Comparative transcriptome profile of embryos at different developmental stages derived from somatic cell nuclear transfer (SCNT) and in-vitro fertilization (IVF) in riverine buffalo (Bubalus bubalis).

Somatic cell nuclear transfer (SCNT) is a very important reproductive technology with many diverse applications, such as fast multiplication of elite animals, the production of transgenic animals and embryonic stem (ES) cells. However, low cloning efficiency, a low live birth rate and the abnormally high incidence of abnormalities in the offspring born are attributed to incomplete or aberrant nuclear reprogramming. In SCNT embryos, the aberrant expression pattern of the genes throughout embryonic development is responsible for the incomplete nuclear reprogramming. The present study was carried out to identify the differential gene expression (DEGs) profile and molecular pathways of the SCNT and IVF embryos at different developmental stages (2 cell, 8 cell and blastocyst stages). In the present study, 1164 (2 cell), 1004 (8 cell) and 530 (blastocyst stage) DEGs were identified in the SCNT embryos as compared to IVF embryos. In addition, several genes such as ZEB1, GDF1, HSF5, PDE3B, VIM, TNNC, HSD3B1, TAGLN, ITGA4 and AGMAT were affecting the development of SCNT embryos as compared to IVF embryos. Further, Gene Ontology (GO) and molecular pathways analysis suggested, SCNT embryos exhibit variations compared to their IVF counterparts and affected the development of embryos throughout the different developmental stages. Apart from this, q-PCR analysis of the GDF1, TMEM114, and IGSF22 genes were utilized to validate the RNA-seq data. These findings contribute valuable insights about the different genes and molecular pathways underlying SCNT embryo development and offer crucial information for improving SCNT efficiency.

Embryonic and larval development of yellow perch (Perca flavescens) and its sensitivity to incubation temperature.

The ontogenetic development in teleost fish is sensitive to temperature, and the developmental rate has a direct relationship with the environmental temperature within a species' thermal tolerance limit. Temperature determines time to and survival at hatching. Yellow perch is a North American species of ecological and commercial importance, and its phenology is vulnerable to climate change. The embryonic development of yellow perch was comparable to closely related members of the family Percidae. Developmental progression was fastest at 18°C and slowest at 12°C, with medial progression at 15°C. Time to hatch and swim-up, feeding onset, and exogenous feeding phases were different across all incubation temperatures regardless of a gradual post-hatch warming of the 12 and 15°C groups to a common garden temperature of 18°C. Incubation temperature may lower the rate of survival to hatch at 15°C and had complex impacts on developmental abnormalities. Temperature had significant effects on the development rate, time of hatch, survival, and incidence of developmental abnormalities. Early ontogenetic thermal history in ectotherms is an important factor determining phenotypic variation. It will be important to link the thermally induced changes in development described here to the physiological and morphological differences and to link the developmental abnormalities to functional performance.

[Content of secondary metabolites and antioxidant activities of Lonicera japonica flowers at different developmental stages from Shandong province].

This study established an ultrasound-assisted extraction-high performance liquid chromatography method for simulta-neously determinining the content of 11 bioactive compounds including iridoids, phenolic acids, and flavonoids in Lonicera japonica flowers. The flowers at six stages from the rice bud stage(ML) to the golden flower stage(JH) of L. japonica varieties 'Sijuhua' and 'Beihua No.1' in two planting bases in Shandong province were collected. The established method was employed to determine the content of 11 target compounds, on the basis of which the dynamics of active components in L. japonica sampels during different development stages was investigated. The correlation analysis was carried out to reveal the correlations of the content of iridoids, phenolic acids, and flavonoids. Furthermore, the antioxidant activities of samples at different developmental stages were determined, and the relationship between antioxidant activity and chemical components was analyzed by the correlation analysis. The results showed that the total content of the 11 components in 'Sijihua' changed in a "W" pattern from the ML to JH, being the highest at the ML and the second at the slight white stage(EB). The total content of 11 compounds in 'Beihua No.1' was the highest at the ML and decreased gra-dually from the ML to JH. The samples of 'Sijihua' had higher content of iridoids and lower content of phenolic acids than those of 'Beihua No.1'. The content of flavonoids and phenolic acids showed a positive correlation(R~2=0.90, P<0.05) in 'Sijihua' but no obvious correlation in 'Beihua No.1'. The antioxidant activity and phenolic acid content showed positive correlations, with the determination coefficients(R~2) of 0.84(P<0.05) in 'Beihua No.1' and 0.73(P<0.05) in 'Sijihua'. The antioxidant activity of both varieties was the strongest at the ML and the second at the EB. This study revealed that the content dynamics of iridoids, phenolic acids, and flavonoids in 'Sijihua' and 'Beihua No.1' cultivated in Shandong province during different developmental stages. The results indicated that the antioxidant activity of L. japonica flowers was significantly correlated with the content of phenolic acids at different deve-lopmental stages, which provided a basis for determining the optimum harvest time of L. japonica flowers.

