Genome-scale clustered regularly interspaced short palindromic repeats screen identifies nucleotide metabolism as an actionable therapeutic vulnerability in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most common malignancy that develops in patients with ataxia-telangiectasia, a cancer-predisposing inherited syndrome characterized by inactivating germline ATM mutations. ATM is also frequently mutated in sporadic DLBCL. To investigate lymphomagenic mechanisms and lymphoma-specific dependencies underlying defective ATM, we applied ribonucleic acid (RNA)-seq and genome-scale loss-offunction clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 screens to systematically interrogate B-cell lymphomas arising in a novel murine model (Atm-/-nu-/-) with constitutional Atm loss, thymic aplasia but residual T-cell populations. Atm-/-nu-/-lymphomas, which phenotypically resemble either activated B-cell-like or germinal center Bcell-like DLBCL, harbor a complex karyotype, and are characterized by MYC pathway activation. In Atm-/-nu-/-lymphomas, we discovered nucleotide biosynthesis as a MYCdependent cellular vulnerability that can be targeted through the synergistic nucleotidedepleting actions of mycophenolate mofetil (MMF) and the WEE1 inhibitor, adavosertib (AZD1775). The latter is mediated through a synthetically lethal interaction between RRM2 suppression and MYC dysregulation that results in replication stress overload in Atm-/-nu-/-lymphoma cells. Validation in cell line models of human DLBCL confirmed the broad applicability of nucleotide depletion as a therapeutic strategy for MYC-driven DLBCL independent of ATM mutation status. Our findings extend current understanding of lymphomagenic mechanisms underpinning ATM loss and highlight nucleotide metabolism as a targetable therapeutic vulnerability in MYC-driven DLBCL.

Enhanced personal protective equipment and dental students' experience and quality of a restorative procedure in a simulated clinical setting.

OBJECTIVES: To evaluate the effects of enhanced personal protective equipment (Enhanced_PPE) on student-operator's experience and restorative procedure. METHODS: Student-operators (N = 29 Year 3 dental students) performed Class II composite restorations (SimpliShade, Kerr) in typodont upper molars (OneDental) equipped with N95 respirators, full-face shields, disposable headwear and gowns (Enhanced_PPE) or surgical masks, protective glasses/goggles and non-disposable gowns (Standard_PPE) 2 weeks later. Cavity dimensions were measured on cone beam computed tomography images. The quality of composite restorations was assessed using selected FDI criteria and Vickers hardness. A questionnaire assessed the operators' discomfort, anxiety, confidence, ability to perform, and procedure outcome. Data were analyzed using paired t-test and McNemar test (alpha = 0.05). RESULTS: Student-operators experienced greater discomfort and anxiety, reduced confidence and ability to perform, and rated the procedure as less satisfactory with Enhanced_PPE (p < 0.05). Differences in proximal box width were marginally significant (Enhanced_PPE 1.8 ± 0.4 mm, Standard_PPE 1.6 ± 0.3 mm) (p = 0.047). Other cavity dimensions were similar between groups (p > 0.05) as were restorations regarding surface luster, anatomical form, marginal adaptation, proximal contour, and contact (p > 0.05). There were no differences in the hardness of composite restorations (top p = 0.349, bottom p = 0.334). CONCLUSIONS: Enhanced_PPE led to student-operator discomfort, anxiety, and reduced confidence, but did not impact the quality of Class II preparation and composite restorations versus Standard_PPE.

Preheating and "snow-plow" composite application technique affect double bond conversion but not bond strength to dentine.

