Search

Human embryonic kidney (HEK) 293 cells are widely used in protein and viral vector production owing to their high transfection efficiency, rapid growth, and suspension growth capability. Given their antiviral, anticancer, and immune-enhancing effects, type I interferons (IFNs) have been used to prevent and treat human and animal diseases. However, the binding of type I IFNs to the IFN-α and-β receptor (IFNAR) stimulates the expression of IFN-stimulated genes (ISGs). This phenomenon induces an antiviral state and promotes apoptosis in cells, thereby impeding protein or viral vector production. In this study, we generated an IFNAR subtype 1 knockout (KO) HEK 293 suspension (IFNAR-KO) cell line by using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) technology. Upon treatment with human IFN-α, the IFNAR-KO cells showed a constant expression of ISGs, including 2ʹ-5ʹ-oligoadenylate synthetase 1 (OAS1), myxovirus resistance 1 (Mx1), protein kinase RNA-activated (PKR), and IFN-induced transmembrane protein 1 (IFITM1), when compared with the wild-type HEK 293 (WT) cells, wherein the ISGs were significantly upregulated. As a result, the titer of recombinant adenovirus expressing porcine IFN-α was significantly higher in the IFNAR-KO cells than in the WT cells. Furthermore, the IFNAR-KO cells continuously produced higher amounts of IFN-α protein than the WT cells. Thus, the CRISPR-Cas9-mediated IFNAR1 KO cell line can improve the production efficiency of proteins or viral vectors related to IFNs. The novel cell line may be used for producing vaccines and elucidating the type I IFN signaling pathway in cells.

Foot-and-mouth disease (FMD) is an acute contagious infectious disease that affects cloven-hoofed animals. Although current emergency FMD vaccines only take effect 7 days after vaccination, antiviral agents, such as quercetin, which is a common flavonoid, could reduce the spread of FMD virus (FMDV) during outbreaks. We investigated the in vitro and in vivo antiviral effects of quercetin against FMDV. Analysis of viral copy numbers showed that quercetin had a dose-dependent inhibitory effect on FMDV at concentrations between 19.5 and 1,250 μM in porcine cells. In addition, we observed a quercetin-induced interferon (IFN)-α protein and interferon-stimulated gene (ISG) upregulation in swine cells. Enzyme-linked immunosorbent assay of sera revealed that quercetin induces the production of IFN-α, IFN-β, IFN-γ, interleukin (IL)-12, and IL-15 in mice. Inoculation of mice with quercetin or a combination of quercetin with an inactivated FMD vaccine enhanced both the survival rate and neutralizing antibody titer. Therefore, we suggest the use of quercetin as a novel and effective antiviral agent for controlling FMDV infection; however, further investigation of its application in livestock is required.

The study examined the effect of reciprocal teaching on students' reading comprehension, reading fluency, and metacognitive knowledge of reading strategies, especially among students with learning difficulties. The special focus was to assess how the increase in reading fluency and metacognitive knowledge during the intervention contributes to the reading comprehension outcome. The sample consisted of 301 Grade 3 students, of whom 77 had learning difficulties. Analyzes of (co)variances were used for estimating the effects of reciprocal teaching on the development of reading comprehension, reading fluency, and metacognitive knowledge. Multigroup path analysis was used for testing the effect of increase in reading fluency and metacognitive knowledge on reading comprehension. We found reciprocal teaching had a positive effect on reading comprehension, especially for students with learning difficulties. Reading fluency and metacognitive knowledge improved significantly, but similarly in both the intervention and control groups. However, the increase in metacognitive knowledge only contributed to reading comprehension in the intervention group, not in the control group. The study sheds light on the cognitive and metacognitive mechanisms underlying students' reading comprehension process, emphasizing metacognitive knowledge and especially the systematic practice of reading strategies as key factors in improving reading comprehension. [ABSTRACT FROM AUTHOR]

The Let's Talk about Children intervention is a tool for parents and professionals to work together to promote children's positive development, resilience, and psychosocial well-being in social and healthcare services, at school, and in day care. The aim of this study was to evaluate the fidelity, parents' experiences, and perceived benefits of using the Let's Talk about Children intervention in a school context. Participants (N = 65 first-grader parents) completed an online questionnaire after the intervention. The results show that the intervention was delivered as designed and conducted with high fidelity. Parents' experiences of the Let's Talk about Children discussions were positive, parents felt that the atmosphere was good during the discussion, and the participants reported benefits from the intervention. [ABSTRACT FROM AUTHOR]

One goal of synthetic biology is to provide well-characterised biological parts that behave predictably in genetic assemblies. To achieve this, each part must be characterised in a time-resolved manner under relevant conditions. Robotic platforms can be used to automate this task and provide sufficiently large and reproducible data sets including provenance. Although robotics can significantly speed up the data collection process, the collation and analysis of the resulting data, needed to reprogram and refine workflows for future iterations, is often a manual process. As a result, even in times of rapidly advancing artificial intelligence, the common design-build-test-learn (DBTL) cycle is still not circular without human intervention. To move towards fully automated DBTL cycles, we developed a software framework to enable a robotic platform to autonomously adjust test parameters. This interdisciplinary work between computer science and biology thus transforms a static robotic platform into a dynamic one. The software framework includes software components such as an importer that retrieves measurement data from the platform’s devices and writes it to a database. This is followed by an optimizer that selects the next measurement points based on a balance between exploration and exploitation. The platform is shown to be able to automatically and autonomously optimize the inducer concentration for a Bacillus subtilis system and the combination of inducer and feed release for a Escherichia coli system. As a target product the readily measurable green fluorescent reporter protein (GFP) is produced over multiple, consecutive iterations of testing. An evaluation of chosen (learning) algorithms for single and dual factor optimization was performed. In this article, we share the lessons learned from the development, implementation and execution of this automated design-build-test-learn cycles on a robotic platform.

