DNA G-quadruplexes in the genome of Trypanosoma cruzi as potential therapeutic targets for Chagas disease: Dithienylethene ligands as effective antiparasitic agents.

Chagas disease is caused by the parasite Trypanosoma cruzi and affects over 7 million people worldwide. The two actual treatments, Benznidazole (Bzn) and Nifurtimox, cause serious side effects due to their high toxicity leading to treatment abandonment by the patients. In this work, we propose DNA G-quadruplexes (G4) as potential therapeutic targets for this infectious disease. We have found 174 PQS per 100,000 nucleotides in the genome of T. cruzi and confirmed G4 formation of three frequent motifs. We synthesized a family of 14 quadruplex ligands based in the dithienylethene (DTE) scaffold and demonstrated their binding to these identified G4 sequences. Several DTE derivatives exhibited micromolar activity against epimastigotes of four different strains of T. cruzi, in the same concentration range as Bzn. Compounds L3 and L4 presented remarkable activity against trypomastigotes, the active form in blood, of T. cruzi SOL strain (IC50 = 1.5-3.3 µM, SI = 25-40.9), being around 40 times more active than Bzn and displaying much better selectivity indexes.

Impact of the COVID-19 pandemic on the self-care and health condition of the older adults. CUIDAMOS+75. A mixed methods study protocol.

Aims: To assess the impact of the COVID-19 pandemic on the health condition of people ≥75 years of age and on their family caregivers in Spain. Design: Multicentric, mixed method concurrent study. Methods: This work, which will be conducted within the primary care setting in 11 administrative regions of Spain, will include three coordinated studies with different methodologies. The first is a population-based cohort study that will use real-life data to analyze the rates and evolution of health needs, care provision, and services utilization before, during, and after the pandemic. The second is a prospective cohort study with 18 months of follow-up that will evaluate the impact of COVID-19 disease on mortality, frailty, functional and cognitive capacity, and quality of life of the participants. Finally, the third will be a qualitative study with a critical social approach to understand and interpret the social, political, and economic dimensions associated with the use of health services during the pandemic. We have followed the SPIRIT Checklist to address trial protocol and related documents. This research is being funded by the Instituto de Salud Carlos III since 2021 and was approved by its ethics committee (June 2022). Discussion: The study findings will reveal the long-term impact of the COVID-19 pandemic on the older adults and their caregivers. This information will serve policymakers to adapt health policies to the needs of this population in situations of maximum stress, such as that produced by the COVID-19 pandemic. Trial Registration: Identifier: NCT05249868 [ClinicalTrials.gov].

Hu14.18K.322A Causes Direct Cell Cytotoxicity and Synergizes with Induction Chemotherapy in High-Risk Neuroblastoma.

The disialoganglioside, GD2, is a promising therapeutic target due to its overexpression in certain tumors, particularly neuroblastoma (NB), with limited expression in normal tissues. Despite progress, the intricate mechanisms of action and the full spectrum of the direct cellular responses to anti-GD2 antibodies remain incompletely understood. In this study, we examined the direct cytotoxic effects of the humanized anti-GD2 antibody hu14.18K322A (hu14) on NB cell lines, by exploring the associated cell-death pathways. Additionally, we assessed the synergy between hu14 and conventional induction chemotherapy drugs. Our results revealed that hu14 treatment induced direct cytotoxic effects in CHLA15 and SK-N-BE1 cell lines, with a pronounced impact on proliferation and colony formation. Apoptosis emerged as the predominant cell-death pathway triggered by hu14. Furthermore, we saw a reduction in GD2 surface expression in response to hu14 treatment. Hu14 demonstrated synergy with induction chemotherapy drugs with alterations in GD2 expression. Our comprehensive investigation provides valuable insights into the multifaceted effects of hu14 on NB cells, shedding light on its direct cytotoxicity, cell-death pathways, and interactions with induction chemotherapy drugs. This study contributes to the evolving understanding of anti-GD2 antibody therapy and its potential synergies with conventional treatments in the context of NB.