OBJECTIVE: To measure degree of conversion (DC) of a flowable composite, microtensile bond strength (MTBS) to dentine in the snow-plow technique with/without preheating and temperature in the preheated composite. MATERIALS AND METHODS: For DC, snow-plow specimens of Filtek Ultimate Flowable (Flow) and Filtek Bulk Fill (Bulk) (3M) were prepared by light-curing composites simultaneously in standardized molds while in control groups light-curing was separate. DC of Flow was measured using micro-Raman spectroscopy. For MTBS, specimens were prepared on dentine of human extracted molars by simultaneous (snow-plow) or separate (control) light-curing. MTBS was measured using a universal testing machine after 24 h and 6 months. Data were analyzed using ANOVA with Tukey's post hoc (α = 0.05). RESULTS: Increased curing time significantly increased DC in snow-plow from 13.4% ± 11.6% (10 s) to 31.8% ± 4.4% (40 s) albeit significantly lower than controls (p < 0.05). Preheated Bulk improved conversion in snow-plow (44.3% ± 1.7%) and control (50.5% ± 2.6%) (p < 0.05). No significant differences occurred in MTBS between groups (p > 0.05). MTBS values ranged between 66.7 ± 8.4 MPa (snow-plow group_21°C_baseline) and 54.1 ± 15.8 MPa (control_21°C_long-term). Temperature in the preheated Bulk dropped to ~38°C after 30 s. CONCLUSIONS: Snow-plow technique, irrespective of preheating, resulted in lower DC of Flow than separate light-curing of composite increments. Snow-plow, irrespective of preheating, resulted in similar initial and long-term MTBS to dentine. CLINICAL SIGNIFICANCE: Clinicians with preference for flowable composite liners in Class II restorations should be aware that the snow-plow technique of simultaneous light-curing of flowable and bulk-fill composite increments affects monomer-to-polymer conversion, albeit no effect on immediate, and long-term bonding to dentine was detected. Preheating sculptable bulk-fill composite improves conversion of the flowable liner in the snow-plow technique but has no detectable effect on bond strength to dentine.

DNA damage response defects in hematologic malignancies: mechanistic insights and therapeutic strategies.

ABSTRACT: The DNA damage response (DDR) encompasses the detection and repair of DNA lesions and is fundamental to the maintenance of genome integrity. Germ line DDR alterations underlie hereditary chromosome instability syndromes by promoting the acquisition of pathogenic structural variants in hematopoietic cells, resulting in increased predisposition to hematologic malignancies. Also frequent in hematologic malignancies are somatic mutations of DDR genes, typically arising from replication stress triggered by oncogene activation or deregulated tumor proliferation that provides a selective pressure for DDR loss. These defects impair homology-directed DNA repair or replication stress response, leading to an excessive reliance on error-prone DNA repair mechanisms that results in genomic instability and tumor progression. In hematologic malignancies, loss-of-function DDR alterations confer clonal growth advantage and adverse prognostic impact but may also provide therapeutic opportunities. Selective targeting of functional dependencies arising from these defects could achieve synthetic lethality, a therapeutic concept exemplified by inhibition of poly-(adenosine 5'-diphosphate ribose) polymerase or the ataxia telangiectasia and Rad 3 related-CHK1-WEE1 axis in malignancies harboring the BRCAness phenotype or genetic defects that increase replication stress. Furthermore, the role of DDR defects as a source of tumor immunogenicity, as well as their impact on the cross talk between DDR, inflammation, and tumor immunity are increasingly recognized, thus providing rationale for combining DDR modulation with immune modulation. The nature of the DDR-immune interface and the cellular vulnerabilities conferred by DDR defects may nonetheless be disease-specific and remain incompletely understood in many hematologic malignancies. Their comprehensive elucidation will be critical for optimizing therapeutic strategies to target DDR defects in these diseases.

SF3B1 hotspot mutations confer sensitivity to PARP inhibition by eliciting a defective replication stress response.

SF3B1 hotspot mutations are associated with a poor prognosis in several tumor types and lead to global disruption of canonical splicing. Through synthetic lethal drug screens, we identify that SF3B1 mutant (SF3B1MUT) cells are selectively sensitive to poly (ADP-ribose) polymerase inhibitors (PARPi), independent of hotspot mutation and tumor site. SF3B1MUT cells display a defective response to PARPi-induced replication stress that occurs via downregulation of the cyclin-dependent kinase 2 interacting protein (CINP), leading to increased replication fork origin firing and loss of phosphorylated CHK1 (pCHK1; S317) induction. This results in subsequent failure to resolve DNA replication intermediates and G2/M cell cycle arrest. These defects are rescued through CINP overexpression, or further targeted by a combination of ataxia-telangiectasia mutated and PARP inhibition. In vivo, PARPi produce profound antitumor effects in multiple SF3B1MUT cancer models and eliminate distant metastases. These data provide the rationale for testing the clinical efficacy of PARPi in a biomarker-driven, homologous recombination proficient, patient population.

