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CST–Polα/Primase maintains telomeres through fill-in synthesis of the C-rich telomeric DNA. We report cryo-EM structures that reveal how human CST is recruited to telomeres by the shelterin subunits POT1 and TPP1. CST–POT1/TPP1 is formed through interactions between POT1 and the Ctc1 subunit of CST. Coats plus syndrome mutations map to the POT1–Ctc1 interface, providing mechanistic insights into this disease. CST–POT1/TPP1 is compatible with the previously reported inactive recruitment complex of CST–Polα/Primase but not with the distinct conformation of active CST–Polα/Primase. We propose that shelterin both recruits and regulates CST–Polα/Primase. Structural and biochemical data indicate that this regulation involves phosphorylation of POT1, which promotes CST–POT1/TPP1 interaction and recruitment, whereas POT1 dephosphorylation releases CST–Polα/Primase for fill-in synthesis.One-Sentence SummaryCryo-EM structures reveal how telomere maintenance factors are recruited and regulated by the shelterin complex.

Introduction Gene expression has been poorly studied in acute pancreatitis (AP). We examined leukocyte differential gene expression (DGE) in fatty acid ethyl ester-induced AP (FAEE-AP), which models alcohol-associated AP. Methods Wild type C57Bl/7 mice were randomly divided into 5 groups of 6 receiving two hourly intraperitoneal injections of either 150 mg/kg palmitoleic acid (POA) and 1.35 g/kg ethanol (FAEE-AP), or 50 mg/kg POA and 1.35 g/kg ethanol (FAEE-AP), or 1.35 g/kg ethanol alone (contrast), or saline (control), or no injections (control). Mice were humanely killed at 12 hours for AP assessment and Agilent two-colour RNA blood microarray, with Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analyses (DGE P 1). Results AP developed in FAEE-AP, more severe with 150 mg/kg POA, but not in either control group, with Conclusion These data suggest appropriateness of either control, with little detectable difference in DGEs between FAEE-AP protocols despite differing AP severity; limited additional data were obtained using an ethanol group. Comparison with other AP models may improve understanding of disease mechanisms.

Introduction Persistent opening of the mitochondrial permeability transition pore (MPTP) is central to acute pancreatitis (AP). Mitochondrial translocation of P53 promotes MPTP opening, which induces necrotic cell death. We sought to determine whether and how mitochondrial P53 inhibition is protective in experimental AP. Methods Mitochondrial translocation of P53 was inhibited by pifithrin-mu (PFT-µ, mitochondrial specific P53 inhibitor) or by Trp53 KO. Mitochondrial membrane potential (ΔΨm) and necrosis of murine pancreatic acinar cells (PACs) were evaluated by confocal microscopy with TMRM and propidium iodide, respectively, in the presence of TLCS. Fluo4 was used for calcium imaging. Caerulein AP (Cer-AP) was induced in male (WT, P53−/+, P53−/−; 10-12 week old) mice by 7 hourly i.p. injections of 50 μg/kg caerulein. PFT-µ (5 mg/kg) was injected after the third injection of caerulein; sacrifice was made 12h after the first injection of caerulein. AP was assessed by standard biomarkers and blinded histopathology. Results TLCS-induced collapse of ΔΨm and cell necrosis of wild type PACs were significantly reduced in the presence of PFTµ or in PACs isolated from Trp53 KO mice; calcium overload was also reduced, indicative of improved calcium clearance, and ATP was maintained. Pharmacological or genetic P53 inhibition resulted in significant reductions of biochemical, immunological, and histopathological indices of severity in the Cer-AP model. Conclusion Inhibition of P53 maintained mitochondrial integrity, reduced PACs necrosis, and ameliorated the severity of Cer-AP. These data suggest that inhibition of mitochondrial translocation of P53 could be an effective strategy for the treatment of AP.

