BACKGROUND: Fine characterization of gene expression patterns is crucial to understand many aspects of embryonic development. The chicken embryo is a well-established and valuable animal model for developmental biology. The period spanning from the third to sixth embryonic days (E3 to E6) is critical for many organ developments. Hybridization chain reaction RNA fluorescent in situ hybridization (HCR RNA-FISH) enables multiplex RNA detection in thick samples including embryos of various animal models. However, its use is limited by tissue opacity.
RESULTS: We optimized HCR RNA-FISH protocol to efficiently label RNAs in whole mount chicken embryos from E3.5 to E5.5 and adapted it to ethyl cinnamate (ECi) tissue clearing. We show that light sheet imaging of HCR RNA-FISH after ECi clearing allows RNA expression analysis within embryonic tissues with good sensitivity and spatial resolution. Finally, whole mount immunofluorescence can be performed after HCR RNA-FISH enabling as exemplified to assay complex spatial relationships between axons and their environment or to monitor GFP electroporated neurons.
CONCLUSIONS: We could extend the use of HCR RNA-FISH to older chick embryos by optimizing HCR RNA-FISH and combining it with tissue clearing and 3D imaging. The integration of immunostaining makes possible to combine gene expression with classical cell markers, to correlate expressions with morphological differentiation and to depict gene expressions in gain or loss of function contexts. Altogether, this combined procedure further extends the potential of HCR RNA-FISH technique for chicken embryology.

During organ development, tissue stem cells first expand via symmetric divisions and then switch to asymmetric divisions to minimize the time to obtain a mature tissue. In the Drosophila midgut, intestinal stem cells switch their divisions from symmetric to asymmetric at midpupal development to produce enteroendocrine cells. However, the signals that initiate this switch are unknown. Here, we identify the signal as ecdysteroids. In the presence of ecdysone, EcR and Usp promote the expression of E93 to suppress Br expression, resulting in asymmetric divisions. Surprisingly, the primary source of pupal ecdysone is not from the prothoracic gland but from dorsal internal oblique muscles (DIOMs), a group of transient skeletal muscles that are required for eclosion. Genetic analysis shows that DIOMs secrete ecdysteroids during mTOR-mediated muscle remodeling. Our findings identify sequential endocrine and mechanical roles for skeletal muscle, which ensure the timely asymmetric divisions of intestinal stem cells.

Environmental deterioration is one of the major problems the globe is facing in the modern period. On the other hand, several groups around the world have endeavored to launch efforts to protect the planet, such as the Sustainable Development Goals. Therefore, the proposed objectives\' primary duty is to strike a balance between development and environmental concerns. This study looked at 65 Belt and Road Initiative (BRI) economies to see how factors, including the economic complexity index, urbanization, ICT, higher education, and green innovation, affected carbon emissions in the presence of sustainable development. Annual time series data from 2000 to 2020 have been used in the analysis. This study employs the CC-EMG to determine the durability of the association between the variables. AMG and quantile GMM regression estimations were used to test the robustness and reproducibility of the results. The results reveal that higher education and green innovation help lower carbon emissions, whereas the economic complexity index and urbanization are beneficial for increasing economic activity and advancing information and communication technologies. The economic complexity index, ICT, and higher education are all negatively impacted by green innovation. Important policy implications of the computed coefficients for the selected and other developing markets in planning a suitable path forward to a sustainable environment are also provided.

The insect steroid hormone ecdysone plays a critical role in insect development. Several recent studies have shown that ecdysone enters cells through Organic Anion Transporting Polypeptides (OATPs) in insects such as flies and mosquitoes. However, the conservation of this mechanism across other arthropods and the role of this transporter in canonical ecdysone pathways are less well studied. Herein we functionally characterized the putative ecdysone importer (EcI) from two major agricultural moth pests: Helicoverpa armigera (cotton bollworm) and Spodoptera frugiperda (fall armyworm). Phylogenetic analysis of OATP transporters across the superphylum Ecdysozoa revealed that EcI likely appeared only at the root of the arthropod lineage. Partial disruption of EcI in S. frugiperda decreased embryo hatching rate and larval survival, suggesting that this gene is essential for development in vivo. Depletion and re-expression of EcI in the lepidoptera cell line RP-HzGUT-AW1(MG) demonstrated this protein\'s ability to control ecdysone mediated signaling in gene regulation, its role in ecdysone mediated cell death, and its sensitivity to rifampicin, a well-known organic anion transporter inhibitor. Overall, this work sheds light on ecdysone uptake mechanisms across insect species and broadens our knowledge of the physiological roles of OATPs in the transportation of endogenous substrates.

