Search Results (134)

Background/Objectives: The clinical potential of antimicrobial peptides (AMPs) against dual threats like antimicrobial resistance (AMR) and cancer is often limited by their high host cell toxicity. Here, we focused on brevinin-2OS (B2OS), a novel peptide from the skin of Odorrana schmackeri with potent haemolytic activity. The objective was to study the structure–activity relationship and optimise the safety via targeted modifications. Methods: A dual-modification strategy involving C-terminal truncation and subsequent N-terminal D-amino acid substitution was employed. The bioactivities and safety profiles of the resulting analogues were evaluated using antimicrobial, haemolysis, and cytotoxicity assays. Result: Removal of the rana box in B2OS(1-22)-NH₂ substantially reduced haemolysis while maintaining bioactivities. Remarkably, the D-leucine substitution in [D-Leu²]B2OS(1-22)-NH₂ displayed a superior HC₅₀ value of 118.1 µM, representing a more than ten-fold improvement compared to its parent peptide (HC₅₀ of 10.44 µM). This optimised analogue also demonstrated faster bactericidal kinetics and enhanced membrane permeabilisation, leading to a greater than 22-fold improvement in its therapeutic index against Gram-positive bacteria. Conclusions: The C-terminal rana box is a primary determinant of toxicity rather than a requirement for activity in the B2OS scaffold. The engineered peptide [D-Leu²]B2OS(1-22)-NH₂ emerges as a promising lead compound, and this dual-modification strategy provides a powerful design principle for developing safer, more effective peptide-based therapeutics. Full article

Memory is a fundamental neurological function, essential for animal survival. Over the course of vertebrate evolution, elaborations in the forebrain telencephalon create new memory mechanisms, meaning basal vertebrates such as amphibians must have a less sophisticated system of memory acquisition, storage, and retrieval than the well-known hippocampal-based circuitry of mammals. Personality also appears to be a fundamental vertebrate trait and is generally defined as consistent individual behavior over time and across life history stages. In anuran amphibians (frogs), personality studies generally ask whether adult frogs retain the personality of their tadpole stage or whether personality shifts with metamorphosis, an idea behavioral ecologists term adaptive decoupling. Using a multidisciplinary perspective and recognizing there are ~7843 species of frogs, each with some molecular, morphological, physiological, or behavioral feature that makes it unique, we review, clarify, and provide perspective on what we collectively know about memory and personality and their mechanisms in anuran amphibians. We propose four working hypotheses: (1) as tadpoles grow, new telencephalic neurons become integrated into functional networks, producing behaviors that become more sophisticated with age; (2) since carnivores tend to be more bold/aggressive than herbivores, carnivorous anuran adults will be more aggressive than herbivorous tadpoles; (3) each amphibian species, and perhaps life history stage, will have a set point on the Shy–Bold Continuum; and (4) around this set point there will be a range of individual responses. We also suggest that several factors are slowing our understanding of the variety and depth of memory and personality possibilities in anurans. These include the scala natura approach to comparative studies (i.e., the idea that one frog represents all frogs); the assumption that amphibians are no more than simple reflex machines; that study species tend to be chosen more for convenience than taxonomic representation; and that studies are designed to prove or disprove a construct. This latter factor is a particular hindrance because what we are really seeking as scientists is not the confirmation or refutation of ideas, but rather what those ideas are intended to produce, which is understanding. Full article

Rana dybowskii, widely distributed and extensively farmed in northeast China, holds significant economic value, particularly for its fallopian tubes, which are used as a traditional Chinese medicinal tonic known as “Oviductus Ranae.” As the light spectrum is a cost-effective regulatory factor in aquaculture, understanding its effects on the tadpole stage of R. dybowskii is critical for optimizing cultivation practices. This study investigated the effects of five light colors (white, red, yellow, blue, and green) on steroid hormone levels and gut microbiota composition in R. dybowskii tadpoles. Steroid hormone levels were measured on days 15, 30, 45, and 60 using high-performance liquid chromatography (HPLC), while gut microbial communities were analyzed through high-throughput 16S rRNA sequencing. Results showed that the testosterone (T) level of frogs in green light (group G) peaked on day 60 (2.62 ± 3.70 ng/g). The estradiol (E2) level in blue light (group B) also peaked on day 60 (2.87 ± 0.71 ng/g). Importantly, sex ratio analysis revealed that the proportion of females was highest under blue light, reaching 61.11%. Meanwhile, the richness and diversity of the gut bacterial community of the tadpoles was highest under yellow light, followed by blue light. These data suggest that hormone levels fluctuated and the composition of the gut flora of R. dybowskii changed under different light colors. Our results advance R. dybowskii physiological knowledge and support aquaculture practices. Full article