Developing a simple and efficient modeling solution for predicting key phenological stages of table grapes in a non-traditional viticulture zone in south Asia.

Phenological shifts are one of the most visible signs of climatic variability and change in the biosphere. However, modeling plant phenological responses has always been a key challenge due to climatic variability and plant adaptation. Grapevine is a phenologically sensitive crop and, thus, its developmental stages are affected by the increase in temperature. The goal of this study was to develop a temperature-based grapevine phenology model (GPM) for predicting key developmental stages for different table grape cultivars for a non-traditional viticulture zone in south Asia. Experiments were conducted in two vineyards at two locations (Chakwal and Islamabad) in the Pothawar region of Pakistan during the 2019 and 2020 growing seasons for four cultivars including Perlette, King's Ruby, Sugraone and NARC Black. Detailed phenological observations were obtained starting in January until harvest of the grapes. The Mitscherlich monomolecular equation was used to develop the phenology model for table grapes. There was a strong non-linear correlation between the Eichhorn and Lorenz phenological (ELP) scale and growing degree days (GDD) for all cultivars with coefficient of determinations (R2) ranging from 0.90 to 0.94. The results for model development indicated that GPM was able to predict phenological stages with high skill scores, i.e., a root mean square (RMSE) of 2.14 to 2.78 and mean absolute error (MAE) of 1.86 to 2.26 days. The prediction variability of the model for the onset timings of phenological stages was up to 3 days. The results also reveal that the phenology model based on GDD approach provides an efficient planning tool for viticulture industry in different grape growing regions. The proposed methodology, being a simpler one, can be easily applied to other regions and cultivars as a predictor for grapevine phenology.

Normal table of embryonic development in the Anji salamander Hynobius amjiensis (Hynobiidae).

We established a normal embryonic development table for the Anji salamander Hynobius amjiensis, a critically endangered tailed amphibian of the family Hynobiidae with a very limited distribution in East China, following the standards set by the early developmental table of vertebrates. Put together 32 embryonic stages for the Anji salamander was defined. The total embryonic period from oviposition to hatching is approximately 30 days at 9 °C. Stages 1-16 represent early development from cleavage to neurulation. Stages 17-32 represent organogenesis documenting later developmental events such as tail, gill, and limb formation, and hatching (Stage 32). We provided a detailed description of the external morphology and color changes of the head, trunk, limbs, tail, external gills, and balancers at various stages from egg-laying to hatching. We also described several cases of abnormal embryonic development. The establishment of the embryonic development table in H. amjiensis contributes to better understanding of the ontogeny in tailed amphibians, distinguishing closely related species, and identifying abnormal embryonic amphibians.

Integrative analysis of the metabolome and transcriptome provides insights into the mechanisms of lignan biosynthesis in Herpetospermum pedunculosum (Cucurbitaceae).