A Randomized Controlled Trial Assessing the Impact of Continuous Glucose Monitoring with a Predictive Hypoglycemia Alert Function on Hypoglycemia in Physical Activity for People with Type 1 Diabetes (PACE).

Background: Uptake of exercise in people with type 1 diabetes (T1D) is low despite significant health benefits. Fear of hypoglycemia is the main barrier to exercise. Continuous glucose monitoring (CGM) with predictive alarms warning of impending hypoglycemia may improve self-management of diabetes around exercise. Aim: To assess the impact of Dexcom G6 real-time CGM system with a predictive hypoglycemia alert function on the frequency, duration, and severity of hypoglycemia occurring during and after regular (≥150 min/week) physical activity in people with T1D. Methods: After 10 days of blinded run-in (Baseline), CGM was unblinded and participants randomized 1:1 to have the "urgent low soon" (ULS) alert switched "on" or "off" for 40 days. Participants then switched alerts "off" or "on," respectively, for a further 40 days. Physical activity, and carbohydrate and insulin doses were recorded. Results: Twenty-four participants (8 men, 16 women) were randomized. There was no difference in change from baseline of hypoglycemia <3.0 and <3.9 mmol/L with the ULS on or off during the 24 h after exercise. With ULS alert "on" time spent below 2.8 mmol/L compared with baseline was significantly (P = 0.04) lower than with ULS "off" in the 24 h after exercise. In mixed effects regression, timing of the exercise and baseline HbA1c independently affected risk of hypoglycemia during exercise; exercise timing also affected hypoglycemia risk after exercise. Conclusion: A CGM device with an ULS alert reduces exposure to hypoglycemia below 2.8 mmol/L overall and in the 24 h after exercise compared with a threshold alert.

SARS-CoV-2 vaccination response in pediatric oncology patients.

BACKGROUND: There remains limited knowledge about the immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination in pediatric oncology patients, which is essential to provide counseling and risk adaptation in this vulnerable population. The goal of this study was to understand immunogenicity after vaccination in pediatric oncology patients, and determine if certain clinical factors impacted response. METHODS: Patients 0-25 years of age with a diagnosis of cancer and actively receiving therapy were enrolled on study. We excluded patients who were completely vaccinated prior to their cancer diagnosis. Blood samples were collected pre-vaccination, as well as 2, 4-6, and 8-12 weeks after vaccination. Healthy children who were fully vaccinated enrolled as controls. Clinical data and complete blood counts around time of vaccination were collected. To study B- and T-cell immunity, we measured neutralizing antibodies by enzyme-linked immunoassay and interferon gamma secretion by enzyme-linked immunospot, respectively. RESULTS: Twenty-six patients enrolled on study, for which 11 were evaluable oncology patients and seven were healthy controls. Adequate B-cell response was seen in 36.4% of patients, and adequate T-cell response in 77.8% of patients. Numbers were too small to detect differences based on malignancy type. There was no differences in immunity based on absolute lymphocyte count (ALC) or intensity of therapy. CONCLUSION: Pediatric oncology patients have a suboptimal immune response to SARS-CoV-2 vaccination. Booster doses will be imperative to provide optimal protection against COVID-19; however, blood counts may not be a useful guide to optimize the time of administration.

A quality assurance framework for routine monitoring of deep learning cardiac substructure computed tomography segmentation models in radiotherapy.

