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Objectives: Clinical heterogeneity among patients in addiction treatment settings represents a challenge as most of the treatment programs are designed to treat substance use disorders (SUD) generally rather than offering more tailored approaches addressing individual patient needs. Systematic characterization of clinical heterogeneity may permit more individualized care paths toward improving outcomes. Methods: Data were collected from a large inpatient SUD treatment program between April 2018 and March 2020 (n = 1519). Latent profile analysis (LPA) was applied to identify latent clusters based on differences in substance use and co-occurring depression, anxiety, and post-traumatic stress disorder. Results: Five distinct profiles emerged: Profile 1 (38%) exhibited the lowest substance use and lowest psychiatric severity (Overall Low); Profile 2 (39%) exhibited high alcohol and psychiatric severity; Profile 3 (13%) exhibited high opioid severity and low psychiatric severity. Profile 4 (8%) exhibited high cannabis use and high psychiatric severity, and profile 5 (3%) exhibited high polysubstance use other than alcohol and cannabis use. The latter two profiles were younger and exhibited higher self-regulatory deficits. The (High Alc/high psych) and the (High Cann/Psych) profiles exhibited differentially higher psychiatric severity. Profiles showing high polysubstance use, as well as high cannabis use and high psychiatric severity, showed significantly higher impulsive behavior than the others. Conclusions: LPA revealed five clusters of patients varying substantially in terms of SUD and psychiatric severity. Addressing common features of clinical heterogeneity for tailored care paths in a personalized treatment approach may improve treatment outcomes. [ABSTRACT FROM AUTHOR]

Non-suicidal self-injury (NSSI) among adolescents continues to be a significant public health concern worldwide. A recent systematic review and meta-analysis found that the global prevalence of NSSI in adolescents aged 12–18 years was 17.2%, with higher rates reported among females (19.7%) than males (14.8%). This behavior has been linked to several negative outcomes, such as depression, anxiety, substance abuse, and suicidal ideation. The present study aimed to classify adolescents based on intrapersonal and interpersonal factors associated with NSSI proposed in Nock's (2009) integrated model of NSSI, to identify distinct clusters targeting specific risk factors. This encompassed negative cognition, emotional vulnerability, poor coping skill, peer-victimization, family adaptability, and perceived stress. A total of 881 adolescents aged 11–16 years in South Korea completed self-reported questionnaires on automatic thoughts, depression, emotional regulation, peer victimization, family adaptability and perceived stress. Latent profile analysis (LPA) revealed three distinct classes: "the severe group", "the moderate group", "the mild group". Class 3 ("severe group": N = 127) exhibited greater severity related to NSSI, including negative cognition, emotional vulnerability, poor coping skills, peer victimization, and perceived stress, with weaker levels of factors that can prevent NSSI compared to class 1 ("mild group": N = 416) and class 2 ("moderated group": N = 338). The present study emphasizes the importance of considering both intrapersonal (e.g., negative automatic thoughts & emotional dysregulation) and interpersonal factors (i.e., peer victimization) when understanding NSSI - among adolescents. These findings can be utilized to develop interventions aimed at reducing the prevalence and severity of NSSI among adolescents. [ABSTRACT FROM AUTHOR]

Previous research has shown that the relationship between a supervisor and a subordinate is crucial in shaping employee voice behaviour. However, where these relationships originate needs to be clarified. Studies on relational schemas suggest that the quality of these relationships depends on the relational schemas held by both leaders and subordinates. Unfortunately, these two research areas have developed independently and have yet to intersect, which limits our understanding of how social interactions between leaders and subordinates impact employee voice behaviour. Based on research from both domains, the current study examines how the supervisor–subordinate relational schema affects employees' voice behaviour. Our proposal, grounded in complexity theory, suggests that the expectations of both leaders and subordinates about their relationship model determine whether employees choose to voice their opinions to their leaders. Analyses of data from a survey of 416 matched pairs of leaders and subordinates in China conducted using fsQCA and LPA largely support our view. The results indicate that the configuration of the supervisor–subordinate relational schema among leaders and subordinates can predict the level of employee voice behaviour. Additionally, this configuration is a natural subset of the population. These findings have important implications for improving employee voice behaviour in organizations. [ABSTRACT FROM AUTHOR]

The present study examined the heterogeneity of Chinese romantic couples' filial piety belief patterns. In addition, we investigated how the configuration of filial piety belief profiles within couples can link to individuals' perceptions of romantic relationship satisfaction and different attachment tendencies. We recruited 297 dyads of Chinese couples currently in heterosexual romantic relationships in the university. The couples completed self‐reported measures of dual filial piety beliefs (reciprocal and authoritarian filial piety), romantic relationship satisfaction, and insecure attachment (attachment anxiety and avoidance). We used latent profile analysis (LPA) to identify couples' dual filial piety belief interaction profiles. Four‐couple profiles emerged: (1) authoritarian couples, (2) balanced couples (while men with slightly higher reciprocal filial piety), (3) balanced couples (while men with lower reciprocal filial piety), and (4) divergent couples (balanced men and non‐filial women). The couples in the four profiles did not show significant differences in their romantic relationship satisfaction, but their attachment styles differed. (1) authoritarian couples experienced the highest levels of attachment anxiety and avoidance; (2) balanced couples experienced relatively lower levels of attachment anxiety and avoidance; and (3) non‐filial women in divergent couple profiles experienced significantly higher levels of attachment avoidance. This was the first study to uncover the possible associations between Chinese young couples' dual filial piety interaction profiles, and their dyadic relationship satisfaction and insecure attachment by adopting the person‐centered approach. We further discussed the theoretical and practical implications of the findings. [ABSTRACT FROM AUTHOR]

Lysophosphatidic acid (LPA) is a simple lipid which is endogenously synthesized from lysophosphatidylcholine (LPC) by autotaxin (ATX). LPA mediates a variety of cellular responses through the binding of G protein-coupled LPA receptors (LPA1 to LPA6). It is considered that LPA receptor-mediated signaling plays an important role in the pathogenesis of human malignancy. Genetic alterations and epigenetic changes of LPA receptors have been detected in some cancer cells as well as LPA per se. Moreover, LPA receptors contribute to the promotion of tumor progression, including cell proliferation, invasion, metastasis, tumorigenicity, and angiogenesis. In recent studies, the activation of LPA receptor-mediated signaling regulates chemoresistance and radiosensitivity in cancer cells. This review provides an updated overview on the roles of LPA receptor-mediated signaling in the regulation of cancer cell functions and its potential utility as a molecular target for novel therapies in clinical cancer approaches. [ABSTRACT FROM AUTHOR]

No data are currently available on the functional role of small conductance Ca2+‐activated K+ channels (SKCa) in ovarian cancer. Here, we characterized the role of SK2 (KCa2.2) in ovarian cancer cell migration and chemosensitivity. Using the selective non‐cell‐permeant SK2 inhibitor Lei‐Dab7, we identified functional SK2 channels at the plasma membrane, regulating store‐operated Ca2+ entry (SOCE) in both cell lines tested (COV504 and OVCAR3). Silencing KCNN2 with short interfering RNA (siRNA), or blocking SK2 activity with Lei‐Dab7, decreased cell migration. The more robust effect of KCNN2 knockdown compared to Lei‐Dab7 treatment suggested the involvement of functional intracellular SK2 channels in both cell lines. In cells treated with lysophosphatidic acid (LPA), an ovarian cancer biomarker of progression, SK2 channels are a key player of LPA pro‐migratory activity but their role in SOCE is abolished. Concerning chemotherapy, SK2 inhibition increased chemoresistance to Taxol® and low KCNN2 mRNA expression was associated with the worst prognosis for progression‐free survival in patients with serous ovarian cancer. The dual roles of SK2 mean that SK2 activators could be used as an adjuvant chemotherapy to potentiate treatment efficacy and SK2 inhibitors could be administrated as monotherapy to limit cancer cell dissemination. [ABSTRACT FROM AUTHOR]

