From Skin to Gut: Understanding Microbial Diversity in Rana amurensis and R. dybowskii.

Amphibians face the threat of decline and extinction, and their health is crucially affected by the microbiota. Their health and ecological adaptability essentially depend on the diverse microbial communities that are shaped by unique host traits and environmental factors. However, there is still limited research on this topic. In this study, cutaneous (C) and gut (G) microbiota in Rana amurensis (A) and R. dybowskii (D) was analyzed through 16S amplicon sequencing. Groups AC and DC significantly differed in alpha diversity, while the gut groups (AG and DG) showed no such differences. Analyses of Bray-Curtis dissimilarity matrix and unweighted UniFrac distances showed significant differences in cutaneous microbiota between groups AC and DC, but not between groups AG and DG. Stochastic processes significantly influenced the assembly of cutaneous and gut microbiota in amphibians, with a notably higher species dispersal rate in the gut. The predominant phyla in the skin of R. amurensis and R. dybowskii were Bacteroidetes and Proteobacteria, respectively, with significant variations in Bacteroidota. Contrarily, the gut microbiota of both species was dominated by Firmicutes, Proteobacteria, and Bacteroidetes, without significant phylum-level differences. Linear discriminant analysis effect size (LEfSe) analysis identified distinct microbial enrichment in each group. Predictive analysis using phylogenetic investigation of communities by reconstruction of unobserved states 2 (PICRUSt2) revealed the significant functional pathways associated with the microbiota, which indicates their potential roles in immune system function, development, regeneration, and response to infectious diseases. This research underscores the critical impact of both host and environmental factors in shaping amphibian microbial ecosystems and emphasizes the need for further studies to explore these complex interactions for conservation efforts.

Impact of wildfire ash on skin and gut microbiomes and survival of Rana dybowskii.

Climate change and human activities escalate the frequency and intensity of wildfires, threatening amphibian habitats and survival; yet, research on these impacts remains limited. Wildfire ash alters water quality, introduces contaminants, and may disrupt microbial communities, impacting gut and skin microbiota; however, the effects on gut and skin microbiota remain unclear. Rana dybowskii were exposed to five concentrations (0 g L-1, 1.25 g L-1, 2.5 g L-1, 5 g L-1, and 10 g L-1) of aqueous extracts of wildfire ashes (AEAs) for 30 days to assess AEAs' metal content, survival, and microbiota diversity via Illumina sequencing. Our results showed that the major elements in ash were Ca > K > Mg > Al > Fe > Na > Mn, while in AEA they were K > Ca > Na > Mg > As > Al > Cu. A significant decrease in amphibian survival rates with increased AEA concentration was shown. The beta diversity analysis revealed distinct shifts in microbiota composition. Notably, bacterial genera associated with potential health risks showed increased abundance in skin microbiota, emphasising the potential for ash exposure to affect amphibian health. Functional prediction analyses revealed significant shifts in metabolic pathways related to health and disease, indicating that wildfire ash exposure may influence amphibian health through changes in microbial functions. This study highlights the urgent need for strategies to mitigate wildfire ash impacts on amphibians, as it significantly alters microbiota and affects their survival and health.

Rice straw ash and amphibian health: A deep dive into microbiota changes and potential ecological consequences.

Rice straw is burned as a result of agricultural practices and technical limitations, generating significant volumes of ash that might have environmental and ecological consequences; however, the effects on organisms have not been researched. Amphibians depend on their gut and skin microbiomes. Ash exposure may cause inflammation and changes in microbial diversity and function in frogs' skin and gut microbiota due to its chemical composition and physical presence, but the implications remain unclear. Rana dybowskii were exposed to five aqueous extracts of ashes (AEA) concentrations for 30 days to study survival, metal concentrations, and microbial diversity, analyzing the microbiota of the cutaneous and gut microbiota using Illumina sequencing. Dominant elements in ash: K > Ca > Mg > Na > Al > Fe. In AEA, K > Na > Ca > Mg > As > Cu. Increased AEA concentrations significantly reduced frog survival. Skin microbiota alpha diversity varied significantly among all treatment groups, but not gut microbiota. Skin microbiota differed significantly across treatments via Bray-Curtis and weighted UniFrac; gut microbiota was only affected by Bray-Curtis. Skin microbiota varied significantly with AEA levels in Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes, while the gut microbiota's dominant phyla, Firmicutes, Bacteroidetes, and Proteobacteria, remained consistent across all groups. Lastly, the functional prediction showed that the skin microbiota had big differences in how it worked and looked, which were linked to different health and environmental adaptation pathways. The gut microbiota, on the other hand, had smaller differences. In conclusion, AEA exposure affects R. dybowskii survival and skin microbiota diversity, indicating potential health and ecological impacts, with less effect on gut microbiota.