BACKGROUND: For autosegmentation models, the data used to train the model (e.g., public datasets and/or vendor-collected data) and the data on which the model is deployed in the clinic are typically not the same, potentially impacting the performance of these models by a process called domain shift. Tools to routinely monitor and predict segmentation performance are needed for quality assurance. Here, we develop an approach to perform such monitoring and performance prediction for cardiac substructure segmentation. PURPOSE: To develop a quality assurance (QA) framework for routine or continuous monitoring of domain shift and the performance of cardiac substructure autosegmentation algorithms. METHODS: A benchmark dataset consisting of computed tomography (CT) images along with manual cardiac substructure delineations of 241 breast cancer radiotherapy patients were collected, including one "normal" image domain of clean images and five "abnormal" domains containing images with artifact (metal, contrast), pathology, or quality variations due to scanner protocol differences (field of view, noise, reconstruction kernel, and slice thickness). The QA framework consisted of an image domain shift detector which operated on the input CT images and a shape quality detector on the output of an autosegmentation model, and a regression model for predicting autosegmentation model performance. The image domain shift detector was composed of a trained denoising autoencoder (DAE) and two hand-engineered image quality features to detect normal versus abnormal domains in the input CT images. The shape quality detector was a variational autoencoder (VAE) trained to estimate the shape quality of the auto-segmentation results. The output from the image domain shift and shape quality detectors was used to train a regression model to predict the per-patient segmentation accuracy, measured by Dice coefficient similarity (DSC) to physician contours. Different regression techniques were investigated including linear regression, Bagging, Gaussian process regression, random forest, and gradient boost regression. Of the 241 patients, 60 were used to train the autosegmentation models, 120 for training the QA framework, and the remaining 61 for testing the QA framework. A total of 19 autosegmentation models were used to evaluate QA framework performance, including 18 convolutional neural network (CNN)-based and one transformer-based model. RESULTS: When tested on the benchmark dataset, all abnormal domains resulted in a significant DSC decrease relative to the normal domain for CNN models ( p < 0.001 $p < 0.001$ ), but only for some domains for the transformer model. No significant relationship was found between the performance of an autosegmentation model and scanner protocol parameters ( p = 0.42 $p = 0.42$ ) except noise ( p = 0.01 $p = 0.01$ ). CNN-based autosegmentation models demonstrated a decreased DSC ranging from 0.07 to 0.41 with added noise, while the transformer-based model was not significantly affected (ANOVA, p = 0.99 $p=0.99$ ). For the QA framework, linear regression models with bootstrap aggregation resulted in the highest mean absolute error (MAE) of 0.041 ± 0.002 $0.041 \pm 0.002$ , in predicted DSC (relative to true DSC between autosegmentation and physician). MAE was lowest when combining both input (image) detectors and output (shape) detectors compared to output detectors alone. CONCLUSIONS: A QA framework was able to predict cardiac substructure autosegmentation model performance for clinically anticipated "abnormal" domain shifts.

Comparative evaluation of genotoxicity and cytotoxicity of flowable, bulk-fill flowable, and nanohybrid composites in human gingival cells using cytome assay: An in vivo study.

Context: Biocompatibility is one of the major prerequisites for safe clinical application of materials. Resin composites release their components into oral environment following restoration which cause adverse reactions. Aims: To evaluate and compare the genotoxicity and cytotoxicity of flowable, bulk-fill flowable, and nanohybrid composites with glass ionomer cement in human gingival cells using epithelial-based cytome assay. Methodology: Sixty healthy patients with noncarious cervical lesions were selected and randomly assigned to four groups (n = 15): Group A, glass ionomer cement; Group B, flowable composite; Group C, bulk-fill flowable composite; and Group D, nanohybrid composite. Class V restorations were done in each group with the respective restorative materials. Samples of epithelial cells were collected from gingiva before (control) (T1) and after 10 and 30 days (T2 and T3) postrestoration and examined for the presence of micronuclei and other nuclear anomalies. Statistical Analysis Used: The results were subjected to statistical analysis using Friedman's test and Kruskal-Wallis test. Results: The highest level of cytotoxicity was noted at T2 time point with a significant decline at T3 time point. Least cytotoxic damage was exhibited by Group A followed by Group D. Highest cytotoxic effect was shown by Group B followed by Group C. There was no significant level of genotoxicity induced by any of the materials tested at different time points. Conclusion: There is significant cytotoxicity induced by the tested composite materials which had no long-term effects and no genotoxicity was induced by any of the restorative materials tested.

Genomic analysis unveils genome degradation events and gene flux in the emergence and persistence of S. Paratyphi A lineages.

