Mechanisms of Vertebrate DNA Interstrand Cross-Link Repair.

DNA interstrand cross-links (ICLs) covalently connect the two strands of the double helix and are extremely cytotoxic. Defective ICL repair causes the bone marrow failure and cancer predisposition syndrome, Fanconi anemia, and upregulation of repair causes chemotherapy resistance in cancer. The central event in ICL repair involves resolving the cross-link (unhooking). In this review, we discuss the chemical diversity of ICLs generated by exogenous and endogenous agents. We then describe how proliferating and nonproliferating vertebrate cells unhook ICLs. We emphasize fundamentally new unhooking strategies, dramatic progress in the structural analysis of the Fanconi anemia pathway, and insights into how cells govern the choice between different ICL repair pathways. Throughout, we highlight the many gaps that remain in our knowledge of these fascinating DNA repair pathways.

PrimPol-mediated repriming facilitates replication traverse of DNA interstrand crosslinks.

DNA interstrand crosslinks (ICLs) induced by endogenous aldehydes or chemotherapeutic agents interfere with essential processes such as replication and transcription. ICL recognition and repair by the Fanconi Anemia pathway require the formation of an X-shaped DNA structure that may arise from convergence of two replication forks at the crosslink or traversing of the lesion by a single replication fork. Here, we report that ICL traverse strictly requires DNA repriming events downstream of the lesion, which are carried out by PrimPol, the second primase-polymerase identified in mammalian cells after Polα/Primase. The recruitment of PrimPol to the vicinity of ICLs depends on its interaction with RPA, but not on FANCM translocase or the BLM/TOP3A/RMI1-2 (BTR) complex that also participate in ICL traverse. Genetic ablation of PRIMPOL makes cells more dependent on the fork convergence mechanism to initiate ICL repair, and PRIMPOL KO cells and mice display hypersensitivity to ICL-inducing drugs. These results open the possibility of targeting PrimPol activity to enhance the efficacy of chemotherapy based on DNA crosslinking agents.

Biological effect of food additive titanium dioxide nanoparticles on intestine: an in vitro study.

Titanium dioxide nanoparticles (TiO2 NPs) are widely found in food-related consumer products. Understanding the effect of TiO2 NPs on the intestinal barrier and absorption is essential and vital for the safety assessment of orally administrated TiO2 NPs. In this study, the cytotoxicity and translocation of two native TiO2 NPs, and these two TiO2 NPs pretreated with the digestion simulation fluid or bovine serum albumin were investigated in undifferentiated Caco-2 cells, differentiated Caco-2 cells and Caco-2 monolayer. TiO2 NPs with a concentration less than 200 µg ml(-1) did not induce any toxicity in differentiated cells and Caco-2 monolayer after 24 h exposure. However, TiO2 NPs pretreated with digestion simulation fluids at 200 µg ml(-1) inhibited the growth of undifferentiated Caco-2 cells. Undifferentiated Caco-2 cells swallowed native TiO2 NPs easily, but not pretreated NPs, implying the protein coating on NPs impeded the cellular uptake. Compared with undifferentiated cells, differentiated ones possessed much lower uptake ability of these TiO2 NPs. Similarly, the traverse of TiO2 NPs through the Caco-2 monolayer was also negligible. Therefore, we infer the possibility of TiO2 NPs traversing through the intestine of animal or human after oral intake is quite low. This study provides valuable information for the risk assessment of TiO2 NPs in food.

AZ91 Magnesium Alloy CMT Cladding Layer Processed Using Friction Stir Processing: Effect of Traverse Speed on Microstructure and Mechanical Properties.

Friction stir processing (FSP) is a solid-state treating method to enhance the mechanical properties of materials by altering their microstructure. In this study, FSP was applied to the AZ91 magnesium alloy cladding layer prepared using cold metal transition (CMT) technology, and the purpose was to investigate the effect of the traverse speed of the H13 steel stirring head under a constant rotation speed on the microstructure and mechanical properties of the cladding layer. The results demonstrated that FSP could effectively decrease the grain size of the cladding layer and lead to the dispersion and dissolution of the coarse ß-Mg17Al12 second phase into the α-Mg matrix. The mechanical characteristics of the processed cladding layer were significantly enhanced compared to the unprocessed cladding layer due to the grain refinement and second-phase strengthening induced by FSP. When the stirring head rotation speed was set at 300 r/min, the average microhardness and tensile properties of the specimens showed a tendency of initially increasing and then dropping as the traverse speed increased. The cladding layer, obtained at a traverse speed of 60 mm/min, displayed optimal mechanical properties with an average microhardness, tensile strength, and elongation of 85.6 HV0.1, 278.5 MPa, and 13.4%, respectively.

