Enrichment Items Research Articles

RRP9 and DDX21 as new biomarkers of colorectal cancer.

Colorectal cancer originates from the epithelium of the large intestine and is a common malignant tumor in the gastrointestinal tract. However, the relationship between RRP9 and DDX21 and colorectal cancer (CRC) remains unclear. GSE134834, GSE206800, and GSE209892 profiles for CRC were downloaded from the gene expression omnibus database generated using GPL20115 and GPL23126. Differentially expressed genes (DEGs) were screened and weighted gene co-expression network analysis was performed. The construction and analysis of protein-protein interaction network. Functional enrichment analysis and gene set enrichment analysis were performed. Gene expression heat map was drawn and immune infiltration analysis was performed. Comparative toxicogenomics database analysis were performed to find the disease most related to the core gene. TargetScan was used to screen miRNAs regulating central DEGs. One thousand three hundred eighty DEGs were identified. According to gene ontology analysis, they were mainly concentrated in signal receptor activity regulation and metal titanase activity. Kyoto encyclopedia of gene and genome analysis showed that they mainly focused on IL17 signal pathway, PPAR signal pathway, protein digestion, and absorption, and the interaction of viral proteins with cytokines and cytokine receptors. The intersection of enrichment items and GOKEGG enrichment items of differentially expressed genes is mainly concentrated in PPAR signal pathway and the interaction of viral proteins with cytokines and cytokine receptors. The protein-protein interaction network obtained 16 core genes (MAD2L1, MELK, TPX2, UBE2C, RFC4, PLK1, RACGAP1, DKC1, DDX21, L Y AR, WDR3, RRP9, WDR43, NOLC1, BRIX1, and GTPBP4). Heat map of gene expression showed that core genes (TPX2, UBE2C, RFC4, PLK1, DKC1, LYAR, WDR3, NOLC1, and BRIX1) were not significantly differentially expressed between CRC and normal tissue samples. Core genes (MAD2L1, MELK, RACGAP1, RRP9, WDR43, DDX21, and GTPBP4) were highly expressed in CRC tissue samples and lowly expressed in normal tissue samples. Comparative toxicogenomics database analysis showed that 7 genes (MAD2L1, MELK, RACGAP1, RRP9, WDR43, DDX21, and GTPBP4) were related to necrosis, inflammation, tumor, precancerous symptoms, hemorrhage, and weightlessness. RRP9 and DDX21 are highly expressed in CRC. The higher the expression level of RRP9 and DDX21, the worse the prognosis.

Role of TOP2A and CDC6 in liver cancer.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with high mortality worldwide, which is characterized by aggressive growth and metastasis. However, the relationship between TOP2A and CDC6 and HCC remains unclear. GSE121248 and GSE101728 profiles for liver cancer were downloaded from the gene expression omnibus database generated using GPL21047and GPL570. Differentially expressed genes (DEGs) were screened and weighted gene co-expression network analysis was performed. The construction and analysis of protein-protein interaction network, functional enrichment analysis, gene set enrichment analysis. Gene expression heat map was drawn and survival analysis was performed. Comparative toxicogenomics database analysis were performed to find the disease most related to the core gene. TargetScan was used to screen miRNAs regulating central DEGs. 885 DEGs were identified. According to gene ontology analysis, they were mainly enriched in organic acid metabolism process, metabolic pathway, p53 signal pathway and PPAR signal pathway. The enrichment items are similar to the GOKEGG enrichment items of differentially expressed genes, mainly in the process of organic acid metabolism, p53 signal pathway and PPAR signal pathway. In the enrichment project of metascape, gene ontology has PIDPLK1 pathway, mitotic cell cycle, tumor retinoblastoma gene. The construction and analysis of protein-protein interaction network obtained 10 core genes (TOP2A, CDK1, ASPM, RACGAP1, ZWINT, CDC6, AURKA, NCAPG, BUB1B, CCNB1), and found that these core genes were highly expressed in tumor tissues and low in normal tissues. Comparative toxicogenomics database analysis showed that 10 genes (TOP2A, CDK1, ASPM, RACGAP1, ZWINT, CDC6, AURKA, NCAPG, BUB1B, CCNB1) were related to necrosis, inflammation, HCC, liver cirrhosis, and adenoid cystic carcinoma. TOP2A and CDC6 are highly expressed in liver cancer, which may become molecular targets for early diagnosis and precise treatment.