IntroductionLupus nephritis (LN), caused by immune complexes produced in situ or deposited from the bloodstream, is one of the most severe features of Systemic Lupus Erythematosus (SLE) leading to an increased morbidity and mortality. Toll like receptors (TLRs), such as TLR3, TLR7 and TLR9, may play a key role in its pathogenesis. Interleukin-32 (IL-32), a cytokine involved in both innate and adaptive immune responses, has been widely considered in autoimmune-inflammatory rheumatic diseases. This study aims to evaluate the IL-32 role in LN, also investigating the effect of LN patients IgG (LN-IgG) on IL-32 production via TLR3.MethodsIn LN patients, IL-32 was detected in sera samples by ELISA KIT and in kidney tissue by immunohistochemistry. HEK293/T3 cells were incubated with LN-IgG and analyzed for TBK1, phospho-p65 NF-κB and IL-32 by Western blot.ResultsWe demonstrated IL-32 presence in LN patients compared to SLE patients without renal involvement, observing a direct correlation between IL-32 serum levels and disease duration (p=0.02; r 0.2978). Moreover, IL-32 was strongly expressed in renal samples of LN patients. Phosphorylation of TBK1 resulting in NF-κB activation and IL-32 increase was observed in HEK293/T3 cells following LN-IgG treatment, TLR3 inhibitor using induced a significant reduction in the expression of these molecules.DiscussionThese results showed that IL-32 is up-regulated in the kidney of LN patients suggesting that in renal tissue IL-32 expression could be induced through TLR3 activation by the LN patients’ antibodies. This study may indicate a possible role for IL-32 in the pathogenesis of LN.

Gestational trophoblastic neoplasms are tumors that occur during pregnancy, while non-gestational trophoblastic tumors have a similar histology but are present outside of gestation. Literature reports several cases of non-gestational trophoblastic tumors of primary pulmonary origin, which pose diagnostic challenges and are associated with a poor prognosis. This report details a case of somatic high-grade carcinoma with trophoblastic differentiation primarily manifesting in the left lung with recurrent pneumothoraces. The tumor was initially diagnosed as a poorly differentiated pleomorphic carcinoma and was treated with paclitaxel and pembrolizumab, followed by the EMA-CO/EP regimen after the detection of liver, lung, and brain metastases. Despite initial treatment responses, the disease progressed with widespread metastases and severe complications, including myelotoxicity, empyema, and subarachnoid bleeding. The disease progressed rapidly, resulting in death within two years of diagnosis, highlighting the aggressive nature of this high-grade carcinoma with trophoblastic differentiation. Non-gestational trophoblastic tumors may represent a distinct disease group with unique clinical characteristics, and genetic analysis could help identify more cases.

BackgroundCancer remains a predominant cause of death worldwide. The advent of effective chemotherapy has enormously decreased the mortality rate and increased the life expectancy of cancer patients. However, the adverse effects allied with chemotherapy contribute to the development of neurotoxicity, anxiety, and depression.ObjectiveThe dual effects of a novel flavanone, 6-hydroxyflavanone (6-HF), were investigated in treating chemotherapy-induced neuropathy along with anxiolytic propensity.MethodsThe anti-neuropathy propensity of 6-HF was evaluated utilizing the cisplatin-induced neuropathy (CIN) model, whereas its anxiolytic activity was evaluated utilizing anxiety models, such as the elevated plus maze test (EPM), the staircase test, and the open-field paradigm.ResultCisplatin administration induced static and dynamic allodynia in the rats. Concomitant administration of 6-HF and cisplatin for four successive weeks remarkably reduced the chemotherapy-induced mechanical allodynia, evident from an elevation in the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL). The anxiolytic-like activity of 6-HF in the EPM apparatus was confirmed by the increased number of entries in the open arm and time spent at the central platform, which was further confirmed by the enhanced head-dipping frequency in the same assay. A decrease in rearing behavior of the mice without suppression of the steps ascended further assured the anxiolytic-like potential of 6-HF. Additionally, the animals under investigation spent more time at the intersection of the open-field apparatus, further confirming the anxiolytic potential of 6-HF.Conclusion6-HF might be considered a potential therapeutic agent for counteracting two common adverse effects of chemotherapy, neurotoxicity and anxiety.

BackgroundThe impact of steroid-sparing immunosuppressive agents (SSIAs) for immune-related adverse events (irAEs) on tumor outcome is not well-known. This systematic review evaluates tumor outcomes for corticosteroid (CS) monotherapy versus CS with SSIA (CS-SSIA) for irAE treatment with a focus on melanoma.MethodsSearch was conducted through 1/5/23 using PubMed, Embase, Cochrane CENTRAL, and Web of Science. We included case series, retrospective/prospective observational studies and interventional clinical trials. Individual-level data was analyzed using KM curves and Cox regression for overall survival (OS) and progression free survival (PFS). Time to SSIA was treated as a time-varying exposure using landmark analysis (landmark timepoint=3 months after irAE) to account for immortal time bias. For group-level data, meta-analysis compared the use of SSIA to No SSIA for irAEs.ResultsOf twenty-two publications with individual-level data, 147 patients with any cancer (57 CS, 90 CS-SSIA) and 65 with melanoma (18 CS, 47 CS-SSIA) underwent landmark analysis. Twenty-two publications underwent group-level evaluation and four were included in the meta-analysis. CS-SSIA versus CS showed higher risk of all-cause mortality and progression (HR 2.75, 95%CI: 1.44-5.27, p

Background and hypothesisA static predictive model relying solely on baseline clinicopathological data cannot capture the heterogeneity in predictor trajectories observed in the progression of chronic kidney disease (CKD). To address this, we developed and validated a dynamic survival prediction model using longitudinal clinicopathological data to predict end-stage kidney disease (ESKD), with death as a competing risk.MethodsWe trained a sequence of random survival forests using a landmarking approach and optimized the model with a pre-specified prediction horizon of 5 years. The predicted cumulative incidence function (CIF) values were used to generate a personalized dynamic prediction plot.ResultsThe model was developed using baseline demographics and 13 longitudinal clinicopathological variables from 4,950 patients. Variable importance analysis for ESKD and death informed the creation of a sequence of reduced models that utilized six key variables: age, serum albumin, bicarbonate, chloride, eGFR, and hemoglobin. The models demonstrated robust predictive performance, with a median concordance index of 84.84% for ESKD and 84.1% for death. The median integrated Brier scores were 0.03 for ESKD and 0.038 for death across all landmark times. External validation with 8,729 patients confirmed these results.ConclusionWe successfully developed and validated a dynamic survival prediction model using common longitudinal clinicopathological data. This model predicts ESKD with death as a competing risk and aims to assist clinicians in dialysis planning for patients with CKD.

Magnesium phosphate cement (MPC) is known as a high-performing rapid repair material for concretes in practice, and has also become a fast-moving research area for other applications, although its application has not yet reached its full potential. MPC has been utilized as a repair material in certain applications, although it is still less commonly used/recommended than the most popular and standardized repair materials. Therefore, it is important to systematically investigate the suitability of this material in the construction sector as a repair material. This review provides an in-depth summary of the engineering properties of magnesium potassium phosphate cements (MKPCs), as a particular example of MPCs that do not suffer from the undesirable release of ammonia which is observed in some other MPC formulations. Key properties relevant to the use of MKPCs as repair materials, from the fresh state to the hardened state are systematically examined. On this basis, the suitability of MKPC as a potential repair material is explored, highlighting the state-of-the-art practices, challenges and drawbacks in engineering applications, and future perspectives.