Paratyphoid fever caused by S. Paratyphi A is endemic in parts of South Asia and Southeast Asia. The proportion of enteric fever cases caused by S. Paratyphi A has substantially increased, yet only limited data is available on the population structure and genetic diversity of this serovar. We examined the phylogenetic distribution and evolutionary trajectory of S. Paratyphi A isolates collected as part of the Indian enteric fever surveillance study "Surveillance of Enteric Fever in India (SEFI)." In the study period (2017-2020), S. Paratyphi A comprised 17.6% (441/2503) of total enteric fever cases in India, with the isolates highly susceptible to all the major antibiotics used for treatment except fluoroquinolones. Phylogenetic analysis clustered the global S. Paratyphi A collection into seven lineages (A-G), and the present study isolates were distributed in lineages A, C and F. Our analysis highlights that the genome degradation events and gene acquisitions or losses are key molecular events in the evolution of new S. Paratyphi A lineages/sub-lineages. A total of 10 hypothetically disrupted coding sequences (HDCS) or pseudogenes-forming mutations possibly associated with the emergence of lineages were identified. The pan-genome analysis identified the insertion of P2/PSP3 phage and acquisition of IncX1 plasmid during the selection in 2.3.2/2.3.3 and 1.2.2 genotypes, respectively. We have identified six characteristic missense mutations associated with lipopolysaccharide (LPS) biosynthesis genes of S. Paratyphi A, however, these mutations confer only a low structural impact and possibly have minimal impact on vaccine effectiveness. Since S. Paratyphi A is human-restricted, high levels of genetic drift are not expected unless these bacteria transmit to naive hosts. However, public-health investigation and monitoring by means of genomic surveillance would be constantly needed to avoid S. Paratyphi A serovar becoming a public health threat similar to the S. Typhi of today.

FOXO-mediated repression of Dicer1 regulates metabolism, stress resistance, and longevity in Drosophila.

The adipose tissue plays a crucial role in metabolism and physiology, affecting animal lifespan and susceptibility to disease. In this study, we present evidence that adipose Dicer1 (Dcr-1), a conserved type III endoribonuclease involved in miRNA processing, plays a crucial role in the regulation of metabolism, stress resistance, and longevity. Our results indicate that the expression of Dcr-1 in murine 3T3L1 adipocytes is responsive to changes in nutrient levels and is subject to tight regulation in the Drosophila fat body, analogous to human adipose and hepatic tissues, under various stress and physiological conditions such as starvation, oxidative stress, and aging. The specific depletion of Dcr-1 in the Drosophila fat body leads to changes in lipid metabolism, enhanced resistance to oxidative and nutritional stress, and is associated with a significant increase in lifespan. Moreover, we provide mechanistic evidence showing that the JNK-activated transcription factor FOXO binds to conserved DNA-binding sites in the dcr-1 promoter, directly repressing its expression in response to nutrient deprivation. Our findings emphasize the importance of FOXO in controlling nutrient responses in the fat body by suppressing Dcr-1 expression. This mechanism coupling nutrient status with miRNA biogenesis represents a novel and previously unappreciated function of the JNK-FOXO axis in physiological responses at the organismal level.

Safety and Efficacy of an Adaptive Bolus Calculator for Type 1 Diabetes: A Randomized Controlled Crossover Study.

Background: The Advanced Bolus Calculator for Type 1 Diabetes (ABC4D) is a decision support system using the artificial intelligence technique of case-based reasoning to adapt and personalize insulin bolus doses. The integrated system comprises a smartphone application and clinical web portal. We aimed to assess the safety and efficacy of the ABC4D (intervention) compared with a nonadaptive bolus calculator (control). Methods: This was a prospective randomized controlled crossover study. Following a 2-week run-in period, participants were randomized to ABC4D or control for 12 weeks. After a 6-week washout period, participants crossed over for 12 weeks. The primary outcome was difference in % time in range (%TIR) (3.9-10.0 mmol/L [70-180 mg/dL]) change during the daytime (07:00-22:00) between groups. Results: Thirty-seven adults with type 1 diabetes on multiple daily injections of insulin were randomized, median (interquartile range [IQR]) age 44.7 (28.2-55.2) years, diabetes duration 15.0 (9.5-29.0) years, and glycated hemoglobin 61.0 (58.0-67.0) mmol/mol (7.7 [7.5-8.3]%). Data from 33 participants were analyzed. There was no significant difference in daytime %TIR change with ABC4D compared with control (median [IQR] +0.1 [-2.6 to +4.0]% vs. +1.9 [-3.8 to +10.1]%; P = 0.53). Participants accepted fewer meal dose recommendations in the intervention compared with control (78.7 [55.8-97.6]% vs. 93.5 [73.8-100]%; P = 0.009), with a greater reduction in insulin dosage from that recommended. Conclusion: The ABC4D is safe for adapting insulin bolus doses and provided the same level of glycemic control as the nonadaptive bolus calculator. Results suggest that participants did not follow the ABC4D recommendations as frequently as control, impacting its effectiveness. Clinical Trials Registration: clinicaltrials.gov NCT03963219 (Phase 5).