Structural enrichment for laboratory mice: exploring the effects of novelty and complexity.

Providing structural enrichment is a widespread refinement method for laboratory rodents and other animals in captivity. So far, animal welfare research has mostly focused on the effect of increased complexity either by accumulating or combining different enrichment items. However, increasing complexity is not the only possibility to refine housing conditions. Another refinement option is to increase novelty by regularly exchanging known enrichment items with new ones. In the present study, we used pair-housed non-breeding female C57BL/6J and DBA/2N mice to investigate the effect of novelty when applying structural enrichment. We used a double cage system, in which one cage served as home cage and the other as extra cage. While the home cage was furnished in the same way for all mice, in the extra cage we either provided only space with no additional enrichment items (space), a fixed set of enrichment items (complexity), or a changing set of enrichment items (novelty). Over 5 weeks, we assessed spontaneous behaviors, body weight, and extra cage usage as indicators of welfare and preference. Our main results showed that mice with access to structurally enriched extra cages (complexity and novelty) spent more time in their extra cages and complexity mice had lower latencies to enter their extra cages than mice with access to the extra cages without any structural enrichment (space). This indicates that the mice preferred the structurally enriched extra cages over the structurally non-enriched space cages. We found only one statistically significant difference between the novelty and complexity condition: during week 3, novelty mice spent more time in their extra cages than complexity mice. Although we did not detect any other significant differences between the novelty and complexity condition in the present study, more research is required to further explore the potential benefits of novelty beyond complexity.

Identification and experimental validation of autophagy-related genes in abdominal aortic aneurysm

AimAutophagy plays essential roles in abdominal aortic aneurysm (AAA) development and progression. The objective of this study was to verify the autophagy-related genes (ARGs) underlying AAA empirically and using bioinformatics analysis.MethodsTwo gene expression profile datasets GSE98278 and GSE57691 were downloaded from the Gene Expression Omnibus (GEO) database, and principal component analysis was performed. Following, the R software (version 4.0.0) was employed to analyze potentially differentially expressed genes related with AAA and autophagy. Subsequently, the candidate genes were screened using protein–protein interaction (PPI), gene ontology (GO) enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Finally, quantitative real-time polymerase chain reaction (RT-qPCR) was performed to detect the RNA expression levels of the top five selected abnormal ARGs in clinical samples obtained from the normal and AAA patients.ResultsAccording to the information contained (97 AAA patients and 10 healthy controls) in the two datasets, a total of 44 differentially expressed autophagy-related genes (6 up-regulated genes and 38 down-regulated genes) were screened. GO enrichment analysis of differentially expressed autophagy-related genes (DEARGs) demonstrated that some enrichment items were associated with inflammation, and PPI analysis indicated interaction between these genes. RT-qPCR results presented that the expression levels of IL6, PPARG, SOD1, and MAP1LC3B were in accordance with the bioinformatics prediction results acquired from the mRNA chip.ConclusionBioinformatics analysis identified 44 potential autophagy-related differentially expressed genes in AAA. Further verification by RT- qPCR presented that IL6, PPARG, SOD1, and MAP1LC3B may affect the development of AAA by regulating autophagy. These findings might help explain the pathogenesis of AAA and be helpful in its diagnosis and treatment.

The value of RPS15 and MRPS27 in ischemic stroke.