IntroductionAs 3D design and printing technology gains popularity, there remains limited evidence on dental students' perceptions in Saudi Arabia regarding its use. This study aims to assess the knowledge and awareness of dental students in Saudi Arabia about 3D design and printing technology.MethodsA cross-sectional questionnaire-based study was conducted among dental students in their third, fourth, and fifth years at multiple universities in Saudi Arabia between February and October 2023. A validated self-administered questionnaire with 15 close-ended items, including demographic and knowledge-related questions about 3D design and printing, was used. Statistical analysis was performed using Chi-square and Fisher exact tests to identify factors associated with knowledge and awareness levels.ResultsA total of 374 dental students participated in the study, with 63.1% being female students. Of the participants, 40.4% identified the cost of equipment as the primary barrier to 3D printing usage in dentistry in Saudi Arabia. The majority (94.4%) recognized the advantages of 3D models for implant placement. Awareness of 3D printing utilization in the field was reported by 69.3% of participants, and 74.3% acknowledged its increasing popularity. Furthermore, 73.8% of participants expressed confidence in the biocompatibility and safety of 3D printed materials. A significant percentage (64.0%) were aware of 3D printing's role in creating Invisalign aligners, and 42.4% believed in the value of 3D printed drill guides for root canal treatment. The study found statistically significant regional differences (p

IntroductionSarcopenia is associated with poor clinical outcomes in chronic diseases. Our study aimed to characterize body composition (BC) parameters in patients with inflammatory bowel disease (IBD) and compare skeletal muscle mass (SMM) parameters with the healthy pediatric population.MethodsBC of healthy controls (HC) and of patients with IBD were measured via multifrequency bioelectrical impedance (InBody 720 device) in a cross-sectional manner. The effect of sex, age, height, weight, and body mass index (BMI) on BC parameters, with a special attention to SMM, was assessed. Reference tables from SMM were generated using a maximum-likelihood curve-fitting technique for calculating Z scores.ResultsBC parameters were associated with age, body size, and sex. SMM was lower in patients with IBD (n = 57, aged 6.71 ± 8.7 years) compared to unadjusted HC (n = 307, aged 9.9–19.3 years; 143 males; SMM: 22.34 ± 8.38 vs. 24.4 ± 6.3 kg; p = 0.03). SMM showed a moderately strong correlation with age, weight, height, and BMI (R = 0.65, 0.9, 0.87, and 0.66; p

BackgroundEarly coronary angiography (CAG) in post-cardiac arrest patients without ST-segment elevation is a topic of debate. This meta-analysis aimed to assess its impact on outcomes.MethodsA search of Medline and Cochrane up to February 2023 was conducted to identify randomized controlled trials and observational studies comparing patients undergoing early CAG vs. delayed/no CAG after experiencing out-of-hospital cardiac arrest. A random-effects model pooled odds ratios (ORs) with 95% confidence intervals (CIs). Meta-regression explored factors modifying effect sizes.ResultsWe identified 16 studies (7 RCTs, 9 observational studies) involving 4,737 patients. Early CAG significantly reduced long-term mortality [OR: 0.66 (0.51–0.85)], and increased favorable cerebral performance category (CPC) 1–2 at discharge [OR: 1.49 (1.09–2.03)]. Observational study subgroup showed decreased short-term mortality, long-term mortality, and CPC 1–2 at discharge, unlike RCT subgroup. Meta-regression revealed type 2 diabetes mellitus and follow-up time influencing short-term mortality and CPC 1–2 at discharge, respectively.ConclusionEarly CAG in post-cardiac arrest patients without ST elevation is associated with long-term clinical benefits, particularly evident in observational studies. Interpretation should be cautious.

Sustainable Indigenous resource use reflects balance between animal populations and levels of human consumption, influenced by natural cycles of faunal abundance, community size and subsistence needs, procurement technologies, and the requirements of trade or commodity production. Sustainability is “epiphenomenal” when animal populations are preserved, and community needs met, without deliberate measures to prevent overharvesting. Alternatively, Indigenous conservation—cultural practices that moderate use of a resource to prevent its depletion—may play a determinative role. In this study from the Tlingit community of Yakutat, Alaska in the Northwest Coast cultural region, we interweave Indigenous and scientific perspectives to trace the use and conservation of harbor seals (Phoca vitulina) from before Western contact through the Russian and American colonial periods to the present. Harbor seals, which concentrate in large numbers at a summer ice floe rookery near Hubbard Glacier, are the community's most important subsistence food and a key to its culture and history. The Smithsonian Institution and Yakutat Tlingit Tribe undertook collaborative research in historical ecology and archaeology in 2011–2014 including oral interviews with elders and subsistence providers, excavations at sealing sites, archaeofaunal analysis, historical and archival research, and consideration of climate cycles and biological regime shifts that influence the harbor seal population in the Gulf of Alaska. We compare technologies and hunting practices before and after Western contact, estimate harvest levels in different periods, and evaluate the effectiveness of traditional conservation practices that included hunting quotas enforced by clan leaders and the seasonal delay of hunting with firearms to prevent abandonment of the rookery by the seal herd.

IntroductionCardiac surgery-associated acute kidney injury (CSA-AKI) is a well-known complication that increases morbidity and mortality rates. The objective of this study was to reduce CSA-AKI through nephrologist intervention in patients awaiting cardiac surgery.MethodsWe performed a single center, open-label, randomized clinical trial including 380 patients who underwent scheduled cardiac surgery at the Hospital de Bellvitge between July 2015 and October 2019. A total of 184 patients were evaluated by the same Nephrologist one month before the surgery to minimize the risk factors for AKI. In addition to assessments at the outpatient clinic, we also collected clinical data during hospitalization and during the first year.ResultsDespite the intervention, no differences were observed between the groups in the incidence of CSA-AKI (intervention group 26.37% vs. standard of care 25.13%, p=0.874), mortality (3.91% vs. 3.59%, p=0.999), length of Intensive Care Unit (ICU) stay (10 days [7.00;15.0] for both groups, p=0.347), or renal function after one year of follow-up (estimated glomerular filtration rate (eGFR) by CKD-EPI: 74.5 ml/min (standard deviation 20.6) vs 76.7 (20.8) ml/min, respectively, p=0.364). A reduction in the need for blood transfusion was observed in the intervention group, although the difference was not statistically significant (37.22% vs. 45.03%, p =0.155).ConclusionIn this clinical trial, nephrologist intervention in the entire population on the cardiac surgery waiting list did not show a nephroprotective benefit.Clinical trial registrationClinicalTrials.gov, identifier (NCT02643745).