Recurring episodes of bundle branch reentry ventricular tachycardia due to aortitis preceded by SARS-CoV-2 infection: a case report.

BACKGROUND: SARS-CoV-2 may trigger both vasculitis and arrhythmias as part of a multisystem inflammatory syndrome described in children as well as in adults following COVID-19 infection with only minor respiratory symptoms. The syndrome denotes a severe dysfunction of one or more extra-pulmonary organ systems, with symptom onset approximately 2-5 weeks after the COVID-19 infection. In the present case, a seemingly intractable ventricular tachycardia preceded by SARS-CoV2 infection was only managed following the diagnosis and management of aortitis. CASE PRESENTATION: A 69-year-old woman was hospitalized due to syncope, following a mild COVID-19 infection. She presented with paroxysmal atrial fibrillation and intermittent ventricular tachycardia interpreted as a septum-triggered bundle branch reentry ventricular tachycardia, unaffected by amiodaron, lidocaine and adenosine. A CT-scan revealed inflammation of the aortic arch, extending into the aortic root. In the following days, the tachycardia progressed to ventricular storm with intermittent third-degree AV block. A temporary pacemaker was implanted, and radiofrequency ablation was performed to both sides of the ventricular septum after which the ventricular tachycardia was non-inducible. Following supplemental prednisolone treatment, cardiac symptoms and arrythmia subsided, but recurred after tapering. Long-term prednisolone treatment was therefore initiated with no relapse in the following 14 months. CONCLUSION: We present a rare case of aortitis complicated with life-threatening ventricular tachycardia presided by Covid-19 infection without major respiratory symptoms. Given a known normal AV conduction prior to the COVID-19 infection, it seems likely that the ensuing aortitis in turn affected the septal myocardium, enabling the reentry tachycardia. Generally, bundle branch reentry tachycardia is best treated with radiofrequency ablation, but if it is due to aortitis with myocardial affection, long-term anti-inflammatory treatment is mandatory to prevent relapse and assure arrhythmia control. Our case highlights importance to recognize the existence of the multisystem inflammatory syndrome in adults (MIS-A) following COVID-19 infection in patients with alarming cardiovascular symptoms. The case shows that the early use of an CT-scan was crucial for both proper diagnosis and treatment option.

Membraneless Organelles and Condensates Orchestrate Innate Immunity Against Viruses.

The cellular defense against viruses involves the assembly of oligomers, granules and membraneless organelles (MLOs) that govern the activation of several arms of the innate immune response. Upon interaction with specific pathogen-derived ligands, a number of pattern recognition receptors (PRRs) undergo phase-separation thus triggering downstream signaling pathways. Among other relevant condensates, inflammasomes, apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) specks, cyclic GMP-AMP synthase (cGAS) foci, protein kinase R (PKR) clusters, ribonuclease L-induced bodies (RLBs), stress granules (SGs), processing bodies (PBs) and promyelocytic leukemia protein nuclear bodies (PML NBs) play different roles in the immune response. In turn, viruses have evolved diverse strategies to evade the host defense. Viral DNA or RNA, as well as viral proteases or proteins carrying intrinsically disordered regions may interfere with condensate formation and function in multiple ways. In this review we discuss current and hypothetical mechanisms of viral escape that involve the disassembly, repurposing, or inactivation of membraneless condensates that govern innate immunity. We summarize emerging interconnections between these diverse condensates that ultimately determine the cellular outcome.