Tumorigenesis of basal muscle invasive bladder cancer was mediated by PTEN protein degradation resulting from SNHG1 upregulation.

BACKGROUND: Phosphatase and tensin homolog deleted on chromosome ten (PTEN) serves as a powerful tumor suppressor, and has been found to be downregulated in human bladder cancer (BC) tissues. Despite this observation, the mechanisms contributing to PTEN's downregulation have remained elusive. METHODS: We established targeted genes' knockdown or overexpressed cell lines to explore the mechanism how it drove the malignant transformation of urothelial cells or promoted anchorageindependent growth of human basal muscle invasive BC (BMIBC) cells. The mice model was used to validate the conclusion in vivo. The important findings were also extended to human studies. RESULTS: In this study, we discovered that mice exposed to N-butyl-N-(4-hydroxybu-tyl)nitrosamine (BBN), a specific bladder chemical carcinogen, exhibited primary BMIBC accompanied by a pronounced reduction in PTEN protein expression in vivo. Utilizing a lncRNA deep sequencing high-throughput platform, along with gain- and loss-of-function analyses, we identified small nucleolar RNA host gene 1 (SNHG1) as a critical lncRNA that might drive the formation of primary BMIBCs in BBN-treated mice. Cell culture results further demonstrated that BBN exposure significantly induced SNHG1 in normal human bladder urothelial cell UROtsa. Notably, the ectopic expression of SNHG1 alone was sufficient to induce malignant transformation in human urothelial cells, while SNHG1 knockdown effectively inhibited anchorage-independent growth of human BMIBCs. Our detailed investigation revealed that SNHG1 overexpression led to PTEN protein degradation through its direct interaction with HUR. This interaction reduced HUR binding to ubiquitin-specific peptidase 8 (USP8) mRNA, causing degradation of USP8 mRNA and a subsequent decrease in USP8 protein expression. The downregulation of USP8, in turn, increased PTEN polyubiquitination and degradation, culminating in cell malignant transformation and BMIBC anchorageindependent growth. In vivo studies confirmed the downregulation of PTEN and USP8, as well as their positive correlations in both BBN-treated mouse bladder urothelium and tumor tissues of bladder cancer in nude mice. CONCLUSIONS: Our findings, for the first time, demonstrate that overexpressed SNHG1 competes with USP8 for binding to HUR. This competition attenuates USP8 mRNA stability and protein expression, leading to PTEN protein degradation, consequently, this process drives urothelial cell malignant transformation and fosters BMIBC growth and primary BMIBC formation.

Characteristics and a comparison of the gut microbiota in two frog species at the beginning and end of hibernation.