BACKGROUND: Herpetospermum pedunculosum (Ser.) C. B. Clarke is a traditional Chinese herbal medicine that heavily relies on the lignans found in its dried ripe seeds (Herpetospermum caudigerum), which have antioxidant and hepatoprotective functions. However, little is known regarding the lignan biosynthesis in H. pedunculosum. In this study, we used metabolomic (non-targeted UHPLC-MS/MS) and transcriptome (RNA-Seq) analyses to identify key metabolites and genes (both structural and regulatory) associated with lignan production during the green mature (GM) and yellow mature (YM) stages of H. pedunculosum. RESULTS: The contents of 26 lignan-related metabolites and the expression of 30 genes involved in the lignan pathway differed considerably between the GM and YM stages; most of them were more highly expressed in YM than in GM. UPLC-Q-TOF/MS confirmed that three Herpetospermum-specific lignans (including herpetrione, herpetotriol, and herpetin) were found in YM, but were not detected in GM. In addition, we proposed a lignan biosynthesis pathway for H. pedunculosum based on the fundamental principles of chemistry and biosynthesis. An integrated study of the transcriptome and metabolome identified several transcription factors, including HpGAF1, HpHSFB3, and HpWOX1, that were highly correlated with the metabolism of lignan compounds during seed ripening. Furthermore, functional validation assays revealed that the enzyme 4-Coumarate: CoA ligase (4CL) catalyzes the synthesis of hydroxycinnamate CoA esters. CONCLUSION: These results will deepen our understanding of seed lignan biosynthesis and establish a theoretical basis for molecular breeding of H. pedunculosum.

Influence of temperature on embryonic development of Pontastacus leptodactylus freshwater crayfish, and characterization of growth and osmoregulation related genes.

Narrow clawed crayfish, Pontastacus (Astacus) leptodactylus, represents an ecologically and economically valuable freshwater species. Despite the high importance of artificial breeding for conservation purpose and aquaculture potential, hatching protocols have not been developed so far in this species. Further, limited knowledge exists regarding the artificial egg incubation, the temperature effect on embryonic development, hatching synchronization and hatching rate. In the present study we investigated the temperature increase (from 17 oC to 22oC) effects in two different embryonic developmental stages of P. leptodactylus. Furthermore, two primer pairs for the Fibroblast Growth Factor Receptor 4 (FGFR4) gene cDNA amplification were successfully designed, characterising for the first time the FGFR4 gene in P. leptodactylus in relation to different developmental stages and temperatures. Apart from the FGFR4 gene, the Na+/K+-ATPase α-subunit expression was also explored. Both the FGFR4 and Na+/K+-ATPase α-subunit expression levels were higher in embryos closer to hatching. Egg incubation at 22oC for seven days led to significant increase of FGFR4 expression in embryos from earlier developmental stages. Nevertheless, temperature increase did not affect FGFR4 expression in eggs from latter developmental stages and Na+/K+-ATPase α-subunit expression in all developmental stages. Temperature increase represents therefore probably a promising strategy for accelerating hatching in freshwater crayfish particularly in early developmental stages. Specifically, our results indicate that FGFR4 expression increased in embryonic stages closer to hatching and that temperature influences significantly its expression in embryos from earlier developmental stages. Overall, these findings can provide a better understanding of artificial egg incubation of P. leptodactylus, and therefore can be employed for the effective management of this species, both for economic and biodiversity retention reasons.

How do Children with Intellectual Disabilities Empathize in Comparison to Typically Developing Children?

OBJECTIVES: Two studies were conducted to better understand how children with intellectual disabilities (ID) empathize with the feelings of others during social interactions. The first study tested hypotheses of developmental delay or difference regarding empathy in 79 children with ID by comparing them with typically developing (TD) children, matched for developmental age or chronological age. The second study examined specific aspects of empathy in 23 children with Down syndrome (DS), compared with 23 nonspecific ID children, matched for developmental age, and TD children, matched for developmental age or chronological age. METHOD: An empathy task was administered to the children while their parents completed the French versions of the Empathy Questionnaire and the Griffith Empathy Measure. RESULTS: The first study showed that ID children showed delayed empathy development but were perceived by their parents as deficient in cognitive empathy. The second study showed that DS children were perceived as being more attentive to the feelings of others than TD children and non-specific ID children, matched for developmental age, and as having affective empathy that was similar to that of TD children matched for chronological age. CONCLUSION: These studies have drawn attention to delays or differences in different dimensions of empathy in children with ID and DS, which need to be taken into account in interventions.