Emotions and motivation are central to learning. Control–value theory (CVT) and expectancy–value theory (EVT) explain how emotions and expectations of success affect students' task engagement. Supported by these two frameworks, this study investigates the emotion profiles for maths among fourth- and fifth-grade students (N = 6778) from three European countries and their links to motivation and achievement. Methods: Using latent profile analysis (LPA), we analysed the emotional profiles in students, as well as their associations with gender, country, grade, motivation and maths achievement. Results: Five profiles emerged in the grade 4 data (Bored, Bored and Anxious, Moderate, Happy and Anxious). All five profiles were visible in the grade 5 data, coupled with an additional sixth profile, which appeared only in grade 5 (i.e., Apprehensive–Happy). Girls were found more commonly in the Anxious profile and were less likely to appear in the Happy profile. Norwegian students were more prominent in the Bored and Moderate profiles. Conversely, Portuguese students stood out more in the Anxious profile and were less present in the Bored and Moderate profiles. The Serbian dataset did not stand out, with a particular pattern observed in grade 4. Nevertheless, Serbian fifth graders were overly visible in the Bored and Anxious profile and less present in the Happy and Apprehensive–Happy groups. The Happy profile had higher scores for all task values except for cost and was, along with the Moderate profile, associated with higher achievement; the Bored and Anxious profile was associated with higher scores of cost value and lower achievement. Conclusions: A person-centred approach allowed for a more diverse view of how students experience emotions. These findings highlight the complex interplay between emotions, motivation and achievement, which is affected by cultural and educational contexts. [ABSTRACT FROM AUTHOR]

Abstract Non-suicidal self-injury (NSSI) among adolescents continues to be a significant public health concern worldwide. A recent systematic review and meta-analysis found that the global prevalence of NSSI in adolescents aged 12–18 years was 17.2%, with higher rates reported among females (19.7%) than males (14.8%). This behavior has been linked to several negative outcomes, such as depression, anxiety, substance abuse, and suicidal ideation. The present study aimed to classify adolescents based on intrapersonal and interpersonal factors associated with NSSI proposed in Nock’s (2009) integrated model of NSSI, to identify distinct clusters targeting specific risk factors. This encompassed negative cognition, emotional vulnerability, poor coping skill, peer-victimization, family adaptability, and perceived stress. A total of 881 adolescents aged 11–16 years in South Korea completed self-reported questionnaires on automatic thoughts, depression, emotional regulation, peer victimization, family adaptability and perceived stress. Latent profile analysis (LPA) revealed three distinct classes: “the severe group”, “the moderate group”, “the mild group”. Class 3 (“severe group”: N = 127) exhibited greater severity related to NSSI, including negative cognition, emotional vulnerability, poor coping skills, peer victimization, and perceived stress, with weaker levels of factors that can prevent NSSI compared to class 1 (“mild group”: N = 416) and class 2 (“moderated group”: N = 338). The present study emphasizes the importance of considering both intrapersonal (e.g., negative automatic thoughts & emotional dysregulation) and interpersonal factors (i.e., peer victimization) when understanding NSSI - among adolescents. These findings can be utilized to develop interventions aimed at reducing the prevalence and severity of NSSI among adolescents.

Background: Tuberculosis (TB) remains a significant public health challenge in developing countries, including India, where children under 15 years account for 12% of all TB cases. The National Tuberculosis Elimination Programme (NTEP) recommends the use of GeneXpert/Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) for TB diagnosis, followed by Line Probe Assay (LPA) for detecting drug resistance. This study aimed to determine the prevalence of TB and rifampicin resistance in pediatric patients and to evaluate the utility of CBNAAT in comparison with LPA and Ziehl-Neelsen staining. Methods: This retrospective study was conducted over one year, from 1st January 2022 to 31st December 2022, at a tertiary care hospital in western Maharashtra. A total of 528 samples from pediatric patients (aged

No data are currently available on the functional role of small conductance Ca2+‐activated K+ channels (SKCa) in ovarian cancer. Here, we characterized the role of SK2 (KCa2.2) in ovarian cancer cell migration and chemosensitivity. Using the selective non‐cell‐permeant SK2 inhibitor Lei‐Dab7, we identified functional SK2 channels at the plasma membrane, regulating store‐operated Ca2+ entry (SOCE) in both cell lines tested (COV504 and OVCAR3). Silencing KCNN2 with short interfering RNA (siRNA), or blocking SK2 activity with Lei‐Dab7, decreased cell migration. The more robust effect of KCNN2 knockdown compared to Lei‐Dab7 treatment suggested the involvement of functional intracellular SK2 channels in both cell lines. In cells treated with lysophosphatidic acid (LPA), an ovarian cancer biomarker of progression, SK2 channels are a key player of LPA pro‐migratory activity but their role in SOCE is abolished. Concerning chemotherapy, SK2 inhibition increased chemoresistance to Taxol® and low KCNN2 mRNA expression was associated with the worst prognosis for progression‐free survival in patients with serous ovarian cancer. The dual roles of SK2 mean that SK2 activators could be used as an adjuvant chemotherapy to potentiate treatment efficacy and SK2 inhibitors could be administrated as monotherapy to limit cancer cell dissemination.

• LPA promotes the migration of skeletal muscle-isolated FAPs through LPA 1 signaling. • YAP activity is required for LPA-induced migration of FAPs. • FAPs isolated from dystrophic muscle possess an enhanced migration response to LPA, behavior which can be blocked by inhibiting LPA 1/3 or YAP activity. Skeletal muscle fibrosis is defined as the excessive accumulation of extracellular matrix (ECM) components and is a hallmark of muscular dystrophies. Fibro-adipogenic progenitors (FAPs) are the main source of ECM, and thus have been strongly implicated in fibrogenesis. In skeletal muscle fibrotic models, including muscular dystrophies, FAPs undergo dysregulations in terms of proliferation, differentiation, and apoptosis, however few studies have explored the impact of FAPs migration. Here, we studied fibroblast and FAPs migration and identified lysophosphatidic acid (LPA), a signaling lipid central to skeletal muscle fibrogenesis, as a significant migration inductor. We identified LPA receptor 1 (LPA 1) mediated signaling as crucial for this effect through a mechanism dependent on the Hippo pathway, another pathway implicated in fibrosis across diverse tissues. This cross-talk favors the activation of the Yes-associated protein 1 (YAP) and Transcriptional coactivator with PDZ-binding motif (TAZ), leading to increased expression of fibrosis-associated genes. This study reveals the role of YAP in LPA-mediated fibrotic responses as inhibition of YAP transcriptional coactivator activity hinders LPA-induced migration in fibroblasts and FAPs. Moreover, we found that FAPs derived from the mdx 4cv mice, a murine model of Duchenne muscular dystrophy, display a heightened migratory phenotype due to enhanced LPA signaling compared to wild-type FAPs. Remarkably, we found that the inhibition of LPA 1 or YAP transcriptional coactivator activity in mdx 4cv FAPs reverts this phenotype. In summary, the identified LPA-LPA 1 -YAP pathway emerges as a critical driver of skeletal muscle FAPs migration and provides insights into potential novel targets to mitigate fibrosis in muscular dystrophies. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) remains the number one cause of adult deaths by a curable infectious disease. Mycobacterium tuberculosis bacilli (MTB) is the most common causative organism isolated from mycobacterial lymphadenitis. Accurate and speedy diagnosis is required, especially in paucibacillary extrapulmonary TB. In this study, we compared the efficacy of CBNAAT with line probe assay and liquid culture in the evaluation of tubercular lymph node abscesses. Materials and Methods: This is a hospital-based observational cross-sectional study conducted at Gandhi Medical College and Hospital, Secunderabad, TS, in patients who presented with lymph node abscesses over a period of 18 months after obtaining clearance from the ethics committee and obtaining informed consent. Each sample was evaluated and analyzed in association with the clinical findings, AFB staining, CBNAAT, LPA, and AFB culture, and the results were compared. Results: Fifty patients with TB lymph node abscess diagnosed through FNAC were included in the present study. AFB smear was positive in 29 samples, CBNAAT was positive in 43 samples, LPA was positive in 44 samples, and liquid culture was positive in 39 samples. CBNAAT could detect TB in 16 smear-negative cases, 10 liquid culture-negative cases, and one LPA-negative case. In our study, CBNAAT was more effective in diagnosing TB lymph node abscess than AFB smear. No significant results were obtained when comparing CBNAAT with LPA and liquid culture. Conclusion: This study demonstrates excellent diagnostic accuracy of the Xpert MTB/RIF test in patients with tuberculous lymphadenitis, with a sensitivity of 86%, specificity of 83.33%, negative predictive value of 95.56%, and positive predictive value of 58.82%. Thus, it can be a rule in testing for lymph node TB. These efforts will contribute to the attainment of the TB elimination goal.