Spodoptera litura Fab. is a polyphagous pest causing damage to many agriculture crops leading to yield loss. Recurrent usage of synthetic pesticides to control this pest has resulted in resistance development. Plant-derived diterpenoid compound andrographolide was isolated from the leaves of Andrographis paniculata. It was analysed by gas chromatography-mass spectroscopy and quantified by HPLC. Nutritional indices and digestive enzymatic profile were evaluated. Third, fourth and fifth instar larvae were treated with different concentrations of andrographolide. At 3, 6 and 9 ppm-treated concentrations the larvae showed decreased RGR, RCR, ECI, ECD values with adverse increase in AD. The digestive enzymes were significantly inhibited when compared with control. Conspicuously, andrographolide showed pronounced mortality of S. litura by inhibition of enzyme secretion and intake of food. The binding ability of andrographolide with CYTP450 showed high affinity with low binding energy. Andrographolide has the potential to be exploited as a biocontrol agent against S. litura as an eco-friendly pesticide.

The control of technological systems by human operators has been the object of study for many decades. The increasing complexity in the digital age has made the optimization of the interaction between system and human operator particularly necessary. In the present thematic issue, ten exemplary articles are presented, ranging from observational field studies to experimental work in highly complex navigation simulators. For the human operator, the processes of attention play a crucial role, which are captured in the contributions listed in this thematic issue by eye-tracking devices. For many decades, eye tracking during car driving has been investigated extensively (e.g. 6; 5). In the present special issue, Cvahte Ojsteršek & Topolšek (4) provide a literature review and scientometric analysis of 139 eye-tracking studies investigating driver distraction. For future studies, the authors recommend a wider variety of distractor stimuli, a larger number of tested participants, and an increasing interdisciplinarity of researchers. In addition to most studies investigating bottom-up processes of covered attention, Tuhkanen, Pekkanen, Lehtonen & Lappi (10) include the experimental control of top-down processes of overt attention in an active visuomotor steering task. The results indicate a bottom-up process of biasing the optic flow of the stimulus input in interaction with the top-down saccade planning induced by the steering task. An expanding area of technological development involves autonomous driving where actions of the human operator directly interact with the programmed reactions of the vehicle. Autonomous driving requires, however, a broader exploration of the entire visual input and less gaze directed towards the road centre. Schnebelen, Charron & Mars (9) conducted experimental research in this area and concluded that gaze dynamics played the most important role in distinguishing between manual and automated driving. Through a combination of advanced gaze tracking systems with the latest vehicle environment sensors, Bickerdt, Wendland, Geisler, Sonnenberg & Kasneci (2021) conducted a study with 50 participants in a driving simulator and propose a novel way to determine perceptual limits which are applicable to realistic driving scenarios. Eye-Computer-Interaction (ECI) is an interactive method of directly controlling a technological device by means of ocular parameters. In this context, Niu, Gao, Xue, Zhang & Yang (8) conducted two experiments to explore the optimum target size and gaze-triggering dwell time in ECI. Their results have an exemplary application value for future interface design. Aircraft training and pilot selection is commonly performed on simulators. This makes it possible to study human capabilities and their limitation in interaction with the simulated technological system. Based on their methodological developments and experimental results, Vlačić, Knežević, Mandal, Rođenkov & Vitsas (11) propose a network approach with three target measures describing the individual saccade strategy of the participants in this study. In their analysis of the cognitive load of pilots, Babu, Jeevitha Shree, Prabhakar, Saluja, Pashilkar & Biswas (3) investigated the ocular parameters of 14 pilots in a simulator and during test flights in an aircraft during air to ground attack training. Their results showed that ocular parameters are significantly different in different flying conditions and significantly correlate with altitude gradients during air to ground dive training tasks. In maritime training the use of simulations is per international regulations mandatory. Mao, Hildre & Zhang (7) performed a study of crane lifting and compared novice and expert operators. Similarities and dissimilarities of eye behavior between novice and expert are outlined and discussed. The study of Atik & Arslan (2) involves capturing and analyzing eye movement data of ship officers with sea experience in simulation exercises for assessing competency. Significant differences were found between electronic navigation competencies of expert and novice ship officers. The authors demonstrate that the eye tracking technology is a valuable tool for the assessment of electronic navigation competency. The focus of the study by Atik (1) is the assessment and training of situational awareness of ship officers in naval Bridge Resource Management. The study shows that eye tracking provides the assessor with important novel data in simulator based maritime training, such as focus of attention, which is a decisive factor for the effectiveness of Bridge Resource Management training. The research presented in the different articles of this special thematic issue cover many different areas of application and involve specialists from different fields, but they converge on repeated demonstrations of the usefulness of measuring attentional processes by eye movements or using gaze parameters for controlling complex technological devices. Together, they share the common goal of improving the potential and safety of technology in the digital age by fitting it to human capabilities and limitations.