The pulmonary microbiome has emerged as a significant factor in respiratory health and diseases. Despite the sterile conditions maintained during ex vivo lung perfusion (EVLP), the use of antibiotics in the perfuse liquid can lead to dynamic changes in the lung microbiome. Here, we present the design of a study that aims to investigate the hypothesis that EVLP alters the lung microbiome and induces tissue inflammation. This pilot, prospective, controlled study will be conducted in two Spanish donor centers and will include seven organ donors after brain death or after controlled cardiac death. After standardized retrieval, the left lung will be preserved in cold storage and the right lung will be perfused with EVLP. Samples from bronchoalveolar lavage, perfusion and preservation solutions, and lung biopsies will be collected from both lungs and changes in lung microbiome and inflammatory response will be compared.

PurposeThe focus of this article is to describe the configuration, testing, and commissioning of a novel gantry-less synchrotron-based proton therapy (PT) facility.Materials and methodsThe described PT system delivers protons with a water equivalent range between 4 and 38 cm in 1800 energy layers. The fixed beam delivery permits a maximum field size of 28 × 30 cm2. The patient positioning and imaging system includes a six-degree-of-freedom robotic arm, a convertible patient chair, a vertical 4DCT, and an orthogonal 2D X-ray imaging system.ResultsThe spot positioning reproducibility was consistent within ±1 mm. The width (σ) of the beam profile at the isocenter was energy dependent and ranged from 2.8 mm to 7.7 mm. Absolute dose reproducibility was measured and deviations were found to be

According to the modern expectancy-value theory, students' task values may differ across domains, manifesting as varying motivational patterns. In middle school, students' motivation becomes increasingly apparent and may direct their future occupational aspirations. Using a person-oriented approach, this study examines students' self-concept, and positive and negative task values (i.e., utility value, intrinsic value, and emotional cost) across Finnish language, math, biology, and physics, and the stability of the identified profiles. Further, the associations of the profiles with students' subsequent academic achievement and math and natural science, technology, engineering, and mathematics (STEM)/health science STEM aspirations, and gendered effects were examined. Longitudinal data was collected through Grades 7 to 9 in 21 middle schools in Helsinki, Finland (N D 1,309, N D 1,179, N D 818, respectively; age 13-15 years; 55.9% female). Latent profile analysis (LPA) identified four task value profiles in Grades 7 and 8: Low motivation high cost STEM (13%/13%) showed low task values with high cost, especially in math and physics; High motivation low cost STEM (7%/8%) showed the highest task values with the lowest cost, especially in math and physics; High motivation high cost (18%/17%) showed high task values and cost across domains; and Moderate motivation and cost (62%/62%) showed moderate task values and cost across domains. The latent transition analysis identified Moderate motivation and cost as the most stable profile across 2 years. In comparison to the other profiles, students with a Low motivation high cost STEM profile were less likely to have STEM aspirations in Grade 9. These results suggests that majority of middle school students are highly to moderately motivated in various domains, however, some students simultaneously experience high cost. It may reflect the increasingly difficult courses and study demands in middle school. [ABSTRACT FROM AUTHOR]

Coping with changes in light intensity is challenging for plants, but welldesigned mechanisms allow them to acclimate to most unpredicted situations. The thylakoid K+/H+ antiporter KEA3 and the voltage-dependent Cl− channel VCCN1 play important roles in light acclimation by fine-tuning electron transport and photoprotection. Good evidence exists that the thylakoid Cl− channel ClCe is involved in the regulation of photosynthesis and state transitions in conditions of low light. However, a detailed mechanistic understanding of this effect is lacking. Here we report that the ClCe loss-offunction in Arabidopsis thaliana results in lower levels of phosphorylated lightharvesting complex II (LHCII) proteins as well as lower levels of the photosystem I-LHCII complexes relative to wild type (WT) in low light conditions. The phosphorylation of the photosystem II core D1/D2 proteins was less affected either in low or high light conditions. In low light conditions, the steady-state levels of ATP synthase conductivity and of the total proton flux available for ATP synthesis were lower in ClCe loss-of-function mutants, but comparable to WT at standard and high light intensity. As a long-term acclimation strategy, expression of the ClCe gene was upregulated in WT plants grown in light-limiting conditions, but not in WT plants grown in standard light even when exposed for up to 8 h to low light. Taken together, these results suggest a role of ClCe in the regulation of the ATP synthase activity which under low light conditions impacts LHCII protein phosphorylation and state transitions. [ABSTRACT FROM AUTHOR]

Bruxism is a complex orofacial behavior that can occur during sleep or wakefulness, characterized by the involuntary grinding or clenching of teeth, involving repetitive activity of the jaw muscles. Its etiology is multifactorial, influenced by genetic, psychological, physiological, and lifestyle factors. While the mild bruxism may not necessitate treatment, severe bruxism can lead to significant consequences, including tooth damage, jaw pain, fatigue, and headaches. The bruxism has been associated with medical conditions, such as stress, anxiety, sleep disorders, and various neurological disorders; however, the exact pathophysiology remains elusive. Although the central nervous system is strongly implicated in the development of bruxism, specific neural substrates have not yet been conclusively established. Furthermore, there is evidence to suggest that individuals with bruxism may exhibit neural plasticity, resulting in the establishment of distinct neural circuitry that control the jaw movements. The application of various neurophysiological techniques in both clinical and pre-clinical studies provides valuable insights into the neural mechanisms underlying bruxism. This review aims to comprehensively examine the current literature on the neural pathways involved in bruxism, with the goal of improving the clinical approach and therapeutics for this condition. A deeper understanding of the neural circuitry controlling bruxism holds the potential to advance future treatment approaches and improve the management of patients with bruxism.