Depression and anxiety co-occur frequently and there is heterogeneity in the co-occurrence of such symptoms; however, few previous studies investigated the heterogeneity based on person-centered perspectives in adolescents. The primary aim of our study was to explore it using latent profile analysis (LPA), a person-centered statistical approach. The Patient Health Questionnaire-9 (PHQ-9) and General Anxiety Disorder-7 (GAD-7) were used to examine depression and anxiety symptoms in 7422 Chinese adolescents from 23 primary and secondary schools. To investigate latent profiles and assess profile validity, we employed Latent Profile Analysis (LPA), multinomial logistic regression, and analysis of variance. A three-profile model was suggested as the optimum: low (69.9 %), moderate (21.6 %), and high depression/anxiety (8.5 %). Female with higher negative cognitive bias and higher emotional regulation difficulty are more likely to be categorized in the high depression/anxiety group. Internet addiction, academic "Lying flat" and involution are significantly and positively linked with the severity of anxiety and depression. Reliance on self-reported measures may lead to response bias; the cross-sectional design limits our ability to study how symptom profiles and category membership change over time. Three latent profiles of the co-occurrence of depression and anxiety presented a parallel pattern, which serves as a poignant reminder of the imperative need to identify Chinese adolescents who may be at elevated risk for depression and/or anxiety, and promoting intervention that are meticulously tailored to address the unique symptom presentations of each individual. • Heterogeneous patterns of the co-occurring of depression and anxiety symptoms were examined. • LPA results identified three symptom profiles. • Latent profiles were predicted by gender, negative cognition bias and emotional regulation difficulty. • Internet addiction, academic "Lying flat" and involution varied for different symptom profiles. • The severity of depression and anxiety shows a parallel pattern. [ABSTRACT FROM AUTHOR]

This article seeks to provide a clearer understanding of Digital Capital in education. It introduces a comprehensive analytical framework that explores the relationship between Digital Capital and Bourdieu's Cultural Capital Theory. Instead of treating digital skills and resources as separate entities, it integrates them into Cultural Capital Theory as complementary elements. This approach helps shed light on the disparities in ICT usage. Data from the 2018 OECD‐PISA survey conducted in Italy are analysed to assess whether Digital Capital can be considered a component of Cultural Capital. The findings indicate that differences in Cultural Capital do not significantly impact the possession and usage of digital assets. Instead, distinctions become apparent through students’ behaviours within the school environment. This underscores the connection between digital competencies and various dimensions of cultural and educational capital. The article posits that status and cultural disparities stem not solely from digital competencies but also from their interplay with social and cultural resources. This offers deeper insights into how the digital divide intersects with broader societal power dynamics. [ABSTRACT FROM AUTHOR]

Nucleic acid-based therapies are being rapidly developed for prevention and management of cardiovascular diseases (CVD). Remarkable advancements have been achieved in the delivery, safety, and effectiveness of these therapeutics in the past decade. These therapies can also modulate therapeutic targets that cannot be sufficiently addressed using traditional drugs or antibodies. Among the nucleic acid-targeted therapeutics under development for CVD prevention are RNA-targeted approaches, including antisense oligonucleotides (ASO), small interfering RNAs (siRNA), and novel genome editing techniques. Genetic studies have identified potential therapeutic targets that are suggested to play a causative role in development and progression of CVD. RNA- and DNA-targeted therapeutics can be particularly well delivered to the liver, where atherogenic lipoproteins and angiotensinogen (AGT) are produced. Current targets in lipid metabolism include proprotein convertase subtilisin/kexin type 9 (PCSK9), apolipoprotein A (ApoA), apolipoprotein C3 (ApoC3), angiopoietin-like 3 (ANGPTL3). Several large-scale clinical development programs for nucleic acid-targeted therapies in cardiovascular prevention are under way, which may also be attractive from a therapy adherence point of view, given the long action of these therapeutics. In addition to genome editing, the concept of gene transfer is presently under assessment in preclinical and clinical investigations as a potential approach for addressing low-density lipoprotein receptor deficiency. Furthermore, ongoing research is exploring the use of RNA-targeted therapies to treat arterial hypertension by reducing hepatic angiotensinogen (AGT) production. This review summarizes the rapid translation of siRNA and ASO therapeutics as well as gene editing into clinical studies to treat dyslipidemia and arterial hypertension for CVD prevention. It also outlines potential innovative therapeutic options that are likely relevant to the future of cardiovascular medicine. [ABSTRACT FROM AUTHOR]

Regional debris flow susceptibility assessment is an effective method to prevent debris flow hazards, and deep learning is emerging as a novel approach in this discipline with the development of computers. However, when debris flow samples are insufficient, there will be problems like overfitting or misclassification. To overcome these problems, this paper proposes a semi-supervised deep neural network model (LPA-DNN) combined with label propagation algorithm (LPA), which utilizes high confidence unlabeled samples as pseudo-samples reasonably in few-label scenarios. Xinzhou, Shanxi Province, was selected as the study area, and a dataset containing 292 debris flow samples and 10 types of impact factors was compiled based on watershed units. Using the dataset and pseudo-samples, the LPA-DNN model was built to get debris flow susceptibility map. Meanwhile, DNN and SVM were set up for comparison to demonstrate that the proposed LPA-DNN model has excellent performance and higher accuracy. LPA-DNN alleviates the problem of low accuracy that caused by samples lacking in deep learning to a certain extent, and obtains great classification results, which proves that it is quite potential in regional debris flow susceptibility assessment. [ABSTRACT FROM AUTHOR]

Fluorescence resonance energy transfer (FRET) biosensors have proven to be an indispensable tool in cell biology and, more specifically, in the study of G-protein signalling. The best method of measuring the activation status or FRET state of a biosensor is often fluorescence lifetime imaging microscopy (FLIM), as it does away with many disadvantages inherent to fluorescence intensity-based methods and is easily quantitated. Despite the significant potential, there is a lack of reliable FLIM-FRET biosensors, and the data processing and analysis workflows reported previously face reproducibility challenges. Here, we established a system in live primary mouse pancreatic ductal adenocarcinoma cells, where we can detect the activation of an mNeonGreen-Gαi3-mCherry-Gγ2 biosensor through the lysophosphatidic acid receptor (LPAR) with 2-photon time-correlated single-photon counting (TCSPC) FLIM. This combination gave a superior signal to the commonly used mTurquoise2-mVenus G-protein biosensor. This system has potential as a platform for drug screening, or to answer basic cell biology questions in the field of G-protein signalling. [ABSTRACT FROM AUTHOR]

