Oxidation-sensitive cysteines drive IL-38 amyloid formation.

Interleukin (IL)-1 family cytokines are essential for host defense at epithelial barriers. The IL-1 family member IL-33 was recently linked to stress granules (SGs). Formation of SGs and other biomolecular condensates is promoted by proteins containing low-complexity regions (LCRs). Computational analysis predicts LCRs in six of the 11 IL-1 family members. Among these, IL-38 contains a long LCR including two amyloid cores. IL-38 localizes to intracellular granules in keratinocytes under oxidative stress (OS) and forms OS-induced amyloid aggregates in cells and in vitro. Interestingly, soluble and aggregated IL-38 are released from keratinocytes in an exosome-enriched extracellular vesicle fraction. Disulfide-bond mapping, in silico modeling, and mutational analysis suggest that oxidation-sensitive cysteines act as redox switches to alter IL-38 conformation and promote its aggregation. Finally, the presence of IL-38 granules in human epidermis facing environmental OS suggests that oxidation-induced amyloidogenesis, as an intrinsic property of IL-38, supports barrier function.

Utilization of dissolved CO2 to control methane and acetate production in methanation reactor.

This study investigated the influence of dissolved CO2 on the selection of metabolic pathway using a methanation membrane bioreactor supplied with H2/CO2. Various ratios of H2/CO2 were applied (3.3, 3.8, 4.0, 4.5, and 5.0 (v/v)) to manipulate dissolved CO2 levels in the medium. The findings revealed a correlation between the concentration of dissolved CO2 and the production of CH4 (positive) and acetate (negative). Specifically, at a dissolved concentration of CO2 above 2.0 ±â€¯0.2 mmol/L, production of CH4 was favored. At the opposite, acetate production was favored at lower dissolved CO2 concentrations, with a maximum concentration of 1.9 g/L observed at 0.9 mmol/L of dissolved CO2. This study demonstrates that the modification of dissolved CO2 levels in a methanation bioreactor can provide a strategy for the selection of metabolic pathways and microbial communities, thereby offering a promising opportunity for optimizing the conversion of CO2 into high-value products such as CH4 and acetate.

30 Years' Experience in Percutaneous Pulmonary Artery Interventions in Transposition of the Great Arteries.

Background: Pulmonary artery (PA) stenosis is common after arterial switch operation (ASO) for transposition of the great arteries (TGA). Differences between balloon angioplasty (BA) and stents on right ventricular (RV) and PA pressures are not well studied. Objectives: The purpose of this study was to analyze percutaneous PA interventions' frequency after ASO, complications, and the effects of BA and stents on RV and PA pressures. Methods: All TGA patients with ASO between 1977 and 2022 in 2 Dutch congenital heart centers were included in this multicenter retrospective study. Peri-operative ASO characteristics and pre-intervention and post-intervention invasive and echocardiographic data were analyzed. Results: ASO was performed in 960 TGA patients, of which 888 survived 30 days and had complete follow-up. Seventy-seven (9%) underwent percutaneous PA interventions. Taussig-Bing anomaly (OR: 2.8; 95% CI: 1.228-6.168; P = 0.014), ASO time era 1990 to 1999 (OR: 4.7; 95% CI: 1.762-12.780; P = 0.002), and 2000 to 2009 (OR: 4.3; 95% CI: 1.618-11.330; P = 0.003) were independently associated with percutaneous PA interventions after ASO. Invasive post-interventional pressures and gradients were lower after stent implantation compared to BA (RV pressure: 47 ± 14 vs 58 ± 11; right PA-PA gradient: 11 ± 11 vs 25 ± 12, P < 0.05; RV/left ventricle pressure ratio: 0.4 ± 0.1 vs 0.6 ± 0.2, P < 0.001). Of the patients with unilateral PA stenosis (left PA: 41%, right PA: 59%), 77% showed increased RV pressure (>30 mm Hg) and RV/left ventricle pressure ratio improved post-intervention (0.5 ± 0.2 vs 0.6 ± 0.2, P < 0.05). Seventeen complications, most minor, were reported (13%). Two post-procedural deaths were reported. Conclusions: Percutaneous PA interventions are common after ASO and can be performed safely but caution for serious complications is warranted. Unilateral PA stenosis can impact RV pressures. Stents may be more successful at treating PA stenosis compared to BA.