Season has been suggested to contribute to variation in the gut microbiota of animals. The complicated relationships between amphibians and their gut microbiota and how they change throughout the year require more research. Short-term and long-term hypothermic fasting of amphibians may affect gut microbiota differently; however, these changes have not been explored. In this study, the composition and characteristics of the gut microbiota of Rana amurensis and Rana dybowskii during summer, autumn (short-term fasting) and winter (long-term fasting) were studied by high-throughput Illumina sequencing. Both frog species had higher gut microbiota alpha diversity in summer than autumn and winter, but no significant variations between autumn and spring. The summer, autumn, and spring gut microbiotas of both species differed, as did the autumn and winter microbiomes. In summer, autumn and winter, the dominant phyla in the gut microbiota of both species were Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria. All animals have 10 OTUs (>90% of all 52 frogs). Both species had 23 OTUs (>90% of all 28 frogs) in winter, accounting for 47.49 ± 3.84% and 63.17 ± 3.69% of their relative abundance, respectively. PICRUSt2 analysis showed that the predominant functions of the gut microbiota in these two Rana were focused on carbohydrate metabolism, Global and overview maps, Glycan biosynthesis metabolism, membrane transport, and replication and repair, translation. The BugBase analysis estimated that among the seasons in the R. amurensis group, Facultatively_Anaerobic, Forms_Biofilms, Gram_Negative, Gram_Positive, Potentially_Pathogenic were significantly different. However, there was no difference for R. dybowskii. The research will reveal how the gut microbiota of amphibians adapts to environmental changes during hibernation, aid in the conservation of endangered amphibians, particularly those that hibernate, and advance microbiota research by elucidating the role of microbiota under various physiological states and environmental conditions.

The study of double-J stent free mode of flexible ureteroscopy and laser lithotripsy: a single centre experience.

OBJECTIVE: To evaluate the effectiveness and safety of flexible ureteroscopy in the treatment of kidney and upper ureteral calculi under double-J stent free mode. METHODS: Data from patients who underwent flexible ureteroscopy and laser lithotripsy between February 2018 and September 2021 were retrospectively and analysed. Cases were grouped according to pre- or postoperative use of the double-J stent (6 Fr): Post-F group (preoperative double-J stent but no postoperative double-J stent); Pre-F group (no preoperative stenting but with postoperative double-J stent); and Routine group (preoperative and postoperative double-J stenting). RESULTS: A total of 554 patients (390 male and 164 female) were included. The mean operation time was similar between the three groups, with no statistically significant difference. Incidence of grade 0-1 ureteral injury was significantly higher in the Pre-F group versus other groups, but there were no significant between-group differences in other operation-related complications. During follow-up, stent-associated complications were observed in the Pre-F and Routine groups, but not in the Post-F group. Stone clearance rates were similar between all groups at 1, 3 and 6 months following surgery. CONCLUSIONS: Flexible ureteroscopy using double-J stent free mode was found to be safe, feasible and effective in treating renal and upper ureteral calculi.

Impact of caregiver burden on caregiver mental health in patients with esophageal cancer: chain mediating effects of benefit finding and rumination, a cross-sectional study.

Background: Cancer caregivers are the main supporter for the tumor patients, they not only need to provide daily nursing to the patients, but also suffering the pressure from economy, emotion and even family members. The mental health of tumor patient is mostly noticed, while not caregivers. The mental health of caregiver greatly affected the nursing quality and even the treatment outcomes. In the current study, the mechanisms underlying the links between caregiver burden, benefit finding, mental health, and rumination in those caring for people with esophageal cancer were examined. Methods: The study was using a convenience sampling, 166 esophageal cancer patients in 2 general hospitals in Jiangsu Province, China, and caregivers of patients were included after excluded the non-conforming patients. Data were collected using investigator-developed questionnaires, the Benefit Finding Scale (BFS), the Event-Related Rumination Inventory (ERRI), the Zarit Burden Interview (ZBI), and the Hospital Anxiety and Depression Scale (HADS) during May 2020 to December 2020. The results were analyzed by SPSS, and the chain mediating effect was analyzed by the the SPSS PROCESS Macro Model. Results: The study comprised 166 caregivers with an average age of (59.96±11.48) years, most of them were female (85.5%). The ZBI was positive correlated with HADS (r=0.882, P<0.01), and negative correlated with BFS (r=-0.873, P<0.01). Intrusive rumination and deliberate rumination in caregivers were negatively correlated (r=-0.901, P<0.01) and positive correlated (r=0.904, P<0.01) with BFS scores, respectively. Furthermore, research have discovered a chain mediation impact of benefit finding and rumination between caregiver burden and psychological well-being among carers of esophageal cancer patients. Conclusions: The findings of this study imply that benefit finding and rumination are crucial components of the coping strategy used to buffer against negative emotion (such as anxiety and depression). Therefore, the mental health of caregivers should also be noticed, and health care professionals should provide targeted interventions to increase the caregiver's level of benefit finding and promote deliberate ruminative thinking.

