Consecutive Stages Research Articles

Relationships between empathy, executive functions, and internalizing and externalizing symptoms in early adolescents

BackgroundPrevious studies have shown associations between internalizing and externalizing symptoms, executive functions, and empathy. However, the mechanisms that affect this relationship remain unclear. This study employed regression analysis to explore the relationships between empathy, executive functions, and internalizing and externalizing symptoms.MethodOur study consisted of three consecutive stages. In the first stage, we assessed 2150 adolescents' parents aged 10–12 using the Strengths and Difficulties Questionnaire to determine which participants should be included in the subsequent stages. In the second stage, we conducted a pilot study with 50 participants to assess the validity of the assessment system and the six film scenes to be used in the following stage. Finally, in the third stage, we obtained 170 participants' state empathy, trait empathy, and executive functioning.ResultsThe regression analysis showed that working memory (β = 0.261) and externalizing symptoms (β = -0.157) predicted cognitive empathy, but only externalizing symptoms predicted affective empathy (β = -0.193). The regression analysis also showed that cognitive empathy (β = -0.513) and affective empathy (β = -0.535) predicted externalizing symptoms, but only inhibition predicted internalizing symptoms (β = 0.158).ConclusionThis population-based study highlights the reciprocal effects of externalizing symptoms on cognitive and affective empathy. No significant relationship was found between internalizing symptoms and empathy. This study also highlights the effect of working memory on cognitive empathy. These findings may inform psychological interventions to improve empathic abilities in early adolescence.

The development of algorithms for safe control of an autonomous ship

The paper presents the results of a research on the development of algorithms for ship’s safe ship trajectory calculation and automatic control of the ship along the determined trajectory. These methods are intended for use in the autonomous navigation system of an Unmanned Surface Vehicle (USV) or a Maritime Autonomous Surface Ship (MASS). The work presents the consecutive stages of a research that must be carried out in order to develop a system that will perform the task of multidimensional ship control in a port. The safe trajectory is calculated with the use of the Ant Colony Optimization (ACO) method. The ship motion control is based on the Linear Matrix Inequalities (LMI) method, implemented using the Mathworks and Yalmip libraries. The results of controlling the ship’s motion in accordance with the calculated trajectory are presented in the paper. With the use of the developed system, the ship can move autonomously based on the information from the DGPS system and a gyrocompass. The presented results concerned a computer simulation of maneuvers in the port of the Blue Lady ship from the Foundation for Shipping Safety and Environmental Protection.

Photocatalysis Meets Copper Catalysis: A New Opportunity for Asymmetric Multicomponent Radical Cross-Coupling Reactions.

ConspectusIn recent years, radical-mediated cross-coupling reactions have emerged as a compelling strategy for achieving a rich diversity in molecular topologies under benign conditions. However, the inherent high reactivity of radicals presents considerable challenges in controlling reaction pathways and selectivity, which often results in a limited range of substrates and a constrained reaction profile. Given the capacity of visible-light photoredox catalysis to generate a wide variety of reactive radicals and radical ions in a controlled manner and the propensity of copper complexes toward radical species, we envisaged that the synergy between chiral copper catalysts and photoactive catalysts would pave the way for developing innovative strategies. This integration is poised to unlock a broad spectrum of enantioselective multicomponent radical cross-coupling reactions.In this Account, we describe our insights and recent efforts in the realm of enantioselective multicomponent radical cross-coupling reactions. These advancements have been achieved through the innovative application of dual photoredox/copper catalysis or bifunctional copper catalysis under visible light irradiation. Our work is systematically divided into two sections based on the activation modes. The first section focuses on photoinduced copper-catalyzed chiral C-C and C-O bond formation through a radical addition/nucleophilic trap sequence. Our discussion of chiral C-C bond formation is particularly concentrated on the asymmetric carbocyanation and carboarylation of vinylarenes, 1,3-enynes, and 1,3-dienes. Our findings underscore that irradiation with visible light can adeptly modulate the pace of radical generation, thus orchestrating consecutive reaction stages and ensuring the attainment of both chemo- and stereoselectivity. In the domain of chiral C-O bond formation, leveraging carboxylic acids as a nucleophilic oxygen source, we introduce a suite of esterification reactions of benzylic, allylic, and propargylic radicals. These radicals are derived from a variety of radical precursors, showcasing the versatility of our approach. The following section highlights our innovative discovery in the field of dual photoredox/copper catalysis, which enables enantioselective three-component radical transformations via the direct activation of aromatic alkenes. This methodology begins with the generation of formal distonic radical anions through the photocatalytic single-electron reduction of aromatic alkenes, thus, enabling orthogonal reactivity. Employing H2O, D2O, and CO2 as external electrophile agents, we have developed three types of radical cyanofunctionalization reactions: hydrocyanation, deuteriocyanation, and cyanocarboxylation. These reactions provide practical access to diversely functionalized chiral nitriles with high enantiomeric excess.Collectively, these synthetic methodologies highlight the immense potential inherent in the synergistic integration of photocatalysis and asymmetric copper catalysis. This Account aspires to deepen our comprehension of the advantages conferred by these catalytic systems, elucidating the crucial role of photocatalysis in facilitating enantioselective multicomponent radical cross-couplings. We anticipate that this Account will provide valuable insights and stimulate the evolution of innovative methodologies within this rapidly expanding field.