Three-Level Chirality Transfer and Amplification in Liquid Crystal Supramolecular Assembly for Achieving Full-Color and White Circularly Polarized Luminescence.

Chiral liquid crystal supramolecular assembly provides an ideal strategy for constructing excellent circularly polarized luminescence (CPL) materials. However, the chirality transfer in chiral liquid crystals normally occurs at two levels from the configurational chirality to the supramolecular phase chirality. The more precise and more levels of chirality transmission are fascinating but remain challenging. The present work reports the first success of three-level chirality transfer and amplification from configurationally point chirality of small molecules to conformationally helical chirality of helical polymers and finally to supramolecular phase chirality of cholesteric liquid crystals composed of chiral nonfluorescent polymers (P46) and nematic liquid crystals. Noticeably, the helical twisting power of P46 is five-fold larger than its monomer. Full-color and white CPL with maximum luminescence dissymmetry factor up to 1.54 and photoluminescence quantum yield up to 63.8% are realized utilizing helical supramolecular assembly combined with selective reflection mechanism. Also significantly, the electrically stimuli-responsive CPL switching device as well as anti-counterfeiting security, information encryption, and chiral logic gate applications are developed. This study deepens the understanding of chirality transfer and amplification across different hierarchical levels.

Long-Term High-Fat Diet Aggravates Absence Seizures and Neurobehavioral Disorders Without Inducing Metabolic Disorders in WAG/Rij Rats: Involvement of Systemic and Central Inflammation.

The consumption of a high-fat diet (HFD) represents a risk factor for diseases such as obesity, diabetes, insulin resistance (IR), and different brain disorders. HFD-induced obesity is linked with systemic and neuroinflammation implicated in the pathogenesis of metabolic impairment and epilepsy. In this study, we studied the negative effects of HFD consumption (16 weeks) on absence epilepsy and behavior comorbidities in WAG/Rij rats, a well-validated idiopathic model of absence epilepsy and comorbidities. Moreover, we investigated how, by restoring a normocaloric diet (NCD; 12 weeks), epileptic seizures and neuropsychiatric comorbidities could improve. We found that the HFD group showed a worsening of absence seizures, aggravation of depressive-like behavior, and performance in learning and memory than the NCD group even in the absence of hyperglycemia and/or obesity. In addition, intestinal villus rupture, inflammatory infiltrate, and intestinal permeability alteration increased after prolonged HFD intake, which could prevent weight gain. Inflammatory protein levels were found higher in the colon of the HFD group than in the NCD group, and also in the cortex and hippocampus, regions involved in absence seizures and behavioral alterations. After replacing HFD with NCD, a reduction in absence seizures and behavioral alterations was observed, and this decrease was well correlated with an improvement in inflammatory pathways. In conclusion, HFD consumption is sufficient to disrupt gut integrity resulting in systemic and brain inflammation contributing to the worsening of absence epilepsy and its comorbidities also without obesity development. These alterations can be improved by switching back the diet to NCD.

CuBe: a geminivirus-based copper-regulated expression system suitable for post-harvest activation.

The growing demand for sustainable platforms for biomolecule manufacturing has fuelled the development of plant-based production systems. Agroinfiltration, the current industry standard, offers several advantages but faces limitations for large-scale production due to high operational costs and batch-to-batch variability. Alternatively, here, we describe the CuBe system, a novel bean yellow dwarf virus (BeYDV)-derived conditional replicative expression platform stably transformed in Nicotiana benthamiana and activated by copper sulphate (CuSO4), an inexpensive and widely used agricultural input. The CuBe system utilizes a synthetic circuit of four genetic modules integrated into the plant genome: (i) a replicative vector harbouring the gene of interest (GOI) flanked by cis-acting elements for geminiviral replication and novelly arranged to enable transgene transcription exclusively upon formation of the circular replicon, (ii) copper-inducible Rep/RepA proteins essential for replicon formation, (iii) the yeast-derived CUP2-Gal4 copper-responsive transcriptional activator for Rep/RepA expression, and (iv) a copper-inducible Flp recombinase to minimize basal Rep/RepA expression. CuSO4 application triggers the activation of the system, leading to the formation of extrachromosomal replicons, expression of the GOI, and accumulation of the desired recombinant protein. We demonstrate the functionality of the CuBe system in N. benthamiana plants expressing high levels of eGFP and an anti-SARS-CoV-2 antibody upon copper treatment. Notably, the system is functional in post-harvest applications, a strategy with high potential impact for large-scale biomanufacturing. This work presents the CuBe system as a promising alternative to agroinfiltration for cost-effective and scalable production of recombinant proteins in plants.