The body index [body mass, body mass index (BMI), waist circumference and waist-height ratio] and functional fitness (aerobic capacity, muscular strength, balance, flexibility and functional ability) of people with Down syndrome (DS) are substantially lower than those in the general population, as DS has an overall negative impact on health.This study explored the effect of a 24-week basketball training programme on the body composition and functional fitness of adults with DS.Twenty-two adults with DS (24 ± 6 years; 25 ± 4 kg/mHip circumference and waist-height ratio were statistically different at baseline between the intervention and control groups. After the 24-week basketball training programme, the results showed significant differences between the exercise and control groups for hip circumference, body mass, BMI, waist circumference, hip circumference, waist-height ratio, flexibility, aerobic capacity, static and dynamic balance, balance and basketball functional ability (P 0.05). The exercise group showed significant improvements in all variables of body composition and functional fitness (P 0.05) except for the waist-hip ratio. There was no statistical significance between all physical composition and functional health variables in the control group except for the standing long jump (P 0.05), with effect sizes ranging from small to large.The basketball training programme improved body composition and functional fitness of adults with DS.

To investigate the changes in relevant anatomical parameters of posterolateral protrusion of the vertebral artery (VA) between head-neck rotational and neutral positions using low-dose three-dimensional computed tomography angiography (3D-CTA).Low-dose 3D-CTA images obtained for various craniocervical diseases in 36 non-dominant VA side patients with neutral, left and right head-neck rotational positions were evaluated. The relevant parameters from superior and inferior views, including external diameter (ED), internal diameter (ID), transverse diameter (TD), heights and diameters of posterolateral protrusion of the VA over the posterior arch of the atlas in the neutral and rotational positions, were recorded and compared.There was no significant differences in the rotational angle (left/right: 31.23 ± 6.60/29.94 ± 6.09°, p0.05). There were no significant differences in heights and diameters of bilateral VA between rotational and neutral positions (all p0.05). The contralateral ID, ED, and TD of the rotational positions were significantly shorter than those of the neutral position (all p0.05), while there were no significant differences in the three ipsilateral diameters (all p0.05).Posterolateral protrusion of the VA is not uncommon in the population, and surgeons should be aware of its presence, especially the increased possibility of injury to the VA caused by head-neck rotation, during the operation; thus, preoperative evaluation by low-dose 3D-CTA should be considered.

Sickle cell disease (SCD) results in severe complications, such as anaemia and pain episodes. Hydroxyurea (HU) is efficacious in SCD, yet adherence remains low.To assess the relationship of HU adherence to health care utilization and patients' characteristics.This is a 5-year retrospective chart review. Patients' demographics and medical history were collected from the electronic medical record (EMR). HU adherence was evaluated using foetal haemoglobin "HbF%", mean corpuscular volume "MCV", and absolute neutrophil count "ANC". Age groups included children (12 years), adolescents (12-17 years), and young adults (≥18 years).A total of 113 SCD patients on HU were included (median age 14 years, IQR 10-20; 50% female; 88% HbSS). Young adults had significantly higher HU adherence compared to adolescents and children, including higher median HbF% (24.2Young adults with SCD had significantly higher HU adherence compared to children and adolescents. Patients with lower HU adherence and/or chronic pain had increased health care utilization. Future studies examining barriers to adherence and evaluating interventions to optimize HU adherence in SCD are warranted.KEY MESSAGESYoung adults with SCD had significantly higher HU adherence, as reflected in their laboratory markers, compared to children and adolescents.Patients with higher HU adherence and/or those without chronic pain had lower or less frequent health care utilization.No significant association of HU adherence to patients' sex, socio-economic status and distance from hospital.