LPA3 receptors were expressed in TREx HEK 293 cells, and their signaling and phosphorylation were studied. The agonist, lysophosphatidic acid (LPA), increased intracellular calcium and ERK phosphorylation through pertussis toxin-insensitive processes. Phorbol myristate acetate, but not LPA, desensitizes LPA3-mediated calcium signaling, the agonists, and the phorbol ester-induced LPA3 internalization. Pitstop 2 (clathrin heavy chain inhibitor) markedly reduced LPA-induced receptor internalization; in contrast, phorbol ester-induced internalization was only delayed. LPA induced rapid β-arrestin–LPA3 receptor association. The agonist and the phorbol ester-induced marked LPA3 receptor phosphorylation, and phosphorylation sites were detected using mass spectrometry. Phosphorylated residues were detected in the intracellular loop 3 (S221, T224, S225, and S229) and in the carboxyl terminus (S321, S325, S331, T333, S335, Y337, and S343). Interestingly, phosphorylation sites are within sequences predicted to constitute β-arrestin binding sites. These data provide insight into LPA3 receptor signaling and regulation. [ABSTRACT FROM AUTHOR]

Lysophosphatidic acid (LPA) is a bioactive lipid molecule that regulates a wide array of cellular functions, including proliferation, differentiation, and survival, via activation of cognate receptors. The LPA5 receptor is highly expressed in the intestinal epithelium, but its function in restoring intestinal epithelial integrity following injury has not been examined. Here, we use a radiation-induced injury model to study the role of LPA5 in regulating intestinal epithelial regeneration. Control mice (Lpar5f/f) and mice with an inducible, epithelial cell-specific deletion of Lpar5 in the small intestine (Lpar5IECKO) were subjected to 10 Gy total body X-ray irradiation and analyzed during recovery. Repair of the intestinal mucosa was delayed in Lpar5IECKO mice with reduced epithelial proliferation and increased crypt cell apoptosis. These effects were accompanied by reduced numbers of OLFM4+ intestinal stem cells (ISCs). The effects of LPA5 on ISCs were corroborated by studies using organoids derived from Lgr5-lineage tracking reporter mice with deletion of Lpar5 in Lgr5+-stem cells (Lgr5Cont or Lgr5ΔLpar5). Irradiation of organoids resulted in fewer numbers of Lgr5ΔLpar5 organoids retaining Lgr5+-derived progenitor cells compared with Lgr5Cont organoids. Finally, we observed that impaired regeneration in Lpar5IECKO mice was associated with reduced numbers of Paneth cells and decreased expression of Yes-associated protein (YAP), a critical factor for intestinal epithelial repair. Our study highlights a novel role for LPA5 in regeneration of the intestinal epithelium following irradiation and its effect on the maintenance of Paneth cells that support the stem cell niche. NEW & NOTEWORTHY: We used mice lacking expression of the lysophosphatidic acid receptor 5 (LPA5) in intestinal epithelial cells and intestinal organoids to show that the LPA5 receptor protects intestinal stem cells and progenitors from radiation-induced injury. We show that LPA5 induces YAP signaling and regulates Paneth cells. [ABSTRACT FROM AUTHOR]

• Skeletal muscle isolated FAPs express higher levels of LPARs and show a stronger response to LPA on the induction of CCN2 than other representative parenchymatous skeletal muscle cell types. • LPA acts mainly through LPA 1 and, to less extent, through LPA 6 , modulating CCN2 expression levels in skeletal muscle FAPs. • ROCK kinase and actin polymerization are required upstream YAP signaling for CCN2 induction in response to LPA in skeletal muscle FAPs. Cellular Communication Network Factor 2, CCN2, is a profibrotic cytokine implicated in physiological and pathological processes in mammals. The expression of CCN2 is markedly increased in dystrophic muscles. Interestingly, diminishing CCN2 genetically or inhibiting its function improves the phenotypes of chronic muscular fibrosis in rodent models. Elucidating the cell-specific mechanisms behind the induction of CCN2 is a fundamental step in understanding its relevance in muscular dystrophies. Here, we show that the small lipids LPA and 2S-OMPT induce CCN2 expression in fibro/adipogenic progenitors (FAPs) through the activation of the LPA 1 receptor and, to a lower extent, by also the LPA 6 receptor. These cells show a stronger induction than myoblasts or myotubes. We show that the LPA/LPARs axis requires ROCK kinase activity and organized actin cytoskeleton upstream of YAP/TAZ signaling effectors to upregulate CCN2 levels, suggesting that mechanical signals are part of the mechanism behind this process. In conclusion, we explored the role of the LPA/LPAR axis on CCN2 expression, showing a strong cytoskeletal-dependent response in muscular FAPs. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) remains the number one cause of adult deaths by a curable infectious disease. Mycobacterium tuberculosis bacilli (MTB) is the most common causative organism isolated from mycobacterial lymphadenitis. Accurate and speedy diagnosis is required, especially in paucibacillary extrapulmonary TB. In this study, we compared the efficacy of CBNAAT with line probe assay and liquid culture in the evaluation of tubercular lymph node abscesses. Materials and Methods: This is a hospital‑based observational cross‑sectional study conducted at Gandhi Medical College and Hospital, Secunderabad, TS, in patients who presented with lymph node abscesses over a period of 18 months after obtaining clearance from the ethics committee and obtaining informed consent. Each sample was evaluated and analyzed in association with the clinical findings, AFB staining, CBNAAT, LPA, and AFB culture, and the results were compared. Results: Fifty patients with TB lymph node abscess diagnosed through FNAC were included in the present study. AFB smear was positive in 29 samples, CBNAAT was positive in 43 samples, LPA was positive in 44 samples, and liquid culture was positive in 39 samples. CBNAAT could detect TB in 16 smear‑negative cases, 10 liquid culture‑negative cases, and one LPA‑negative case. In our study, CBNAAT was more effective in diagnosing TB lymph node abscess than AFB smear. No significant results were obtained when comparing CBNAAT with LPA and liquid culture. Conclusion: This study demonstrates excellent diagnostic accuracy of the Xpert MTB/RIF test in patients with tuberculous lymphadenitis, with a sensitivity of 86%, specificity of 83.33%, negative predictive value of 95.56%, and positive predictive value of 58.82%. Thus, it can be a rule in testing for lymph node TB. These efforts will contribute to the attainment of the TB elimination goal. [ABSTRACT FROM AUTHOR]

The improvement of in vitro embryo development is a gateway to enhance the output of assisted reproductive technologies. The Wnt and Hippo signaling pathways are crucial for the early development of bovine embryos. This study investigated the development of bovine embryos under the influence of a Hippo signaling agonist (LPA) and a Wnt signaling inhibitor (DKK1). In this current study, embryos produced in vitro were cultured in media supplemented with LPA and DKK1. We comprehensively analyzed the impact of LPA and DKK1 on various developmental parameters of the bovine embryo, such as blastocyst formation, differential cell counts, YAP fluorescence intensity and apoptosis rate. Furthermore, single-cell RNA sequencing (scRNA-seq) was employed to elucidate the in vitro embryonic development. Our results revealed that LPA and DKK1 improved the blastocyst developmental potential, total cells, trophectoderm (TE) cells and YAP fluorescence intensity and decreased the apoptosis rate of bovine embryos. A total of 1203 genes exhibited differential expression between the control and LPA/DKK1-treated (LD) groups, with 577 genes upregulated and 626 genes downregulated. KEGG pathway analysis revealed significant enrichment of differentially expressed genes (DEGs) associated with TGF-beta signaling, Wnt signaling, apoptosis, Hippo signaling and other critical developmental pathways. Our study shows the role of LPA and DKK1 in embryonic differentiation and embryo establishment of pregnancy. These findings should be helpful for further unraveling the precise contributions of the Hippo and Wnt pathways in bovine trophoblast formation, thus advancing our comprehension of early bovine embryo development. [ABSTRACT FROM AUTHOR]