Breeding of new kiwifruit (Actinidia chinensis) cultivars with yellow (golden) fleshed and superior characteristics

The most widely cultivated species globally is Actinidia deliciosa cv. ‘Hayward’. However, in recent years, consumers have shown greater demand for new varieties with novel flesh colour, flavour and appearance in international markets. To meet these expectations, one breeding study was started in 2016 at the University of Guilan in Iran to develop new kiwifruit cultivars with superior characteristics. In this breeding program, 201 female and 534 male hybrid genotypes were obtained from combinations of six different parents using the controlled cross-breeding technique. The evaluations were carried out over three consecutive stages. In the first stage, 201 female hybrid genotypes and control (‘Golden’) were evaluated based on 25 phenological and pomological traits. In the second stage, hybrids passed the desired threshold values in 7 key attributes were selected. Sensory testing in the next stage evaluated these selected hybrid genotypes more. As a result of the first stage, 30 hybrid genotypes with fruit weight (90 ≥ g) were selected. Then, in the second stage, 21 hybrids that showed acceptable dry matter, hue angle, flesh firmness, soluble solid content, acidity, and ascorbic acid were selected. Finally, 13 hybrid genotypes received high scores in sensory testing, and just two hybrid genotypes represented a 100% of novelty with positive texture. These genotypes were selected as potential cultivar candidates. In future studies, fruit yield, disease/pest resistance, and post-harvest performance trials of the 13 selected candidates will be evaluated at different sites. Thus, the ones that give the best results will be introduced to kiwifruit growers.

Petrology of Tourmaline-Bearing Blueschist from SW Tianshan, China, and Its Implication in B-Rich Fluid Migration in Subduction Zone

Abstract Boron geochemistry can track fluid–rock interaction during subduction zone metamorphism. Rare tourmaline-bearing blueschists, which are associated with ultrahigh-pressure (UHP) serpentinites are first recognized in SW Tianshan, China. Detailed petrology, whole-rock and mineral chemistry, B isotope analysis, and modeling characterized two consecutive stages of tourmaline crystallization (Tur-I, Tur-II). Tourmaline included in, or intergrown with, garnet and the cores of tourmaline in rock matrixes and veins are Tur-I, which grew during prograde metamorphism at 430°C to 460°C/470°C, ~1.9–2.1 GPa. The rims of tourmaline in rock matrixes and veins are Tur-II, which formed during initial exhumation at 460°C to 490°C, ~2.1–1.7 GPa. Variable δ11B values of tourmaline (+8‰, Tur-I to −2‰, Tur-II) point to a 11B-rich signature of the fluid infiltrating at Stage I. With progressing metamorphism, δ11B decreased in the fluid. The high-δ11B Tur-I (up to +8‰) could not have crystallized from fluid released from the high-pressure metapelites (−12‰ to −7‰) and metabasites (−15‰ to −5‰) surrounding the tourmaline host rocks given the lower δ11B values. Modeling of B isotope fractionation yields the δ11B values of −9‰ to −5‰, −11‰ to −1‰, and +8‰ to +17‰ for the fluids equilibrium with the restitic metapelites, metabasites, and serpentinites, respectively. The tourmaline and whole-rock B isotope data, along with the tourmaline compositions, point to the associated serpentinites as source of the fluid that infiltrated the metamorphic rocks. This fluid was released by the partial dehydration of serpentinites through the reaction antigorite + brucite = olivine + water at forearc depth. We propose that metabasites in subduction zones can acquire 11B-rich signatures through interaction with serpentinite-derived fluids, leading to the formation of robust tourmaline minerals at shallow levels. As a new reservoir of heavy boron, these metabasites can then transport this signature to greater depths.