Detecting heart failure using novel bio-signals and a knowledge enhanced neural network.

BACKGROUND: Clinical decisions about Heart Failure (HF) are frequently based on measurements of left ventricular ejection fraction (LVEF), relying mainly on echocardiography measurements for evaluating structural and functional abnormalities of heart disease. As echocardiography is not available in primary care, this means that HF cannot be detected on initial patient presentation. Instead, physicians in primary care must rely on a clinical diagnosis that can take weeks, even months of costly testing and clinical visits. As a result, the opportunity for early detection of HF is lost. METHODS AND RESULTS: The standard 12-Lead ECG provides only limited diagnostic evidence for many common heart problems. ECG findings typically show low sensitivity for structural heart abnormalities and low specificity for function abnormalities, e.g., systolic dysfunction. As a result, structural and functional heart abnormalities are typically diagnosed by echocardiography in secondary care, effectively creating a diagnostic gap between primary and secondary care. This diagnostic gap was successfully reduced by an AI solution, the Cardio-HART™ (CHART), which uses Knowledge-enhanced Neural Networks to process novel bio-signals. Cardio-HART reached higher performance in prediction of HF when compared to the best ECG-based criteria: sensitivity increased from 53.5% to 82.8%, specificity from 85.1% to 86.9%, positive predictive value from 57.1% to 70.0%, the F-score from 56.4% to 72.2%, and area under curve from 0.79 to 0.91. The sensitivity of the HF-indicated findings is doubled by the AI compared to the best rule-based ECG-findings with a similar specificity level: from 38.6% to 71%. CONCLUSION: Using an AI solution to process ECG and novel bio-signals, the CHART algorithms are able to predict structural, functional, and valve abnormalities, effectively reducing this diagnostic gap, thereby allowing for the early detection of most common heart diseases and HF in primary care.

Signatures of TOP1 transcription-associated mutagenesis in cancer and germline.

The mutational landscape is shaped by many processes. Genic regions are vulnerable to mutation but are preferentially protected by transcription-coupled repair1. In microorganisms, transcription has been demonstrated to be mutagenic2,3; however, the impact of transcription-associated mutagenesis remains to be established in higher eukaryotes4. Here we show that ID4-a cancer insertion-deletion (indel) mutation signature of unknown aetiology5 characterized by short (2 to 5 base pair) deletions -is due to a transcription-associated mutagenesis process. We demonstrate that defective ribonucleotide excision repair in mammals is associated with the ID4 signature, with mutations occurring at a TNT sequence motif, implicating topoisomerase 1 (TOP1) activity at sites of genome-embedded ribonucleotides as a mechanistic basis. Such TOP1-mediated deletions occur somatically in cancer, and the ID-TOP1 signature is also found in physiological settings, contributing to genic de novo indel mutations in the germline. Thus, although topoisomerases protect against genome instability by relieving topological stress6, their activity may also be an important source of mutations in the human genome.

Adjacent Joint Restriction Differentially Influences Intra- and Inter-rater Reliability and Agreement of Goniometric Measurements.