Ischemic stroke is caused by insufficient blood supply to the brain. It has acute onset, often disturbance of consciousness, and high mortality and disability rate. However, relationship between ribosomal proteins (RP)-S15 and mitochondrial ribosomal proteins (MRP)-S27 and ischemic stroke remains unclear. The ischemic stroke datasets GSE22255, GSE16561, and GSE199435 were downloaded from gene expression omnibus generated by GPL6883, GPL11154, and GPL570. Differentially expressed genes (DEGs) were screened, and the construction and analysis of protein-protein interaction network, functional enrichment analysis and gene set enrichment analysis were performed. The gene expression heat map was drawn. Comparative toxicogenomics database analysis were performed to find the disease most related to core gene. TargetScan screened miRNAs that regulated central DEGs. Five hundred DEGs were identified. According to gene ontology analysis, they were mainly enriched in leukocyte activation, myoid cell activation involved in immune response, cell membrane, mitochondria, secretory vesicles, catalytic activity, enzyme binding, ribonucleic acid binding, splicing. Gene set enrichment analysis showed that the enrichment items are similar to the enrichment items of differentially expressed genes. And 20 core genes were obtained. Comparative toxicogenomics database analysis showed that 6 genes (RPS15, RPS2, RPS3, MRPS27, POLR2A, MRPS26) were found to be associated with chemical and drug-induced liver injury, necrosis, delayed prenatal exposure, nephropathy, hepatomegaly and tumor. RPS15 and MRPS27 are the core genes of ischemic stroke and play an important role in ischemic stroke.

Research progress on animal environment and welfare

AbstractAnimal welfare closely depends on its ability to adapt and thrive in a harmonious relationship with its environment, ensuring both physical and psychological well‐being. Over the years, the welfare of farm animals has gathered global attention and has become increasingly important to the general public and scientific communities. The connection between the environment and animal welfare is primarily established through the provision of suitable and controllable environment for animals. However, it is essential to recognize that the impact of environment extends beyond merely ensuring freedom from discomfort. The environment plays a crucial role in shaping an animal's response to challenges such as disease, stress, and pathogen. While animals may be housed in controlled environments that provide optimal conditions for health, production, and welfare, it is important to acknowledge that specific scenarios can significantly affect and alter the environmental requirements. Even with access to fresh air, certain factors can have a substantial impact on the well‐being of animals. Furthermore, providing appropriate environmental conditions goes beyond meeting basic needs and can greatly contribute to allowing animals to engage in their natural behaviors. It serves as a relevant tool for ensuring and maintaining adequate welfare standards. This review takes a comprehensive approach to environmental welfare by considering the welfare of animals managed in different stocking systems, considering environmental stress, stocking systems, and the provision of environmental enrichment items. By examining these factors, a broader understanding of the relationship between environment and welfare is achieved and recommendations for future research are outlined.

CD3D and CD247 are the molecular targets of septic shock.

Septic shock is a serious systemic disease with circulatory failure and abnormal cell metabolism caused by sepsis. However, the relationship between CD3D and CD247 and septic shock remains unclear. The septic shock datasets GSE33118 and GSE142255 profiles were generated from the gene expression omnibus databases GPl570, GPl17586. Differentially expressed genes (DEGs) were screened and weighted gene co-expression network analysis was performed. The construction and analysis of protein-protein interaction (PPI) network, functional enrichment analysis, gene set enrichment analysis (GSEA) were performed. Gene expression heat map was drawn. Immune infiltration analysis was performed. Comparative toxicogenomics database (CTD) analysis were performed to find the disease most related to the core gene. Targets can was used to screen miRNAs regulating the hub DEGs. 467 DEGs were identified. According to the gene ontology analysis, they were mainly enriched in the regulation of immune response, cell activation, signaling receptor activity, enzyme binding. Kyoto encyclopedia of genes and genomes analysis showed that they were mainly enriched in the TCR signaling pathway, Fc epsilon RI signaling pathway. GSEA showed that the DEGs were mainly enriched in immune response regulation, cell activation, TCR signaling pathway, Fc epsilon RI signaling pathway. Positive regulation of Fc receptor signaling pathway, PID IL12 2 pathway, immune response was observed in go enrichment items in the enrichment items of metascape. PPI networks got 5 core genes. Gene expression heat map showed that 5 core genes (CD247, Lck, cd3e, cd3d, ITK) were lowly expressed in the sepsis shock samples and highly expressed in the normal samples. CTD analysis showed that 5 core genes (CD247, Lck, cd3e, cd3d, ITK) were found to be associated with hemorrhage and necrosis. Low expression of cd3d, CD247 was observed in septic shock, and the lower the level of cd3d, CD247, the worse the prognosis.