EXPRESS: Advancing an account of hierarchical dual-task control: A focused review on abstract higher-level task representations in dual-task situations.

Dual tasks are a common phenomenon in everyday life. In dual-task contexts, we perform two component tasks in temporal overlap, which usually results in impaired performance in one or both of these component tasks relative to single-task contexts. Numerous studies have examined dual-task interference at the level of response selection, but only few studies addressed the cognitive representation of a dual task and the cognitive mechanisms controlling these representations. The present review outlines recent empirical findings and theoretical developments concerning these two issues. In detail, the review focuses on different components of a cognitive dual-task representation, including the representation of component-task specific information (i.e., information about the goal and stimulus-response mapping of a component task), the representation of component-task order information (i.e., information about the order in which the component tasks have to executed), and the representation of dual-task identity information (i.e., information about which two component tasks have to be performed). A particular emphasis is placed on the cognitive representation of dual-task identity information, which is examined in a recent research line employing the task-pair switching logic as an empirical approach. By conceptualizing a dual-task representation as a hierarchical multi-component representation, the review integrates the research line on the cognitive representation of dual-task identity information with those on the representation of component-task specific information and of component-task order information. Based on this conceptualization, the review provides a new theoretical contribution to dual-task research and highlights an integrative perspective on the different components of cognitive dual-task representations.

Clinical Outcomes of Oral Antibiotic Switch in Children with Staphylococcus aureus Bacteremia.

Staphylococcus aureus is one of the leading causes of bacteremia in children. In this study, we aimed to evaluate our center's experience on the etiology, management, and outcomes of pediatric Staphylococcus aureus bacteremia (SAB) with particular focus on transitioning to oral antibiotic therapy. This retrospective cohort study included children aged ≤ 19 years diagnosed with SAB over a 5-year period. The main outcome was poor clinical outcome related to SAB defined as (1) recurrence of SAB within 30 days after discontinuation of SAB treatment and (2) any-cause mortality within 30 days after detection of SAB. Over a 5-year period, 88 SAB episodes of 76 unique patients were included. The most common source of SAB attributed to central line (n = 34), followed by osteoarticular (n = 24), infections. All patients received at least one day of intravenous (IV) antibiotics and treatment was switched to an oral agent in 45.5% of SAB episodes. Sources of SAB in the oral switch group were osteoarticular (n = 21), skin and soft tissue (n = 7), central line (n = 3), thrombophlebitis (n = 2), head and neck infection (n = 1), and unknown (n = 6). 30-day mortality and SAB recurrence within 30 days after initial treatment completion occurred in 3 and 5 SAB episodes, respectively. None of the patients in oral switch group had poor clinical outcomes. Our study results indicate that 30-day any-cause mortality and SAB-related mortality is low in children. Similar to growing adult literature, oral switch in SAB treatment was not associated with poor SAB outcomes in selected patients.

Examining modal and amodal language processing in proficient bilinguals: evidence from the modality-switch paradigm.