N6-methyladenosine (m6A) is a eukaryotic messenger RNA modification catalyzed by methyltransferase-like 3 (METTL3), which is involved in various developmental and disease processes. However, the connection between the epigenetic modification of m6A and glucose metabolism during osteogenesis is still unclear. Here, we show that interference with METTL3 in dental pulp stem cells (DPSCs) inhibits cell proliferation and osteogenic differentiation. Moreover, transcriptome sequencing and metabolic testing were used to explore the mechanism between glucose metabolism and m6A modification in METTL3-knockdown DPSCs. Methylated RNA immunoprecipitation–quantitative polymerase chain reaction and RNA stability assays were used to determine the target genes of METTL3. Mechanistically, METTL3 directly interacts with ATP citrate lyase (ACLY) and a mitochondrial citrate transporter (SLC25A1) and then further affects the glycolytic pathway. M6A-mediated ACLY and SLC25A1 stability depends on the m6A readers IGF2BP2 and IGF2BP2/3, respectively. Our experiments uncovered the potential molecular mechanism of epigenetic modification in osteogenic differentiation, providing new ideas for the clinical application of stem cells and the intervention of metabolic bone diseases.

The mammalian telomeric shelterin complex—comprised of TRF1, TRF2, Rap1, TIN2, TPP1, and POT1—blocks the DNA damage response at chromosome ends and interacts with telomerase and the CST complex to regulate telomere length. The evolutionary origins of shelterin are unclear, partly because unicellular organisms have distinct telomeric proteins. Here, we describe the evolution of metazoan shelterin, showing that TRF1 emerged in vertebrates upon duplication of a TRF2-like ancestor. TRF1 and TRF2 diverged rapidly during vertebrate evolution through the acquisition of new domains and interacting factors. Vertebrate shelterin is also distinguished by the presence of an HJRL domain in the split C-terminal OB fold of POT1, whereas invertebrate POT1s carry inserts of variable nature. Importantly, the data reveal that, apart from the primate and rodent POT1 orthologs, all metazoan POT1s are predicted to have a fourth OB fold at their N termini. Therefore, we propose that POT1 arose from a four-OB-fold ancestor, most likely an RPA70-like protein. This analysis provides insights into the biology of shelterin and its evolution from ancestral telomeric DNA-binding proteins.

INTRODUCTION Cor triatriatum sinister (CTS) is a rare congenital heart defect characterized by fibromuscular septation of the left atrium associated with atrial fibrillation (AF). The incidence of hemodynamically insignificant CTS in the AF ablation population and effect on ablation success are not known. Furthermore, little is known about the potential effect of CTS on arrhythmogenic substrate. OBJECTIVE We define the incidence of hemodynamically insignificant CTS in patients undergoing AF ablation with RF and cryoballoon ablation, the technical challenges created by the left atrial partitioning, and the potentially arrhythmogenic effects of the membrane. We also review the literature of CA in patients with CTS. METHODS First-time AF ablation cases at our institution over a 10-year period were screened to identify patients with CTS. Retrospective review was performed to obtain clinical characteristics and ablation data. RESULTS Of the 3953 consecutive patients undergoing initial AF ablation during the study period, four patients (0.10%) had CTS. Ablation was successful acutely in all patients. One patient had recurrent AF and required repeat ablation for a single procedure success rate of 75% and multi-procedure success rate of 100%. The CTS membrane was associated with low voltage zones in the two patients in whom it was measured and with substrate for macro-reentrant atrial tachycardia in one of these patients. CONCLUSION The incidence of hemodynamically insignificant CTS in patients undergoing CA for AF is very low, but does not serve as a significant barrier to successful ablation as long as directed access to the superoposterior chamber is obtained.

IntroductionTrypanosoma cruzi is the protozoan parasite causing Chagas disease, a Neglected Tropical Disease that affects 8 million people and causes 12,000 deaths per year, primarily because of cardiac pathology. Effective vaccination for T. cruzi remains an elusive goal. The use of a live vaccine vector, especially one that mimics the pathogen target, may be superior to the use of recombinant protein or DNA vaccine formulations.MethodsWe generated recombinant Leishmania major, a related trypanosomatid parasite, as a vaccine vehicle to express the immunogenic T. cruzi trans-sialidase (TS) antigen. The induction of T cell and antibody responses, as well as T. cruzi protective immunity generated by these vaccines were assessed in vivo.ResultsWe demonstrate that mice inoculated with these recombinant TS-expressing L. major parasites mount T cell and antibody responses directed against TS and are protected against future T. cruzi infection. We also show that the partially attenuated dhfr-ts- CC1 L. major strain, previously found to induce protective immunity to virulent L. major infection without causing pathology, can also be engineered to express the TS antigen. This latter recombinant may represent a safe and effective option to explore for ultimate use in humans.DiscussionAltogether, these data indicate that L. major can stably express a T. cruzi antigen and induce T. cruzi-specific protective immunity, warranting further investigation of attenuated Leishmania parasites as vaccine.

