Capture-based targeted sequencing using a T-cell control in myeloid malignancies and idiopathic cytopenias.

We report on a study of next-generation sequencing in 257 patients undergoing investigations for cytopenias. We sequenced bone marrow aspirates using a target enrichment panel comprising 82 genes and used T cells from paired blood as a control. One hundred and sixty patients had idiopathic cytopenias, 81 had myeloid malignancies and 16 had lymphoid malignancies or other diagnoses. Forty-seven of the 160 patients with idiopathic cytopenias had evidence of somatic pathogenic variants consistent with clonal cytopenias. Only 39 genes of the 82 tested were mutated in the 241 patients with either idiopathic cytopenias or myeloid neoplasms. We confirm that T cells can be used as a control to distinguish between germline and somatic variants. The use of paired analysis with a T-cell control significantly reduced the time molecular scientists spent reporting compared to unpaired analysis. We identified somatic variants of uncertain significance (VUS) in a higher proportion (24%) of patients with myeloid malignancies or clonal cytopenias compared to less than 2% of patients with non-clonal cytopenias. This suggests that somatic VUS are indicators of a clonal process. Lastly, we show that blood depleted of lymphocytes can be used in place of bone marrow as a source of material for sequencing.

The role of medical-grade maggots in facilitating healing of diabetic foot ulcers in Kano, northern Nigeria: a case series.

OBJECTIVE: Approximately 13% of people living with diabetes develop one or more ulcers during the course of the disease, and diabetic foot ulcer (DFU) is responsible for >60% of lower limb amputations worldwide. This case series aimed to demonstrate the effectiveness of medical-grade maggots on DFUs in promoting wound healing and reducing related hospital stays in northern Nigeria. METHOD: Maggot debridement therapy (MDT) was applied to the DFUs of patients who consented to this treatment between January-August 2021 at the Orthopaedic Unit of the Aminu Kano Teaching Hospital (AKTH), Kano, Nigeria. Sterile first instar larvae of Lucilia sericata were obtained indigenously and applied using the confinement (free-range) method under aseptic procedure. RESULTS: A total of 15 patients with DFUs of Wagner classification grades III (33.3%) and IV (66.7%), were seen and documented. The patients included 10 (66.7%) females and five (33.3%) males, giving a female:male ratio of 2:1. The mean age (±standard deviation) of the respondents was 51.6±10.8 years. The surface area of the wounds ranged from 24-140cm2, with a median value of 75cm2. Among the patients, 60% had two cycles of MDT, with a median duration of five days. Most of the wounds (53.3%) were debrided within five days; 13.3% (two wounds) took seven days to be fully debrided. A paired sample t-test showed a statistically significant association between wound surface area and therapy duration (t=8.0; p=0.000) and between wound surface area and cycles of therapy (t=8.3; p=0.000). Before maggot application, 14 (93.3%) DFUs were completely (100%) covered with slough and only one (6.7%) was 95% covered with slough. After maggot application, 10 (66.7%) wounds were completely debrided while five (33.3%) wounds required bedside surgical debridement to achieve complete debridement. A paired sample t-test showed statistically significant difference between the pre- and post-MDT slough covering the wounds (t=45.1; p=0.000). CONCLUSION: In this case series, MDT was successfully used in the debridement of DFUs, which facilitated the healing process with an encouraging clinical outcome.

Results for the London investigation into dielectric scanning of lesions study of the MARIA® M6 breast imaging system.

OBJECTIVES: Determine the proportion of breast cancers for which MARIA® findings correspond to the cancer, with stratification by breast density and histological type. Investigate performance in symptomatic lesions. Gain patient feedback on experience with MARIA®. METHODS: MARIA® uses a radio frequency antennae array to measure signal attenuation and back scatter to build up a 3D map of tissue dielectric values. The study was a prospective, single-centre, interventional, post-approval device study. RECRUITMENT: Patients were eligible if they were attending symptomatic breast clinic or had confirmed or suspected breast cancer from any referral source. Recruitment between May 2018 and March 2020. READING: Regions of higher signal compared to background or distinct by shape were considered candidates for lesion correspondence. Up to 4 candidate regions per breast were annotated in likelihood order for representing a true lesion. RESULTS: 389 patients were recruited, and 6 excluded. 114 patients recruited with breast cancers (2 bilateral, 5 multicentric). 57 (47%) malignant lesions showed correspondence between the MARIA® signal and the cancer. Higher correspondence was seen in invasive (50%) than in situ (29%) disease. There was no reduction in correspondence at higher breast density. Reduced signal correspondence in the central scan volume and for small lesions. MARIA® scanning was well tolerated. CONCLUSIONS: We believe that MARIA® signal corresponds to a malignant lesion in 47% of breast cancers examined. ADVANCES IN KNOWLEDGE: Our study, the largest to date for this type of technology, demonstrates successes and limitations of this MARIA® M6 version.

Predicting radiotherapy response, Toxicities and quality-of-life related functional outcomes in soft tissue sarcoma of the extremities (PredicT) using dose-volume constraints development: a study protocol.

INTRODUCTION: Radiotherapy improves local tumour control in patients with soft tissue sarcoma of the extremities (STSE) but it also increases the probability of long-term toxicities such as tissue fibrosis, joint stiffness and lymphoedema. The use of radiation dose and volume thresholds, called dose constraints, may potentially reduce the development of toxicities in STSE. The aim of this study is to determine predictors of radiotherapy-related side effects for STSE. METHODS AND ANALYSIS: Predicting radiotherapy response, Toxicities and quality-of-life related functional outcomes in soft tissue sarcoma of the extremities (PredicT) is a multicentre observational study comprising two cohorts (PredicT A and B). PredicT A, a retrospective analysis of the UK VorteX (NCT00423618) and IMRiS clinical trials (NCT02520128), is aimed at deriving a statistical model for development of dose-volume constraints. This model will use receiving operator characteristics and multivariate analysis to predict radiotherapy side effects and patient-reported outcomes. PredicT B, a prospective cohort study of 150 patients with STSE, is aimed at testing the validity of those dose-volume constraints. PredicT B is open and planned to complete recruitment by September 2024. ETHICS AND DISSEMINATION: PredicT B has received ethical approval from North West - Liverpool Central Research Ethics Committee (20/NW/0267). Participants gave informed consent to participate in the study before taking part. We will disseminate our findings via publications, presentations, national and international conference meetings and engage with local charities. TRIAL REGISTRATION NUMBER: NCT05978024.