Benchmarking Enrichment Efforts in the US & Canada Across Species and Enrichment Categories.

Enrichment is important for animal welfare and data quality. Provision of enrichment opportunities varies between species and enrichment category. However, data benchmarking these differences does not exist. Our objective was to characterize enrichment provision and associated factors across species in the US and Canada. Personnel who work with research animals (n = 1098) in the US and Canada voluntarily responded to online promotions and completed a survey about enrichment used for the species they worked with most, their control of and wish for more enrichment, stress or pain in the animals they worked the most with, and demographics. All participants (except those working with rats) received the same questionnaire regardless of species to allow objectivity, as the effects of many enrichment items on some species have not yet been determined. The questionnaire asked about enrichments that were beneficial to at least one species. The provision of enrichment was allocated into 2 outcome variables: diversity and frequency per enrichment category. Results showed a significant interaction between enrichment category and species. Generally, physical, nutritional, and sensory enrichments were provided less often than social enrichment. In addition, nonhuman primates received more diverse and more frequent enrichment than did other species (twice as much as rats and mice). Enrichment was provided less frequently by personnel who wished they could do more than the status quo. Both enrichment frequency and diversity were higher in respondents from Canada, those who had more control over provision, and those who had been in the field longer. While our results cannot be used to determine the quality of enrichment provided to various species, they do provide information on current enrichment practices in the US and Canada and identify differences in implementation by species and enrichment category. The data also indicate provision of enrichment is influenced by factors such as country and individual control over enrichment. This information can also be used to identify areas for greater enrichment efforts for some species (for example, rats and mice) and categories, with the ultimate goal of improving animal welfare.

Analytical Validation and Assessment of Baseline Fecal Glucocorticoid Metabolites in Northern Sea Otters (Enhydra lutris kenyoni) in Human Care

Fecal glucocorticoid metabolites (FGMs) have been used as a non-invasive and indirect measurement of the complex stress response in a variety of species. Animals in facilities under managed care allow for the longitudinal study of FGMs in a controlled environment. Animal histories often include environmental, husbandry, and medical notes that can be matched to FGM concentrations to aid in the physiological validation of adrenal stimulation and response. The goal of this study was to demonstrate analytical validations using two enzyme-linked immunosorbent assays (EIA) to measure FGMs from northern sea otters (Enhydra lutris kenyoni) under human care (Seattle Aquarium, Seattle, WA, USA) and to determine baseline and stress response spike levels for individual sea otters. Individual variation was found among the four subjects in the study with fecal baseline levels ranging from 20.2 to 83.7 ng/g for cortisol-immunoreactive metabolites and 52.3 to 102 ng/g for corticosterone-immunoreactive metabolites. As a retrospective study, 39 percent of hormone peaks were associated with notes and most FGM spikes were associated with veterinary procedures or days in which enrichment items were provided and produced an excitatory response. Monitoring baseline FGMs levels and events associated with hormone peak values may provide insight into effective husbandry management to improve the overall welfare of sea otters and other marine mammals.

Genome-wide transcriptomic analysis identifies candidate genes involved in jasmonic acid-mediated salt tolerance of alfalfa.