Background: Drug-resistant tuberculosis (DR-TB) is difficult to treat, and its timely diagnosis is an important factor in successful treatment outcomes. Modern technology has made it possible to diagnose DR-TB more quickly than before when it took a long time to identify the presence of resistance Mycobacterium. Objective: To evaluate the efficacy of the Xpert MTB/XDR to test the susceptibility of MTB towards anti-TB drugs by using sputum samples from infected people. Methodology: A cross-sectional study was conducted at the Programmatic Management of Drug-Resistant TB Unit (PMDT), Lady Reading Hospital, Peshawar, from March 2021 to March 2023. This study included samples from patients identified as rifampicin-resistant patients. The Xpert® MTB/XDR test's diagnosis accuracy was compared to that of the MGIT960, the reference drug susceptibility testing (DST) techniques, and the Hain Genotype® MTBDR plus and MDRsl assays (LPA). Results: The present study included 180 patients. These patients were divided into three groups for study purposes based on technique used, i.e., Pre-LPA or DST, Post-LPA, and Ten color Xpert MTB/XDR, and followed up for their culture conversion. Gene-Xpert MTB/RIF showed 93% of cases were positive, and on DST all cases showed positive results. For drug resistance in DST, LPA, and Gene-Xpert MTB/XDR, 61.1% of patients were resistant to three first-line drugs, and 11.1% were resistant to the second line. The overall median time from the identification of patients suspected of MDR-TB to the initiation of treatment in DST was 93 days. In LPA it was 25 and 5 days by Ten color Gene Xpert MTB/XDR. Conclusion: This study concludes that based on the timely initiation of treatment and culture conversion reports, DST and LPA are less efficient than the Ten-color Gene Xpert MTB/XDR assay. [ABSTRACT FROM AUTHOR]

Abstract Background Mixed M. tuberculosis (MTB) infection occurs when one is infected with more than one clonally distinct MTB strain. This form of infection can assist MTB strains to acquire additional mutations, facilitate the spread of drug-resistant strains, and boost the rate of treatment failure. Hence, the presence of mixed MTB infection could affect the performance of some rapid molecular diagnostic tests such as Line Probe Assay (LPA) and GeneXpert MTB/RIF (Xpert) assays. Methods This was a cross-sectional study that used sputum specimens collected from participants screened for STREAM 2 clinical trial between October 2017 and October 2019. Samples from 62 MTB smear-positive patients and rifampicin-resistant patients from peripheral health facilities were processed for Xpert and LPA as screening tests for eligibility in the trial. From November 2020, processed stored sputum samples were retrieved and genotyped to determine the presence of mixed-MTB strain infection using a standard 24-locus Mycobacterial Interspersed Repetitive Unit–Variable Number Tandem-Repeat (MIRU-VNTR). Samples with at least 20/24 MIRU-VNTR loci amplified were considered for analysis. Agar proportional Drug Susceptibility Test (DST) was performed on culture isolates of samples that had discordant results between LPA and Xpert. The impact of the presence of mixed-MTB strain on Xpert and LPA test interpretation was analyzed. Results A total of 53/62 (85%) samples had analyzable results from MIRU-VNTR. The overall prevalence of mixed-MTB infection was 5/53 (9.4%). The prevalence was highest among male’s 3/31 (9.7%) and among middle-aged adults, 4/30 (33.3%). Lineage 4 of MTB contributed 3/5 (60.0%) of the mixed-MTB infection prevalence. Having mixed MTB strain infection increased the odds of false susceptible Xpert test results (OR 7.556, 95% CI 0.88–64.44) but not for LPA. Being HIV-positive (P = 0.04) independently predicted the presence of mixed MTB infection. Conclusions The presence of mixed-MTB strain infection may affect the performance of the GeneXpert test but not for LPA. For patients with high pre-test probability of rifampicin resistance, an alternative rapid method such as LPA should be considered.

Ruifeng Bai,1 Jianqiu Pei,1 Shengqiang Pei,1 Xiangfeng Cong,1 Jerold Chun,2 Fang Wang,1,3,4 Xi Chen1,3 1State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People’s Republic of China; 2Neuroscience Drug Discovery, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA; 3Diagnostic Laboratory Service, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People’s Republic of China; 4Department of Clinical Laboratory, Fuwai Yunnan Cardiovascular Hospital, Kunming, People’s Republic of ChinaCorrespondence: Xi Chen; Fang Wang, Email chenxifw@pumc.edu.cn; fray.163@163.comBackground: Increased endothelial permeability of pulmonary vessels is a primary pathological characteristic of septic acute lung injury (ALI). Previously, elevated lysophosphatidic acid (LPA) levels and LPA2 (an LPA receptor) expression have been found in the peripheral blood and lungs of septic mice, respectively. However, the specific role of LPA2 in septic ALI remains unclear.Methods: A lipopolysaccharide (LPS)-induced model of sepsis was established in wild-type (WT) and global LPA2 knockout (Lpar2−/−) mice. We examined mortality, lung injury, assessed endothelial permeability through Evans blue dye (EBD) assay in vivo, and transendothelial electrical resistance (TEER) of mouse lung microvascular endothelial cells (MLMECs) in vitro. Enzyme-linked immunosorbent assay (ELISA), histopathological, immunofluorescence, immunohistochemistry, and Western blot were employed to investigate the role of LPA2 in septic ALI.Results: Lpar2 deficiency increased vascular endothelial permeability, impaired lung injury, and increased mortality. Histological examination revealed aggravated inflammation, edema, hemorrhage and alveolar septal thickening in the lungs of septic Lpar2−/− mice. In vitro, loss of Lpar2 resulted in increased permeability of MLMECs. Pharmacological activation of LPA2 by the agonist DBIBB led to significantly reduced inflammation, edema and hemorrhage, as well as increased expression of the vascular endothelial tight junction (TJ) protein zonula occludens-1 (ZO-1) and claudin-5, as well as the adheren junction (AJ) protein VE-cadherin. Moreover, DBIBB treatment was found to alleviate mortality by protecting against vascular endothelial permeability. Mechanistically, we demonstrated that vascular endothelial permeability was alleviated through LPA-LPA2 signaling via the PLC-PKC-FAK pathway.Conclusion: These data provide a novel mechanism of endothelial barrier protection via PLC-PKC-FAK pathway and suggest that LPA2 may contribute to the therapeutic effects of septic ALI.Keywords: septic ALI, LPA, vascular endothelial permeability, LPA2, lung microvascular endothelial cells

Emotions and motivation are central to learning. Control–value theory (CVT) and expectancy–value theory (EVT) explain how emotions and expectations of success affect students’ task engagement. Supported by these two frameworks, this study investigates the emotion profiles for maths among fourth- and fifth-grade students (N = 6778) from three European countries and their links to motivation and achievement. Methods: Using latent profile analysis (LPA), we analysed the emotional profiles in students, as well as their associations with gender, country, grade, motivation and maths achievement. Results: Five profiles emerged in the grade 4 data (Bored, Bored and Anxious, Moderate, Happy and Anxious). All five profiles were visible in the grade 5 data, coupled with an additional sixth profile, which appeared only in grade 5 (i.e., Apprehensive–Happy). Girls were found more commonly in the Anxious profile and were less likely to appear in the Happy profile. Norwegian students were more prominent in the Bored and Moderate profiles. Conversely, Portuguese students stood out more in the Anxious profile and were less present in the Bored and Moderate profiles. The Serbian dataset did not stand out, with a particular pattern observed in grade 4. Nevertheless, Serbian fifth graders were overly visible in the Bored and Anxious profile and less present in the Happy and Apprehensive–Happy groups. The Happy profile had higher scores for all task values except for cost and was, along with the Moderate profile, associated with higher achievement; the Bored and Anxious profile was associated with higher scores of cost value and lower achievement. Conclusions: A person-centred approach allowed for a more diverse view of how students experience emotions. These findings highlight the complex interplay between emotions, motivation and achievement, which is affected by cultural and educational contexts.