IntroductionAlthough Campylobacter spp., including Campylobacter coli, have emerged as important zoonotic foodborne pathogens globally, the understanding of the genomic epidemiology of C. coli of porcine origin is limited.MethodsAs pigs are an important reservoir of C. coli, we analyzed C. coli genomes that were isolated (n = 3) from pigs and sequenced (this study) them along with all other C. coli genomes for which pig intestines, pig feces, and pigs were mentioned as sources in the NCBI database up to January 6, 2023. In this paper, we report the pan-genomic features, the multi-locus sequence types, the resistome, virulome, and mobilome, and the phylogenomic analysis of these organisms that were obtained from pigs.Results and discussionOur analysis revealed that, in addition to having an open pan-genome, majority (63%) of the typeable isolates of C. coli of pig origin belonged to a single clonal complex, ST-828. The resistome of these C. coli isolates was predominated by the genes tetO (53%), blaOXA-193 (49%), and APH (3′)-IIIa (21%); however, the virulome analysis revealed a core set of 37 virulence genes. Analysis of the mobile genetic elements in the genomes revealed wide diversity of the plasmids and bacteriophages, while 30 transposons were common to all genomes of C. coli of porcine origin. Phylogenomic analysis showed two discernible clusters comprising isolates originating from Japan and another set of isolates comprising mostly copies of a type strain stored in three different culture collections.

IntroductionAccurate taxonomic profiling of the human microbiome composition is crucial for linking microbial species to health outcomes. Therefore, we created the Clinical Microbiomics Human Microbiome Profiler (CHAMP), a comprehensive tool designed for the profiling of prokaryotes, eukaryotes, and viruses across all body sites.MethodsCHAMP uses a reference database derived from 30,382 human microbiome samples, covering 6,567 prokaryotic and 244 eukaryotic species, as well as 64,003 viruses. We benchmarked CHAMP against established profiling tools (MetaPhlAn 4, Bracken 2, mOTUs 3, and Phanta) using a diverse set of in silico metagenomes and DNA mock communities.ResultsCHAMP demonstrated unparalleled species recall, F1 score, and significantly reduced false positives compared to all other tools benchmarked. The false positive relative abundance (FPRA) for CHAMP was, on average, 50-fold lower than the second-best performing profiler. CHAMP also proved to be more robust than other tools at low sequencing depths, highlighting its application for low biomass samples.DiscussionTaken together, this establishes CHAMP as a best-in-class human microbiome profiler of prokaryotes, eukaryotes, and viruses in diverse and complex communities across low and high biomass samples. CHAMP profiling is offered as a service by Clinical Microbiomics A/S and is available for a fee at https://cosmosidhub.com.

IntroductionThe coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has posed extraordinary challenges to global health systems and economies. The virus’s rapid evolution has resulted in several variants of concern (VOCs), including the highly transmissible Omicron variant, characterized by extensive mutations. In this study, we investigated the genetic diversity, population differentiation, and evolutionary dynamics of the Omicron VOC during the fifth wave of COVID-19 in Pakistan.MethodsA total of 954 Omicron genomes sequenced during the fifth wave of COVID-19 in Pakistan were analyzed. A Bayesian framework was employed for phylogenetic reconstructions, molecular dating, and population dynamics analysis.ResultsUsing a population genomics approach, we analyzed Pakistani Omicron samples, revealing low within-population genetic diversity and significant structural variation in the spike (S) protein. Phylogenetic analysis showed that the Omicron variant in Pakistan originated from two distinct lineages, BA.1 and BA.2, which were introduced from South Africa, Thailand, Spain, and Belgium. Omicron-specific mutations, including those in the receptor-binding domain, were identified. The estimated molecular evolutionary rate was 2.562E-3 mutations per site per year (95% HPD interval: 8.8067E-4 to 4.1462E-3). Bayesian skyline plot analysis indicated a significant population expansion at the end of 2021, coinciding with the global Omicron outbreak. Comparative analysis with other VOCs showed Omicron as a highly divergent, monophyletic group, suggesting a unique evolutionary pathway.ConclusionsThis study provides a comprehensive overview of Omicron’s genetic diversity, genomic epidemiology, and evolutionary dynamics in Pakistan, emphasizing the need for global collaboration in monitoring variants and enhancing pandemic preparedness.

BackgroundAngiotensin-converting enzyme inhibitors (ACEIs) are prescribed for individuals with high cardiovascular (CV) risk; however, persistent cough limits the use of ACEIs in a large number of patients. The current study aimed to identify the genetic variants in the SLCO1B1 gene that might be associated with ACEI-related cough in a Pakistani hypertensive population.MethodsA prospective cohort study was conducted at a tertiary care hospital in Pakistan. A total of 74 patients who had been treated with ACEIs were recruited through a convenient sampling method. The study was approved by the Institutional Review Board & Ethics Committee of the Shifa International Hospital, Islamabad. Patients provided 2 ml of blood for sequencing after signing informed consent. Partial gene sequencing of SLCO1B1 was carried out to find single nucleotide polymorphisms (SNPs) and haplotypes.ResultsIt was found, through a structured questionnaire, that thirty-eight (38) patients experienced cough within 2 weeks of ACEI administration and were considered as a case group (cough), and thirty-six (36) patients were considered as a control group (no cough). The incidence of cough was 51%. We found six different SNPs and 9 haplotypes in the partial gene sequences of SLCO1B1. Haplotype H4 was associated significantly with cough after adjusting for sex and smoking status. Other SNPs and haplotypes were not significantly associated with ACE-Is-induced cough.ConclusionThese findings emphasize the significance of SLCO1B1 genetic variants, specifically H4, as a potential predictor of ACEI-induced cough. It could be included in clinical practice as a possible risk factor for ACEI-induced cough once confirmed in larger clinical trials with bigger sample sizes. The replication of these findings in larger and more diverse populations is likely to contribute to the therapeutic use of ACEIs by predicting ACEI-induced cough.

This methods paper describes the methodological and statistical underpinnings of the highly adaptive testing design (HAT), which was developed for the Programme for International Student Assessment (PISA). The aim of HAT is to allow for a maximum of adaptivity in selecting items while taking the constraints of PISA into account with appropriate computer algorithms. HAT combines established methods from the area of computerized adaptive testing (a) to improve item selection when items are nested in units, (b) to make use of the correlation between the dimensions measured, (c) to efficiently accomplish constraint management, (d) to control for item position effects, and (e) to foster students’ test-taking experience. The algorithm is implemented using the programming language R and readers are provided with the necessary code. This should facilitate future implementations of the HAT design and inspire other adaptive testing designs that aim to maximize adaptivity while meeting constraints.