Patterns of Gender Development Across Intersections of Age, Gender, and Ethnicity-Race.

Two components of gender identity are gender similarity, how one's self-concept relates to the major gender collectives (i.e., female, male), and felt pressure to conform to gender norms. The development of these components across ages and contexts has been understudied. The focus of this study was to examine variations in gender similarity and felt pressure across multiple intersecting contexts: developmental stage, gender, and ethnic-racial group. Six data sets were harmonized and means were compared across 2628 participants (51% girls, 49% boys) from four different developmental cohorts (childhood n = 678, early adolescence n = 1322, adolescence n = 415, and young adulthood n = 213) from diverse ethnic-racial backgrounds (45% White, 23% Latinx/Hispanic, 11% Black/African-American, 7% Asian-American, 5% American Indian, and 5% Multiracial). Results revealed nuanced patterns: Gender intensification was supported in early adolescence, primarily for boys. Young adult men reported lower levels of pressure and gender typicality than younger boys, but young adult women's levels were generally not different than younger girls. Surprisingly, young adult women's levels of own-gender similarity and pressure from parents were higher than adolescent girls. Expectations of gender differences in gender typicality and felt pressure were supported for all ages except young adults, with higher levels for boys. Finally, there were more similarities than differences across ethnic-racial groups, though when there were differences, minoritized participants reported heightened gender typicality and pressure (largely accounted for by higher scores for Black and Latinx participants and lower scores for White and Multiracial participants). These results add to what is understood about contextually dependent gender development.

In silico identification and expression analyses of peroxidases in Tenebrio molitor.

Human advancements in agriculture, urbanization, and industrialization have led to various forms of environmental pollution, including heavy metal pollution. Insects, as highly adaptable organisms, can survive under various environmental stresses, which induce oxidative damage and impair antioxidant systems. To investigate the peroxidase (POX) family in Tenebrio molitor, we characterized two POXs, namely TmPOX-iso1 and TmPOX-iso2. The full-length cDNA sequences of TmPox-iso1 and TmPox-iso2 respectively consisted of an open reading frame of 1815 bp encoding 605 amino acids and an open reading frame of 2229 bp encoding 743 amino acids. TmPOX-iso1 and TmPOX-iso2 homologs were found in five distinct insect orders. In the phylogenetic tree analysis, TmPOX-iso1 was clustered with the predicted POX protein of T. castaneum, and TmPOX-iso2 was clustered with the POX precursor protein of T. castaneum. During development, the highest expression level of TmPox-iso1 was observed in the pre-pupal stage, while that of TmPox-iso2 expression were observed in the pre-pupal and 4-day pupal stages. TmPox-iso1 was primarily expressed in the early and late larval gut, while TmPox-iso2 mRNA expression was higher in the fat bodies and Malpighian tubules. In response to cadmium chloride treatment, TmPox-iso1 expression increased at 3 hours and then declined until 24 hours, while in the zinc chloride-treated group, TmPox-iso1 expression peaked 24 hours after the treatment. Both treated groups showed increases in TmPox-iso2 expression 24 hours after the treatments.

Comprehensive transcriptomic analysis revealing the regulatory dynamics and networks of the pituitary-testis axis in sheep across developmental stages.