Recent discussions have emphasized the significance of embodied processing in language comprehension. Nevertheless, continuous debates persist regarding the relative contribution of modal (embodied) and amodal (abstract) processing of language. The current study investigated the contribution of modal processing in the first (L1) and second (L2) language, hypothesizing higher level of abstract amodal symbol processing in L2 than L1, since the correspondence of L1 and L2 (i.e., the symbol-to-symbol assignment) is in the foreground when learning L2. We employed the modality-switch paradigm (Pecher et al., 2003) in both German and French versions with proficient sequential German and French bilinguals (N = 79). Participants were presented with noun-adjective pairs (e.g., keys - jingling) in both languages and decided whether the adjective could be applied to the noun. This task repeatedly requires switching modality between trials, (e.g., from auditory [keys - jingling] to olfactory [soap - perfumed]), typically causing switch costs on response latency as compared to maintaining the modality. Contrary to the hypothesis, we observed modality switch effects (MSE) in both L1 and L2. This result suggests that embodied language processing occurs not only in the first language but also extends to the second language thus challenging the assumption that L2 processing predominantly involves abstract amodal symbol processing. Notably, however, significant L1 and L2 MSEs were found for French, whereas for German already the L1 effect was rather weak (though significant); the corresponding L2 effect was not significant. Thus, the results hinted at differences between languages regarding the relative role of modal and amodal processing.

REAL life study of subcutaneous SEMaglutide in patients with type 2 diabetes in SPain: Ambispective, multicenter clinical study. Results in the GLP1-experienced cohort.

OBJECTIVE: To evaluate the efficacy of switching to once-weekly subcutaneous semaglutide in patients with type 2 diabetes mellitus (T2DM) who were previously treated with other glucagon-like peptide-1 receptor agonists (GLP-1 RAs) in a real-world setting in Spain. METHODS: The REAL Life study of SEMaglutide in Patients with Type 2 diabetes in Spain (REALSEM-SP) was conducted in four endocrinology departments in Madrid, Spain. Adult patients with T2DM who were prescribed once-weekly (OW) subcutaneous semaglutide and had been previously treated with other GLP-1 RAs were included. Baseline characteristics, including demographic, anthropometric, and laboratory variables, were recorded at baseline and at 6 ± 3 and 12 ± 3 months of follow-up. The primary outcome was the change in HbA1c at 12 ± 3 months of follow-up, with secondary outcomes including changes in weight, BMI, and other glycemic parameters. RESULTS: A total of 267 patients were included in the analysis, with a mean age of 61.6 years and a mean T2DM duration of 11.3 years. The majority of patients had grade 1 or 2 obesity at baseline. Switching to OW-semaglutide was associated with a significant reduction in HbA1c from baseline to 13 months (-0.35 % ± 0.81). Patients who reached the 1.0 mg OW-dose showed a significant reduction in HbA1c compared to those on the ≤0.5 mg OW-dose. Significant reductions in weight, BMI, and fasting plasma glucose were also observed. Adverse events were mostly gastrointestinal and led to treatment withdrawal in few cases. CONCLUSION: Switching to OW-subcutaneous semaglutide in patients with T2DM previously treated with other GLP-1 RAs was associated to improvements in glycemic control and weight management in a real-world setting in Spain. These findings support the use of OW-semaglutide as an effective option for patients with T2DM who require additional glycemic control and weight management.

Long non-coding RNA GRASLND links melanoma differentiation and interferon-gamma response.

Melanoma is a highly malignant tumor, that stands as the most lethal form of skin cancer and is characterized by notable phenotypic plasticity and intratumoral heterogeneity. Melanoma plasticity is involved in tumor growth, metastasis and therapy resistance. Long non-coding RNAs (lncRNAs) could influence plasticity due to their regulatory function. However, their role and mode of action are poorly studied. Here, we show a relevance of lncRNA GRASLND in melanoma differentiation and IFNγ signaling. GRASLND knockdown revealed switching of differentiated, melanocytic melanoma cells towards a dedifferentiated, slow-proliferating and highly-invasive cell state. Interestingly, GRASLND is overexpressed in differentiated melanomas and associated with poor prognosis. Accordingly, we found GRASLND expressed in immunological "cold" tumors and it negatively correlates with gene signatures of immune response activation. In line, silencing of GRASLND under IFNγ enhanced the expression of IFNγ-stimulated genes, including HLA-I antigen presentation, demonstrating suppressive activity of GRASLND on IFNγ signaling. Our findings demonstrate that in differentiated melanomas elevated expression of GRASLND interferes with anti-tumor effects of IFNγ, suggesting a role of GRASLND in tumor immune evasion.