Value acquisition, value co-creation: The impact of perceived organic grocerant value on customer engagement behavior through brand trust.

Affected by COVID-19, there is a growing trend toward healthy lifestyles and organic food consumption. The literature on organic foods focuses on the factors that influence buying behavior. A thriving organic business requires both sustained consumption and consumer contributions beyond the purchase-customer engagement behavior. The purpose of this study is to examine the factors that may drive member customers to engage with organic grocerants. This study surveyed 280 Chinese member customers of an organic grocerant to explore how to drive customer engagement behavior. Based on value co-creation theory and the customer engagement literature, this study proposed a "value acquisition-value co-creation" framework to explore the relationship between perceived value, brand trust, and customer engagement behavior. The results show that emotional and social value can directly and effectively motivate customer engagement behavior in organic grocerants. However, consumers' perceived quality value and price value will not directly affect customer engagement behavior but instead indirectly affect it through brand trust. Furthermore, improving the perceived value of emotion, quality and price can strengthen brand trust in organic grocerants. The study confirms that brand trust is critical to organic grocerant and customer engagement. Our findings provide a new perspective for understanding the relationship between the value customers receive from organic food consumption and value co-creation through customer engagement behavior.

Increase consumers' willingness to pay a premium for organic food in restaurants: Explore the role of comparative advertising.

Offering organic food is a new trend in the hospitality industry seeking sustainable competitiveness. Premiums and information barriers impede continued growth in organic consumption. This study aims to explore the role of comparative advertising (CA) in organic food communication. Three empirical studies were used to verify the effect of CA vs. non-comparative advertising (NCA) on consumers' willingness to pay a premium (WTPP) for organic food, examining how benefit appeals (health vs. environmental) and consumers' organic skepticism affects CA. The results indicate that matching CA and health appeals increase consumers' WTPP, while environmental appeals have no significant differences between the CA and NCA groups (Study 1). Information persuasiveness mediates the interaction between CA and benefit appeal on WTPP (Study 2). CA increases WTPP among consumers with high organic skepticism, while the interaction between CA and health appeal is only effective for low skepticism consumers (Study 3). The findings unravel and explain the mechanics of how CA works in organic products, which can help restaurants, retailers and tourist destinations advertise organic food to increase consumers' WTPP.

Changes in the gut microbiota diversity of brown frogs (Rana dybowskii) after an antibiotic bath.

BACKGROUND: Captive amphibians frequently receive antibiotic baths to control bacterial diseases. The potential collateral effect of these antibiotics on the microbiota of frogs is largely unknown. To date, studies have mainly relied on oral administration to examine the effects of antibiotics on the gut microbiota; in contrast, little is known regarding the effects of bath-applied antibiotics on the gut microbiota. The gut microbiota compositions of the gentamicin, recovery, and control groups were compared by Illumina high-throughput sequencing, and the functional profiles were analysed using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). Furthermore, the relationship between the structure and predicted functional composition of the gut microbiota was determined. RESULTS: The alpha diversity indices were significantly reduced by the gentamicin bath, illustrating that this treatment significantly changed the composition of the gut microbiota. After 7 days, the gut microbiota of the recovery group was not significantly different from that of the gentamicin group. Forty-four indicator taxa were selected at the genus level, comprising 42 indicators representing the control group and 2 indicators representing the gentamicin and recovery groups. Potential pathogenic bacteria of the genera Aeromonas, Citrobacter, and Chryseobacterium were significantly depleted after the gentamicin bath. There was no significant positive association between the community composition and functional composition of the gut microbiota in the gentamicin or control frogs, indicating that the functional redundancy of the gut bacterial community was high. CONCLUSIONS: Gentamicin significantly changed the structure of the gut microbiota of R. dybowskii, and the gut microbiota exhibited weak resilience. However, the gentamicin bath did not change the functional composition of the gut microbiota of R. dybowskii, and there was no significant correlation between the structural composition and the functional composition of the gut microbiota.