FANCM branchpoint translocase: Master of traverse, reverse and adverse DNA repair.

FANCM is a multifunctional DNA repair enzyme that acts as a sensor and coordinator of replication stress responses, especially interstrand crosslink (ICL) repair mediated by the Fanconi anaemia (FA) pathway. Its specialised ability to bind and remodel branched DNA structures enables diverse genome maintenance activities. Through ATP-powered "branchpoint translocation", FANCM can promote fork reversal, facilitate replication traverse of ICLs, resolve deleterious R-loop structures, and restrain recombination. These remodelling functions also support a role as sensor of perturbed replication, eliciting checkpoint signalling and recruitment of downstream repair factors like the Fanconi anaemia FANCI:FANCD2 complex. Accordingly, FANCM deficiency causes chromosome fragility and cancer susceptibility. Other recent advances link FANCM to roles in gene editing efficiency and meiotic recombination, along with emerging synthetic lethal relationships, and targeting opportunities in ALT-positive cancers. Here we review key properties of FANCM's biochemical activities, with a particular focus on branchpoint translocation as a distinguishing characteristic.

Testosterone therapy and the risk of cardiovascular disease in older, hypogonadal men.

The debate over the cardiovascular (CV) implications of testosterone therapy (TT) have resulted in diverging safety recommendations and clinical guidelines worldwide. This narrative review synthesizes and critically evaluates long-term studies examining the effects of TT within the context of aging, obesity, and endogenous sex hormones on CV disease (CVD) risk to support informed clinical decision-making. Observational studies have variably linked low endogenous testosterone with increased CVD risk, while randomized controlled trials (RCTs) demonstrate that TT yields cardiometabolic benefits without increasing short-term CV risk. The TRAVERSE trial, as the first RCT powered to assess CVD events, did not show increased major adverse cardiac events (MACE) incidence; however, its limitations - specifically the maintenance of testosterone at low-normal levels, a high participant discontinuation rate, and short follow-up - warrant a careful interpretation of its results. Furthermore, findings from the TTrials cardiovascular sub-study, which showed an increase in non-calcified plaque, indicate the need for ongoing research into the long-term CV impact of TT. The decision to initiate TT should consider the current evidence gaps, particularly for older men with known CVD. The CV effects of maintaining physiological testosterone levels through exogenous means remain to be fully explored. Until more definitive evidence is available, clinical practice should prioritize individualized care and informed discussions on the potential CV implications of TT.

Experimental and Numerical Study of Al2219 Powders Deposition on Al2219-T6 Substrate by Cold Spray: Effects of Spray Angle, Traverse Speed, and Standoff Distance.

Cold spray (CS) is an emerging technology for repairing and 3D additive manufacturing of a variety of metallic components using deformable metal powders. In CS deposition, gas type, gas pressure, gas temperature, and powder feed rate are the four key process parameters that have been intensively studied. Spray angle, spray gun traverse speed, and standoff distance (SoD) are the other three process parameters that have been less investigated but are also important, especially when depositing on uneven substrates or building up 3D freeform structures. Herein, the effects of spray angle, traverse speed, and SoD during CS deposition have been investigated holistically on a single material system (i.e., Al2219 powders on Al2219-T6 substrate). The coatings' mass gain, thickness, porosity, and residual stress have been characterized, and the results show that spray angle and traverse speed exercise much more effects than SoD in determining coatings' buildup. Finite element method (FEM) modeling and computational fluid dynamic (CFD) simulation have been carried out to understand the effects of these three parameters for implementing CS as repairing and additive manufacturing using aluminum-based alloy powders.

Dupilumab Reduces Oral Corticosteroid Use in Patients With Corticosteroid-Dependent Severe Asthma: An Analysis of the Phase 3, Open-Label Extension TRAVERSE Trial.