Previous reproductive success and environmental variation influence nest-site fidelity of a subarctic-nesting goose.

Nest-site fidelity is a common strategy in birds and is believed to be adaptive due to familiarity with local conditions. Returning to previously successful nest sites (i.e., the win-stay lose-switch strategy) may be beneficial when habitat quality is spatially variable and temporally predictable; however, changes in environmental conditions may constrain dispersal decisions despite previous reproductive success. We used long-term (2000-2017) capture-mark-reencounter data and hierarchical models to examine fine-scale nest-site fidelity of emperor geese (Anser canagicus) on the Yukon-Kuskokwim Delta in Alaska. Our objectives were to quantify nest-site dispersal distances, determine whether dispersal distance is affected by previous nest fate, spring timing, or major flooding events on the study area, and determine if nest-site fidelity is adaptive in that it leads to higher nest survival. Consistent with the win-stay lose-switch strategy, expected dispersal distance for individuals that failed their nesting attempt in the previous year was greater (207.7 m, 95% HPDI: 151.1-272.7) than expected dispersal distance for individuals that nested successfully in the previous year (125.5 m, 95% HPDI: 107.1-144.9). Expected dispersal distance was slightly greater following years of major flooding events for individuals that nested successfully, although this pattern was not observed for individuals that failed their nesting attempt. We did not find evidence that expected dispersal distance was influenced by spring timing. Importantly, dispersal distance was positively related to daily survival probability of emperor goose nests for individuals that failed their previous nesting attempt, suggesting an adaptive benefit to the win-stay lose-switch strategy. Our results highlight the importance of previous experience and environmental variation for informing dispersal decisions of a long-lived goose species. However, it is unclear if dispersal decisions based on previous experience will continue to be adaptive as variability in environmental conditions increases in northern breeding areas.

Efficacy, Safety, and Tolerability of Switching From Bictegravir/Emtricitabine/Tenofovir Alafenamide to Dolutegravir/Lamivudine Among Adults With Virologically Suppressed HIV: The DYAD Study.

Background: In TANGO and SALSA, switching to dolutegravir/lamivudine (DTG/3TC) was noninferior to continuing a baseline regimen among adults who were treatment experienced, although few switched from bictegravir (B) / emtricitabine (F) / tenofovir alafenamide (TAF). Here, we present the efficacy and safety of switching to DTG/3TC as compared with continuing with B/F/TAF among adults with virologic suppression. Methods: DYAD is an open-label clinical trial that randomized adults with HIV-1 RNA <50 copies/mL and no prior virologic failure (2:1) to switch to once-daily fixed-dose DTG/3TC or maintain B/F/TAF. The primary end point is the proportion with HIV-1 RNA ≥50 copies/mL at week 48 (Food and Drug Administration Snapshot algorithm, intention-to-treat exposed population, 6% noninferiority margin). Results: Overall, 222 adults were randomized (16% women, 51% aged ≥50 years, 28% Black). At week 48, 6 (4%) with DTG/3TC and 5 (7%) with B/F/TAF had HIV-1 RNA ≥50 copies/mL (treatment difference, -2.8%; 95% CI, -11.4% to 3.1%), meeting noninferiority criteria. Through week 48, 18 participants (12 with DTG/3TC, 6 with B/F/TAF) met confirmed virologic withdrawal (CVW) criteria, and 2 of 18 had resistance: 1 with B/F/TAF developed M184M/I and G140G/S at week 12, and 1 with DTG/3TC had M184V at week 12. One participant with DTG/3TC and non-CVW developed M184V and K65R at week 12. Drug-related adverse events (AEs) and withdrawals due to AEs occurred in 31 (21%) and 6 (4%) participants with DTG/3TC and 2 (3%) and 0 participants with B/F/TAF, respectively. Conclusions: Switching to DTG/3TC was noninferior to continuing B/F/TAF among adults with virologic suppression at week 48. Drug-related AEs and withdrawals were higher in the DTG/3TC arm, which is likely consistent with the open-label nature of this switch study.