Development of a porcine model of skin and soft-tissue infection caused by Staphylococcus aureus, including methicillin-resistant strains suitable for testing topical antimicrobial agents.

In view of the ever-increasing representation of Staphylococcus spp. strains resistant to various antibiotics, the development of invivo models for evaluation of novel antimicrobials is of utmost importance. In this article, we describe the development of a fully immunocompetent porcine model of extensive skin and soft tissue damage suitable for testing topical antimicrobial agents that matches the real clinical situation. The model was developed in three consecutive stages with protocols for each stage amended based on the results of the previous one. In the final model, 10 excisions of the skin and underlying soft tissue were created in each pig under general anesthesia, with additional incisions to the fascia performed at the base of the defects and immediately inoculated with Staphylococcus aureus suspension. One pig was not inoculated and used as the negative control. Subsequently, the bandages were changed on Days 4, 8, 11, and 15. At these time points, a filter paper imprint technique (FPIT) was made from each wound for semi-quantitative microbiological evaluation. Tissue samples from the base of the wound together with the adjacent intact tissue of three randomly selected defects of each pig were taken for microbiological, histopathological, and molecular-biological examination. The infection with the inoculated S. aureus strains was sufficient during the whole experiment as confirmed by both FPIT and from tissue samples. The dynamics of the inflammatory markers and clinical signs of infection are also described. A successfully developed porcine model is suitable for invivo testing of novel short-acting topical antimicrobial agents.

An Artificial Intelligent System for Prostate Cancer Diagnosis in Whole Slide Images

In recent years a significant demand to develop computer-assisted diagnostic tools to assess prostate cancer using whole slide images has been observed. In this study we develop and validate a machine learning system for cancer assessment, inclusive of detection of perineural invasion and measurement of cancer portion to meet clinical reporting needs. The system analyses the whole slide image in three consecutive stages: tissue detection, classification, and slide level analysis. The whole slide image is divided into smaller regions (patches). The tissue detection stage relies upon traditional machine learning to identify WSI patches containing tissue, which are then further assessed at the classification stage where deep learning algorithms are employed to detect and classify cancer tissue. At the slide level analysis stage, entire slide level information is generated by aggregating all the patch level information of the slide. A total of 2340 haematoxylin and eosin stained slides were used to train and validate the system. A medical team consisting of 11 board certified pathologists with prostatic pathology subspeciality competences working independently in 4 different medical centres performed the annotations. Pixel-level annotation based on an agreed set of 10 annotation terms, determined based on medical relevance and prevalence, was created by the team. The system achieved an accuracy of 99.53% in tissue detection, with sensitivity and specificity respectively of 99.78% and 99.12%. The system achieved an accuracy of 92.80% in classifying tissue terms, with sensitivity and specificity respectively 92.61% and 99.25%, when 5x magnification level was used. For 10x magnification, these values were respectively 91.04%, 90.49%, and 99.07%. For 20x magnification they were 84.71%, 83.95%, 90.13%.