As sustainability becomes increasingly central to organizational strategy, social economy organizations (SEOs) are rethinking their business models. This study employs stakeholder analysis using the value mapping (VM) tool developed by Short, Rana, Bocken, and Evans for the development of the VOLTO JÁ project. The objective of the VOLTO JÁ project is to operationalize a senior exchange programme between SEOs. The VM approach extends beyond conventional customer value propositions to prioritize sustainability for all stakeholders and identify key drivers of sustainable business model (SBM) innovation. The multi-stakeholder methodology comprises the following elements: (1) sequential focus groups aimed at enhancing sustainable business thinking; (2) semi-structured interviews; and (3) workshop to facilitate qualitative analysis and co-create the VM. The findings are then categorized into four value dimensions: (1) value captured—improved participant well-being, enhanced reputational capital, mitigation of social asymmetries, and affordable service experiences; (2) value lost—underused community assets; (3) value destroyed—institutional and systemic barriers to innovation; and (4) new value opportunities—knowledge sharing, service diversification, and open innovation to foster collaborative networks. The study demonstrates that the application of VM in SEOs supports SBM development by generating strategic insights, enhancing resource efficiency, and fostering the delivery of socially impactful services. Full article

The Chinese forest frog (Rana chensinensis) has high ecological and economic value and is an ecologically important species that is very sensitive to environmental changes. However, climate change and increasing human activity are posing growing threats to its natural habitat. To address these challenges, this study aimed to predict the current and future geographic distribution of the Chinese forest frog and to identify the environmental drivers influencing its habitat suitability. Here, the optimized MaxEnt and Biomod2 were used to assess 127 species occurrence records and 22 environmental variables and model and analyze changes in the geographic distribution of the Chinese forest frog in different periods. The environmental factors underlying geographical distribution changes and migration trends in distribution areas under climate change were assessed. The optimized MaxEnt model generated the best predictions, showing that the current most highly suitable areas are located in Chongqing, Sichuan, and Gansu provinces. The optimized MaxEnt model demonstrated exceptionally high predictive accuracy, with mean AUC values of 0.968 ± 0.000 and TSS values of 0.800 ± 0.089. The total area of the current potentially suitable habitat was estimated at 426 × 10⁴ km². The principal ecological factors influencing the distribution of the Chinese forest frog are Bio9, Bio10, and human activity. Under future scenarios, the potentially suitable habitat for the Chinese forest frog is projected to shift westward and southward. Compared to the current situation, the potentially suitable habitat for the Chinese forest frog is expected to significantly shrink across all four future scenarios (2050s-SSP126, 2050s-SSP585, 2090s-SSP126, and 2090s-SSP585), posing serious threats to its survival. This study not only identifies the key ecological factors limiting the potential distribution of the Chinese forest frog but also provides a scientific basis and data support for the development of conservation strategies and habitat restoration efforts. Full article

Background: As a broad-spectrum fluoroquinolone, enrofloxacin (ENR) is commonly employed to manage bacterial infections in aquatic species. Nevertheless, there have been no documented pharmacokinetic and residue studies conducted on Dybowski’s frog (Rana dybowskii). Therefore, the objective of our study was to characterize the pharmacokinetics (PK) of ENR and its metabolite ciprofloxacin (CIP) in R. dybowskii, establish withdrawal times, and evaluate the physiological effects associated with ENR administration. Methods: Adult Rana dybowskii (120 individuals; 60 males and 60 females) were sex-separated and acclimated in four tanks. Prior to dosing, three males and three females were randomly selected as untreated controls (without ENR administration). Following the oral gavage of ENR (10 mg/kg), blood, liver, and kidney tissues were collected at 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24, 36, 48, and 72 h (n = 6) for pharmacokinetic analysis. Muscle and oviduct tissues were additionally sampled at 1, 3, 7, 15, and 30 days post-dose (n = 6) for ENR content determination. Serum/tissue ENR concentrations were measured via Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS) and analyzed using a non-compartmental model (WinNonLin 6.1 software) to calculate PK parameters including peak time (T_max), peak concentration (C_max), and area under the curve (AUC_0−t). In studying the physiology effects of ENR administration, biochemical enzyme activities and gene expressions in the liver and intestine were assessed post-ENR administration. Results: ENR demonstrated rapid absorption and extensive distribution in R. dybowskii. The withdrawal periods were determined to be over 33 days for females and 34 days for males in R. dybowskii. Following ENR administration, there was an increase in immune enzymes (AKP (alkaline phosphatase) and ACP (acid phosphatase)) as well as glycolytic enzymes (HK (hexokinase), PK (pyruvate kinase), PFK (phosphofructokinase)). Antioxidant enzyme levels, specifically SOD (superoxide dismutase) and CAT (catalase), peaked at 1.5 h post-ENR administration but subsequently declined by the 8 h mark. Additionally, following ENR treatment, IGF1, PI3K, and Akt exhibited up-regulation, whereas Keap1 and GYS1 showed down-regulation. Conclusions: The administration of ENR at a dosage of 10 mg/kg significantly enhances the activities of AKP and ACP, promotes glycolysis, and activates the Keap1/Nrf2 and PI3K-Akt signaling pathways in R. dybowskii. These findings establish a foundation for the rational application of ENR and the determination of withdrawal times in frog aquaculture. Full article