Interactive feedback of interface elements and low level of spatial accuracy are two main key points for the interaction research in the Eye-computer interaction system. This study tried to solve these two problems from the perspective of human-computer interactions and ergonomics. Two experiments were conducted to explore the optimum target size and gaze-triggering dwell time of the eye-computer interaction (ECI) system. Experimental Series 1 was used as the pre-experiment to identify the size that has a greater task completion rate. Experimental Series 2 was used as the main experiment to investigate the optimum gaze-triggering dwell time by using a comprehensive evaluation of the task completion rate, reaction time, and NASA-TLX (Task Load Index). In Experimental Series 1, the optimal element size was determined to be 256 × 256p x 2. The conclusion of Experimental Series 2 was that when the dwell time is set to 600 ms, the efficiency of the interface is the highest, and the task load of subjects is minimal as well. Finally, the results of Experiment Series 1 and 2 have positive effects on improving the usability of the interface. The optimal control size and the optimal dwell time obtained from the experiments have certain reference and application value for interface design and software development of the ECI system.

Major depressive disorder (MDD) is characterized by pervasive mood disturbance as well as deficits in emotional processing, reactivity, and regulation. There is accumulating evidence that MDD is characterized by emotional patterns consistent with environmental disengagement, as reflected in attenuated positive and negative emotional reactivity, consistent with Emotion Context Insensitivity (ECI) theory. However, MDD individuals vary considerably in the extent to which they exhibit specific alterations in patterns of emotional responding. Emotions are complex, multicomponent processes that invoke responses across multiple functional domains and levels of analysis, including subjective experience, behavior, autonomic regulation, cognition, and neural processing. In this article, I review the current state of the literature on emotional responding and MDD from the lens of ECI. I focus on the importance of assessing emotional indices from multiple levels of analysis across development and contexts. I also discuss methodological and measurement issues that may contribute to inconsistent findings. In particular, I emphasize how psychophysiological measures can help elucidate emotional processes that underlie the pathophysiology of MDD as part of an integrated and contextualized approach.