Ductal stenting for retraining the left ventricle in patients with transposition of great arteries with intact ventricular septum: a single-centre experience.

INTRODUCTION: Ductal stenting in late presenters with transposition of great arteries with intact ventricular septum retrains the left ventricle before arterial switch operation. However, the experience is limited for its efficacy and safety. This study aims to highlight the efficacy and safety of ductal stenting for retraining the left ventricle. METHODS: Eight children with transposition of great arteries-intact ventricular septum and regressed left ventricle underwent ductal stenting. Serial echocardiographic measurements of left ventricle shape, mass, volume, free wall thickness, and function were done, and arterial switch operation was performed once the left ventricle was adequately prepared. Post-operative outcome in terms of duration of mechanical ventilation, ICU stay, and improvement in left ventricle function were monitored. RESULTS: The procedure was successful in all patients. Babies were divided into two groups on basis of age at ductal stenting (group 1 age less than 90 days and group 2 age more than 90 days) and were evaluated for the degree of left ventricle retraining as evidenced by echocardiographic parameters and post-operative variables. The left ventricle posterior wall thickness and mass index after ductal stenting increased significantly in both the groups. Postoperatively, one baby of group two expired after seven days due to severe left ventricle dysfunction. Rest babies had an uneventful post-operative ICU stay with no statistical difference in the duration of invasive mechanical ventilation or ICU stay. On six-month follow-up, all surviving babies were doing well with normal left ventricle function. CONCLUSION: Ductal stenting is a good alternative measure as compared to surgical procedures for left ventricle retraining in transposition of great arteries with regressed left ventricle.

Has Ghana's Rotavirus Vaccine Switch Met Programmatic Expectations? An Analysis of National Surveillance Data; 2018-2022.

Background: Ghana introduced a 2-dose schedule rotavirus vaccine, Rotarix, into childhood immunization in 2012 but switched to a 3-dose schedule vaccine, Rotavac, in 2020 on account of programmatic advantages offered by the latter, including lower cost per fully immunized child and lower cold chain volume requirement. The objective of the study was to assess the effect of the vaccine switch on the trends of rotavirus vaccine uptake and health facility outpatient department (OPD) attendance due to diarrhea among children aged 1-11 months. Methods: A retrospective analysis was conducted on childhood immunization and diarrhea surveillance data for 2018-2022. The uptake of the different rotavirus vaccine products and the proportion of health facility OPD attendance attributed to diarrhea, respectively, were compared between the pre- and postswitch study periods. Results: The uptake of rotavirus vaccine was sustained following the switch. There were no significant differences in vaccination coverages (rota1, Rotarix coverage [94.3%], vs rota1, Rotavac coverage [95.3%]; P = .757; rota2, Rotarix coverage [91.3%], vs rota2, Rotavac coverage [92.7%]; P = .789). The proportions of health facility OPD attendance due to diarrhea were comparable (preswitch [12.4%] vs postswitch [12.1%]; P = .838). Conclusions: Ghana's rotavirus vaccine switch yielded expected programmatic benefits without any untoward effects. The trends of vaccine uptake and reduction in diarrhea morbidity were sustained. These experiences and lessons from the rotavirus vaccine switch are vital for potential switches for other vaccines in the current immunization schedule to mitigate the annual vaccine expenditure.

Characterization of Interchanging Incretin Analogs in Clinical Practice: A Descriptive Report.