Amphibian conservation concerns frequently center on the idea of ‘saving’ them, with the underlying assumption they are the passive victims of anthropogenic environmental change. But this approach ignores the physiological, biochemical, and behavioral flexibility amphibians have employed since they first evolved ~365 million years ago. One overlooked advantage amphibians possess in the struggle for survival, and one humans might use in their efforts to conserve them, is their brains share the same blueprint as human brains, which allows them to acquire knowledge and understanding through experiences—in other words, amphibians have cognitive capabilities that assist them in their effort to survive. Here, we use four examples from our work on the behavioral ecology of Crawfish Frogs (Rana areolata) to form hypotheses about how cognition affects amphibian reaction to environmental and social change. The first two examples describe Crawfish Frog responses to seasonality and reproductive status, the third details their reaction to ecological disturbance, and the fourth describes how their response to the same stimulus changes with growth/age. In each example, we detail the neuronal circuitry thought to be involved and hypothesize the role of cognition. We propose that as one component of our fight to conserve amphibians, researchers should consider the full range of anatomical, physiological, biochemical, and behavioral features amphibians themselves employ in their defense, which are features responsible for their historical evolutionary success up until the Anthropocene. Further, we submit that acknowledging amphibians possess cognitive abilities can enrich interpretations of not only behavioral and ecological observations but also of neuroanatomical and neurophysiological results. Full article

Devices that measure the presence of instability in the pulse shapes in trains of ultrashort laser pulses do not exist, so this task necessarily falls to pulse-measurement devices, like Frequency-Resolved Optical Gating (FROG) and its variations, which have proven to be a highly reliable class of techniques for measuring stable trains of ultrashort laser pulses. Fortunately, multi-shot versions of FROG have also been shown to sensitively distinguish trains of stable from those of unstable pulse shapes by displaying readily visible systematic discrepancies between the measured and retrieved traces in the presence of unstable pulse trains. However, the effects of pulse-shape instability and algorithm stagnation can be indistinguishable, so a never-stagnating algorithm—even when instability is present—is required and is generally important. In previous work, we demonstrated that our recently introduced Retrieved-Amplitude N-grid Algorithmic (RANA) approach produces highly reliable (100%) pulse-retrieval in the second-harmonic-generation (SHG) version of FROG for thousands of sample trains of pulses with stable pulse shapes. Further, it does so even for trains of unstable pulse shapes and thus both reliably distinguishes between the two cases and provides a rough measure of the degree of instability as well as a reasonable estimate of most typical pulse parameters. Here, we perform the analogous study for the polarization-gating (PG) and transient-grating (TG) versions of FROG, which are often used for higher-energy pulse trains. We conclude that PG and TG FROG, coupled with the RANA approach, also provide reliable indicators of pulse-shape instability. In addition, for PG and TG FROG, the RANA approach provides an even better estimate of a typical pulse in an unstable pulse train than SHG FROG does, even in cases of significant pulse-shape instability. Full article