Background: Mixed M. tuberculosis (MTB) infection occurs when one is infected with more than one clonally distinct MTB strain. This form of infection can assist MTB strains to acquire additional mutations, facilitate the spread of drug-resistant strains, and boost the rate of treatment failure. Hence, the presence of mixed MTB infection could affect the performance of some rapid molecular diagnostic tests such as Line Probe Assay (LPA) and GeneXpert MTB/RIF (Xpert) assays. Methods: This was a cross-sectional study that used sputum specimens collected from participants screened for STREAM 2 clinical trial between October 2017 and October 2019. Samples from 62 MTB smear-positive patients and rifampicin-resistant patients from peripheral health facilities were processed for Xpert and LPA as screening tests for eligibility in the trial. From November 2020, processed stored sputum samples were retrieved and genotyped to determine the presence of mixed-MTB strain infection using a standard 24-locus Mycobacterial Interspersed Repetitive Unit–Variable Number Tandem-Repeat (MIRU-VNTR). Samples with at least 20/24 MIRU-VNTR loci amplified were considered for analysis. Agar proportional Drug Susceptibility Test (DST) was performed on culture isolates of samples that had discordant results between LPA and Xpert. The impact of the presence of mixed-MTB strain on Xpert and LPA test interpretation was analyzed. Results: A total of 53/62 (85%) samples had analyzable results from MIRU-VNTR. The overall prevalence of mixed-MTB infection was 5/53 (9.4%). The prevalence was highest among male's 3/31 (9.7%) and among middle-aged adults, 4/30 (33.3%). Lineage 4 of MTB contributed 3/5 (60.0%) of the mixed-MTB infection prevalence. Having mixed MTB strain infection increased the odds of false susceptible Xpert test results (OR 7.556, 95% CI 0.88–64.44) but not for LPA. Being HIV-positive (P = 0.04) independently predicted the presence of mixed MTB infection. Conclusions: The presence of mixed-MTB strain infection may affect the performance of the GeneXpert test but not for LPA. For patients with high pre-test probability of rifampicin resistance, an alternative rapid method such as LPA should be considered. [ABSTRACT FROM AUTHOR]

This article discusses the process of replicative senescence in human embryonic bone marrow mesenchymal stem cells (MSCs). The study focuses on changes in cell motility and the organization of the actin cytoskeleton during replicative senescence. The researchers found that there are alterations in the localization of certain proteins and changes in cell morphology and motility during this process. The study suggests that Rho signaling pathways may be involved in replicative senescence, but further research is needed to understand the underlying mechanisms. The article also provides a list of references for further reading on related topics in cell biology and biochemistry. [Extracted from the article]

Pain associated with bone cancer remains poorly managed, and chemotherapeutic drugs used to treat cancer usually increase pain. The discovery of dual-acting drugs that reduce cancer and produce analgesia is an optimal approach. The mechanisms underlying bone cancer pain involve interactions between cancer cells and nociceptive neurons. We demonstrated that fibrosarcoma cells express high levels of autotaxin (ATX), the enzyme synthetizing lysophosphatidic acid (LPA). Lysophosphatidic acid increased proliferation of fibrosarcoma cells in vitro. Lysophosphatidic acid is also a pain-signaling molecule, which activates LPA receptors (LPARs) located on nociceptive neurons and satellite cells in dorsal root ganglia. We therefore investigated the contribution of the ATX–LPA–LPAR signaling to pain in a mouse model of bone cancer pain in which fibrosarcoma cells are implanted into and around the calcaneus bone, resulting in tumor growth and hypersensitivity. LPA was elevated in serum of tumorbearing mice, and blockade of ATX or LPAR reduced tumor-evoked hypersensitivity. Because cancer cell–secreted exosomes contribute to hypersensitivity and ATX is bound to exosomes, we determined the role of exosome-associated ATX–LPA–LPAR signaling in hypersensitivity produced by cancer exosomes. Intraplantar injection of cancer exosomes into naive mice produced hypersensitivity by sensitizing C-fiber nociceptors. Inhibition of ATX or blockade of LPAR attenuated cancer exosome-evoked hypersensitivity in an ATX–LPA–LPAR-dependent manner. Parallel in vitro studies revealed the involvement of ATX–LPA–LPAR signaling in direct sensitization of dorsal root ganglion neurons by cancer exosomes. Thus, our study identified a cancer exosomemediated pathway, which may represent a therapeutic target for treating tumor growth and pain in patients with bone cancer. [ABSTRACT FROM AUTHOR]

Background: China is one of the countries with the fastest growing prevalence of diabetes mellitus (DM) in the world. This study intended to investigate the association of single nucleotide polymorphisms (SNPs) of FHL5 and LPA with DM risk in the Chinese population. Methods: This case-control study involved 1,420 Chinese individuals (710 DM patients and 710 controls). Four candidate loci (rs2252816/rs9373985 in FHL5 and rs3124784/rs7765781 in LPA) were successfully screened. The association of SNPs with DM risk was assessed by logistic regression analysis. Differences in clinical characteristics among subjects with different genotypes were analyzed by one-way analysis of variance. Results: Overall analysis indicated that rs3124784 was associated with an increased risk of DM. Stratification analysis showed that rs3124784 significantly increased DM risk in different sub-groups (male, non-smoking, non-drinking, and BMI > 24), while rs7765781 increased DM risk only in participants with BMI ≤ 24. Rs2252816 was associated with the course of DM. We also found that rs2252816 GG genotype and rs9373985 GG genotype were linked to the increased cystatin c in DM patients. Conclusion: The genetic polymorphisms of LPA may be associated with DM risk in the Chinese population, which will provide useful information for the prevention and diagnosis of DM. [ABSTRACT FROM AUTHOR]

Purpose: To assess health-related quality of life (HRQOL) among adolescents and young adults (AYAs) with chronic conditions. Methods: AYAs (N = 872) aged 14–20 years completed NIH's Patient-Reported Outcomes Measurement Information System® (PROMIS®) measures of physical function, pain interference, fatigue, social health, depression, anxiety, and anger. Latent profile analysis (LPA) was used to group AYAs into HRQOL profiles using PROMIS T-scores. The optimal number of profiles was determined by model fit statistics, likelihood ratio test, and entropy. Multinomial logistic regression models were used to examine how LPA's HRQOL profile membership was associated with patient demographic and chronic conditions. The model prediction accuracy on profile membership was evaluated using Huberty's I index with a threshold of 0.35 for good effect. Results: A 4-profile LPA model was selected. A total of 161 (18.5%), 256 (29.4%), 364 (41.7%), and 91 (10.4%) AYAs were classified into Minimal, Mild, Moderate, and Severe HRQOL Impact profiles. AYAs in each profile had distinctive mean scores with over a half standard deviation (5-points in PROMIS T-scores) of difference between profiles across most HRQOL domains. AYAs who were female or had conditions such as mental health condition, hypertension, and self-reported chronic pain were more likely to be in the Severe HRQOL Impact profile. The Huberty's I index was 0.36. Conclusions: Approximately half of AYAs with a chronic condition experience moderate to severe HRQOL impact. The availability of risk prediction models for HRQOL impact will help to identify AYAs who are in greatest need of closer clinical care follow-up. [ABSTRACT FROM AUTHOR]