We generated recombinantWe demonstrate that mice inoculated with these recombinant TS-expressingAltogether, these data indicate that

Aim Method 44 orthogeriatric patients’ records who underwent NOF surgery between December 2019 and March 2020 were examined in the first round. The following data were extracted from the electronic records: age, pre- and post-operative haemoglobin, uptake of pre- and post-operative haematinics, blood, and iron transfusions throughout the surgical period. Subsequently, two more rounds of data collection were undertaken, and they followed the same method. These involved 58 patients between January and April 2021 and the 26 patients between May and June 2021, respectively. Results Our first cycle saw uptake of pre-operative haematinics increase by 31% (from 20% to 51%). The final round saw uptake of pre-operative haematinics decrease to 27%. In addition, more than 1/3 patients (37%) who received blood had not met transfusion criteria according to NICE guidelines. Almost half of patients (46%) who received iron transfusion did not have confirmed iron deficiency anaemia. Conclusions Pre-operative anaemia continues to be poorly investigated despite an initial increase in haematinics uptake. This could be attributed to the changeover of teams in April. To sustain the level of previous improvement, a hospital-wide educational session for clinicians is needed and this is best incorporated into the induction programme.

To identify the valuable predictors of grade≥2 radiation pneumonitis (RP) in patient treated with radiotherapy after pneumonectomy for non-small cell lung cancer (NSCLC); and to construct a nomogram predicting the incidence of grade≥2 RP in such patients.We reviewed 82 patients with NSCLC received radiotherapy after pneumonectomy from 2008 to 2018. The endpoint was grade≥2 RP. Univariate and multivariate regression analysis were conducted to evaluate significant factors of grade≥2 RP. Receiver operating characteristic (ROC) curve was used to establish optimal cutoff values and the nomogram was built to make the predictive model visualized. Descriptive analysis was performed on 5 patients with grade 3 RP.A total of 22(26.8%) patients developed grade 2 RP and 5(6.1%) patients were grade 3 RP. V5, V10, V20, V30, MLD, PTV, and PTV/TLV were associated with the occurrence of grade≥2 RP in univariate analysis, while none of the clinical factors was significant; V5(OR,1.213;95%CI,1.099-1.339; P0.001) and V20(OR,1.435;95%CI,1.166-1.765; P=0.001) were the independent significant predictors by multivariate analysis and were included in the nomogram. The ROC analysis for the cutoff values for predicting grade≥2 RP were V523% (AUC=0.819, sensitivity:0.701, specificity:0.832) and V208% (AUC=0.812, sensitivity:0.683, specificity:0.811). Additionally, grade≥3 RP did not occur when V530%, V2013% and MLD751.2cGy, respectively.Our study showed that V5 and V20 were independent predictors for grade≥2 RP in NSCLC patients receiving radiotherapy after pneumonectomy. Grade 3 RP did not occur whenV530%, V2013% and MLD751.2cGy, respectively. In addition, patient underwent right pneumonectomy may have a lower tolerance to radiation compared to left pneumonectomy.