OBJECTIVE: To characterize the tolerability associated with incretin analog interchanges to equipotent or higher strengths based on an interchange process in an adult outpatient setting. METHODS: This was a retrospective chart review of adult patients receiving care through a participating family medicine or endocrinology clinic between January 1, 2022, and November 30, 2022 at a major academic medical center. An incretin analog equivalency table and protocol for interchange was created in response to on-going shortages and need for therapy adjustments to different medications within the same class. Patients were included if a recommended incretin analog interchange was initiated, and a tolerability assessment was conducted. Patients were excluded if they did not meet inclusion criteria or if they were unreachable for tolerability assessments for interchanged agents. RESULTS: There were 156 patients included for characterization and response to tolerability of interchange. It was determined that 96% of patients tolerated the incretin analog interchange. A dose escalation occurred in 58% of patients, 41% were transitioned to an equipotent dose, and 1 patient was considered a dose decrease. Prior authorizations were required 74% of the time for the new therapy. The most common interchanges were dulaglutide 4.5 mg to tirzepatide 7.5 mg, dulaglutide 4.5 mg to tirzepatide 10 mg, and dulaglutide 3 mg to tirzepatide 7.5 mg. These interchanges made up 37.3% of the total population and were observed to have 93% tolerability. Seven patients did not tolerate incretin analog interchange. Five experienced gastrointestinal effects and two experienced injection site reactions. The interchange of incretin analog was estimated to reduce time to maximum dose by a median of three months. During this study, no patients experienced interruption of therapy defined as missing a dose of incretin analog. CONCLUSIONS: This characterization report demonstrates an effective approach to addressing incretin analog interchanges. A high level of tolerability is evident with the defined interchange process. Future studies should continue to confirm effective and safe interchanges of incretin analogs from outcomes and tolerability reports.

Dissociation of the nuclear WWOX/TRAF2 switch renders UV/cold shock-mediated nuclear bubbling cell death at low temperatures.

BACKGROUND: Normal cells express functional tumor suppressor WW domain-containing oxidoreductase (WWOX), designated WWOXf. UV irradiation induces WWOXf cells to undergo bubbling cell death (BCD) - an event due to the accumulation of nuclear nitric oxide (NO) gas that forcefully pushes the nuclear and cell membranes to form one or two bubbles at room temperature (22 °C) and below. In contrast, when WWOX-deficient or -dysfunctional (WWOXd) cells are exposed to UV and/or cold shock, the cells undergo nuclear pop-out explosion death (POD). We aimed to determine the morphological and biochemical changes in WWOXf cells during BCD versus apoptosis. METHODS: WWOXf and WWOXd cells were exposed to UV followed by measuring BCD or POD by time-lapse microscopy and/or time-lapse holographic microscopy at 4, 22, or 37 °C to visualize morphological changes. Live cell stains were used to measure the kinetics of nitric oxide (NO) production and Ca2+ influx. Extent of cell death was measured by uptake of propidium iodide and by internucleosomal DNA fragmentation using agarose gel electrophoresis. RESULTS: WWOXf cells were exposed to UV and then cold shock, or cold shock and then UV, and cultured at 4, 10, and 22 °C, respectively. Initially, UV induced calcium influx and NO production, which led to nuclear bubbling and final death. Cold shock pretreatment completely suppressed UV-mediated bubbling at 37 °C, so the UV/cold shock-treated cells underwent apoptosis. Without cold shock, UV only induced bubbling at all temperatures, whereas the efficiency of bubbling at 37 °C was reduced by greater than 50%. Morphologically, the WWOXf cell height or thickness was significantly increased during cell division or apoptosis, but the event did not occur in BCD. In comparison, when WWOXd cancer cells received UV or UV/cold shock, these cells underwent NO-independent POD. UV/cold shock effectively downregulated the expression of many proteins such as the housekeeping α-tubulin (> 70%) and ß-actin (< 50%), and cortactin (> 70%) in WWOXf COS7 cells. UV/cold shock induced relocation of α-tubulin to the nucleus and nuclear bubbles in damaged cells. UV induced co-translocation of the WWOX/TRAF2 complex to the nuclei, in which the prosurvival TRAF2 blocked the proapoptotic WWOX via its zinc finger domain. Without WWOX, TRAF2 did not relocate to the nuclei. Cold shock caused the dissociation of the WWOX/TRAF2 complex in the nucleus needed for BCD. In contrast, the formation of the WWOX/TRAF2 complex, plus p53, was strengthened at 37 °C required for apoptosis. CONCLUSIONS: The temperature-sensitive nuclear WWOX/TRAF2 complex acts as a molecular switch, whose dissociation favors BCD at low temperatures, and the association supports apoptosis at 37 °C in UV-treated WWOXf cells.