BackgroundAcute Type A aortic dissection (ATAAD) with supra-aortic branch (SAB) malperfusion remains a formidable clinical challenge, often resulting in high mortality and complex treatment dilemmas. The introduction of the AMDS represents a significant innovation, designed to stabilize the aortic arch, and manage malperfusion effectively.MethodsThis case study evaluates the utility of AMDS in the treatment of a 63-year-old male with hypertension, who presented with severe, acute chest pain. Diagnosed with a DeBakey type I ATAAD involving SAB, the patient underwent cardiopulmonary bypass, aortic root replacement, aortic arch repair with AMDS implantation, and subsequent endovascular stenting for severe left common carotid artery malperfusion that developed postoperatively. The AMDS was instrumental in facilitating crucial aortic arch reconstruction and addressing the initial severe malperfusion. Despite postoperative cerebral malperfusion, targeted endovascular stenting resulted in a rapid and substantial neurological recovery. The patient was discharged to a rehabilitation facility on postoperative day 20, free of neurological deficits.ConclusionsThe use of AMDS in managing ATAAD with SAB involvement is transformative, enabling less invasive surgical techniques and offering immediate, effective correction of malperfusion. This case underscores the essential role of integrating advanced endovascular strategies to enhance outcomes in high-risk aortic surgeries, marking a pivotal advancement in the therapeutic approach to complex aortic dissections.

Objective: This study investigated the effects of a school-based skill-training intervention in attention control and executive functions for pupils with hyperactivity-impulsivity (H-I) and cognitive control (CC) deficits. The main aim was to examine whether the intervention differently influenced H-I and CC, and whether cognitive abilities or conduct problems moderated response to the intervention. Method: Elementary school pupils from 41 schools participated the study and were divided into an intervention group (n = 71) and a waitlist control group (n = 77). Intervention outcomes were assessed with an inventory assessing executive function difficulties (including H-I and CC) completed by classroom teachers. Results: Significant intervention effects and positive changes were detected in CC but not in H-I. Significant intervention effects were found mainly among pupils with low levels of conduct problems. Conclusions: The results suggest that a skill-training intervention has specific positive effects on CC, but conduct problems may diminish response to intervention. [ABSTRACT FROM AUTHOR]

To better understand the gender gap in science, technology, engineering and math (STEM) aspiration, the article examines the critical role of domain-specific motivation (i.e., expectancy and task values). Using longitudinal data from 5th and 6th grade (~11–12-year-old) students (n = 360, 55% girls), person-oriented analyses was applied to understand the gendered motivational profiles and their longitudinal influence on achievement and STEM aspiration. Specifically, we aimed to (1) derive motivational belief profiles regarding science, mathematics, and language (Finnish), (2) analyze the stability and change in the profiles between the 5th and 6th grade, (3) assess the relationship between motivational profiles and achievement and STEM aspiration, and (4) test for gender differences. We derived four motivational profiles for both years: high motivation in all subjects (~21%), high mathematics motivation (~46%), low mathematics motivation (~11%), and low motivation in all subjects (~8%). Latent transition analysis revealed that most students remained in the same profile throughout the 2 years. We found evidence of gendered differences in the motivational profiles and the chance of transitioning between profiles. More girls are characterized by low math motivation, while boys are more likely to transition to higher math motivation in 6th grade. The motivational difference is reflected in their achievement, although not strongly coupled with their STEM aspiration. The findings suggest that at this developmental stage, Finnish students have not developed a strong association between (gendered) STEM aspiration and their domain-specific motivation, although their motivation may have influenced their achievement. Interpretation and practical implications are discussed. [ABSTRACT FROM AUTHOR]

Previous studies have examined the neural correlates of receiving negative feedback from peers during virtual social interaction in young people. However, there is a lack of studies applying platforms adolescents use in daily life. In the present study, 92 late-adolescent participants performed a task that involved receiving positive and negative feedback to their opinions from peers in a Facebook-like platform, while brain activity was measured using functional magnetic resonance imaging (fMRI). Peer feedback was shown to activate clusters in the ventrolateral prefrontal cortex (VLPFC), medial prefrontal cortex (MPFC), superior temporal gyrus and sulcus (STG/STS), and occipital cortex (OC). Negative feedback was related to greater activity in the VLPFC, MPFC, and anterior insula than positive feedback, replicating previous findings on peer feedback and social rejection. Real-life habits of social media use did not correlate with brain responses to negative feedback. [ABSTRACT FROM AUTHOR]

Proteomes of an oxygenic photosynthetic cyanobacterium, Synechocystis sp. PCC 6803, were analyzed under photoautotrophic (low and high CO2, assigned as ATLC and ATHC), photomixotrophic (MT), and light-activated heterotrophic (LAH) conditions. Allocation of proteome mass fraction to seven sub-proteomes and differential expression of individual proteins were analyzed, paying particular attention to photosynthesis and carbon metabolism–centered sub-proteomes affected by the quality and quantity of the carbon source and light regime upon growth. A distinct common feature of the ATHC, MT, and LAH cultures was low abundance of inducible carbon-concentrating mechanisms and photorespiration-related enzymes, independent of the inorganic or organic carbon source. On the other hand, these cells accumulated a respiratory NAD(P)H dehydrogenase I (NDH-11) complex in the thylakoid membrane (TM). Additionally, in glucose-supplemented cultures, a distinct NDH-2 protein, NdbA, accumulated in the TM, while the plasma membrane-localized NdbC and terminal oxidase decreased in abundance in comparison to both AT conditions. Photosynthetic complexes were uniquely depleted under the LAH condition but accumulated under the ATHC condition. The MT proteome displayed several heterotrophic features typical of the LAH proteome, particularly including the high abundance of ribosome as well as amino acid and protein biosynthesis machinery-related components. It is also noteworthy that the two equally light-exposed ATHC and MT cultures allocated similar mass fractions of the total proteome to the seven distinct sub-proteomes. Unique trophic condition-specific expression patterns were likewise observed among individual proteins, including the accumulation of phosphate transporters and polyphosphate polymers storing energy surplus in highly energetic bonds under the MT condition and accumulation under the LAH condition of an enzyme catalyzing cyanophycin biosynthesis. It is concluded that the rigor of cell growth in the MT condition results, to a great extent, by combining photosynthetic activity with high intracellular inorganic carbon conditions created upon glucose breakdown and release of CO2, besides the direct utilization of glucose-derived carbon skeletons for growth. This combination provides the MT cultures with excellent conditions for growth that often exceeds that of mere ATHC. [ABSTRACT FROM AUTHOR]

IntroductionWhile Immune checkpoint inhibition (ICI) therapy shows significant efficacy in metastatic melanoma, only about 50% respond, lacking reliable predictive methods. We introduce a panel of six proteins aimed at predicting response to ICI therapy.MethodsEvaluating previously reported proteins in two untreated melanoma cohorts, we used a published predictive model (EaSIeR score) to identify potential proteins distinguishing responders and non-responders.ResultsSix proteins initially identified in the ICI cohort correlated with predicted response in the untreated cohort. Additionally, three proteins correlated with patient survival, both at the protein, and at the transcript levels, in an independent immunotherapy treated cohort.DiscussionOur study identifies predictive biomarkers across three melanoma cohorts, suggesting their use in therapeutic decision-making.