Spermatogenesis is a complex process intricately regulated by the hypothalamic-pituitary-testis (HPT) axis. However, research on the regulatory factors governing the HPT axis remains limited. This study addresses this gap by conducting a comprehensive analysis of transcriptomes from the pituitary and testis tissues across various developmental stages, encompassing embryonic day (E120), neonatal period (P0), pre-puberty (P90), and post-puberty day (P270). Utilizing edgeR and WGCNA, we identified stage-specific genes in both the pituitary and testis throughout the four developmental stages. Notably, 380, 242, 34, and 479 stage-specific genes were identified in the pituitary, while 886, 297, 201, and 3,678 genes were identified in the testis. Subsequent analyses unveiled associations between these stage-specific genes and crucial pathways such as the cAMP signaling pathway, GnRH secretion, and male gamete generation. Furthermore, leveraging single-cell data from the pituitary and testis, we identified some signaling pathways involving BMP, HGF, IGF, and TGF-ß, highlighting mutual regulation between the pituitary and testis at different developmental stages. This study sheds light on the pivotal role of the pituitary-testis axis in the reproductive process of sheep across four distinct developmental stages. Additionally, it delves into the intricate regulatory networks governing reproduction, offering novel insights into the dynamics of the pituitary-testis axis within the reproductive system.

Body Composition and Anthropometric Measurements in Children and Adolescents with Autism Spectrum Disorder: A Case-Control Study in Lebanon.

The occurrence of overweight and obesity among individuals with Autism Spectrum Disorder (ASD) has become a worldwide epidemic. However, there is limited research on this topic in the Lebanese population. Therefore, this study aimed to assess the differences in anthropometric measurements and body composition variables among Lebanese children, pre-adolescents, and adolescents diagnosed with ASD in contrast to typically developing peers across various developmental stages. Additionally, it aimed to investigate the prevalence of overweight and obesity within this population. A total of 86 participants with ASD and 86 controls were involved in this case-control study, conducted between June 2022 and June 2023. Anthropometric measurements and body composition variables were assessed, followed by statistical analyses to examine the differences between these two groups. The results revealed a significantly higher prevalence of overweight and obesity among individuals with ASD, particularly evident during childhood and pre-adolescence. Additionally, this group exhibited a higher body fat mass and total body fat percentage compared to controls. However, there were no significant differences observed between the two groups during adolescence. These findings emphasize the significance of monitoring and addressing weight status in individuals with ASD to improve their overall health outcomes. Future research directions could focus on investigating the underlying mechanisms contributing to the heightened prevalence of overweight and obesity in this population, ultimately enhancing their quality of life and well-being.

Profile of the gut microbiota of Pacific white shrimp under industrial indoor farming system.

The gut microbial communities interact with the host immunity and physiological functions. In this study, we investigated the bacterial composition in Litopenaeus vannamei shrimp's gut and rearing water under different host (developmental stage: juvenile and adult; health status: healthy and diseased) and environmental factors (temperature 25 °C and 28 °C; and light intensity: low and high). The PCoA analysis showed that all water samples were clustered together in a quarter, whereas the gut samples spread among three quarters. In terms of functional bacteria, gut samples of adult shrimp, healthy adult shrimp, adult shrimp raised at 28 °C, and juvenile shrimp under high light intensity exhibited a higher abundance of Vibrionaceae compared to each other opposite group. Gut samples of juvenile shrimp, infected adult shrimp, juvenile shrimp with low light intensity, and adult shrimp with a water temperature of 25 °C showed a higher abundance of Pseudoaltromonadaceae bacteria compared to each other opposite group. Gut samples of juvenile shrimp, healthy adult shrimp, adult shrimp raised at a water temperature of 28 °C, and juvenile shrimp with high light intensity showed the higher abundance of Firmicutes/Bacteroidota ratio compared to each other opposite group. Our results showed that L. vannamei juveniles are more sensitive to bacterial infections; besides, water temperature of 28 °C and high light intensity groups were both important conditions improving the shrimp gut bacterial composition under industrial indoor farming systems. KEY POINTS: • Bacteria diversity was higher among shrimp intestinal microbiota compared to the rearing water. • Shrimp juveniles are more sensitive to bacterial infection compared to adults. • Water temperature of 28 °C and high light intensity are recommended conditions for white shrimp aquaculture.

Evaluation of suitable reference genes for expression profile analyses of target genes in the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae).