Development and Evaluation of a Digital Health Intervention to Prevent Type 2 Diabetes in Primary Care: The PREDIABETEXT Study Protocol for a Randomised Clinical Trial.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a highly prevalent disease associated with an increased risk of comorbidities, premature death, and health costs. Prediabetes is a stage of glucose alteration previous to T2DM, that can be reversed. The aim of the study is to develop and evaluate a low-intensity, multifaceted, digital intervention to prevent T2DM. The intervention comprises: (1) the use of mobile health technology to send tailored text messages promoting lifestyle changes to people at risk of T2DM and (2) the provision of online education to primary healthcare physicians and nurses about management of prediabetes. METHODS: In stages 1-4 we will design, develop and pilot-test the intervention. In Stage 5 we will conduct a phase II, six-month, three-arm, cluster randomized, clinical trial with 42 primary care professionals and 420 patients at risk of T2DM. Patients will be allocated to a control group (usual care), intervention A (patient messaging intervention), or intervention B (patient messaging intervention plus online education to their primary healthcare professionals). The primary outcome will be glycated haemoglobin. All the procedures obtained ethical approval in June 2021 (CEI-IB Ref No: IB4495/21PI). DISCUSSION: Digital health interventions can effectively prevent T2DM and reduce important T2DM risk factors such as overweight or hypertension. In Spain, this type of intervention is understudied. Moreover, there is controversy regarding the type of digital health interventions that are more effective. Findings from this study may contribute to address T2DM prevention, through a low-cost and easily implementable intervention.

Selective gene expression profiling contributes to a better understanding of the molecular pathways underlying the histological changes observed after RHMVL.

BACKGROUND: In previous studies, five vasoactive drugs were investigated for their effect on the recovery process after extended liver resection without observing relevant improvements. We hypothesized that an analysis of gene expression could help to identify potentially druggable pathways and could support the selection of promising drug candidates. METHODS: Liver samples obtained from rats after combined 70% partial hepatectomy and right median hepatic vein ligation (n = 6/group) sacrificed at 0 h, 24 h, 48 h, and 7days were selected for this study. Liver samples were collected from differentially perfused regions of the median lobe (obstruction-zone, border-zone, normal-zone). Gene expression profiling of marker genes regulating hepatic hemodynamics, vascular remodeling, and liver regeneration was performed with microfluidic chips. We used 3 technical replicates from each sample. Raw data were normalized using LEMming and differentially expressed genes were identified using LIMMA. RESULTS: The strongest differences were found in obstruction-zone at 24 h and 48 h postoperatively compared to all other groups. mRNA expression of marker genes from hepatic hemodynamics pathways (iNOS,Ptgs2,Edn1) was most upregulated. CONCLUSION: These upregulated genes suggest a strong vasoconstrictive effect promoting arterial hypoperfusion in the obstruction-zone. Reducing iNOS expression using selective iNOS inhibitors seems to be a promising approach to promote vasodilation and liver regeneration.

Heart Sparing Radiotherapy Techniques in Breast Cancer: A Focus on Deep Inspiration Breath Hold.

Adjuvant radiation therapy is a critical component of breast cancer management. However, when breast cancer patients receive incidental radiation to the heart, there is an increased risk of cardiac disease and mortality. This is most common for patients with left-sided breast cancers and those receiving nodal irradiation as part of treatment. The overall risk of cardiac toxicity increases 4-16% with each Gray increase in mean heart radiation dose, with data suggesting that no lower limit exists which would eliminate cardiac risk entirely. Radiation techniques have improved over time, leading to lower cardiac radiation exposure than in the past. This decline is expected to reduce the incidence of radiation-induced heart dysfunction in patients. Deep inspiration breath hold (DIBH) is one such technique that was developed to reduce the risk of cardiac death and coronary events. DIBH is a non-invasive approach that capitalizes on the natural physiology of the respiratory cycle to increase the distance between the heart and the therapeutic target throughout the course of radiation therapy. DIBH has been shown to decrease the mean incidental radiation doses to the heart and left anterior descending coronary artery by approximately 20-70%. In this review, we summarize different techniques for DIBH and discuss recent data on this technique.