Soil salinity imposes a major threat to plant growth and agricultural productivity. Despite being one of the most common fodder crops in saline locations, alfalfa is vulnerable to salt stress. Jasmonic acid (JA) is a phytohormone that influences plant response to abiotic stimuli such as salt stress. However, key genes and pathways by which JA-mediated salt tolerance of alfalfa are little known. A comprehensive transcriptome analysis was performed to elucidate the underlying molecular mechanisms of JA-mediated salt tolerance. The transcripts regulated by salt (S) compared to control (C) and JA+salt (JS) compared to C were investigated. Venn diagram and expression pattern of DEGs indicated that JS further altered a series of genes expression regulated by salt treatment, implying the roles of JA in priming salt tolerance. Enrichment analysis revealed that DEGs exclusively regulated by JS treatment belonged to primary or secondary metabolism, respiratory electron transport chain, and oxidative stress resistance. Alternatively, splicing (AS) was induced by salt alone or JA combined treatment, with skipped exon (SE) events predominately. DEGs undergo exon skipping involving some enriched items mentioned above and transcription factors. Finally, the gene expressions were validated using quantitative polymerase chain reaction (qPCR), which produced results that agreed with the sequencing results. Taken together, these findings suggest that JA modulates the expression of genes related to energy supply and antioxidant capacity at both the transcriptional and post-transcriptional levels, possibly through the involvement of transcription factors and AS events.

Ruff Morning? The Use of Environmental Enrichment during an Acute Stressor in Kenneled Shelter Dogs.

The length of stay for some animals has increased with the recent reduction of animals euthanized in US shelters and animal control facilities. Research examining the effectiveness of different types of enrichment on buffering the effects of acute daily stressors in the shelter environment, such as kennel cleaning, is lacking. In addition, daily known stressors can result in undesirable behaviors that could lead to a need for euthanasia. Ways to effectively reduce the effects of daily stressors while optimizing strained resources is currently a high priority. In this study, we presented shelter dogs with food, tactile, and scent enrichment items to increase (arousing) or decrease (calming) activity during the daily stressor of morning kennel cleaning. We found that calming, rather than arousing, enrichment items were associated with body position scores indicative of lower stress in dogs, with calming scent enrichment (lavender) producing the most significant benefit. In contrast, items that showed the greatest reduction in vocalization were arousing (ball) compared to other arousing conditions. Our findings suggest that different unwanted behaviors in the kennel environment often associated with stress can be reduced using specific types of enrichment during a daily stressful event. Further, the results illustrate that enrichment items other than food might be more effective at decreasing certain undesirable behaviors. Overall, this study provides insight into how shelter workers might effectively use enrichment items during an unavoidable acute stressor. With many shelters keeping dogs longer, addressing events that might cause repeated stress in this population may indirectly help with adopting and lowering euthanasia rates due to unwanted behavior that develops due to repeated exposure to this necessary but acute stressor of morning cleaning.

The Effect of Enrichment on Leopard Geckos (Eublepharis macularius) Housed in Two Different Maintenance Systems (Rack System vs. Terrarium).

The aim of this study was to test the usefulness of environmental enrichment for Eublepharis macularius depending on the maintenance method (terrarium vs. rack system). The hypothesis was that reptiles kept in an extremely low-stimulus environment (rack system) would be more prone to interact with environmental enrichment items than those kept in a biotope terrarium. During the study, 21 female geckos were kept in two types of captive enclosures: 9 in terrariums, and 12 in rack system boxes in groups of 3 animals per enclosure. During the enrichment sessions, geckos were observed for 45 min while enrichment items (dry and wet hides, a new feeding method, a new object) were present in the enclosure. All geckos showed interest in enrichment items that enabled hiding and climbing. Animals kept in the rack system showed significantly lower latency in approaching enrichment items and a higher frequency of enrichment interactions than lizards in biotope terrariums. However, no significant differences were found in the total time spent interacting with enrichment items between geckos in the two settings.