Ways of self-renewal of kindergarten teachers in forest kindergartens change

ABSTRACT The study examined the self-renewal and conceptual change kindergarten teachers in Israel, underwent from a professional career perspective in the process of implementing the transition to a forest kindergarten model. Educator’s career consists of four consecutive stages: entry, building, mid-career, and the late career. In education, self-renewal is a phenomenon characteristic in the mid-career stage. Self-renewal has scarcely been studied in early childhood educators. This article examines self-renewal based on interpretations of interviews with 19 kindergarten teachers in Israel who responded to an institutional initiative to create forest kindergartens. Results show three ways that mid-career kindergarten teachers used to update their pedagogical knowledge and reframe their educational perceptions. Self-renewal reflects the professional kindergarten teachers’ background based on their past experiences and aspects of entrepreneurial behavior. The article can help policymakers improve early childhood education and launch new enterprises.

Stream of variation modeling and monitoring for heterogeneous profiles in multi-stage manufacturing processes

ABTRACT Advanced manufacturing systems typically involve multiple operating stages, where in-process data are observed in the form of profiles that contain rich information for process monitoring. Emerging methodologies have been developed for multi-stage profile monitoring. However, the cases where different stages’ profiles are heterogeneous and variations propagate over stages have not been addressed so far. To this end, this article proposes a heterogeneous profiles-based stream of variation (HP-SoV) framework for multi-stage manufacturing process monitoring. It uses functional decomposition to extract features of heterogeneous profiles from different stages, where the decomposition coefficients are regarded as latent states that propagate along consecutive stages to capture the variation propagation. HP-SoV includes many existing models as its special cases, and enjoys an efficient inference algorithm via maximum likelihood estimation. Based on the one-step-ahead forecast errors of HP-SoV, a group monitoring scheme is further developed for online monitoring. Extensive numerical studies explore the modeling robustness and monitoring effectiveness of HP-SoV under different settings. A case study demonstrates the applicability of HP-SoV in multi-stage process monitoring for heterogeneous profiles.

Development of a new patient-reported outcome measure for patients with multiple sclerosis: the Multiple Sclerosis Autonomy Scale (MSAS)

BackgroundIn multiple sclerosis (MS), the measure of the loss of autonomy appears to be particularly relevant to provide adapted and personalized responses to improve the quality of care in routine clinical practice. In this context, this research aimed to develop a new patient-reported outcome measure (PROM) evaluating MS patients’ autonomy, in order to provide an easy-to-use tool in the context of the relations between healthcare professionals and patients with MS, and to be used in future clinical trials for treatment assessment. MethodsThis research was conducted in two consecutive stages. First, a preliminary questionnaire was generated using an innovative sociological approach for MS (after literature review, patient interviews, experts’ opinion, and patient focus groups). This questionnaire was then completed by patients with MS, before the reduction of the scale while maintaining relevant information, using a principal component analysis. The internal consistency reliability was assessed using the Cronbach's alpha coefficient. The external validity was evaluated using an analysis of variance to estimate the relation between the reduced questionnaire dimension scores and disease severity classes assessed by the SymptoMScreen questionnaire. ResultsThe first qualitative step of the research led to provide a definition of disease-related autonomy as perceived by patients (to be able to carry out the roles the patient thinks the most important whether or not he/she receives assistance) as well as an associated taxonomy. On this basis, a preliminary questionnaire of 131 items grouped into 13 social dimensions was generated (seven dimensions with 63 questions concerning all the patients, and six dimensions with 68 questions concerning specific patients). This questionnaire was completed on a web platform by 653 analyzable patients with MS. Their main characteristics were as follows: female patients: 83.6 %, mean age at MS diagnosis: 34.8 ± 10.5 years, age ≥40 years at data collection: 68.1 %, MS duration ≥5 years: 68.4 %, severe MS (SymptoMScreen score ≥30): 36.8 %. On the basis on completed 131-item questionnaires, it was reduced in a 36-item short form of 10 social dimensions (five dimensions with 19 questions concerning all the patients, and five dimensions with 17 questions concerning specific patients). The internal consistency of the final questionnaire was good for all the dimensions, as the Cronbach's alpha coefficient ranged from 0.77 to 0.85 depending on dimensions. The construct validity of the questionnaire was also confirmed. ConclusionOur research allowed to build the first PROM designed to evaluate the autonomy of patients suffering from MS: the Multiple Sclerosis Autonomy Scale (MSAS). A confirmatory study, conducted in patients with MS using this validated questionnaire, is currently conducted.