Microplastic (MP) pollution is a major environmental problem, but a comparative study of the toxicological effects of different MPs remains lacking. To explore the toxicological effects of three different microplastics, namely, polypropylene (PP), polystyrene (PS) and polyethylene (PE), Zhenhai brown frog (Rana zhenhaiensis) tadpoles were used as the model animal. The results showed that exposure to PE and PS significantly reduced the metamorphosis rate of the tadpoles. Compared with the control group, the body weight of tadpoles in all MP treatments was significantly reduced compared with that of the control group. In addition, exposure to PE reduced the body length and hind limb length of tadpoles. The number of pigment cells increased and intercellular spaces expanded in the liver tissues of tadpoles receiving PS and PE treatments. The composition and function of the intestinal microbiota in the PP treatment and control groups were similar, whereas between the PS treatment and control, they differed. Liver transcriptome sequencing revealed significant alterations in key genes associated with oxidative stress, energy metabolism, immune response, and apoptosis signaling pathways with PS treatment and PP treatment. In summary, MPs may have harmed tadpoles to varying degrees by interfering with related signaling pathways. The negative effects of PE and PS were greater than those of PP. Full article

Geographical variation in morphological traits represents a significant ecological phenomenon. According to Allen’s rule, animals inhabiting cooler environments typically exhibit shorter extremities compared to those in warmer regions. While Allen’s rule has been extensively validated along latitudinal gradients, its applicability to altitudinal variation in morphological traits remains less well understood. In this study, we analyzed morphological data—including forelimb length and hindlimb length—from 492 individuals of Rana kukunoris from 26 populations to assess whether relative limb size in both sexes declines with altitude, aligning with Allen’s rule. However, this pattern exhibited geographical regional variation. In the northern and central regions, relative limb length consistently followed the predictions of Allen’s rule. In contrast, the southern region showed no significant altitudinal variation in limb proportions. These results highlight that the applicability of Allen’s rule may be modulated by regional environmental factors and altitude vertical differences, underscoring the need for a nuanced understanding of how local contexts influence morphological adaptations. Full article

Prion diseases are fatal neurodegenerative diseases that can be transmitted by infectious protein particles, PrP^Scs, encoded by the endogenous prion protein gene (PRNP). The origin of prion seeds is unclear, especially in non-human hosts, and this identification is pivotal to preventing the spread of prion diseases from host animals. Recently, an abnormally high amyloid propensity in prion proteins (PrPs) was found in a frog, of which the genetic variations in the PRNP gene have not been investigated. In this study, genetic polymorphisms in the PRNP gene were investigated in 194 Dybowski’s frogs using polymerase chain reaction (PCR) and amplicon sequencing. We carried out in silico analyses to predict functional alterations according to non-synonymous single nucleotide polymorphisms (SNPs) using PolyPhen-2, PANTHER, SIFT, and MutPred2. We used ClustalW2 and MEGA X to compare frog PRNP and PrP sequences with those of prion-related animals. To evaluate the impact of the SNPs on protein aggregation propensity and 3D structure, we utilized AMYCO and ColabFold. We identified 34 novel genetic polymorphisms including 6 non-synonymous SNPs in the frog PRNP gene. The hydrogen bond length varied at codons 143 and 207 according to non-synonymous SNPs, even if the electrostatic potential was not changed. In silico analysis predicted S143N to increase the aggregation propensity, and W6L, C8Y, R211W, and L241F had damaging effects on frog PrPs. The PRNP and PrP sequences of frogs showed low homology with those of prion-related mammals. To the best of our knowledge, this study was the first to discover genetic polymorphisms in the PRNP gene in amphibians. Full article

Freeze tolerance is a remarkable adaptive trait exhibited by wood frogs (Rana sylvatica) during their hibernation period. To show the epigenetic mechanisms that contribute to kidney protection during freezing stress, this present study provides the first investigation of the role and dynamics of histone arginine methylation and the expression of protein arginine methyltransferases (PRMTs) in a freeze-tolerant vertebrate. Kidney samples from three groups were assessed: (a) control frogs acclimated at 5 °C, (b) 24 h frozen frogs, and (c) 8 h thawed frogs. Our findings revealed significant downregulation of PRMT1, PRMT3, and PRMT5 in kidneys from frozen wood frogs compared to the control group. This downregulation indicates a potential role for PRMT enzymes in the regulation of arginine methylation under freezing stress. In addition, we observed distinct changes in histone marks. H3R17me2a showed significant upregulation after 24 h of freezing, potentially indicating its involvement in the activation of genes related to freezing survival. By contrast, H3R26me2a was downregulated after both 24 h freezing and 8 h thawing, whereas H3R8me2a showed sustained levels after freezing but was downregulated after thawing. These findings highlight the dynamic nature of histone arginine methylation and PRMT expression in wood frog kidneys during freezing–thawing. Our results indicate that epigenetic modifications play a crucial role in shaping the adaptive responses of wood frog kidneys to freezing stress and contribute new information on the underlying biochemical modifications that support vertebrate freeze tolerance. Full article