BACKGROUND: Reliability and agreement of goniometric measurements can be altered by variations in measurement technique such as restricting adjacent joints to influence bi-articular muscles. It is unknown if the influence of adjacent joint restriction is consistent across different range of motion (ROM) tests, as this has yet to be assessed within a single study. Additionally, between-study comparisons are challenged by differences between methodology, participants and raters, obscuring the development of a conceptual understanding of the extent to which adjacent joint restriction can influence goniometric ROM measurements. PURPOSE: To quantify intra- and inter-rater reliability and levels of agreement of goniometric measurements across five ROM tests, with and without adjacent joint restriction. STUDY DESIGN: Descriptive reliability study. METHODS: Three trained and experienced raters made two measurements of bilateral ankle dorsiflexion, first metatarsophalangeal dorsiflexion, hip extension, hip flexion, and shoulder flexion, with and without adjacent joint restriction. Intraclass correlation coefficient (ICC), standard error of measurement (SEM), along with participant, measurement/rater and random error variance were estimated. RESULTS: Eleven females (age 21.4 ±2.3 years) and 19 males (age 22.1 ±2.8 years) participated. Adjacent joint restriction did not influence the reliability and agreement in a consistent way across the five ROM tests. Changes in the inter-rater reliability and agreement were more pronounced compared to the intra-rater reliability and agreement. Assessing variance components (participant, measurement/rater and random error variance) that are used to calculate the ICC and SEM, improved interpretation of ICC and SEM scores. CONCLUSION: The effects of adjacent joint restriction on reliability and agreement of goniometric measurements depend on the ROM test and should be considered when comparing measurements between multiple raters. Reporting variance components that are used to calculate the ICC and SEM can improve interpretation and may improve between-study comparisons, towards developing a conceptual framework to guide goniometric measurement technique. LEVEL OF EVIDENCE: 3b.

Novel tech throws knock-out punch to ECG improving GP referral decisions to cardiology.

PURPOSE: In a comparator study, designed with assistance from the Food and Drug Administration, a State-of-the-Art (SOTA) ECG device augmented with automated analysis, the comparator, was compared with a breakthrough technology, Cardio-HART (CHART). METHODS: The referral decision defined by physician reading biosignal-based ECG or CHART report were compared for 550 patients, where its performance is calculated against the ground truth referral decision. The ground truth was established by cardiologist consensus based on all the available measurements and findings including echocardiography (ECHO). RESULTS: The results confirmed that CHART analysis was far more effective than ECG only analysis: CHART reduced false negative rates 15.8% and false positive (FP) rates by 5%, when compared with SOTA ECG devices. General physicians (GP's) using CHART saw their positive diagnosis rate significantly increased, from ~10% to ~26% (260% increase), and the uncertainty rate significantly decreased, from ~31% to ~1.9% (94% decrease). For cardiology, the study showed that in 98% of the cases, the CHART report was found to be a good indicator as to what kind of heart problems can be expected (the 'start-point') in the ECHO examination. CONCLUSIONS: The study revealed that GP use of CHART resulted in more accurate referrals for cardiology, resulting in fewer true negative or FP-healthy or mildly abnormal patients not in need of ECHO confirmation. The indirect benefit is the reduction in wait-times and in unnecessary and costly testing in secondary care. Moreover, when used as a start-point, CHART can shorten the echocardiograph examination time.

Correction: Kwok et al. Targeting the p53 Pathway in CLL: State of the Art and Future Perspectives. Cancers 2021, 13, 4681.

The authors wish to make the following corrections to their paper [1] [...].

Functional Movement Screen Task Scores and Joint Range-of-motion: A Construct Validity Study.

Little is known about the construct validity of the Functional Movement Screen (FMS). We aimed to assess associations between FMS task scores and measures of maximum joint range-of-motion (ROM) among university varsity student-athletes from 4 sports (volleyball, basketball, ice hockey, and soccer). Athletes performed FMS tasks and had their maximum ankle, hip and shoulder ROM measured. Multivariable linear regression was used to estimate associations between FMS task scores and ROM measurements. 101 university student-athletes were recruited (52 W/49 M; mean age 20.4±1.9 years). In general, athletes with higher FMS task scores had greater ROM compared to those with lower task scores. For example, athletes who scored 2 on the FMS squat task had 4° (95% CI, 1° to 7°) more uni-articular ankle dorsiflexion ROM compared with those who scored 1, while those who scored 3 on the FMS squat task had 10° (4° to 17°) more uni-articular ankle dorsiflexion ROM compared with those who scored 1. Large variation in ROM measurements was observed. In sum, substantial overlap in joint ROM between groups of athletes with different FMS task scores weakens the construct validity of the FMS as an indicator of specific joint ROM.