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is a major destructive insect pest of coffee, which impacts the coffee crops negatively. As a draft genome has been completed for this insect, most molecular studies on gene transcriptional levels under different experimental conditions will be conducted using real-time reverse-transcription quantitative polymerase chain reactions (RT-qPCR). However, the lack of suitable internal reference genes will affect the accuracy of RT-qPCR results. In this study, the expression stability of nine candidate reference genes was evaluated under different developmental stages, temperature stress, and Beauveria bassiana infection. Data analyses were completed by four commonly used programs, BestKeeper, NormFinder, geNorm, and RefFinder. The result showed that RPL3 and EF1α combination were recommended as the most stable reference genes for developmental stages. EF1α and RPS3a combination were the top two stable reference genes for B. bassiana infection. RPS3a and RPL3 combination performed as the optimal reference genes both in temperature stress and all samples. Our results should provide a good foundation for the expression profile analyses of target genes in the future, especially for molecular studies on insect genetic development, temperature adaptability, and immune mechanism to entomogenous fungi in H. hampei.

Characterization of dicaffeoylspermidine derivatives related glucosyltransferases during fruit development of goji berry.

The fruit of Lycium barbarum (Lb), known as red goji berry, is a "superfruit" due to its abundance of bioactive compounds. Among these compounds, dicaffeoylspermidine derivatives (DCSPDs) have anti-oxidant and anti-Alzheimer's Disease activity. This study employed ultra-high-performance liquid chromatography with tandem mass spectrometry to investigate metabolic changes during the development and ripening stages of red goji berries. Totally 97 compounds, including 51 DCSPDs, were tentatively identified. Correlation analysis of these DCSPDs revealed that glycosyltransferases (GTs) play an important role in the formation of glycosylated DCSPDs. In vitro experiments characterized 3 novel GTs could add a glucosyl moiety to N1-caffeoyl-N10-dihydrocaffeoyl spermidine. Homologous GTs from L. ruthenicum (Lr) exhibited similar activity, despite the absence of abundant glycosylated DCSPDs in Lr. These findings provide insights into the metabolic changes and interconnections among active compounds in red goji berries. The identified GTs hold potential for metabolic engineering of DCSPDs and functional food development.

Evolution of a tidal channel network in the Yellow River Delta, China, and simulation of optimization scenarios.

Tidal channel networks, which characterize all river deltas, control the exchange of water and nutrients (hydrological connectivity) between the ocean and the delta area. Therefore, a tidal channel network in optimal conditions ensures the maintenance of the diversity and stability of the deltaic ecosystem. However, the developmental status of channel networks in the Yellow River Delta, China, has not been clearly determined. Here, we selected a typical tidal channel network in this delta that showed different spatial patterns (e.g., connectivity attributes) in the past three decades and explored its evolution using entropy as an index of connectivity. Seven scenarios were set up to determine the optimal status of the tidal channel network by optimizing its structure. The optimization effect was evaluated by comparing the connectivity attributes of the channel network before and after optimization. The results showed that the network experienced two obviously different developmental phases: an evolution before 2005 and a regression after 2005. Mann-Kendall analysis indicated that the channel network achieved dynamic stability before 2014 and became unstable thereafter. The simulations conducted to optimize the system showed that adding outlets changed the current patterns of the network' structural and functional connectivity. As the optimization proceeded, structural connectivity increased while functional connectivity decreased, and the tidal channel network tended to be dynamically stable. Our study elucidated the quantitative relationship between outlet number and stability within tidal channel networks, providing reference information that could be incorporated into future projects for the restoration and management of river deltas.

Method for the molecular and quantitative identification of oocytes and their developmental stage in teleost fish gonads.

