Induced PSIG expression by Herbacetin contributes to suppressing the proliferation, migration, and invasion of melanoma cells.

Melanoma is a very common malignant tumor with poor prognosis. Herbacetin is a flavonol compound with outstanding anti-tumor effects. Our work investigated the biological effects and mechanism of Herbacetin in melanoma. In our study, the mRNA and protein expressions were assessed using qRT-PCR, Western blot and IHC. MSP was performed to evaluated PGIS promoter methylation level. Cell viability, migration and invasion were examined by MTT assay, transwell migration and invasion assay, respectively. Our results revealed that DNMT3B was markedly upregulated in melanoma, while PGIS was lowly expressed. Herbacetin treatment could not only inhibit the proliferation, migration, invasion of melanoma cells and inhibit the growth of melanoma in vivo. Herbacetin could also restore the abnormal expressions of DNMT3B and PGIS in melanoma cells and tumor tissues. PGIS silencing neutralized the inhibitory effects of Herbacetin on the malignant behaviors of melanoma cells. Besides, DNMT3B knockdown promoted PGIS expression via reducing PGIS promoter methylation level in melanoma cells, thereby inhibiting malignant behaviors of melanoma cells. And as expected, the inhibitory effects of Herbacetin on malignant behaviors of melanoma cells were all abolished by DNMT3B overexpression. Collectively, Herbacetin reduced DNMT3B expression to upregulate PGIS in melanoma cells and participated in suppressing the proliferation, migration, and invasion of melanoma cells.

Comparing patient global impression of severity and patient global impression of change to evaluate test-retest reliability of depression, non-small cell lung cancer, and asthma measures.

PURPOSE: Score reproducibility is an important measurement property of fit-for-purpose patient-reported outcome (PRO) measures. It is commonly assessed via test-retest reliability, and best evaluated with a stable participant sample, which can be challenging to identify in diseases with highly variable symptoms. To provide empirical evidence comparing the retrospective (patient global impression of change [PGIC]) and current state (patient global impression of severity [PGIS]) approaches to identifying a stable subgroup for test-retest analyses, 3 PRO Consortium working groups collected data using both items as anchor measures. METHODS: The PGIS was completed on Day 1 and Day 8 + 3 for the depression and non-small cell lung cancer (NSCLC) studies, and daily for the asthma study and compared between Day 3 and 10. The PGIC was completed on the final day in each study. Scores were compared using an intraclass correlation coefficient (ICC) for participants who reported "no change" between timepoints for each anchor. RESULTS: ICCs using the PGIS "no change" group were higher for depression (0.84 vs. 0.74), nighttime asthma (0.95 vs. 0.53) and daytime asthma (0.86 vs. 0.68) compared to the PGIC "no change" group. ICCs were similar for NSCLC (PGIS: 0.87; PGIC: 0.85). CONCLUSION: When considering anchor measures to identify a stable subgroup for test-retest reliability analyses, current state anchors perform better than retrospective anchors. Researchers should carefully consider the type of anchor selected, the time period covered, and should ensure anchor content is consistent with the target measure concept, as well as inclusion of both current and retrospective anchor measures.

The first report of prostacyclin and its physiological roles in insects.