BACKGROUND: Many patients with severe asthma require chronic corticosteroid treatment to maintain asthma control. RESEARCH QUESTION: Are the reduction in oral corticosteroid (OCS) use and the clinical efficacy observed with dupilumab treatment maintained long-term in patients with severe OCS-dependent asthma? STUDY DESIGN AND METHODS: The LIBERTY ASTHMA TRAVERSE study (ClinicalTrials.gov identifier: NCT02134028) was a multinational, multicenter, single-arm, open-label extension study in patients ≥ 12 years of age with asthma who participated in previous dupilumab studies. Treatment consisted of dupilumab 300 mg every 2 weeks for up to 96 weeks. In this analysis, we present the data from patients who initially enrolled in the LIBERTY ASTHMA VENTURE study (ClinicalTrials.gov identifier: NCT02528214), a 24-week placebo-controlled study of dupilumab in patients with OCS-dependent severe asthma, and continued in the TRAVERSE study. The subgroups analyzed were: those who received dupilumab in both (dupilumab/dupilumab group) and those who received placebo in the VENTURE study and dupilumab in the TRAVERSE study (placebo/dupilumab group). Outcomes included OCS use, exacerbation rates, and measures of lung function and asthma control. RESULTS: Ninety patients treated with dupilumab/dupilumab and 97 patients treated with placebo/dupilumab in the VENTURE study were enrolled and treated in the TRAVERSE study, with a mean OCS dosage of 11.0 mg/d (dupilumab) and 11.6 mg/d (placebo) at VENTURE study baseline. At TRAVERSE week 0, the mean daily OCS dosage was 3.1 mg/d and 6.4 mg/d (percentage decrease from the VENTURE study baseline, 68.8% and 41.3%) for the dupilumab/dupilumab group and placebo/dupilumab group, respectively, and decreased to 2.2 mg/d and 4.9 mg/d (78.3% and 53.4%) at week 48 and to 1.2 mg/d and 3.0 mg/d (89.3% and 74.4%) at week 96, respectively. Exacerbation rates were low during the TRAVERSE study. Further improvements from the VENTURE to TRAVERSE studies also were seen in FEV1 and 5-item Asthma Control Questionnaire scores. Safety findings were consistent with the known dupilumab safety profile. INTERPRETATION: In the open-label TRAVERSE study, dupilumab demonstrated the ability to sustain the OCS dosage reduction from the parent OCS-sparing study, while maintaining a low exacerbation rate and improved lung function. TRIAL REGISTRY: ClinicalTrials.gov; Nos.: NCT02134028 (TRAVERSE) and NCT02528214 (VENTURE); URL: www. CLINICALTRIALS: gov.

Analysis of Several Physical Phenomena Measured on the Metallic Materials Cut by Abrasive Water Jets (AWJ).

Cutting using an abrasive water jet is a complex process involving several physical phenomena. This research studies some of them, mostly the influence of selected variables on the measured forces and vibrations. The traverse speed represents one of the key parameters when cutting using the AWJ. In the presented research, a set of experiments was performed on twelve different metal samples, while the force sensor measured the exerted forces and accelerometers measured the vibrations. Ten different types of steel samples of the same dimensions were cut applying five different traverse speeds. The data obtained during these measurements show that an increase in the traverse speed leads to an increase in the measured forces and vibrations. An analogous experiment performed on bronze and duralumin samples of the same dimensions, having applied higher speeds to compensate for the difference in the material structure and properties, completes the presented data. The most important results of the research are that exerted forces in the z-axis are higher than those in the x-axis, whereas measured vibrations are higher in the x-axis. According to our research, the elemental structure, especially the carbide formation, affects the measured forces and vibrations substantially.

Visualizing replication fork encounters with DNA interstrand crosslinks.

Replication forks encounter numerous challenges as they move through eu- and hetero-chromatin during S phase in mammalian cells. These include a variety of impediments to the unwinding of DNA by the replicative helicase such as alternate DNA structures, transcription complexes and R-loops, DNA-protein complexes, and DNA chemical adducts. Much of our knowledge of these events is based on analysis of markers of the replication stress and DNA Damage Response that follow stalling of replisomes. To examine consequences for the replisomes more directly, we developed an approach for imaging collisions of replication forks with the potent block presented by an interstrand crosslink (ICL). The strategy is based on the visualization on DNA fibers of the encounter of replication tracts and an antigen tagged ICL. Our studies revealed an unexpected restart of DNA synthesis past an intact ICL. In addition, and also unexpected, we found two distinct versions of the replisome, one biased toward euchromatin and the other more prominent in heterochromatin. Here, we present details of our experimental procedures that led to these observations.

