagement-mediated amplification (TEMA) technology, where oligonucleotide-conjugated drugs are used to visualize and measure target engagement in situ, amplified via rolling-circle replication of circularized oligonucleotide probes. We illustrate the TEMA technique using dasatinib and gefitinib, two kinase inhibitors with distinct selectivity profiles. In vitro binding by the dasatinib probe to arrays of displayed proteins accurately reproduced known selectivity profiles, while their differential binding to fixed adherent cells agreed with expectations from expression profiles of the cells. We also introduce a proximity ligation variant of TEMA to selectively investigate binding to specific target proteins of interest. This form of the assay serves to improve resolution of binding to on- and off-target proteins. In conclusion, TEMA has the potential to aid in drug development and clinical routine by conferring valuable insights in drug-target interactions at spatial resolution in protein arrays, cells and in tissues., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Background: Rectal washout (RW) is routinely performed during anterior resection (AR) for rectal cancer to reduce local recurrence (LR), although is sometimes not performed during minimally invasive surgery (MIS) procedures due to technical challenges and time consumption. The aim was to investigate the impact of RW on the oncological outcome after AR for rectal cancer in a registry cohort., Methods: Data on patients registered in the Swedish Colorectal Cancer Registry who had undergone elective radical (R0) AR for TNM stage I-III rectal cancer between 2007 and 2017 with a 3-year follow-up were analysed. Multivariable analyses were performed and the primary endpoint was LR at 3 and 5 years after AR. The occurrence of distant metastasis (DM) and overall recurrence (OAR), overall survival, and relative survival were also analysed as a secondary aim. A subgroup analysis was performed for the same outcomes in patients treated with MIS., Results: Out of 6186 patients (1923 with TNM stage I, 1907 with TNM stage II, and 2356 with TNM stage III), RW was performed in 5706 (92.2 per cent). The median age of the cohort was 67 years. RW did not impact the 3-year risk of LR. LR within 5 years occurred in 104 of 4583 patients (2.3 per cent) in the RW group compared with 16 of 408 patients (3.9 per cent) in the no RW group (P = 0.037). In multivariable analysis of the LR risk, the HR was 0.53 (95 per cent c.i. 0.31 to 0.90), favouring RW. There were no differences in rates of DM and OAR, overall survival, and relative survival. A subgroup analysis of the 1410 patients undergoing MIS did not demonstrate any differences between the groups, given, however, the low rate of LR., Conclusions: RW in AR for rectal cancer does not impact the 3-year oncological outcome; however, after the 5-year follow-up a reduction in LR risk was observed after RW., (© The Author(s) 2022. Published by Oxford University Press on behalf of BJS Society Ltd.)

Objective: The aim of this study was to explore the associations between inadequate glycemic control of diabetes and psychological, anthropometric, and lifestyle variables in a population-based cohort of type 1 diabetes patients., Design: Cross-sectional study., Methods: In this study, 292 patients with type 1 diabetes, aged 1859 years, participated. psychological data were assessed by self-report instruments: Hospital Anxiety and Depression Scale and Toronto Alexithymia Scale-20. Anthropometrics, blood analyses, data from medical records, and data from the Swedish National Diabetes Registry were collected., Results: Self-reported depression (adjusted odds ratio (AOR) 4.8), obesity (AOR 4.3), and smoking (AOR 3.0) were independently associated with inadequate glycemic control of diabetes (HbA1c>8.6%). Gender-stratified analyses showed that self-reported depression (AOR 19.8) and obesity (AOR 7.0) in women and smoking in men (AOR 4.2) were associated with HbA1c>8.6%. Alexithymia, antidepressant medication, and physical inactivity were associated with HbA1c>8.6% only in bivariate analyses. Alexithymia, self-rated anxiety, physical inactivity, and absence of abdominal obesity were associated with self-reported depression., Conclusions: Depression was the only psychological factor independently associated with HbA1c>8.6%. The association was of comparable importance as obesity and smoking, well-known risk factors for inadequate glycemic control and diabetes complications. The association between depression and HbA1c>8.6% was particularly strong for women. Alexithymia, which is a relatively stable personality trait, was associated with depression. In the future care of patients with diabetes, psychological aspects should be considered alongside anthropometrics and lifestyle factors in order to achieve the goals for HbA1c.

A new thin transmission target technique for fast dose delivery using narrow scanned photon beams has been developed. High-energy, 50-100 MeV, electron beams of low emittance incident on thin low-Z targets produce narrow and intense high-energy bremsstrahlung beams. However, electrons transmitted through the target are bent from the therapeutic beam by a purging magnet and have to be effectively absorbed in a dedicated electron collector. The electron-photon transport through a treatment head has been studied using the Monte Carlo simulation toolkit Geant4. The Geant4 electromagnetic physics processes have been compared with experimental data of radial dose profiles. The differences between calculated and measured radial dose distributions are approximately 2-10%. Preliminary investigations of the collector design have been carried out in order to minimise secondary electron and photon contamination of the therapeutic beam. The toolkit presented here is promising for further development of narrow photon beam therapy.

Bacillus anthracis plasmids pX01 and pX02, harboured by the Sterne and Pasteur strains, respectively, have been sequenced by random 'shotgun' cloning and high throughout sequence analysis. These sequences have been assembled (Sequencher) to generate a circulate pX01 plasmid containing 181 656 bp and a single linear (gapped) pX02 contig containing at least 93.479 bp. Initial annotation suggests that the two plasmids combined contain at least 200 potential open reading frames (ORFs) with < 40% having significant similarity to sequences registered in open databases. Collectively, only 118 566 bp of the pX01 DNA (65%) represent predicted coding regions. This value is similar to published gene densities for other plasmids and is indicative of the larger intergenic spaces in plasmids vs those found in the chromosomes of the parental microbes (85-93% gene density). A 70 kbp region including the toxin genes (cya, lef and pag) is distinct from the remainder of the pX01 sequence: (1) it has a lower gene density (58 vs 70%) than the remaining 111 kbp; (2) it contains all but one of the co-regulated transcriptional fusions identified by transposon mutagenesis (Hoffmaster & Koehler 1997) and (3) it contains a significantly higher proportion of positive BLAST scores (62 vs 20%) for putative ORFs. These data suggest different origins for the two regions of pX01.

In an open trial, the efficacy of intravenously administered ceftazidime was evaluated in 106 adult patients with urinary or respiratory tract infections, soft tissue infections, osteitis and septicaemia. The most commonly isolated pathogens were Escherichia coli, Pseudomonas spp. and Staphylococcus aureus. Of 85 organisms isolated before treatment, 79% were cleared and 15% were cleared but later relapsed often because of the underlying conditions. Clinical cure was achieved in 70%, improvement occurred in 25% of the patients and 6% failed to respond. Marked increases in serum creatinine occurred in three patients with pre-existing renal impairment treated with ceftazidime in unmodified dosage. Exanthema, diarrhoea or local thrombophlebitis was noted in 13 patients.