Prostaglandins (PGs) mediate physiological processes of insects as well as mammals. Prostaglandin I2 (PGI2) is a relatively well-known eicosanoid with potent hormone-like actions on various tissues of vertebrates, however, its presence and biosynthetic pathway have not been described in insects. This study demonstrated that fat bodies of the lepidopteran species, Spodoptera exigua, contained ~ 3.6 pg/g PGI2. To identify its biosynthetic pathway, a PGI2 synthase gene of S. exigua (Se-PGIS) was predicted from a transcriptome of S. exigua; 25.6% homology with human PGIS was demonstrated. Furthermore, a predicted three-dimensional structure of Se-PGIS was demonstrated to be 38.3% similar to the human PGIS ortholog, including catalytic residues. Se-PGIS was expressed in all developmental stages of S. exigua and most abundant larval and adult stages; immune challenging of larvae significantly up-regulated these expression levels. The inducible expression of Se-PGIS expression was followed by a greater than four-fold increase in the concentration of PGI2 in fat bodies 10 h after immune challenge. RNA interference (RNAi) against Se-PGIS was performed by injecting double-stranded RNA (dsRNA). Under these RNAi conditions, cellular immune responses (e.g., hemocyte-spreading behavior, nodulation, phenoloxidase activity) were not affected by bacterial challenge. The addition of PGI2 to larvae treated with an eicosanoid biosynthesis inhibitor did not rescue the immunosuppression. Interestingly, PGI2 injection significantly suppressed nodule formation in response to bacterial challenge. In addition to the negative effect of PGI2 against immunity, the Se-PGIS-RNAi treatment significantly interfered with immature development and severely impaired oocyte development in female adults; the addition of PGI2 to RNAi-treated females significantly recovered oocyte development. Se-PGIS RNAi treatment also impaired male fertility by reducing fecundity after mating with untreated females. These results suggest that PGI2 acts as a negative regulator of immune responses initiated by other factors and mediates S. exigua development and reproduction.

Psychometric properties and validation of two global impression questionnaires (PGI-S, PGI-I) for stress incontinence in a German-speaking female population.

AIMS: The Patient Global Index of Severity (PGI-S) and the Patient Global Index of Improvement (PGI-I) are global impression questionnaires developed in English and validated in women with stress urinary incontinence (SUI). This validation study tested the psychometric properties of German-language versions of the two questionnaires in German-speaking women with SUI. METHODS: The German-language PGI-S and PGI-I were psychometrically tested and validated using the SF-12 questionnaire, the Kinǵs Health Questionnaire (KHQ), clinical parameters, incontinence episode frequency and pad use in 311 patients before and 3 months after receiving a TVT-O or TVT tape for SUI. RESULTS: At baseline and 3 months postoperatively there was a positive correlation between PGI-S response categories and clinical parameters, IEF and pad use, and nearly all KHQ subscales. There were no correlations between response categories of PGI-S at baseline and PGI-I at 3 months and the SF-12 scales PCS-12 and MCS-12. CONCLUSION: Our results demonstrated good psychometric properties of the German-language PGI-S and PGI in German-speaking women with SUI.

A Reciprocal Triangulation Process For Identifying And Mapping Potential Land Use Conflict.

Land use in many areas is highly contested. An understanding of the nature of such conflicts, and of spatial variation in their intensity, is required to develop planning solutions. We present a novel process for attaining these outcomes which involves mapping of values and potential conflict between stakeholders determined using participatory GIS (PGIS) processes. Our starting point was an a priori identification of the values that were potentially in conflict. We produced quantitative and qualitative maps of each of the values that formed a basis for workshop discussion among small stakeholder groups. Each participant was asked to complete a questionnaire to determine their values and their attitudes to land use and to map the places that they would not be prepared to lose. Principal components analysis was used to identify the major independent axes in values and attitudes among all participants. We then used repeatable descriptive quantitative procedures to identify attitude groups. These analyses allowed us to identify potential conflicts between values that could be expressed in land use, spatial variation in attachment of groups and the intensity of potential conflict. In our test of the process in the Tarkine region of Tasmania, Australia, we found that land use conflict was multidimensional, involving incompatible recreational activities and incompatibility between nature conservation and economic production. Two-fifths of the area was shown to be not in contest, with considerable spatial variation in the intensity of conflict potential in the remainder. This latter variation could facilitate a process of land use compromise.

[Assessment of severity of neurogenic bowel dysfunction in chronic patients with a simple 1-item questionnaire (PGI-S)]. / Sévérité des troubles colorectaux associés aux maladies neurologiques et évaluation pratique par questionnaire à un item (PGI-S).