Second Generation Small Molecule Inhibitors of Gankyrin for the Treatment of Pediatric Liver Cancer.

Background: Gankyrin, a member of the 26S proteasome, is an overexpressed oncoprotein in hepatoblastoma (HBL) and hepatocellular carcinoma (HCC). Cjoc42 was the first small molecule inhibitor of Gankyrin developed; however, the IC50 values of >50 µM made them unattractive for clinical use. Second-generation inhibitors demonstrate a stronger affinity toward Gankyrin and increased cytotoxicity. The aim of this study was to characterize the in vitro effects of three cjoc42 derivatives. Methods: Experiments were performed on the HepG2 (HBL) and Hep3B (pediatric HCC) cell lines. We evaluated the expression of TSPs, cell cycle markers, and stem cell markers by Western blotting and/or real-time quantitative reverse transcription PCR. We also performed apoptotic, synergy, and methylation assays. Results: The treatment with cjoc42 derivatives led to an increase in TSPs and a dose-dependent decrease in the stem cell phenotype in both cell lines. An increase in apoptosis was only seen with AFM-1 and -2 in Hep3B cells. Drug synergy was seen with doxorubicin, and antagonism was seen with cisplatin. In the presence of cjoc42 derivatives, the 20S subunit of the 26S proteasome was more available to transport doxorubicin to the nucleus, leading to synergy. Conclusion: Small-molecule inhibitors for Gankyrin are a promising therapeutic strategy, especially in combination with doxorubicin.

Delta variant SARS-CoV-2 infections in pediatric cases during the second wave in India.

BACKGROUND: During October 2020, Delta variant was detected for the first time in India and rampantly spread across the globe. It also led to second wave of pandemic in India which affected millions of people. However, there is limited information pertaining to the SARS-CoV-2 strain infecting the children in India. METHODS: Here, we assessed the SARS-CoV-2 lineages circulating in the pediatric population of India during the second wave of the pandemic. Clinical and demographic details linked with the nasopharyngeal/oropharyngeal swabs (NPS/OPS) collected from SARS-CoV-2 cases (n = 583) aged 0-18 year and tested positive by real-time RT-PCR were retrieved from March to June 2021. RESULTS: Symptoms were reported among 37.2% of patients and 14.8% reported to be hospitalized. The E gene CT value had significant statistical difference at the point of sample collection when compared to that observed in the sequencing laboratory. Out of these 512 sequences 372 were VOCs, 51 were VOIs. Most common lineages observed were Delta, followed by Kappa, Alpha and B.1.36, seen in 65.82%, 9.96%, 6.83% and 4.68%, respectively in the study population. CONCLUSION: Overall, it was observed that Delta strain was the leading cause of SARS-CoV-2 infection in Indian children during the second wave of the pandemic. We emphasize on the need of continuous genomic surveillance in SARS-CoV-2 infection even amongst children.

Patient and Treatment Factors Associated With Improved Local Control and Survival in Oligometastatic Bone Disease: Results From a Large Single-Institution Experience Using Stereotactic Body Radiation Therapy.