Content Evaluation of Pruritus, Skin Pain, and Sleep Disturbance Patient-Reported Outcome Measures in Adolescents and Adults With Moderate-to-Severe Atopic Dermatitis: Qualitative Interviews.

Pruritus, skin pain, and sleep disturbance are a significant burden to individuals with moderate-to-severe atopic dermatitis (AD) and negatively impact quality of life. Fit-for-purpose patient-reported outcome measures (PROMs) that assess AD-related pruritus, skin pain, and sleep disturbance are important for evaluating the effectiveness of new AD treatments. To evaluate content validity of five AD-related PROMs in adolescents and adults with moderate-to-severe AD (the Worst Pruritus Numeric Rating Scale [NRS], the AD Skin Pain NRS; the Sleep Disturbance NRS, the skin pain-specific Patient Global Impression of Change [PGIC], and the skin pain-specific Patient Global Impression of Severity [PGIS]) and to assess patient-reported experience with pruritus, skin pain, and sleep disturbance. A qualitative study in adolescents (aged 12-17 years) and adults with moderate-to-severe AD was conducted in two consecutive stages. In each stage, two iterative rounds of individual interviews were conducted by experienced interviewers. All interviews included concept elicitation and cognitive debriefing components. Data were analysed using thematic analysis. A total of 27 adults and 20 adolescents with moderate-to-severe AD participated in the initial content evaluation (Stage 1) of the Worst Pruritus NRS [1.0] and AD Skin Pain NRS [1.0] (n = 26; 16 adults, 10 adolescents) and in the subsequent content evaluation (Stage 2) of the revised Worst Pruritus NRS [1.1], revised AD Skin Pain NRS [1.1], Sleep Disturbance NRS, skin pain-specific PGIC, and skin pain-specific PGIS (n = 21; 11 adults, 10 adolescents). Results were generally aligned and consistent for both adult and adolescent participants. Additionally, we found that sleep disturbance is relevant and important for evaluation in adults and adolescents with moderate-to-severe AD (Stage 2), while also providing further confirmation of this for pruritus and skin pain (Stages 1 and 2). Our findings support the content validity of the revised Worst Pruritus NRS [1.1], revised AD Skin Pain NRS [1.1], Sleep Disturbance NRS, skin pain-specific PGIC, and skin pain-specific PGIS in individuals aged ≥12 years with moderate-to-severe AD. Furthermore, the findings support sleep disturbance, skin pain, and pruritus as being relevant and important features for both adolescents and adults with moderate-to-severe AD.

Reliability and usability of a novel inertial sensor-based system to test craniocervical flexion movement control

BackgroundNeck pain has a significant global impact, ranking as the fourth leading cause of disability. Recurrent neck pain often leads to impaired sensorimotor control, particularly in craniocervical flexion (CFF). The Craniocervical Flexion Test (CCFT) has been widely investigated to assess the performance of deep cervical flexor muscles. However, its use requires skilled assessors who need to subjectively detect compensations, progressive increases in range of motion (ROM) or excessive superficial flexor activation during the test. The aim of this study was to design and develop a novel Craniocervical Flexion Movement Control Test (CFMCT) based on inertial sensor technology and real-time computer feedback and to evaluate its safety and usability, as well as inter and intra-rater reliability in both healthy individuals and patients with neck pain.MethodsWe used inertial sensor technology associated with new software that provides real-time computer feedback to assess CCF movement control through two independent test protocols, the progressive consecutive stages protocol (including progressive incremental stages of ROM) and the random stages protocol (providing dynamic and less predictable movement patterns). We determined intra and inter-rater reliability and standard error of the measurement for both protocols. The participants rated their usability was analysed through the System Usability Scale (SUS) and possible secondary effects associated with the tests were registered.ResultsThe progressive consecutive stages protocol and the random stages protocol were safe and easy to use (SUS scores of 82.00 ± 11.55 in the pain group and 79.56 ± 13.36 in the asymptomatic group). The progressive consecutive stages protocol demonstrated good inter-rater reliability (intraclass correlation coefficient [ICC] ≥ 0.75) and moderate to good intra-rater reliability (ICC 0.62–0.80). However, the random stages protocol exhibited lower intra-rater reliability, especially in the neck pain group, where the reliability values were poor in some cases (ICC 0.48–0.72).ConclusionThe CFMCT (progressive consecutive stages protocol) is a promising instrument to evaluate CCF motor control in patients with chronic neck pain. It has potential for telehealth assessment and easy adherence for exercise prescription and seems to be a safe and usable tool for patients with pain and asymptomatic individuals. Its use as a test or for exercise needs to be further investigated to facilitate its transfer to clinical practice.