INTRODUCTION: Bowel symptoms (constipation and incontinence) are frequent in patients with a neurologic disease, but rarely assessed in rehabilitation centres. AIM: To study the prevalence of neurogenic bowel dysfunction (NBD) in those patients, and to assess its severity with the Patient Global Impression of Severity (PGI-S). MATERIAL: Prospective study by questionnaires, with the Neurogenic Bowel Dysfunction Score (0-47) and the PGI-S, a 1-item questionnaire (absent, mild, moderate, severe) for the severity of the bowel symptoms, and the Bristol Stool Chart for stool consistency. All patients presenting a chronic (>2months) neurological disease were included. RESULTS: Inclusion of 169 patients, 97 with spinal cord injury, 42 with multiple sclerosis and 30 with hemiplegia. In each population, prevalence of constipation was 67 %, 45 % and 17 %, of pelvic floor dyssynergia 82 %, 45 % and 27 %, and leakages (gas or stools) de 74 %, 48 % and 43 %, respectively. Moderate to severe bowel symptoms were seen in 61 % of spinal cord injury, 43 % of multiple sclerosis and 23 % of hemiplegic patients, with NBD scores of 11.9±6.5, 5.7±4.9 and 3.7±4.2, respectively (P<0.01). There was a significant relation between PGI-S and NBD score (P<0.01). Significant lower NBD scores were associated with normal stool consistency (Bristol type 3 or 4) (P<0.01). In case of severe bowel symptoms, the use of transanal irrigation was hampered by patients' motivation and acceptation, and their autonomy. CONCLUSION: PGI-S and Bristol Stool Chart are reliable tools to assess the presence of bowel symptoms in clinical practice.

A structural model of PpoA derived from SAXS-analysis-implications for substrate conversion.

In plants and mammals, oxylipins may be synthesized via multi step processes that consist of dioxygenation and isomerization of the intermediately formed hydroperoxy fatty acid. These processes are typically catalyzed by two distinct enzyme classes: dioxygenases and cytochrome P450 enzymes. In ascomycetes biosynthesis of oxylipins may proceed by a similar two-step pathway. An important difference, however, is that both enzymatic activities may be combined in a single bifunctional enzyme. These types of enzymes are named Psi-factor producing oxygenases (Ppo). Here, the spatial organization of the two domains of PpoA from Aspergillus nidulans was analyzed by small-angle X-ray scattering and the obtained data show that the enzyme exhibits a relatively flat trimeric shape. Atomic structures of the single domains were obtained by template-based structure prediction and docked into the enzyme envelope of the low resolution structure obtained by SAXS. EPR-based distance measurements between the tyrosyl radicals formed in the activated dioxygenase domain of the enzyme supported the trimeric structure obtained from SAXS and the previous assignment of Tyr374 as radical-site in PpoA. Furthermore, two phenylalanine residues in the cytochrome P450 domain were shown to modulate the specificity of hydroperoxy fatty acid rearrangement.

Thromboxane A2 synthase inhibitors prevent production of infectious hepatitis C virus in mice with humanized livers.

BACKGROUND & AIMS: A 3-dimensional (3D) culture system for immortalized human hepatocytes (HuS-E/2 cells) recently was shown to support the lifecycle of blood-borne hepatitis C virus (HCV). We used this system to identify proteins that are active during the HCV lifecycle under 3D culture conditions. METHODS: We compared gene expression profiles of HuS-E/2 cells cultured under 2-dimensional and 3D conditions. We identified signaling pathways that were activated differentially in the cells, and analyzed their functions in the HCV lifecycle using a recombinant HCV-producing cell-culture system, with small interfering RNAs and chemical reagents. We investigated the effects of anti-HCV reagents that altered these signaling pathways in mice with humanized livers (carrying human hepatocytes). RESULTS: Microarray analysis showed that cells cultured under 2-dimensional vs 3D conditions expressed different levels of messenger RNAs encoding prostaglandin synthases. Small interfering RNA-mediated knockdown of thromboxane A2 synthase (TXAS) and incubation of hepatocytes with a TXAS inhibitor showed that this enzyme is required for production of infectious HCV, but does not affect replication of the HCV genome or particle release. The TXAS inhibitor and a prostaglandin I2 receptor agonist, which has effects that are opposite those of thromboxane A2, reduced serum levels of HCV and inhibited the infection of human hepatocytes by blood-borne HCV in mice. CONCLUSIONS: An inhibitor of the prostaglandin synthase TXAS inhibits production of infectious HCV particles in cultured hepatocytes and HCV infection of hepatocytes in mice with humanized livers. It therefore might be therapeutic for HCV infection.