Dopamine and histamine receptors D2R and H3R are G protein-coupled receptors (GPCRs) which can establish physical receptor–receptor interactions (RRIs), leading to homo/hetero-complexes in a dynamic equilibrium. Although D2R and H3R expression has been detected within the carotid body (CB), their possible heterodimerization has never been demonstrated. The aim of this work was to verify D2R and H3R colocalization in the CB, thus suggesting a possible interplay that, in turn, may be responsible of specific D2R–H3R antagonistic functional implications. The CBs of both Sprague–Dawley rats (n = 5) and human donors (n = 5) were dissected, and immunolocalization of D2R and H3R was performed; thereafter, in situ proximity ligation assay (PLA) was developed. According to experimental evidence (immunohistochemistry and double immunofluorescence), all the samples displayed positive D2R/H3R elements; hence, PLA assay followed by confocal microscopy analysis was positive for D2R–H3R RRIs. Additionally, D2R–H3R heterodimers were mainly detected in type I cells (βIII-tubulin-positive cells), but type II cells’ involvement cannot be excluded. RRIs may play a role in functional modulation of CB cells; investigating RRIs in the CB may guide toward the comprehension of its plastic changes and fine regulatory role while also unveiling their possible clinical implications.

The Health Impacts of Artificial Reef Advancement (HIARA; in the Malagasy language, “together”) study cohort was set up in December 2022 to assess the economic and nutritional importance of seafood for the coastal Malagasy population living along the Bay of Ranobe in southwestern Madagascar. Over the course of the research, which will continue until at least 2026, the primary question we seek to answer is whether the creation of artificial coral reefs can rehabilitate fish biomass, increase fish catch, and positively influence fisher livelihoods, community nutrition, and mental health. Through prospective, longitudinal monitoring of the ecological and social systems of Bay of Ranobe, we aim to understand the influence of seasonal and long-term shifts in marine ecological resources and their benefits to human livelihoods and health. Fourteen communities (12 coastal and two inland) were enrolled into the study including 450 households across both the coastal (n = 360 households) and inland (n = 90 households) ecosystems. In the ecological component, we quantify the extent and health of coral reef ecosystems and collect data on the diversity and abundance of fisheries resources. In the social component, we collect data on the diets, resource acquisition strategies, fisheries and agricultural practices, and other social, demographic and economic indicators, repeated every 3 months. At these visits, clinical measures are collected including anthropometric measures, blood pressure, and mental health diagnostic screening. By analyzing changes in fish catch and consumption arising from varying distances to artificial reef construction and associated impacts on fish biomass, our cohort study could provide valuable insights into the public health impacts of artificial coral reef construction on local populations. Specifically, we aim to assess the impact of changes in fish catch (caused by artificial reefs) on various health outcomes, such as stunting, underweight, wasting, nutrient intake, hypertension, anxiety, and depression.

The persistent expansion in world energy and synthetic compounds requires the improvement of renewable alternatives in contrast to non-sustainable energy wellsprings. Lignocellulose is an encouraging feedstock to be utilized in biorefineries for its conversion into value-added products, including biomaterials, biofuels and several bio-based synthetic compounds. Aside from all categories, biofuel, particularly bioethanol is the most substantial fuel derived from lignocellulosic biomass and can be obtained through microbial fermentation. Generally, extreme settings are required for lignocellulosic pretreatment which results in the formation of inhibitors during biomassdegradation. Occasionally, lignin polymers also act as inhibitors and are left untreated during the pretreatment, engendering inefficient hydrolysis. The valorization of lignocellulosic biomass by laccases can be viewed as a fundamental trend for improving bioethanol production. However, one of the main obstacles for developing commercially viable biofuel industries is the cost of enzymes, which can be resolved by utilizing laccases derived from microbial sources. Microbial laccases have been considered an exceptionally integral asset for delignification and detoxification of pretreated LCB, which amplify the resultant fermentation and saccharification processes. This review provides a summary of microbial laccases and their role in valorizing LCB to bioethanol, compelling enthralling applications in bio-refining industries all across the globe.

This study provides a brief review of the adaptation of the short MacArthur-Bates Communicative Development Inventories (CDI) instruments into Basque. The study aims to provide the scientific community with several standardized instruments designed to measure the vocabulary size of young children (aged 8–50 months) who are learning Basque, a language spoken in Spain and France. Data from over 2,400 children, obtained using the Basque adaptations of the short CDI parental questionnaires, the BCDI-1s(hort), BCDI-2s, and BCDI-3, revealed the capacity of these instruments to identify the effect of age on the language development of preschool-age children exposed to this language, as has been attested in other (minority and non-minority) languages. The study also examined the effect size of age, sex, and language input. The results showed that sex had a null or very small effect on both lexical development and the rest of the scales, whereas the effect of language input increased as children grew older and was even stronger than the effect of age from 30 months onwards. These findings provide solid ground for discussing the relevance of various factors that affect young children's language acquisition.

IntroductionThe main obstacle in treating cancer patients is drug resistance. Lenvatinib treatment poses challenges due to loss of response and the common dose-limiting adverse events (AEs). The Constrained-disorder-principle (CDP)-based second-generation artificial intelligence (AI) systems introduce variability into treatment regimens and offer a potential strategy for enhancing treatment efficacy. This proof-of-concept clinical trial aimed to assess the impact of a personalized algorithm-controlled therapeutic regimen on lenvatinib effectiveness and tolerability.MethodsA 14-week open-label, non-randomized trial was conducted with five cancer patients receiving lenvatinib—an AI-assisted application tailored to a personalized therapeutic regimen for each patient, which the treating physician approved. The study assessed changes in tumor response through FDG-PET-CT and tumor markers and quality of life via the EORTC QLQ-THY34 questionnaire, AEs, and laboratory evaluations. The app monitored treatment adherence.ResultsAt 14 weeks of follow-up, the disease control rate (including the following outcomes: complete response, partial response, stable disease) was 80%. The FDG-PET-CT scan-based RECIST v1.1 and PERCIST criteria showed partial response in 40% of patients and stable disease in an additional 40% of patients. One patient experienced a progressing disease. Of the participants with thyroid cancer, 75% showed a reduction in thyroglobulin levels, and 60% of all the participants showed a decrease in neutrophil-to-lymphocyte ratio during treatment. Improvement in the median social support score among patients utilizing the system supports an ancillary benefit of the intervention. No grade 4 AEs or functional deteriorations were recorded.SummaryThe results of this proof-of-concept open-labeled clinical trial suggest that the CDP-based second-generation AI system-generated personalized therapeutic recommendations may improve the response to lenvatinib with manageable AEs. Prospective controlled studies are needed to determine the efficacy of this approach.