Temporal Gene Expression Profiles From Pollination to Seed Maturity in Sorghum Provide Core Candidates for Engineering Seed Traits.

Sorghum (Sorghum bicolor (L.) Moench) is a highly nutritional multipurpose millet crop. However, the genetic and molecular regulatory mechanisms governing sorghum grain development and the associated agronomic traits remain unexplored. In this study, we performed a comprehensive transcriptomic analysis of pistils collected 1-2 days before pollination, and developing seeds collected -2, 10, 20 and 30 days after pollination of S. bicolor variety M35-1. Out of 31 337 genes expressed in these stages, 12 804 were differentially expressed in the consecutive stages of seed development. These exhibited 10 dominant expression patterns correlated with the distinct pathways and gene functions. Functional analysis, based on the pathway mapping, transcription factor enrichment and orthology, delineated the key patterns associated with pollination, fertilization, early seed development, grain filling and seed maturation. Furthermore, colocalization with previously reported quantitative trait loci (QTLs) for grain weight/size revealed 48 differentially expressed genes mapping to these QTL regions. Comprehensive literature mining integrated with QTL mapping and expression data shortlisted 25, 17 and 8 core candidates for engineering grain size, starch and protein content, respectively.

Importance of Shatkriya Kala in the Manifestation and Management of Amavata

“Ayurveda, the ancient Indian system of medicine, elucidates a comprehensive framework for understanding the pathogenesis and progression of diseases. A critical concept within this framework is the “Shatkriya kala”, explained by Acharya Sushruta which describes the six stages of disease development. These stages- Sanchaya, Prakopa, Prasara, Sthana Samshraya, Vyakta, Bheda- offer insight into the dynamic process of disease evaluation and guide timely intervention. It shows the stepwise progression of Dosha and Rogas. A clear understanding about the consecutive stages of disease, early diagnosis, prognosis, to adopt preventive and curative measures in Ayurvedic Pathology, the Shatkriya kala becomes essential. Likewise, understanding and inervening at the appropriate stages of Shatkriya Kala is crucial for the effective management and treatment of Amavata. In Ayurveda, Amavata is a debilitating condition compared to Rheumatoid Arthritis in modern, characterised by the accumulation of Ama(toxins) and the aggravation of Vata dosha. So, through this article, an attempt is made to highlight the importance of Shatkriya kala in the context of Amavata.

Proteomic analysis during seed development provides insight into the early establishment of seed dormancy in Xanthium strumarium.

This experiment was carried out to provide a comprehensive insight into the protein activities involved in dormancy establishment in seeds of common cocklebur (Xanthium strumarium), an annual plant with two dimorphic seeds contained in one casing known as a burr. These consist of a smaller dormant seed and a larger non-dormant seed. The proteome profile was compared between developing dormant and non-dormant seeds of Xanthium strumarium at five consecutive stages including three, 10, 20, 30, and 45 days after burr emergence (stages 1 to 5). We identified 6524 proteins in total, and approximately 3.6% of these were differentially abundant proteins (DAPs) between the two seed types. Both seed types showed fundamental changes in developmental programs during the examined stages. More than 38% of all DAPs were observed at the first stage, supporting the importance of the early developmental stage in seed fate determination. The detected DAPs at stage 1 were mainly associated with the cell division phase, which showed a delay in the dormant seeds. Over-representation of proteins responsible for cell wall biosynthesis, cytokinesis, and seed development were detected for non-dormant seeds at the first stage, while dormancy-associated proteins showed less abundance. Stage 3 was the critical stage for switching processes toward seed maturation and abscisic acid (ABA) signaling. Interestingly, higher abundance proteins in the mature non-dormant seed were mainly involved in the facilitation of seed germination. Taken together, the temporal pattern of the accumulated proteins in developing dormant seeds demonstrated a delay in the initiation of active cell division, enriched response to ABA, and defects in seed maturation. Moreover, stored proteins in the mature dormant seed delay germination but not dormancy induction. Finally, our results suggest that dormancy may be established at a stage of seed development earlier than previously thought.