Quantifying Acute Behavioral Reactions of Bali Mynas (Leucopsar rothschildi) to Environmental and Progressively Challenging Enrichment

Animals use specific behaviors and skills to overcome challenges and access resources. Environmental enrichment is provided to animals in human care to both promote species-appropriate behaviors and reduce undesired behaviors. Feather pecking in birds is an undesired behavior without a clear cause. The Saint Louis Zoo houses three pairs of young Bali mynas (Leucopsar rothschildi) who pluck neck feathers from conspecifics. To reduce this behavior, animal care staff presented the birds with seven enrichment items from four categories, presenting each item twice. The enrichment included a modifiable, progressively challenging bamboo tube device at multiple levels of difficulty. While plucking was not affected by any enrichment item, we observed significant increases in locomotion and decreases in autopreening, allogrooming, and head bobbing. Leafy greens produced the greatest changes when compared to other enrichment types. Overall engagement with the progressively challenging enrichment increased with the change from the first to the second level of difficulty, and interaction with the device was highest for the third and most difficult version. These increases suggest that no habituation to the progressively challenging device occurred, while a possible neophobic effect declined with multiple uses and increased familiarity.

Network pharmacology research of mechanism of Maxing Shigan Decoction in treating COVID‐19

BackgroundThe COVID‐19 has a huge negative impact on people’s health. Traditional Chinese Medicine (TCM) has a good effect on viral pneumonia. It is of great practical significance to study its pharmacology.MethodsThe ingredients and targets of each herb in Maxing Shigan Decoction which obtained from Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, and the related targets of COVID‐19 were screened by GeneCards database based on the network pharmacology. Venn was used to analyze the intersection target between active ingredients and diseases. Cytoscape software was used to construct an active ingredient‐disease target network. The Protein‐Protein Interaction network was constructed by STRING database and Cytohubba was used to screen out the key targets. Gene Ontology (GO) functional enrichment analysis and KEGG pathway analysis were performed by David database.ResultsIn this study, a total of 134 active ingredients and 229 related targets, 198 targets of COVID‐19 and 48 common targets of drug‐disease were chosen. Enrichment items and pathways were obtained through GO and KEGG pathway analysis. The predicted active ingredients were quercetin, kaempferol, luteolin, naringenin, glycyrol, and the key targets involved IL6, MAPK3, MAPK8, CASP3, IL10, etc. The results showed that the active ingredients of Maxing Shigan Decoction acted on multiple targets which played roles in the treatment of COVID‐19 by regulating inflammation, immune system and other pathways.ConclusionsThe main contribution of this paper is to use data to mine the principles of the treatment of COVID‐19 from the pharmacology of these prescriptions, and the results can be provided theoretical reference for medical workers.

Long-Lasting Chews Elicit Positive Emotional States in Dogs during Short Periods of Social Isolation

Dogs may experience negative emotional states when isolated from human caregivers and conspecifics. This study aimed to evaluate how dogs interact with different enrichments during a short period of social isolation, as a first step towards identifying methods for improving their emotional wellbeing. Using a cross-over design, dogs (n = 20) at the Waltham Petcare Science Institute were exposed to four different food-based enrichments while left alone in a familiar room for 20 min: long-lasting chew (Chew), kibble in a treat-dispensing toy (Toy), and kibble dispensed through a smart treat-dispensing device with (Device + Voice) and without (Device) a person talking to the dog. Time spent engaging with each enrichment item and emotional valence and arousal (7-point scale collected every 5-min) were scored from videos. The results of linear mixed models indicated Chew was the most successful enrichment, with dogs having lower arousal scores (p < 0.05 vs. Device and Toy) during the first five minutes of isolation, higher positive valence scores (p < 0.05 vs. all) during the second five minutes of isolation, and spending the most total time engaged (p < 0.01 vs. all). Based on these findings, long-lasting chews should be further explored to assess their impact on dog emotional wellbeing.