Assessment of natural radioactivity levels and hazard indicators in Tarakeshwor Municipality, Nepal through in-situ technique and multivariate analysis.

The evaluation of primordial radionuclide concentrations in rapidly urbanized and concrete-laden areas through the importation of construction materials from various regions of Nepal is both important and essential. This study utilized a portable gamma-ray spectrometer (PGIS 2) to analyze the distribution of three natural radionuclides: uranium (238U), thorium (232Th), and potassium (40K) in Tarakeshwor Municipality, Kathmandu, Nepal. The measured dose rates ranged from 70.22 nSv hr-1 to 163.66 nSv hr-1, with an average of 124.65±20.29 nSv hr-1, surpassing the global average of 59 nSv hr-1. The activity concentrations of 40K, 238U, and 232Th exceeded global averages, indicating relatively higher natural radioactivity concentrations in the region. Specifically, the average values for 40K, 238U, and 232Th were 935.26±172.30 Bq kg-1, 80.47±15.53 Bq kg-1, and 80.44±18.58 Bq kg-1, respectively. The calculated radium equivalent (Raeq) ranged from 132.26 to 351.22 Bq kg-1, and the annual gonadal equivalent dose (AGED) varied from 372.61 to 1028.81 µSv yr-1. The annual effective dose rates for indoor and outdoor environments were 0.54±0.09 mSv yr-1 and 0.15±0.03 mSv yr-1, respectively, both exceeding the global average. The representative level index (RLI) within the study area averaged 1.96±0.32, indicating an elevated radiation risk. The excess lifetime cancer risk (ELCR) values for outdoor and indoor environments were 0.52×10-3 ±0.09 ×10-3 and 1.87 ×10-3 ±0.31×10-3, respectively, surpassing the world average. Additionally, external hazard indices (Hex) ranged from 0.36 to 0.59, while internal hazard indices (Hin) ranged from 0.38 to 1.20, both indicating values higher than UNSCEAR recommendations. These findings underscore the necessity for further experimental analysis employing ex-situ equipment. The data generated in this study can provide a valuable baseline for future assessments and interventions in radiation risk management guidelines within the country.

Personal growth initiative across the life span: a systematic review protocol of quantitative studies using the Personal Growth Initiative Scale-II.

BACKGROUND: Personal Growth Initiative (PGI) a multi-dimensional construct, conceptualised as a skill set that helps individuals to intentionally grow is considered an important construct throughout the life span. Coping with the challenges, transitions, experiences and stressors of life requires an active growth orientation. In previous empirical research, the construct has been measured by either the PGIS-I or PGIS-II, of which only the latter takes account of the theoretically established multi-dimensionality of the construct. This paper describes the protocol for conducting a systematic review of published peer-reviewed empirical research articles on the multi-dimensional construct of PGI. The aim of this review is threefold: (1) to better understand the multi-dimensional construct PGI in different contexts and populations, (2) to improve our understanding of the reliability and validity of the PGIS-II in various research populations and (3) to obtain an overview of its associations with relevant psychosocial factors. METHODS: For the development of this protocol, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols (PRISMA-P) reporting guidelines were used. Four databases and one registry will be searched using a predetermined search strategy for relevant studies. Studies will be screened, by two reviewers independently, against the established inclusion criteria. During the data extraction process, the quality of the included studies will be assessed using the Quality Assessment for Survey Studies in Psychology (Q-SSP). The collected data will then be analysed and reported in both narrative and tabular form according to the PRISMA 2020 statement guidelines and flow diagram. DISCUSSION: The findings of this study will increase our understanding of the dynamics of PGI throughout the lifespan, its associations with other psychosocial factors and the psychometric properties of the PGIS-II. It will also clarify where additional research is needed. The objectives of the proposed review can provide a basis for the development of practical applications and interventions. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42022377342.