BackgroundMicrovascular resistance reserve (MRR) is a recently introduced specific index of coronary microcirculation. MRR calculation can utilize parameters deriving from coronary flow reserve (CFR) assessment, provided that intracoronary pressure data are also available. The previously proposed pressure-bounded CFR (CFRpb) defines the possible CFR interval on the basis of resting and hyperemic pressure gradients in the epicardial vessel, however, its correlation to the Doppler wire measurement was reported to be rather poor without the correction for hydrostatic pressure.PurposeWe aimed to determine the pressure-bounded coronary MRR interval with hydrostatic pressure correction according to the previously established equations of CFRpb adapted for the MRR concept. Furthermore, we also aimed to design a prediction model using the actual MRR value within the pressure-bounded interval and validate the results against the gold-standard Doppler wire technique.MethodsHydrostatic pressure between the tip of the catheter and the sensor of the pressure wire was calculated by height difference measurement from a lateral angiographic view. In the derivation cohort the pressure-bounded MRR interval (between MRRpbmin and MRRpbmax) was determined solely from hydrostatic pressure-corrected intracoronary pressure data. The actual MRR was calculated by simple hemodynamic equations incorporating the anatomical data of the three-dimensionally reconstructed coronary artery (MRRp−3D). These results were analyzed by regression analyses to find relations between the MRRpb bounds and the actual MRRp−3D.ResultsIn the derivation cohort of 23 measurements, linear regression analysis showed a tight relation between MRRpbmax and MRRp−3D (r2 = 0.74, p

Brain death triggers a systemic inflammatory response. Whether systemic inflammation is different in lung donors after brain- (DBD) or circulatory-death (DCD) is unknown, but this may potentially increase the incidence of primary graft dysfunction (PGD) after lung transplantation. We compared the plasma levels of interleukin (IL)-6, IL-8, IL-10 and TNF-α in BDB and DCD and their respective recipients, as well as their relationship with PGD and mortality after LT. A prospective, observational, multicenter, comparative, cohort-nested study that included 40 DBD and 40 DCD lung donors matched and their respective recipients. Relevant clinical information and blood samples were collected before/during lung retrieval in donors and before/during/after (24, 48 and 72 h) LT in recipients. Incidence of PGD and short-term mortality after LT was recorded. Plasma levels of all determined cytokines were numerically higher in DBD than in DCD donors and reached statistical significance for IL-6, IL-10 and IL-8. In recipients with PGD the donor’s plasma levels of TNF-α were higher. The post-operative mortality rate was very low and similar in both groups. DBD is associated with higher systemic inflammation than DCD donors, and higher TNF-α plasma levels in donors are associated with a higher incidence of PGD.

BackgroundMultiple Sclerosis (MS) is a common neurological disease among white populations of European origin. Frequencies among Latin Americans continue to be studied, however, epidemiologic, and clinical characterization studies lack from Central American and Caribbean countries. Ethnicity in these countries is uniformly similar with a prevalent Mestizo population.Methods and resultsData from January 2014 to December 2019 from Guatemala, El Salvador, Honduras, Nicaragua, Costa Rica, Panama, Dominican Republic, and Aruba on demographic, clinical, MRI and phenotypic traits were determined in coordinated studies: ENHANCE, a population-based, retrospective, observational study on incidence and clinical characteristics, and from the subgroup with MS national registries (Aruba, Dominican Republic, Honduras, and Panama), data on prevalence, phenotypes and demographics. Expanded Disability Status Scale (EDSS), and therapeutic schemes were included. ENHANCE data from 758 patients disclosed 79.8% of Mestizo ethnicity; 72.4% female; median age at onset 31.0 years and 33.2 at diagnosis. The highest incidence rate was from Aruba, 2.3–3.5 × 100,000 inhabitants, and the lowest, 0.07–0.15 × 100,000, from Honduras. Crude prevalence rates per 100,000 inhabitants fluctuated from 27.3 (Aruba) to 1.0 (Honduras). Relapsing MS accounted for 87.4% of cases; EDSS

Climate change has emerged across many regions. Some observed regional climate changes, such as amplified Arctic warming and land-sea warming contrasts have been predicted by climate models. However, many other observed regional changes, such as changes in tropical sea surface temperature and monsoon rainfall are not well simulated by climate model ensembles even when taking into account natural internal variability and structural uncertainties in the response of models to anthropogenic radiative forcing. This suggests climate model predictions may not fully reflect what our future will look like. The discrepancies between models and observations are not well understood due to several real and apparent puzzles and limitations such as the “signal-to-noise paradox” and real-world record-shattering extremes falling outside of the possible range predicted by models. Addressing these discrepancies, puzzles and limitations is essential, because understanding and reliably predicting regional climate change is necessary in order to communicate effectively about the underlying drivers of change, provide reliable information to stakeholders, enable societies to adapt, and increase resilience and reduce vulnerability. The challenges of achieving this are greater in the Global South, especially because of the lack of observational data over long time periods and a lack of scientific focus on Global South climate change. To address discrepancies between observations and models, it is important to prioritize resources for understanding regional climate predictions and analyzing where and why models and observations disagree via testing hypotheses of drivers of biases using observations and models. Gaps in understanding can be discovered and filled by exploiting new tools, such as artificial intelligence/machine learning, high-resolution models, new modeling experiments in the model hierarchy, better quantification of forcing, and new observations. Conscious efforts are needed toward creating opportunities that allow regional experts, particularly those from the Global South, to take the lead in regional climate research. This includes co-learning in technical aspects of analyzing simulations and in the physics and dynamics of regional climate change. Finally, improved methods of regional climate communication are needed, which account for the underlying uncertainties, in order to provide reliable and actionable information to stakeholders and the media.