Vegetarian vs. Carnivore Feeding Enrichment in a Pack of Captive Iberian Wolves (Canis lupus signatus): Towards Individual and Species-typical Needs?

The aim of this study was to compare two different types of feeding enrichment (vegetarian vs. carnivore) through the daily activity, space use and inter-individual distances in a captive Iberian wolf (Canis lupus signatus) pack housed at the Barcelona Zoo. Multifocal sampling methods were used for data collection and instantaneous scans were made at 15-min intervals during sessions of varying duration. The enrichment sessions were carried out once a day, three days per week –Monday, Wednesday, and Friday- repeating the delivered schedule of items every two weeks. Thus, both feeding phases included six different enrichment sessions; the vegetarian phase included chopped fruit – apples inside a burlap sack, scattered bananas, peaches, frozen pears, and oranges, and kiwis inside a frozen water block; the carnivore phase included animal products -beef raw lean meat inside a burlap sack or inside a frozen water block, horse leg, live carp, frozen rabbit skin and live grasshoppers-. There were individual differences in the response to both feeding enrichments. Only the carnivore enrichment provoked statistically significant differences in exploration, locomotion, inactivity and not visible. Only the indeterminate zone showed changes during both enrichment phases. The inter-individual distance between Iberian wolves was statistically significantly lower during both feeding enrichment phases –especially during the carnivore as compared to the vegetarian phase- than during the baseline phase. Enrichment items designed more according to the natural history of these animals seem to have a greater capacity to improve the wolf welfare. Future studies on this species are necessary to improve husbandry techniques, welfare, and conservation programs.

Rating enrichment items by female group-housed laboratory mice in multiple binary choice tests using an RFID-based tracking system.

Laboratory mice spend most of their lives in cages, not experiments, so improving housing conditions is a first-choice approach to improving their welfare. Despite the increasing popularity of enrichment, little is known about the benefits from an animal perspective. For a detailed analysis, we categorized enrichment items according to their prospective use into the categories 'structural', 'housing', and 'foraging'. In homecage-based multiple binary choice tests 12 female C57BL/6J mice chose between enrichment items within the respective categories over a 46-hour period. A new analyzing method combined the binary decisions and ranked the enrichment items within each category by calculating worth values and consensus errors. Although there was no unequivocal ranking that was true in its entire rank order for all individual mice, certain elements (e.g. lattice ball, second plane) were always among the top positions. Overall, a high consensus error in ranking positions reflects strong individual differences in preferences which could not be resolved due to the relatively small sample size. However, individual differences in the preference for enrichment items highlights the importance of a varied enrichment approach, as there does not seem to be one item that satisfies the wants and needs of all individuals to the same degree. An enrichment concept, in which the needs of the animals are central, contributes to a more specific refinement of housing conditions.

Cognitive Foraging Enrichment (but Not Non-Cognitive Enrichment) Improved Several Longer-Term Welfare Indicators in Bottlenose Dolphins.

Bottlenose dolphins are the most common cetacean kept globally in zoos and aquaria (hereafter zoos), and are gregarious animals with a mostly opportunistic, generalist feeding strategy in the wild. In zoos, they have limited to no opportunities to express natural foraging behaviours as they receive their daily food ration of dead fish in a series of training sessions. Enrichment provision has increased in recent years, but items are still predominantly simple and floating in nature, and do not always target the animals' problem-solving or food-acquisition behaviours. These discrepancies run concurrently with the intense debate about dolphin welfare in zoos and how to improve it. The current study used a within-subject design on 11 bottlenose dolphins at Kolmårdens Djurpark and measured how several welfare indicators differed between two treatments of "cognitive" and "non-cognitive" food-based enrichment. The treatments were provided on an alternating basis for eight consecutive weeks: during cognitive enrichment weeks, the animals received items which stimulated their problem-solving and foraging behaviours, and during non-cognitive enrichment weeks, they received simple items paired with fish (to eliminate bias due to food value). Data were taken related to several multidisciplinary welfare parameters during enrichment provision and training sessions, and to activity budget behaviours throughout the week. During the cognitive as opposed to non-cognitive enrichment weeks, the dolphins engaged more with the enrichment, were more motivated to participate in training sessions and performed less anticipatory and stereotypic behaviours, suggesting that cognitive enrichment improved several indicators of bottlenose dolphin welfare. Valuable lines of further investigation would be to understand how individual differences and different types of cognitive enrichment impact potential welfare benefits. Our results suggest that enrichment items promoting cognitive foraging behaviours may improve dolphin welfare, and therefore zoos might prioritise giving cognitive enrichment to this species as well as considering the same for other species with similar cognitive skills and foraging ecologies.