Animal studies have demonstrated that myeloid cell leukemia-1 (Mcl-1) gene deficiency leads to premature ovarian failure and decreased reproductive ability in mice. This study investigated the relationship between MCL-1 gene variation and idiopathic premature ovarian insufficiency (POI) in Chinese women.A total of 200 idiopathic POI patients and 100 healthy controls were recruited for this study, and peripheral blood was collected. First, genomic DNA was extracted from peripheral leukocytes. Then, the entire coding region and splice sites of the MCL-1 gene were amplified by polymerase chain reaction. Chi-squared tests were used to compare the genotype distribution and allele frequency of single nucleotide polymorphisms between the POI and control groups.Three mutations of the MCL-1 gene (c.-36C T, c.-131C T and c.78C T) were identified. After data analysis, c.-36C T and c.-131C T in the 5'-untranslated region were both found in the POI group and the control group. No difference was found in the genotype distribution or allelic frequency of either variant between the POI group and the control group (MCL-1 gene mutation may not be associated with idiopathic POI in Chinese women.

The dynamic mechanical behaviour induced by adiabatic temperature rise of Fe–Mn–Al–C twinning-induced plasticity steel was investigated via the dynamic tensile test and a dynamic constitutive model...

Ultrasonic metal welding (UMW) has been widely applied as a high throughput solid-state joining technology for multilayers of sheet metal. During a typical UMW process, multilayer work materials are mechanically compressed by a knurl-patterned horn (also known as a sonotrode) onto an anvil tool, and a simultaneous in-plane sliding is applied to the horn at an ultrasonic frequency (20 kHz or higher) to help form the weld at the material interfaces. There is a great challenge in modeling and simulating the dynamic behavior of the work material and the whole weld formation process is subject to ultrasonic mechanical loadings imposed by the knurl-patterned horn tool. In this work, finite element (FE) models are developed to simulate the multilayer UMW process using knurl-patterned tools by directly applying the ultrasonic vibration as a model input. For a short weld duration of 0.1∼0.5 s, a high-fidelity FE modeling approach is developed using ABAQUS/Explicit to simulate the dynamic material response under the 20 kHz horn vibration. For an extended long welding duration of approximately 1.0 s, a computationally efficient hybrid approach is developed using both ABAQUS/Explicit and DEFORM-3D in order to leverage the strengths of each software package. The developed models are validated using experimental data of dynamic welding force, temperature, and weld geometry from in-situ process measurements of UMW. The 3D FE models developed in this study are the most comprehensive solution to date to simulate the complex material response subject to UMW process conditions and provide engineering guidance for the design of UMW applications.

Introduction/Objective Patients with hematological cancer routinely receive red blood cell (RBC) transfusions in the outpatient infusion clinical setting as part of their clinical therapy. Typically, RBCs are delivered to the outpatient infusion clinic via the pneumatic tube (P-tube). However, due to the recent expansion of the hospital footprint and the new location of the transfusion services, the P-tube encountered challenges that impacted the reliability of the delivery of RBC, which can delay patient care and affect patient satisfaction. This study’s purpose is to evaluate the pre-and post-implementation of the Haemobank-20 (HB20), remote blood storage and dispensing device, by reviewing the turn-around time (TAT) and the number of orders fulfilled outside of the transfusion service. Methods/Case Report Blood products are stocked to the HB in the infusion center. An electronic transfusion schedule is reviewed the night before scheduled transfusions to ensure appropriate inventory. When a patient is ready for the transfusion, the clinical nurse will use the device to retrieve blood products for the patient from the HB20. The average timed delivery time from the transfusion service to the outpatient infusion center is about 11 minutes. Data was retrospectively gathered from a 10-month interval before and after HB20 implementation to evaluate the number of orders fulfilled by the HB20. Results (if a Case Study enter NA) The HB has helped maintain nurse-to-patient ratios, reduced traffic at the blood bank issue window, and significantly sped up the turnaround time of RBCs from 11 minutes to less than 60 seconds. Before HB implementation, staff at the blood bank received 2853 units of RBCs through the blood bank window. This has been decreased by 61.2 percent to 1136 RBCs. Conclusion Implementing the Haemobank-20 in the outpatient infusion center has successfully reduced the turnaround time and the workload within the Transfusion Services. The current results study demonstrate that the remote blood-release system is safe and helpful in improving the efficiency of blood issue for patients in remote outpatient locations.