The effect of COVID-19 pandemic on final year dental students' self-confidence level in performing clinical procedures.

BACKGROUND: The outbreak of COVID-19 pandemic in 2020 has dramatically changed teaching approach in dental schools due to the switch to distance learning and the lack of practice training in direct contact with patients with possible impact on clinical skills of students. The aim of the study was to assess the level of the 2020 final year dental students' self-confidence in performing different dental procedures through specially designed questionnaire and compare it to self-confidence of the 2019 final year students. MATERIALS AND METHODS: An anonymous questionnaire consisting of 40 questions regarding self-confidence level in performing 40 different dental procedures and based on five points Likert-like scale was distributed during November 2020 to final year dental students whose studies were disrupted by the COVID-19 pandemic lockdown in one semester. The study group comprised their answers. The comparison was done with the control group that consisted of students' answers on the same questionnaire from previous 2019 year conducted as a part of regular internal educational evaluation. RESULTS: Response rate was 74.2% in study group and 89.3% in control group. Mean level of self-confidence reported by 115 students in study group was significantly lower than that reported by 100 students in control group (3.28±1.08 vs. 3.58±0.88, respectively) and the distribution of self-confidence scores were different in observed groups. Graduates from study group felt less confident than those from control in 8 clinical skills. CONCLUSION: Abrupt changes in teaching modalities caused by COVID-19 pandemic had significant impact on final year dental students' self-confidence indicating additional educational needs in postgraduate period.

Targeting the p53 Pathway in CLL: State of the Art and Future Perspectives.

The p53 pathway is a desirable therapeutic target, owing to its critical role in the maintenance of genome integrity. This is exemplified in chronic lymphocytic leukemia (CLL), one of the most common adult hematologic malignancies, in which functional loss of p53 arising from genomic aberrations are frequently associated with clonal evolution, disease progression, and therapeutic resistance, even in the contemporary era of CLL targeted therapy and immunotherapy. Targeting the 'undruggable' p53 pathway therefore arguably represents the holy grail of cancer research. In recent years, several strategies have been proposed to exploit p53 pathway defects for cancer treatment. Such strategies include upregulating wild-type p53, restoring tumor suppressive function in mutant p53, inducing synthetic lethality by targeting collateral genome maintenance pathways, and harnessing the immunogenicity of p53 pathway aberrations. In this review, we will examine the biological and clinical implications of p53 pathway defects, as well as our progress towards development of therapeutic approaches targeting the p53 pathway, specifically within the context of CLL. We will appraise the opportunities and pitfalls associated with these therapeutic strategies, and evaluate their place amongst the array of new biological therapies for CLL.

Effects of whitening gels on color and surface properties of a microhybrid and nanohybrid composite.

The aim of this study was to compare color changes, surface roughness and gloss of a microhybrid and nanohybrid composite whitened in a simulated in-office or at-home procedure using 40% hydrogen peroxide or 16% carbamide peroxide, respectively. CIELab coordinates were measured before, during and after treatment using VITA EasyShade V (VITA) and ∆E00 calculated. Surface roughness was measured using a surface roughness tester (SJ210; Mitutoyo). Gloss was measured using a gloss checker (IG-331; Horiba). At-home whitening resulted in ∆E00 of 1.23±0.49 (microhybrid) and 1.01±0.76 (nanohybrid). In-office exposure resulted in ∆E00 of 0.69±0.38 (microhybrid) and 0.72±0.50 (nanohybrid). There were no significant differences in ∆E00, surface roughness and gloss between whitening protocols (p>0.05). Color changes of a microhybrid and nanohybrid composite following simulated in-office or at-home whitening reached CIEDE2000 50:50% perceptibility but not acceptability threshold. Surface roughness and gloss of the microhybrid and nanohybrid composite were not affected by whitening.