Összefoglaló. A malignus csonttumorok sebészi ellátása során kialakuló szegmentális csontdefektusok pótlása fontos szempont a végtagmegtartó sebészetben. Felnőttkorban a megoldás rendszerint tumorprotézis beültetése, 10 évesnél fiatalabb gyermekeknél azonban ez nehezen alkalmazható módszer a kis csontméret és az igen magas várható szövődményarány miatt. A bemutatott, hazánkban még ritkán alkalmazott beavatkozás, a tumoros csontszegmentum műtét alatti sugárkezelése, visszaültetése megfelelő rekonstrukciós lehetőség a végtag funkciójának megtartása mellett, csökkentve a későbbi reoperációk, protézisrevíziók számát. Célkitűzés: A hazánkban eddig az ismertetett módon végzett műtétek bemutatása, az eredmények összevetése nemzetközi irodalmi adatokkal. Módszer: Magyaroszágon eddig 12 alkalommal végeztünk végtagmegtartó műtétet malignus csonttumor miatt 12 évesnél fiatalabb gyermekeknél, és e műtétek során biológiai rekonstrukciós módszerként extracorporalis irradiatiót, autograft-reimplantatiót (ECRT, ECI) végeztünk. A műtétek mindegyike primer malignus csonttumor (Ewing-sarcoma 7 esetben, osteosarcoma 4 esetben, chondrosarcoma 1 esetben) miatt történt. Betegeink átlagéletkora 9 (3-12) év volt, az átlagos utánkövetési idő 32,5 (2-73) hónap. A felmérés során fizikális vizsgálat, valamint minden alkalommal röntgenfelvételek készítése történt, szükség esetén szövettani mintavétellel, vérelemzéssel vagy egyéb képalkotással kiegészítve. Eredményeinket nemzetközi irodalmi adatokkal hasonlítottuk össze. Eredmények: Lokális tumorkiújulást egyetlen esetben sem észleltünk, másik csontot érintő skip (\'ugró\') metastasis miatt egy alkalommal amputatiót végeztünk. A szövődmények tekintetében eredményeink megfelelnek a nemzetközi irodalomban leírtaknak. Két esetben jelentkezett szeptikus szövődmény (16,7%). A resectiós sík radiológiai átépülése 3-9 hónap alatt történt meg az esetek 60%-ában. Álízület, grafttörés, graft részleges elhalása miatt 3 esetben végeztünk reoperációt, kétszer újabb allograft felhasználásával, egy esetben pedig tumorprotézis beültetésével. Betegeink szubjektív véleménye a módszerről pozitív, a legtöbb esetben megőrizték jó fizikai aktivitásukat, az esetleges reoperációk ellenére végtagjukat terhelik, akár sporttevékenységet is végeznek. Következtetések: Vizsgálatunk alapján az extracorporalis irradiatio hasznos biológiai rekonstrukciós módszer 12 évesnél fiatalabb gyermekek esetében szegmentális csontdefektusok pótlására tumoros indikációval. A felmerülő szövődmények aránya alatta marad a hasonló korban beültetett tumorprotézisek szövődményarányainak, ideális esetben pedig több, további kiterjesztett műtét elkerülhető vele. Szövődmény esetén a későbbiekben tumorprotézis-beültetés mint végtagmegtartó vészmegoldás még mindig elvégezhető. Orv Hetil. 2020; 161(45): 1914-1919.

Reconstruction of massive segmental bone defects is a crucial point of limb salvage surgeries after malignant bone tumor resections. Megaendoprostheses implantation is a commonly used method for adult patients, but hardly usable for children below 12 years old, because of the small size of the host bone and multiple mechanic complications.
Extracorporeal irradiation and allograft reimplantation (ECRT, ECI) are promising methods for these young children for limb salvage, reducing the number of prostheses revisions, reoperations.
In Hungary, we performed limb salvage surgery for malignant bone tumor in 12 cases in children under 12 years old, using extracorporeally irradiated autografts as biological reconstruction. All cases were primary bone tumors (Ewing\'s sarcoma: 7, osteosarcoma: 4, chondrosarcoma: 1). The average age of our patients was 9 (3-12) years, the average follow-up was 32.5 (2-73) months. At follow-up, we performed physical examination, X-ray, and other imaging methods if they were necessary. Our results were compared to international publications.
We observed no local recurrence, but in one case we had to perform above-knee amputation, due to a skip metastasis in the proximal tibia. The complication rates were similar to those reported in other papers. Septic complications were treated in two cases (16.7%). Total union of the resection lines was observed at 3-9 months in 60% of all cases. Reoperations were performed due to nonunion, or allograft fracture/partly desorganization in 3 cases, twice with new allograft, and with prostheses in one case. The subjective opinion of our patients is positive, they walk with full weightbearing, some of them do even light sporting activities.
We found extracorporeal irradiation a useful and safe method for children under 12 years old for biological reconstruction after malignant bone tumor resection. Complication rate stays under the rate of growing prostheses complications, in optimal cases further radical, and extensive surgeries may be avoided. In the case of major complications, megaendoprostheses implantation later on is still an option as salvage procedure with limb salvage. Orv Hetil. 2020; 161(45): 1914-1919.

Families with early concerns about infant symptoms of ASD have limited access to experienced professionals for screening and guidance. Telehealth has been used to reduce access disparities in other pediatric populations and has shown promise in parent-implemented interventions for ASD. We investigated the feasibility of a novel level-2 telehealth assessment of infants\' early social communication and ASD symptoms, the Telehealth Evaluation of Development for Infants (TEDI). Parents of eleven infants aged 6-12 months were coached to administer specific semi-structured behavioral probes. Initial feasibility, reliability, and acceptability benchmarks were met. These findings suggest the feasibility of screening infants via telehealth, and are supportive of further large-scale efforts to validate this method for longitudinal monitoring of symptomatic infants in community settings.