MKI67 an potential oncogene of oral squamous cell carcinoma via the high throughput technology

Oral squamous cell carcinoma is a malignant tumor that occurs in the oral cavity, with poor prognosis and easy recurrence. However, the relationship between MKI67 and oral squamous cell carcinoma remains unclear. The oral squamous cell carcinoma datasets GSE138206, GSE146483 and GSE184616 were downloaded from the gene expression omnibus database, and the differentially expressed genes (DEGs) were screened. The protein-protein interaction network was constructed and analyzed by search tool for the retrieval of interacting genes database and Cytoscape software. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used for functional enrichment analysis. GO and KEGG analyses were performed on the whole genome, as formulated by gene set enrichment analysis. comparative toxicogenomics database was used to identify the diseases most associated with the core genes. TargetScan was used to screen miRNA regulating central DEGs. A total of 1472 DEGs were identified. GO analysis showed that the differentially expressed genes were mainly enriched in the tissues of extracellular matrix, type i interferon signaling pathway, human papillomavirus infection, adhesion spot, hepatitis C and ECM-receptor interaction. Enrichment items were similar to GO and KEGG enrichment items of differentially expressed genes. 10 core genes were obtained, and their expression was different between oral squamous cell carcinoma and normal tissue samples. MKI67 is highly expressed in oral squamous cell carcinoma and may be an oncogene in oral squamous cell carcinoma.

Single-cell analysis reveals distinct functional heterogeneity of CD34+ cells in anagen wound and diabetic wound

Wound healing is a complex biological process involving multiple cell types with their critical functions. The diabetic wounds show delayed wound healing, while the anagen wounds display accelerated wound closure. However, the mechanisms underlying the effect of cellular heterogeneity on wound healing are still unclear. CD34+ cells exhibit high heterogeneity in wound skins and improve wound healing. Herein, we investigated the phenotypic and functional heterogeneity of CD34+ cells in normal, anagen, and diabetic wounds. We obtained CD34 lineage tracing mice, constructed distinct wound models, collected CD34+ cells from wound edges, and performed single-cell RNA sequencing. We identified 10 cell clusters and 6 cell types of CD34+ cells, including endothelial cells, fibroblasts, keratinocytes, neutrophils, macrophages, and T cells. 5 subclusters were defined as fibroblasts. The CD34+ fibroblasts C2 highly expressed papillary fibroblastic markers took up the largest proportion in anagen wounds and were associated with inflammation and extracellular matrix. Increased CD34+ endothelial cells, fibroblasts C4, and neutrophils as well as decreased fibroblasts C1 were discovered in diabetic wounds. We also filtered out differentially expressed genes (DEGs) of each cell cluster in anagen wounds and diabetic wounds. Functional enrichment analysis was performed on these DEGs to figure out the enriched pathways and items for each cell cluster. Pseudotime analysis of CD34+ fibroblasts was next carried out indicating fibroblast C4 mainly with low differentiation. Our results have important implications for understanding CD34+ cell type-specific roles in anagen and diabetic wounds, provide the possible mechanisms of wound healing from a new perspective, and uncover potential therapeutic approaches to treating wounds.