Design, synthesis and characterization of lead compounds as anti-inflammatory drugs targeting mPGES-1 via enzymelink screening.

Aim: The objective of this study was to synthesize and validate a set of compounds that selectively inhibit mPGES-1, with the potential to be developed into a novel anti-inflammatory drug. Methods: The synthesized compounds were characterized using 1H NMR spectroscopy and LC-MS to confirm their structure. Cellular and enzymatic assays were used to demonstrate their inhibitory activity on prostaglandin E2 production. Results: Docking studies revealed that compounds containing fluoro-, chloro- and methyl- groups displayed strong inhibitory activity against prostaglandin E2. The inhibitory activity of synthesized trimethyl and trifluoro was further validated using enzymatic and cell migration assays. Conclusion: The findings demonstrated that the synthesized compounds possess significant potential as a new generation of nonsteroidal anti-inflammatory drugs that selectively target mPGES-1 with fewer side effects.

Recommendations to guide sampling effort for polygon-based participatory mapping used to identify perceived ecosystem services hotspots.

Participatory mapping is increasingly used to map spatial variation in people's perceptions about ecosystem services. It has growing use in the identification of locations where places perceived to be important converge. Few recommendations have been published to navigate decisions about sampling effort in participatory mapping research when polygon data is collected, although one recommendation is for ≥ 25 participants assuming each participant maps c. 4-5 polygons per ecosystem service. Underlying data informing this recommendation reflects a particular context: collected using postal questionnaires to map a vast spatial area in southern Australia. Although not intended as definitive or suited to all contexts, the 25 participant (or 100-125 polygon) minimum sometimes informs participatory mapping research. Our empirical work, undertaken using face-to-face questionnaires in a small Vietnamese coastal study area, suggests the recommendation may not be appropriate in all contexts. We propose a modified stepwise approach which:•Prioritises spatial agreement (polygon overlap) rather than polygon count and participant numbers to assess data sufficiency•Uses narratives to triangulate outputs generated from participatory mapping data to reduce uncertainty related to low polygon counts.

Understanding and modifying Fabry disease: Rationale and design of a pivotal Phase 3 study and results from a patient-reported outcome validation study.

The use of available treatments for Fabry disease (FD) (including enzyme replacement therapy [ERT]) may be restricted by their limited symptom improvement and mode of administration. Lucerastat is currently being investigated in the MODIFY study as oral substrate reduction therapy for the treatment of FD. By reducing the net globotriaosylceramide (Gb3) load in tissues, lucerastat has disease-modifying potential to improve symptoms and delay disease progression. MODIFY is a multicenter, double-blind, randomized, placebo-controlled, parallel-group Phase 3 study (ClinicalTrial.gov: NCT03425539); here we present the rationale and design of this study. Eligible adults with a genetically confirmed diagnosis of FD and FD-specific neuropathic pain entered screening. Patients were randomized (2:1) to receive either oral lucerastat twice daily or placebo for 6 months; treatment allocation was stratified according to sex and ERT treatment status. The main objectives of MODIFY are to assess the effects of lucerastat on neuropathic pain, gastrointestinal (GI) symptoms, FD biomarkers, and determine its safety and tolerability. Neuropathic pain and GI symptoms are key features of FD that have a significant impact on quality of life. Despite various tools available to assess pain and GI symptoms, there are currently limited tools available to assess neuropathic and GI symptoms in FD, validated according to health authority guidelines. Based on FDA recommendations, we undertook a patient-reported outcome (PRO) validation study, using a novel eDiary-based PRO tool to assess the validity of evaluating neuropathic pain as a primary efficacy endpoint in MODIFY. Results from the PRO validation study are included. To date, MODIFY is the largest Phase 3 clinical study conducted in patients with FD. Enrollment to MODIFY is now complete, with 118 patients randomized. Results will be presented in a separate publication. Long-term effects of lucerastat are being assessed in the ongoing open-label extension study (NCT03737214).

Treatment Satisfaction of Galcanezumab in Japanese Patients with Episodic Migraine: A Phase 2 Randomized Controlled Study.