Fish display diverse reproductive strategies and their gametogenesis is influenced by numerous genetic, physiological and environmental factors. The analysis of 5S rRNA expression levels in gonads has been proposed as useful method for the molecular identification of the presence of oocytes in fish tissues. The present method provides an easy and unbiased approach to analyse the expression of tRNAs and 5S rRNA in teleost gonads and stablish the presence and developmental stage of oocytes. Total RNA extracted from gonads is analysed through capillary electrophoresis in a Bioanalyzer 2100 (Agilent Technologies) using Small RNA Assays. Electropherograms allow quantifying the concentrations of tRNAs, 5S rRNA and 5.8S rRNA per sample and calculate their tRNA/5.8S rRNA and 5S/5.8S rRNA indices. Both indices clearly differentiate ovaries from testes and can be used to identify testes that present oocytes due to exposure to environmental xenoestrogens. The tRNA/5.8S and 5S/5.8S indices show the highest values in ovaries in previtellogenic stage, values decreasing as they advance towards maturity.•Detailed molecular method to sex fish and quantitatively identify the maturity stage of females.•tRNA levels in gonads can help in the study of teleost reproduction (female fecundity assessment, molecular gonad sexing) and environmental health assessment.

Transcriptional patterns of the cortical Morphometric Inverse Divergence in first-episode, treatment-naïve early-onset schizophrenia.

Early-onset Schizophrenia (EOS) is a profoundly progressive psychiatric disorder characterized by both positive and negative symptoms, whose pathogenesis is influenced by genes, environment and brain structure development. In this study, the MIND (Morphometric Inverse Divergence) network was employed to explore the relationship between morphological similarity and specific transcriptional expression patterns in EOS patients. This study involved a cohort of 187 participants aged between 7 and 17 years, consisting of 97 EOS patients and 90 healthy controls (HC). Multiple morphological features were used to construct the MIND network for all participants. Furthermore, we explored the associations between MIND network and brain-wide gene expression in EOS patients through partial least squares (PLS) regression, shared genetic predispositions with other psychiatric disorders, functional enrichment of PLS weighted genes, as well as transcriptional signature assessment of cell types, cortical layers, and developmental stages. The MIND showed similarity differences in the orbitofrontal cortex, pericalcarine cortex, lingual gyrus, and multiple networks in EOS patients compared to HC. Moreover, our exploration revealed a significant overlap of PLS2 weighted genes linking to EOS-related MIND differences and the dysregulated genes reported in other psychiatric diseases. Interestingly, genes correlated with MIND changes (PLS2-) exhibited a significant enrichment not only in metabolism-related pathways, but also in specific astrocytes, cortical layers (specifically layer I and III), and posterior developmental stages (late infancy to young adulthood stages). However, PLS2+ genes were primarily enriched in synapses signaling-related pathways and early developmental stages (from early-mid fetal to neonatal early infancy) but not in special cell types or layers. These findings provide a novel perspective on the intricate relationship between macroscopic morphometric structural abnormalities and microscopic transcriptional patterns during the onset and progression of EOS.

Single-cell profiling of the pig cecum at various developmental stages.

The gastrointestinal tract is essential for food digestion, nutrient absorption, waste elimination, and microbial defense. Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding of cellular diversity, functional heterogeneity, and their importance in intestinal tract development and disease. Although such profiling has been extensively conducted in humans and mice, the single-cell gene expression landscape of the pig cecum remains unexplored. Here, single-cell RNA sequencing was performed on 45 572 cells obtained from seven cecal samples in pigs at four different developmental stages (days (D) 30, 42, 150, and 730). Analysis revealed 12 major cell types and 38 subtypes, as well as their distinctive genes, transcription factors, and regulons, many of which were conserved in humans. An increase in the relative proportions of CD8 + T and Granzyme A (low expression) natural killer T cells (GZMA low NKT) cells and a decrease in the relative proportions of epithelial stem cells, Tregs, RHEX + T cells, and plasmacytoid dendritic cells (pDCs) were noted across the developmental stages. Moreover, the post-weaning period exhibited an up-regulation in mitochondrial genes, COX2 and ND2, as well as genes involved in immune activation in multiple cell types. Cell-cell crosstalk analysis indicated that IBP6 + fibroblasts were the main signal senders at D30, whereas IBP6 - fibroblasts assumed this role at the other stages. NKT cells established interactions with epithelial cells and IBP6 + fibroblasts in the D730 cecum through mediation of GZMA-F2RL1/F2RL2 pairs. This study provides valuable insights into cellular heterogeneity and function in the pig cecum at different development stages.