Background: Increased endothelial permeability of pulmonary vessels is a primary pathological characteristic of septic acute lung injury (ALI). Previously, elevated lysophosphatidic acid (LPA) levels and LPA2 (an LPA receptor) expression have been found in the peripheral blood and lungs of septic mice, respectively. However, the specific role of LPA2 in septic ALI remains unclear.Methods: A lipopolysaccharide (LPS)-induced model of sepsis was established in wild-type (WT) and global LPA2 knockout (Lpar2−/−) mice. We examined mortality, lung injury, assessed endothelial permeability through Evans blue dye (EBD) assay in vivo, and transendothelial electrical resistance (TEER) of mouse lung microvascular endothelial cells (MLMECs) in vitro. Enzyme-linked immunosorbent assay (ELISA), histopathological, immunofluorescence, immunohistochemistry, and Western blot were employed to investigate the role of LPA2 in septic ALI.Results:Lpar2 deficiency increased vascular endothelial permeability, impaired lung injury, and increased mortality. Histological examination revealed aggravated inflammation, edema, hemorrhage and alveolar septal thickening in the lungs of septic Lpar2−/− mice. In vitro, loss of Lpar2 resulted in increased permeability of MLMECs. Pharmacological activation of LPA2 by the agonist DBIBB led to significantly reduced inflammation, edema and hemorrhage, as well as increased expression of the vascular endothelial tight junction (TJ) protein zonula occludens-1 (ZO-1) and claudin-5, as well as the adheren junction (AJ) protein VE-cadherin. Moreover, DBIBB treatment was found to alleviate mortality by protecting against vascular endothelial permeability. Mechanistically, we demonstrated that vascular endothelial permeability was alleviated through LPA-LPA2 signaling via the PLC-PKC-FAK pathway.Conclusion: These data provide a novel mechanism of endothelial barrier protection via PLC-PKC-FAK pathway and suggest that LPA2 may contribute to the therapeutic effects of septic ALI. [ABSTRACT FROM AUTHOR]

Background and aims: India is a high-burden tuberculosis (TB) region and a drug-resistance hotspot. Despite numerous reports of pulmonary tuberculosis (PTB) cases, there needs to be more literature available on the importance of diagnostic methods in the case of extrapulmonary tuberculosis (EPTB). A prospective study was performed from July 2017 to June 2018 to compare the efficacy of conventional and molecular methods in detecting PTB and EPTB cases. Methods: Overall, 1000 presumptive PTB and 412 EPTB cases were subjected to staining (Ziehl- Neelsen [ZN] and fluorescent staining), culture, GeneXpert, and line probe assay (LPA). Results: The sensitivity, specificity, and strength of association [i.e., kappa (k) value of light-emitting diode-fluorescent microscopy, ZN, and GeneXpert] were calculated using standard formulae using solid culture as the gold standard. The sensitivity of GeneXpert in smear-positive/culture-positive PTB was comparable with the smear-negative/culture-positive PTB cases (95.7% vs. 87.5%) with an overall sensitivity and specificity of 90.5% and 90.1% in EPTB cases, respectively. However, sensitivity was lower for pleural fluid (75%) and tissues (85.7%). In pulmonary instances, 10% (6.7% Rifampicin [RIF] + isoniazid [INH] resistant and 3.3% INH monoresistant) drug resistance was observed, and no drug resistance was found in extra-pulmonary samples. Conclusion: Among conventional methods, fluorescent staining is more sensitive than ZN staining, while the sensitivity of GeneXpert varies w.r.t type of sample using culture positivity as the gold standard. In general, the present study suggests the promotion of universal drug susceptibility testing (DST) for all individuals with TB to control drug-resistant TB.

Fluorescence resonance energy transfer (FRET) biosensors have proven to be an indispensable tool in cell biology and, more specifically, in the study of G-protein signalling. The best method of measuring the activation status or FRET state of a biosensor is often fluorescence lifetime imaging microscopy (FLIM), as it does away with many disadvantages inherent to fluorescence intensity-based methods and is easily quantitated. Despite the significant potential, there is a lack of reliable FLIM-FRET biosensors, and the data processing and analysis workflows reported previously face reproducibility challenges. Here, we established a system in live primary mouse pancreatic ductal adenocarcinoma cells, where we can detect the activation of an mNeonGreen-Gαi3-mCherry-Gγ2 biosensor through the lysophosphatidic acid receptor (LPAR) with 2-photon time-correlated single-photon counting (TCSPC) FLIM. This combination gave a superior signal to the commonly used mTurquoise2-mVenus G-protein biosensor. This system has potential as a platform for drug screening, or to answer basic cell biology questions in the field of G-protein signalling.

LPA3 receptors were expressed in TREx HEK 293 cells, and their signaling and phosphorylation were studied. The agonist, lysophosphatidic acid (LPA), increased intracellular calcium and ERK phosphorylation through pertussis toxin-insensitive processes. Phorbol myristate acetate, but not LPA, desensitizes LPA3-mediated calcium signaling, the agonists, and the phorbol ester-induced LPA3 internalization. Pitstop 2 (clathrin heavy chain inhibitor) markedly reduced LPA-induced receptor internalization; in contrast, phorbol ester-induced internalization was only delayed. LPA induced rapid β-arrestin–LPA3 receptor association. The agonist and the phorbol ester-induced marked LPA3 receptor phosphorylation, and phosphorylation sites were detected using mass spectrometry. Phosphorylated residues were detected in the intracellular loop 3 (S221, T224, S225, and S229) and in the carboxyl terminus (S321, S325, S331, T333, S335, Y337, and S343). Interestingly, phosphorylation sites are within sequences predicted to constitute β-arrestin binding sites. These data provide insight into LPA3 receptor signaling and regulation.

Background: Nontuberculous mycobacteria (NTM) are generally free-living and have a potential to cause opportunistic infection. More than 190 species of NTM have been identified (http://www.bacterio.net/mycobacterium.html), up to 60 species of them are pathogenic. The distribution is geographically specific for some species and others are cosmopolitan. Understanding their diversity has practical value for the treatment and management of NTM disease. Except for isolated reports, there is no accurate information about NTM spread among Bulgarian patients. Material and Methods: We retrospectively analysed the data on the NTM isolated from patients throughout the country for the period from January 2010 to December 2017. Identification to the species level was done by Line Probe Assay (LPA). Results: 586 NTM strains were identified. We calculated the rate of NTM isolation in Bulgaria and NTM disease incidence by applying microbiological criteria defined by the American Thoracic Society (ATS). As a result, the laboratory isolation rate amounted to 1/100 000 people for the period, and when applying only the ATS criteria, the prevalence of NTM disease was 0,23 per 100 000 people. In both cases, we reported that NTM disease incidence remains low as compared to tuberculosis, though with an increasing trend. The prevalence of NTM varied significantly between northern and southern Bulgaria, as well as between the capital and the rest of the country's regions. Conclusions: Slow growing NTM species predominate in Bulgaria but rapidly growing ones have isolation levels higher than the average for Europe and closer to that of Asian countries. Given the demographic situation in Bulgaria and the changing climatic factors, NTM infections need special attention.