Interleukin-1ß modulates lymphoid differentiation of Flt3-positive multipotent progenitors after transplantation.

Myeloablative pre-conditioning facilitates the differentiation of transplanted hematopoietic stem and progenitor cells (HSPCs). However, the factors in the stress environment that regulate HSPC behavior remain elusive. Here, we investigated the mechanisms that shaped the cell fates of transplanted murine multipotent progenitors (MPPs) expressing the Fms-related receptor tyrosine kinase 3 gene (Flt3). Using lineage tracing, clonal analysis, and single-cell RNA sequencing (RNA-seq), we showed that the myeloablative environment increased lymphoid priming of Flt3+ MPPs and that their efficient B cell output required intact interleukin 1 (IL-1) signaling. The Flt3+ MPPs with short-term exposure to IL-1ß underwent a myeloid-biased to lymphoid-biased cell fate switch and produced more lymphoid-biased progeny with a stronger B lymphopoiesis capacity in vitro. Correspondingly, a brief exposure to IL-1ß facilitated the B cell output of transplanted Flt3+ MPPs in vivo. Together, our study demonstrated an unrecognized function of IL-1ß in promoting B lymphopoiesis and highlighted a latent effect of IL-1ß in regulating MPP cell fate dynamics.

Insights into the distinct membrane targeting mechanisms of WDR91 family proteins.

WDR91 and SORF1, members of the WD repeat-containing protein 91 family, control phosphoinositide conversion by inhibiting phosphatidylinositol 3-kinase activity on endosomes, which promotes endosome maturation. Here, we report the crystal structure of the human WDR91 WD40 domain complexed with Rab7 that has an unusual interface at the C-terminus of the Rab7 switch II region. WDR91 is highly selective for Rab7 among the tested GTPases. A LIS1 homology (LisH) motif within the WDR91 N-terminal domain (NTD) mediates self-association and may contribute partly to the augmented interaction between full-length WDR91 and Rab7. Both the Rab7 binding site and the LisH motif are indispensable for WDR91 function in endocytic trafficking. For the WDR91 orthologue SORF1 lacking the C-terminal WD40 domain, a C-terminal amphipathic helix (AH) mediates strong interactions with liposomes containing acidic lipids. During evolution the human WDR91 ancestor gene might have acquired a WD40 domain to replace the AH for endosomal membrane targeting.

Effects of Switching FSH Preparations on Sperm Parameters and Pregnancy: A Prospective Controlled Study.

Objective: To study the effect of switching to a follicle-stimulating hormone (FSH) preparation other than that to which infertile male patients have not had an effective response. Patients and methods: Seventy-four normogonadotropinemic, non-obstructive, oligozoospermic patients who were poor responders to the administration of highly purified FSH (hpFSH) (Group 1 (n = 22) and Group 3 (n = 15)) or to recombinant human FSH (rhFSH) (Group 2 (n = 22) and Group 4 (n = 15)) were selected for this prospective study. After 3 months of washout from treatment with the first FSH preparation of choice, rhFSH was administered to patients in Groups 1 and 4 and hpFSH to those in Groups 2 and 3. Serum luteinizing hormone, FSH, total testosterone levels, conventional sperm parameters, testicular volume, and the number of pregnancies were evaluated at study entry and after the first and second treatment cycles. Results: Comparing treatment groups, the greatest improvement in sperm parameters was recorded in the groups of patients prescribed the switch in FSH preparation. Group 1 had the greatest benefit from therapy, with the highest pregnancy rate after the second treatment cycle. Indeed, eight couples achieved pregnancy (36.4%), compared to Groups 2 (n = 4; 18.2%), 3 (n = 1; 6.7%), and 4 (n = 2; 13.3%) (p = 0.04). Conclusions: The results of this study suggest that a therapeutic scheme involving the "switching" of the FSH preparation yields better results than a protocol using the same FSH preparation for six months. These findings, if confirmed by further studies, will help us better design a treatment strategy with FSH for infertile patients with oligozoospermia.