INTRODUCTION: This analysis evaluated the treatment satisfaction of Japanese patients receiving galcanezumab (GMB) as a preventive medication for episodic migraine (4-14 monthly migraine headache days). METHODS: This phase 2, randomized, double-blind, placebo-controlled study enrolled patients aged 18-65 years at 40 centers in Japan. Patients were randomized 2:1:1 to receive monthly subcutaneous injections of placebo (PBO, n = 230), GMB 120 mg (n = 115), or GMB 240 mg (n = 114) for 6 months. Patients' experience with treatment was measured using the Patient Global Impression of Severity (PGI-S), Patient Global Impression of Improvement (PGI-I), and Patient Satisfaction with Medication Questionnaire-Modified (PSMQ-M) scales. PGI-S was administered at baseline and months 1-6, PGI-I at months 1-6, and PSMQ-M at months 1 and 6. Prespecified analyses were differences between GMB and PBO in PGI-I and the change from baseline in PGI-S, and evaluating positive responses for the PGI-I and PSMQ-M. RESULTS: Average change ± SE from baseline across months 1-6 was - 0.09 ± 0.05 (PBO), - 0.17 ± 0.07 (GMB 120 mg, p = 0.33), and - 0.30 ± 0.07 (GMB 240 mg, p = 0.013) for PGI-S. Average PGI-I across months 1-6 was 3.39 ± 0.05 (PBO), 2.55 ± 0.07 (GMB 120 mg, p < 0.05), and 2.71 ± 0.07 (GMB 240 mg, p < 0.05). Reductions of 2.8-3.0 monthly migraine headache days corresponded to 25-31% higher positive PGI-I response rates with GMB compared with PBO. Positive PSMQ-M response rates for satisfaction and preference were statistically significantly higher for GMB compared with PBO (odds ratio [95% confidence interval], all p < 0.05 vs. PBO): satisfaction GMB 120 mg (3.142 [1.936-5.098]) and GMB 240 mg (3.924 [2.417-6.369]), and preference GMB 120 mg (3.691 [2.265-6.017]) and GMB 240 mg (3.510 [2.180-5.652]). CONCLUSION: Japanese patients with episodic migraine receiving preventive treatment with GMB are significantly more satisfied than those receiving PBO. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02959177 (registered November 7, 2016).

Engineering 'Enzymelink' for screening lead compounds to inhibit mPGES-1 while maintaining prostacyclin synthase activity.

Aim: This study investigated our Enzymelinks, COX-2-10aa-mPGES-1 and COX-2-10aa-PGIS, as cellular cross-screening targets for quick identification of lead compounds to inhibit inflammatory PGE2 biosynthesis while maintaining prostacyclin synthesis. Methods: We integrated virtual and wet cross-screening using Enzymelinks to rapidly identify lead compounds from a large compound library. Results: From 380,000 compounds virtually cross-screened with the Enzymelinks, 1576 compounds were identified and used for wet cross-screening using HEK293 cells that overexpressed individual Enzymelinks as targets. The top 15 lead compounds that inhibited mPGES-1 activity were identified. The top compound that specifically inhibited inflammatory PGE2 biosynthesis alone without affecting COX-2 coupled to PGI2 synthase (PGIS) for PGI2 biosynthesis was obtained. Conclusion: Enzymelink technology could advance cyclooxygenase pathway-targeted drug discovery to a significant degree.

[Interaction of prostacyclin synthase with cytochromes P450]. / Vzaimodeistvie prostatsiklinsintazy s tsitokhromami P450.

Biosensor experiments on investigation of interaction between prostacyclin synthase (PGIS) and different proteins of the cytochrome P450 monooxygenase systems were perfomed. Interaction of PGIS with microsomal (CYP21A2, CYP2E1) and mitochondrial (CYP27A1, CYP11B1, CYP11B2, CYP11A1) cytochrome P450s was detected. Kinetic and equilibrium parameters of protein complexes formation were determined. Data obtained suggest an essential role of these hemoproteins interaction in regulation of prostacyclin and thromboxane A2 biosynthesis.