Impact of Preparation of Titanium Alloys on Their Abrasive Water Jet Machining.

Several titanium alloys, i.e., grade 2 Ti, Ti6Al4V and NiTi alloy, prepared by selected deformation procedures were subjected to abrasive water jet (AWJ) cutting and subsequently analysed. The study describes samples' preparations and respective material structures. The impact of deformation processing of the selected alloys on the declination angle during cutting, and the results of measurements of surface wall quality performed for the selected samples at the Department of Physics of Faculty of Electrical Engineering and Computer Science at VSB-Technical University of Ostrava, are presented and discussed, as are also the influences of structural features of the processed titanium alloys on surface qualities of the investigated samples. The results showed that the highest resistance to AWJ machining exhibited the Ti6Al4V alloy prepared by forward extrusion. Its declination angle (recalculated to the thickness 10 mm to compare all the studied samples) was 12.33° at the traverse speed of 100 mm/min, pumping pressure of 380 MPa, and abrasive mass flow rate of 250 g/min.

Investigation of Significant Parameters during Abrasive Waterjet Turning.

This paper presents an investigation of abrasive waterjet turning (AWJT). The purpose of the article was to investigate significant parameters of the turning process and to evaluate their impact on the turning product. The influence of the traverse speed, the rotational speed, and the relative position of the jet to the specimen (lateral jet shift) were investigated. Based on the previous research done in this field, the multi-pass tangential turning method was selected. Rotational speed does not seem to have a significant impact on the AWJ turning process. However, the relative position of the jet is a key parameter for improving the efficiency of the process. Increasing the lateral jet shift causes the volume of the material removed to increase until the optimal impact angle is reached. These findings need to be extended in order to adjust AWJT. Without these improvements, a comparison of jet to traditional technologies is inappropriate.

Response Surface Methods Used for Optimization of Abrasive Waterjet Machining of the Stainless Steel X2 CrNiMo 17-12-2.

Abrasive waterjet machining (AWJM) has a particularly high potential for the machining of stainless steels. One of the main optimization objectives of the machining of X2 CrNiMo 17-12-2 stainless steel is obtaining a minimal surface roughness. This entails selecting an optimum configuration of the main influencing factors of the machining process. Optimization of the machining system was achieved by intervening on four selected input quantities (traverse speed, waterjet pressure, stand-off distance, and grit size), with three set points considered for each. The effects of modifying the set-points of each input parameter on the surface roughness were studied. By means of response surface methodology (RSM) the combination of factor set points was determined that ensures a minimum roughness of the machined surface. The main benefit of RSM is the reduced time needed for experimenting.

[Thoracic Outlet Syndrome: Rare, Often Missed or Over-Diagnosed?] / Das Thoracic-Outlet-Syndrom: selten, vielmals verpasst oder zu häufig diagnostiziert?

Thoracic Outlet Syndrome: Rare, Often Missed or Over-Diagnosed? Abstract. The thoracic outlet syndrome (TOS) presents with various symptoms caused by compression of the neurovascular bundle in the region of the upper thoracic aperture. Since the pathogenesis also determines the therapy of TOS, the classification according to the affected structure into neurogenic, venous and arterial TOS (nTOS, vTOS and aTOS) is useful. However, mixed forms are often to be assumed, which are then usually also classified under the term 'non-specific or disputed TOS' in the group of nTOS. In the absence of a gold standard diagnostic test, accurate history taking and clinical examination continue to be of great importance. Diagnostic experience and therapeutic advances have led to hopeful possibilities in the challenging management of this condition.

Standoff Distance in Ultrasonic Pulsating Water Jet.

The water hammer effect is the basis of technologies which is artificially responsible for the decay of continuous jets. A recently developed technique enhances the pressure fluctuations using an acoustic chamber, leading to enhanced erosion effects for various water volume flow rates. The optimum standoff distance for an ultrasonic enhanced water jet is not appropriately estimated using an inclined trajectory. The objective of this study is to comprehend the true nature of the interaction of the standoff distance following the stair trajectory and traverse speed of the nozzle on the erosion depth. Additionally, it also critically compares the new method (staircase trajectory) that obeys the variation in frequency of the impingements for defined volume flow rates with the inclined trajectory. In this study, at constant pressure (p = 70 MPa), the role of impingement distribution with the variation of traverse speed (v = 5-35 mm/s) along the centerline of the footprint was investigated. The maximum erosion depth corresponding to each traverse speed is observed at approximately same standoff distance (65 ± 5 mm) and decreases with the increment in traverse speed (h= 1042 and 47 µm at v = 5 and 35 mm/s, respectively). The results are attributed to the variation in the number of impingements per unit length. The surface and morphology analysis of the cross-section using SEM manifested the presence of erosion characteristics (micro-cracks, cavities, voids, and upheaved surface). By varying the water cluster, different impingement densities can be achieved that are suitable for technological operations such as surface peening, material disintegration, or surface roughening.