A single-cell atlas of spatial and temporal gene expression in the mouse cranial neural plate.

The formation of the mammalian brain requires regionalization and morphogenesis of the cranial neural plate, which transforms from an epithelial sheet into a closed tube that provides the structural foundation for neural patterning and circuit formation. Sonic hedgehog (SHH) signaling is important for cranial neural plate patterning and closure, but the transcriptional changes that give rise to the spatially regulated cell fates and behaviors that build the cranial neural tube have not been systematically analyzed. Here we used single-cell RNA sequencing to generate an atlas of gene expression at six consecutive stages of cranial neural tube closure in the mouse embryo. Ordering transcriptional profiles relative to the major axes of gene expression predicted spatially regulated expression of 870 genes along the anterior-posterior and mediolateral axes of the cranial neural plate and reproduced known expression patterns with over 85% accuracy. Single-cell RNA sequencing of embryos with activated SHH signaling revealed distinct SHH-regulated transcriptional programs in the developing forebrain, midbrain, and hindbrain, suggesting a complex interplay between anterior-posterior and mediolateral patterning systems. These results define a spatiotemporally resolved map of gene expression during cranial neural tube closure and provide a resource for investigating the transcriptional events that drive early mammalian brain development.

Routing and scheduling of trains and engines in a railway marshalling station yard

In a busy railway marshalling station, train and engine traffic management in the receiving/departure yard plays a crucial role in efficient and stable operations. Traditionally, a track assignment problem (TAP) is solved to assign tracks to trains for berthing at a receiving/departure yard. However, the TAP does not encompass shunting operations in the yard (e.g., engine replacement and train disassembly), which can result in additional scheduling challenges for dispatchers and route conflicts between operations. This paper investigates a train and engine routing and scheduling problem (TERSP) in a receiving/departure yard of railway marshalling stations, which involves simultaneously assigning routes and scheduling route-setting start times for both train and shunting operations to be conducted in the yard. By introducing the concepts of task, activity, and pattern, we transform the original problem into assigning pre-generated patterns incorporating both route and route-setting start time alternatives to activities. The transformed problem is formulated into a compact binary integer linear programming model with a linear number of constraints and the objective of minimizing the total time deviation of all involved tasks. An improved technique that relies on listing all maximal (bi)cliques in a constructed graph is designed to effectively model the time coherence and track section occupation constraints. A heuristic that gradually expands the patterns for the identified key activities by adding more start time alternatives is applied to remedy an infeasible model caused by potential route conflicts. In addition, a rolling horizon algorithm that decomposes the original problem into consecutive smaller stages using either a time-rolling or a train-rolling rule is developed to efficiently solve instances. Finally, numerical experiments based on the physical layouts and real timetables of a receiving yard and a departure yard of a large marshalling station in China are conducted to assess the performance and applicability of our proposed approaches. The results demonstrate that our approaches typically find (near-)optimal solutions within several minutes for the investigated instances by simultaneously addressing different classes of yard operations and resources.