Routine genetic testing in hypercholesterolemia patients reveals a causative monogenic variant in less than 50% of affected individuals. Incomplete genetic characterization is partly due to polygenic factors influencing low‐density‐lipoprotein‐cholesterol (LDL‐C). Additionally, functional variants in the LPA gene affect lipoprotein(a)‐associated cholesterol concentrations but are difficult to determine due to the complex structure of the LPA gene. In this study we examined whether complementing standard sequencing with the analysis of genetic scores associated with LDL‐C and Lp(a) concentrations improves the diagnostic output in hypercholesterolemia patients. 1.020 individuals including 252 clinically diagnosed hypercholesterolemia patients from the FH Register Austria were analyzed by massive‐parallel‐sequencing of candidate genes combined with array genotyping, identifying nine novel variants in LDLR. For each individual, validated genetic scores associated with elevated LDL‐C and Lp(a) were calculated based on imputed genotypes. Integrating these scores especially the score for Lp(a) increased the proportion of individuals with a clearly defined disease etiology to 68.8% compared to 46.6% in standard genetic testing. The study highlights the major role of Lp(a) in disease etiology in clinically diagnosed hypercholesterolemia patients, of which parts are misclassified. Screening for monogenic causes of hypercholesterolemia and genetic scores for LDL‐C and Lp(a) permits more precise diagnosis, allowing individualized treatment. [ABSTRACT FROM AUTHOR]

Triglycerides are associated with cardiovascular risk largely mediated by their final product of catabolism, the cholesterol-rich remnant, which may come from the liver or the intestine. There is a direct association between fasting and postprandial triglycerides and cardiovascular risk. ApoB is associated with the number of particles and the amount of RC, which explains its determining role in CVD. Non-HDL cholesterol represents atherogenic lipoproteins, with correlation and concordance with apoB; both are cardiovascular risk markers. Lipoprotein (a) is a cardiovascular risk factor not only because of the cholesterol and apoB it contributes, but also because of its proinflammatory and prothrombotic activity. Viewing these factors holistically, it is important to note that it is primarily a reduction in apoB, remnant cholesterol and non-HDL cholesterol that reduces cardiovascular events. A 10% weight loss improves TG levels without reducing apoB or cardiovascular risk. Of the classic lipid-lowering agents, high-dose statins and ethyl eicosapentanoate reduce cardiovascular risk through apoB, RC, and non-HDL reduction. To date, we do not have lipid-lowering agents aimed at reducing Lp(a), which is only slightly reduced by PCSK9 inhibitors and markedly reduced by LDL apheresis. These residual risk markers serve not only to stratify cardiovascular risk but also to establish follow-up goals. [ABSTRACT FROM AUTHOR]

The α-arrestin domain containing protein 3 (ARRDC3) is a tumor suppressor in triple-negative breast carcinoma (TNBC), a highly metastatic subtype of breast cancer that lacks targeted therapies. Thus, understanding the mechanisms and targets of ARRDC3 in TNBC is important. ARRDC3 regulates trafficking of protease-activated receptor 1 (PAR1, also known as F2R), a G-protein-coupled receptor (GPCR) implicated in breast cancer metastasis. Loss of ARRDC3 causes overexpression of PAR1 and aberrant signaling. Moreover, dysregulation of GPCR-induced Hippo signaling is associated with breast cancer progression. However, the mechanisms responsible for Hippo dysregulation remain unknown. Here, we report that the Hippo pathway transcriptional co-activator TAZ (also known as WWTR1) is the major effector of GPCR signaling and is required for TNBC migration and invasion. Additionally, ARRDC3 suppresses PAR1-induced Hippo signaling via sequestration of TAZ, which occurs independently of ARRDC3-regulated PAR1 trafficking. The ARRDC3 C-terminal PPXY motifs and TAZ WW domain are crucial for this interaction and are required for suppression of TNBC migration and lung metastasis in vivo. These studies are the first to demonstrate a role for ARRDC3 in regulating GPCR-induced TAZ activity in TNBC and reveal multi-faceted tumor suppressor functions of ARRDC3. This article has an associated First Person interview with the first author of the paper.

Lysophosphatidic acid (LPA) is a small molecule glycerophospholipid, which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development. In this study, sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment. At first, the maturation medium containing estrus female sheep serum and synthetic oviduct fluid (SOF) were optimized for sheep IVF, and then the effects of LPA were investigated. From 0.1 to 10 μmol L–1, LPA had no significant effect on the cleavage rate (P>0.05), but the maturation rate and blastocyst rate increased dependently with LPA concentration (P

Laser-ion acceleration with ultra-short pulse, petawatt-class lasers is dominated by non-thermal, intra-pulse plasma dynamics. The presence of multiple ion species or multiple charge states in targets leads to characteristic modulations and even mono-energetic features, depending on the choice of target material. As spectral signatures of generated ion beams are frequently used to characterize underlying acceleration mechanisms, thermal, multi-fluid descriptions require revision for predictive capabilities and control in next-generation particle beam sources. We present an analytical model with explicit inter-species interactions, supported by extensive ab initio simulations. This enables us to derive important ensemble properties from the spectral distribution resulting from these multi-species effects for arbitrary mixtures. We further propose a potential experimental implementation with a novel cryogenic target, delivering jets with variable mixtures of hydrogen and deuterium. Free from contaminants and without strong influence of hardly controllable processes such as ionization dynamics, this would allow a systematic realization of our predictions for the multi-species effect.

The improvement of in vitro embryo development is a gateway to enhance the output of assisted reproductive technologies. The Wnt and Hippo signaling pathways are crucial for the early development of bovine embryos. This study investigated the development of bovine embryos under the influence of a Hippo signaling agonist (LPA) and a Wnt signaling inhibitor (DKK1). In this current study, embryos produced in vitro were cultured in media supplemented with LPA and DKK1. We comprehensively analyzed the impact of LPA and DKK1 on various developmental parameters of the bovine embryo, such as blastocyst formation, differential cell counts, YAP fluorescence intensity and apoptosis rate. Furthermore, single-cell RNA sequencing (scRNA-seq) was employed to elucidate the in vitro embryonic development. Our results revealed that LPA and DKK1 improved the blastocyst developmental potential, total cells, trophectoderm (TE) cells and YAP fluorescence intensity and decreased the apoptosis rate of bovine embryos. A total of 1203 genes exhibited differential expression between the control and LPA/DKK1-treated (LD) groups, with 577 genes upregulated and 626 genes downregulated. KEGG pathway analysis revealed significant enrichment of differentially expressed genes (DEGs) associated with TGF-beta signaling, Wnt signaling, apoptosis, Hippo signaling and other critical developmental pathways. Our study shows the role of LPA and DKK1 in embryonic differentiation and embryo establishment of pregnancy. These findings should be helpful for further unraveling the precise contributions of the Hippo and Wnt pathways in bovine trophoblast formation, thus advancing our comprehension of early bovine embryo development.

Housework is a key area of research across many academic fields as it represents the intersection of micro- and macro-level gender dynamics. Despite many shifts in both women's and men's economic activities, and men's changing gender beliefs, women remain largely responsible for the management and performance of domestic labor. Given the relationship between paid employment and household work, this research describes patterns of women's and men's housework before, during, and after the Great Recession. Using American Time Use Survey data, I perform latent profile analysis to document the distributions of housework tasks and time for women and men across these three time periods. While women perform the majority of housework across the time frame, women and men converge in their time during the Recession. Further, men's time becomes more varied and more similar to women's Post-Recession. The findings in this research brief highlight the connections between macro-level change and micro-level behavior.

This study aimed to examine the impact of COVID-19 on the pattern of social-emotional development and learning engagement of Korean students. It employs latent profile analysis to categorize middle and high school students based on their social-emotional development (action-orientation, optimism, perseverance, relationship with adults) and learning engagement (cooperative learning, and self-directed learning). The analysis, conducted during the fall semester of 2019 and the spring semester of 2020 amid COVID-19 school closures, explores nuanced patterns and effects. The findings reveal distinct student groups with discernible differences across all factors both before and during school closure. Notably, the research suggests that early social-emotional development may influence subsequent developmental stages and that a student's social-emotional skills correlate with cooperative and self-directed learning. Furthermore, the study highlights the pandemic's varied impact on student groups, indicating that those with advanced social-emotional competencies and established learning practices were resilient to school closures. In contrast, 'average' students faced challenges in cooperative and active learning during lockdown. The study underscores the need for targeted educational measures, particularly for at-risk students, and suggests proactive preparation for future pandemics. [ABSTRACT FROM AUTHOR]