PURPOSE: Limited data exist to guide optimal patient selection and treatment of bone metastases with curative intent despite the increasing application of stereotactic body radiation therapy (SBRT) for oligometastatic (OM) disease control and reirradiation (Re-RT). METHODS AND MATERIALS: Clinical characteristics for 434 patients consecutively treated with bone SBRT at a single institution from March 2011 to June 2020 were analyzed by OM, spine, and nonspine bone using Cox regression to determine association with local control (LC), progression-free survival (PFS), and overall survival (OS), and the Kaplan-Meier method to estimate PFS and OS. RESULTS: Most patients had prostate (39%) or breast/lung (21%) cancer and 1 to 3 lesions (96%), with 651 lesions (spine 63%) treated for Re-RT (12%) or OMD (88%), including synchronous (10%), metachronous (28%), repeat (27%), or induced (23%) states as defined by The European Society for Radiotherapy and Oncology and European Organisation for Research and Treatment of Cancer criteria. Biologically effective dose (BED10) ≥50 (hazard ratio, 0.68; 95% confidence interval, 0.48-0.96; P < .03) predicted improved LC among OM lesions and planning target volume (PTV) ≥150 cc (hazard ratio, 1.94; 95% confidence interval, 1.02-3.70; P < .04) predicted worse LC for nonspine bone. Prostate histology, performance status (PS) 0 to 1, and metastasis-free interval ≥2 year predicted improved PFS and OS (P < .05). Metachronous, synchronous, or repeat OM had higher PFS and OS (P ≤ .001) than induced OM. With median follow-up 25.7 months, 1- and 2-year PFS was 63% and 47% for OM and 36% and 25% for Re-RT; 1- and 2-year OS was 87% and 73% for OM and 58% and 43% for Re-RT. Acute toxicities included grade 1 to 2 pain flare (9%) and fatigue (14%). Late toxicities included fracture (1%) for OM and myelopathy (2.5%) or nerve pain (1.2%) for Re-RT. CONCLUSIONS: BED10 ≥50 for OM and PTV <150 cc for nonspine bone lesions was associated with improved LC. Prostate histology, PS 0 to 1, metastasis-free interval ≥2 years, and metachronous, synchronous, or repeat presentations per The European Society for Radiotherapy and Oncology and European Organisation for Research and Treatment of Cancer criteria predicted improved PFS and OS among OM patients treated with bone SBRT.

Humoral and Cellular Immune Response in Asymptomatic Dogs with Visceral Leishmaniasis: A Review.

Visceral leishmaniasis is one of the deadliest parasitic diseases in the world and affects both humans and dogs. The host immune response to Leishmania infection plays a critical role in the evolution of canine visceral leishmaniasis (CVL) and consequently in the manifestation of clinical signs. The asymptomatic form of the disease is a major concern in the diagnosis of CVL and in the transmission control of Leishmania infection. Asymptomatic dogs are found in large proportions in endemic areas and are an unquantifiable source of infection. The present review analyzes the possible relationship between the activation of the antigen-specific immune response of the host and resistance or susceptibility to CVL. The review focuses on works that address the characterization of the humoral and cellular immune response profile, at both the functional and phenotypic levels, in infected dogs. Most studies relate the absence of clinical symptomatology to an increased proliferative response and a Th1 cytokine profile. Despite the numerous findings pointing to a differential immune response in asymptomatic dogs, the contradictory results reported in this review highlight the importance of establishing a precise clinical classification of the disease, performing more longitudinal studies, and including a higher number of animals in trials.

Smaug1 membrane-less organelles respond to AMPK and mTOR and affect mitochondrial function.

Smaug is a conserved translational regulator that binds numerous mRNAs, including nuclear transcripts that encode mitochondrial enzymes. Smaug orthologs form cytosolic membrane-less organelles (MLOs) in several organisms and cell types. We have performed single-molecule fluorescence in situ hybridization (FISH) assays that revealed that SDHB and UQCRC1 mRNAs associate with Smaug1 bodies in U2OS cells. Loss of function of Smaug1 and Smaug2 (also known as SAMD4A and SAMD4B, respectively) affected both mitochondrial respiration and morphology of the mitochondrial network. Phenotype rescue by Smaug1 transfection depends on the presence of its RNA-binding domain. Moreover, we identified specific Smaug1 domains involved in MLO formation, and found that impaired Smaug1 MLO condensation correlates with mitochondrial defects. Mitochondrial complex I inhibition upon exposure to rotenone, but not strong mitochondrial uncoupling upon exposure to CCCP, rapidly induced the dissolution of Smaug1 MLOs. Metformin and rapamycin elicited similar effects, which were blocked by pharmacological inhibition of AMP-activated protein kinase (AMPK). Finally, we found that Smaug1 MLO dissolution weakens the interaction with target mRNAs, thus enabling their release. We propose that mitochondrial respiration and the AMPK-mTOR balance controls the condensation and dissolution of Smaug1 MLOs, thus regulating nuclear mRNAs that encode key mitochondrial proteins. This article has an associated First Person interview with the first authors of the paper.