Sustainable experimental cultivation of the Greek endemic Helichrysum amorginum Boiss. and Orph.: Assessment of the total phenolic content at different flower harvest stages

Helichrysum amorginum Boiss. and Orph. constitutes a range-restricted, endemic species of Greece with known pharmaceutical potency. The current work presents the results of a targeted evaluation of the total phenolic content (TPC) of methanolic extracts of H. amorginum flower heads under cultivation conditions at three consecutive harvest stages in two separate cultivation locations across a period of two years (2018 and 2019). The harvested inflorescence tissue was assessed for TPC via the Folin–Ciocalteu method in three and four-year old H. amorginum individuals. Older plants, during the second evaluation year, generally showed higher flower TPC content throughout. The pooled analysis of the results indicated significant differences in TPC among samples of extracts prepared with plant material from different harvest stages with the early flowering stage (A) presenting higher TPC in both experimental cultivation locations (142.6-156 mg GAE g-1 extract on average among the two cultivation locations) followed by the full bloom stage (B) (139-145.7 mg GAE g-1 extract on average) and the late bloom or early post-anthesis stage (C) which showed the lowest TPC in all cases (130.3-131 mg GAE g-1 extract on average). The current results provide for the first-time basic information on the optimum inflorescence harvest stage during the prolonged flowering period of cultivated H. amorginum in terms of TPC. The proposed work can be incorporated into the establishment of a sustainable cultivation protocol for achieving polyphenol-rich extracts and ultimately contributes to the utilization of H. amorginum in the pharmaceutical/cosmetic sectors.

The SPARC-related modular calcium binding 1 (Smoc1) regulated by androgen is required for mouse gubernaculum development and testicular descent.

Testicular descent occurs in two consecutive stages: the transabdominal stage and the inguinoscrotal stage. Androgens play a crucial role in the second stage by influencing the development of the gubernaculum, a structure that pulls the testis into the scrotum. However, the mechanisms of androgen actions underlying many of the processes associated with gubernaculum development have not been fully elucidated. To identify the androgen-regulated genes, we conducted large-scale gene expression analyses on the gubernaculum harvested from luteinizing hormone/choriogonadotropin receptor knockout (Lhcgr KO) mice, an animal model of inguinoscrotal testis maldescent resulting from androgen deficiency. We found that the expression of secreted protein acidic and rich in cysteine (SPARC)-related modular calcium binding 1 (Smoc1) was the most severely suppressed at both the transcript and protein levels, while its expression was the most dramatically induced by testosterone administration in the gubernacula of Lhcgr KO mice. The upregulation of Smoc1 expression by testosterone was curtailed by the addition of an androgen receptor antagonist, flutamide. In addition, in vitro studies demonstrated that SMOC1 modestly but significantly promoted the proliferation of gubernacular cells. In the cultures of myogenic differentiation medium, both testosterone and SMOC1 enhanced the expression of myogenic regulatory factors such as paired box 7 (Pax7) and myogenic factor 5 (Myf5). After short-interfering RNA-mediated knocking down of Smoc1, the expression of Pax7 and Myf5 diminished, and testosterone alone did not recover, but additional SMOC1 did. These observations indicate that SMOC1 is pivotal in mediating androgen action to regulate gubernaculum development during inguinoscrotal testicular descent.

Reactive extraction of muconic acid by hydrophobic phosphonium ionic liquids - Experimental, modelling and optimisation with Artificial Neural Networks

Muconic acid is a six-carbon dicarboxylic acid with conjugated double bonds that finds extensive use in the food (additive), chemical (production of adipic acid, monomer for functional resins and bio-plastics), and pharmaceutical sectors. The biosynthesis of muconic acid has been the subject of recent industrial and scientific attention. However, because of its low concentration in aqueous solutions and high purity requirement, downstream separation presents a significant problem. Artificial Neural Networks and Differential Evolution were used to optimize process parameters for the recovery of muconic acid from aqueous streams in a system with n-heptane as an organic diluent and ionic liquids as extractants. The system using 120 g/L tri-hexyl-tetra-decyl-phosphonium decanoate dissolved in n-heptane, pH of the aqueous phase 3, 20 min contact time, and 45 °C temperature assured a muconic acid extraction efficiency of 99,24 %. Low stripping efficiency compared to extraction efficiency was observed for the optimum conditions on the extraction step (120 g/L ionic liquids dissolved in heptane). However, re-extraction efficiencies obtained for the recycled organic phase in three consecutive stages were close to the first extraction stage. The mechanism analysis proved that the analysed phosphonium ionic liquids (PILSs) extracts only undissociated molecules of muconic acid through H-bonding.