Influence of Material Structure on Forces Measured during Abrasive Waterjet (AWJ) Machining.

Material structure is one of the important factors influencing abrasive waterjet (AWJ) machining efficiency and quality. The force measurements were performed on samples prepared from two very similar steels with different thicknesses and heat treatment. The samples were austenitized at 850 °C, quenched in polymer and tempered at various temperatures between 20 °C and 640 °C. The resulting states of material substantially differed in strength and hardness. Therefore, samples prepared from these material states are ideal for testing of material response to AWJ. The force measurements were chosen to test the possible influence of material structure on the material response to the AWJ impact. The results show that differences in material structure and respective material properties influence the limit traverse speed. The cutting to deformation force ratio seems to be a function of relative traverse speed independently on material structure.

Effect of FSW Traverse Speed on Mechanical Properties of Copper Plate Joints.

The paper describes the influence of the friction stir welding travel speed on the mechanical properties of the butt joints of copper plates. The results of static and fatigue tests of the base material (Cu-ETP R220) and welded specimens produced at various travel speeds were compared, considering a loading applied both parallel and perpendicularly to the rolling direction of the plates. The mechanical properties of the FSW joints were evaluated with respect to parameters of plates' material in the delivery state and after recrystallisation annealing. The strength parameters of friction stir welding joints were compared with the data on tungsten inert gas welded joints of copper plates available in the literature. The results of microhardness tests and fractographic analysis of tested joints are also presented. Based on the above test results, it was shown that although in the whole range of considered traverse speeds (from 40 to 80 mm/min), comparable properties were obtained for FSW copper joints in terms of their visual and microstructural evaluation, their static and especially fatigue parameters were different, most apparent in the nine-fold greater observed average fatigue life. The fatigue tests turned out to be more sensitive criteria for evaluation of the FSW joints' qualities.

Investigation on Effect of Strain Rate and Heat Generation on Traverse Force in FSW of Dissimilar Aerospace Grade Aluminium Alloys.

The emergence of the aerospace sector requires efficient joining of aerospace grade aluminium alloys. For large-scale industrial practices, achievement of optimum friction stir welding (FSW) parameters is chiefly aimed at obtaining maximum strain rate in deforming material with least application of traverse force on the tool pin. Exact computation of strain rate is not possible due to complex and unexposed material flow kinematics. Estimation using micro-structural evolution serves as one of the very few methods applicable to analyze the yet unmapped interdependence of strain rate and traverse force. Therefore, the present work assessed strain rate in the stir zone using Zener Holloman parameter. The maximum and minimum strain rates of 6.95 and 0.31 s-1 were obtained for highest and least traverse force, respectively.

Remodeling of Interstrand Crosslink Proximal Replisomes Is Dependent on ATR, FANCM, and FANCD2.

Eukaryotic replisomes are driven by the mini chromosome maintenance (MCM [M]) helicase complex, an offset ring locked around the template for leading strand synthesis by CDC45 (C) and GINS (G) proteins. Although the CDC45 MCM GINS (CMG) structure implies that interstrand crosslinks (ICLs) are absolute blocks to replisomes, recent studies indicate that cells can restart DNA synthesis on the side of the ICL distal to the initial encounter. Here, we report that restart requires ATR and is promoted by FANCD2 and phosphorylated FANCM. Following introduction of genomic ICLs and dependent on ATR and FANCD2 but not on the Fanconi anemia core proteins or FAAP24, FANCM binds the replisome complex, with concomitant release of the GINS proteins. In situ analysis of replisomes proximal to ICLs confirms the ATR-dependent release of GINS proteins while CDC45 is retained on the remodeled replisome. The results demonstrate the plasticity of CMG composition in response to replication stress.