Microplastics pollution in freshwater systems is attracting increasing attention. However, our knowledge of its combined toxicity with heavy metals is scarce. In this study, Rana zhenhaiensis was used as the model animal to study the combined poisoning mechanism of cadmium or microplastics on the digestive systems of tadpoles in freshwater. Results showed that the exposure to cadmium and polystyrene increased the mortality and metamorphosis rate of R. zhenhaiensis tadpoles, and delayed their growth and development. Cadmium was detected in the livers and intestines, while polystyrene mainly accumulated in the gills and intestines of tadpoles. The individual exposure of cadmium or polystyrene can cause pathological damage to liver tissue, induce oxidative stress in liver, and change gene expression. Cadmium co-exposure with polystyrene can reduce the cadmium accumulation in the liver. While polystyrene can slightly increase cadmium accumulation in the intestine. Exposure to cadmium and polystyrene altered the abundance and community structure of intestinal microbiota, and polystyrene increased the dysregulation of the gut microbiome. In this study, the combined exposure of microplastics and cadmium had a negative impact on R. zhenhaiensis tadpoles, but the introduction of microplastics on the toxicity of cadmium on the tadpoles needs further investigation, due to the different characteristics of microplastics. Full article

Background/Objective: Amphibian skin is a valuable source of host defense peptides (HDPs). This study aimed to identify HDPs with novel amino acid sequences from the skin of Rana tagoi yakushimensis and analyze their functions. Methods: cDNAs encoding HDP precursors were cloned and sequenced using RT-PCR and 3′-RACE. The novel HDPs were synthesized to evaluate their antimicrobial activity, antioxidant activity, and cytotoxicity. Antimicrobial activity was evaluated by way of broth microdilution and endotoxin- and β-glucan-binding capacity using an enzyme-linked endotoxin binding assay (ELEBA) and a modified ELEBA, respectively. Results: Nine cDNAs encoding precursors for various HDP families, including temporin, ranatuerin-2, brevinin-1, amurin-9, and a novel yakushimin peptide, were identified. Brevinin-1TYa exhibited antibacterial activity against Staphylococcus aureus, and brevinin-1TYa and amurin-9TYa induced morphological changes in Escherichia coli and S. aureus. Yakushimin-TYa, amurin-9TYa, and brevinin-1TYa showed concentration-dependent antibacterial effects against the plant pathogens Xanthomonas oryzae pv. oryzae and Clavibacter michiganensis subsp. michiganensis. Amurin-9TYa demonstrated strong binding affinity to lipopolysaccharide, lipoteichoic acid, and β-glucan, exhibited antioxidant activity, and lacked cytotoxicity, making it a promising therapeutic candidate. Moreover, brevinin-1TYa showed strong cytotoxicity, whereas yakushimin-TYa exhibited weak cytotoxicity. Conclusions: These findings highlight the potential of these peptides, particularly amurin-9TYa, for future applications as antimicrobial and therapeutic agents. Full article

Background: Freeze-tolerant animals undergo significant physiological and biochemical changes to overcome challenges associated with prolonged whole-body freezing. In wood frog Rana sylvatica (now Lithobates sylvaticus), up to 65% of total body water freezes in extracellular ice masses and, during this state of suspended animation, it is completely immobile and displays no detectable brain, heart, or respirometry activity. To survive such extensive freezing, frogs integrate various regulatory mechanisms to ensure quick and smooth transitions into or out of this hypometabolic state. One such rapid and reversible regulatory molecule capable of coordinating many aspects of biological functions is microRNA. Herein, we present a large-scale analysis of the biogenesis and regulation of microRNAs in wood frog liver over the course of a freeze–thaw cycle (control, 24 h frozen, and 8 h thawed). Methods/Results: Immunoblotting of key microRNA biogenesis factors showed an upregulation and enhancement of microRNA processing capacity during freezing and thawing. This was followed with RT-qPCR analysis of 109 microRNA species, of which 20 were significantly differentially expressed during freezing and thawing, with the majority being upregulated. Downstream bioinformatics analysis of miRNA/mRNA targeting coupled with in silico protein–protein interactions and functional clustering of biological processes suggested that these microRNAs were suppressing pro-growth functions, including DNA replication, mRNA processing and splicing, protein translation and turnover, and carbohydrate metabolism. Conclusions: Our findings suggest that this enhanced miRNA maturation capacity might be one key factor in the vital hepatic miRNA-mediated suppression of energy-expensive processes needed for long-term survival in a frozen state. Full article