Metabolic Adaption of Flexor Carpi Radialis to Amplexus Behavior in Asiatic Toads (Bufo gargarizans).

Amplexus is a type of mating behavior among toads that is essential for successful external fertilization. Most studies have primarily focused on the behavioral diversity of amplexus, and less is known regarding the metabolic changes occurring in amplectant males. The aim of this study was to compare the metabolic profiles of amplectant Asiatic toad (Bufo gargarizans) males in the breeding period (BP group) and the resting males in the non-breeding period (NP group). A metabolomic analysis was conducted on the flexor carpi radialis (FCR), an essential forelimb muscle responsible for clasping during courtship. A total of 66 differential metabolites were identified between the BP and NP groups, including 18 amino acids, 12 carbohydrates, and 8 lipids, and they were classified into 9 categories. Among these differential metabolites, 13 amino acids, 11 carbohydrates, and 7 lipids were significantly upregulated in the BP group compared to the NP group. In addition, a KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis identified 17 significant metabolic pathways, including ABC transporters, aminoacyl-tRNA biosynthesis, arginine biosynthesis, pantothenate and CoA biosynthesis, and fructose and mannose metabolism. These results suggest that amplectant male toads are metabolically more active than those during the non-breeding period, and this metabolic adaptation increases the likelihood of reproductive success.

Histopathological validation of safe margin for nephron-sparing surgery based on individual tumor growth pattern.

BACKGROUND: To evaluate the clinicopathologic value of morphological growth patterns of small renal cell carcinoma (sRCC) and determine the actual demand for taking a rim of healthy parenchyma to avoid positive SM. METHODS: Data was collected from 560 sRCC patients who underwent laparoscopic surgeries from May 2010 to October 2017. One hundred forty-nine cases received nephron-sparing surgery (NSS) and others received radical nephrectomy (RN). All specimens were analyzed separately by two uropathologists, and three morphological growth patterns were identified. The presence of pseudocapsule (PC), surgical margins (SM), and other routine variables were recorded. The relationship between growth patterns and included variables was measured by the χ2 test and Fisher's exact probability test. Survival outcomes were evaluated by Kaplan-Meier method and the log-rank test. RESULTS: The median age of patients was 63.2 years old and the mean tumor diameter was 3.0 cm. Four hundred eighty (85.7%) cases were clear cell RCC and 541 (96.6%) cases were at the pT1a stage. Peritumoral PC was detected in 512 (92.5%) specimens, and the ratio of tumor invasion in PC in infiltration pattern increased obviously than that of the other growth patterns. Similarly, the pT stage was significantly correlated with the infiltration pattern as well. One hundred forty-nine patients underwent NSS and 3 (2.0%) of them showed positive SM after operation. Statistical differences of the 5-year overall survival (OS) and the cancer-specific survival (CSS) existed between different morphological growth patterns, PC status, and pT stages. CONCLUSIONS: Morphological growth patterns of sRCC might be used as a potential biomarker to help operate NSS to avoid the risk of positive SM. How to distinguish different morphological growth patterns before operation and the effectiveness of the growth pattern as a novel proposed parameter to direct NSS in sRCC patients deserves further exploration.

MiR-375 Mediates CRH Signaling Pathway in Inhibiting E2 Synthesis in Porcine Ovary.

The corticotropin-releasing hormone (CRH) signaling system is involved in numbers of stress-related physiological and pathological responses，including its inhibiting effects on estradiol (E2) synthesis and follicular development in the ovary. In addition, there are reports that microRNAs (miRNAs) can control the function of animal reproductive system. The aim of present study was to investigate the functions of miR-375 and the relationship between miR-375 and CRH signaling molecules in the porcine ovary. First, our common PCR results show that miR-375 and the CRH receptor 1 (CRHR1) are expressed in porcine ovary, whereas CRH receptor 2 (CRHR2) is not detected. We further have located the cell types of miR-375 and CRHR1 by in situ hybridization (ISH), and the results show that miR-375 is located only in the granulosa cells, whereas CRHR1 is positive in all of granulosa cells and oocytes, inferring that miR-375 and CRHR1 are co-localized in granulosa cells. Second, we show that overexpression of miR-375 in cultured granulosa cells suppresses the E2 production, while miR-375 knockdown demonstrates the opposite result. Besides, our in vitro results demonstrate that miR-375 mediates the signaling pathway of CRH inhibiting E2 synthesis. Finally, our data show that the action of miR-375 is accomplished by directly binding to the 3'UTR of specificity protein1 (SP1) mRNA to decrease the SP1 protein level. Thus, we conclude that miR-375 is a key factor in regulating E2 synthesis by mediating the CRH signaling pathway.

The paddy frog genome provides insight into the molecular adaptations and regulation of hibernation in ectotherms.

Amphibians, like the paddy frog (Fejervarya multistriata), have played a critical role in the transition from water to land. Hibernation is a vital survival adaptation in cold environments with limited food resources. We decoded the paddy frog genome to reveal the molecular adaptations linked to hibernation in ectotherms. The genome contained 13 chromosomes, with a significant proportion of repetitive sequences. We identified the key genes encoding the proteins of AANAT, TRPM8, EGLN1, and VEGFA essential for circadian rhythms, thermosensation, and hypoxia during hibernation by comparing the hibernator and non-hibernator genomes. Examining organ changes during hibernation revealed the central regulatory role of the brain. We identified 21 factors contributing to hibernation, involving hormone biosynthesis, protein digestion, DNA replication, and the cell cycle. These findings provide deeper insight into the complex mechanisms of ectothermic hibernation and contribute to our understanding of the broader significance of this evolutionary adaptation.

Predicting Conservation Status of Testudoformes under Climate Change Using Habitat Models.

Climate change promotes variations in distribution ranges, potentially leading to biodiversity loss and increased extinction risks for species. It is crucial to investigate these variations under future climate change scenarios for effective biodiversity conservation. Here, we studied the future distribution ranges of 268 Testudoformes species under climate change using habitat models, specifically species distribution models (SDMs), to assess their conservation status. Our results have indicated that over half of species are projected to experience declines in their potential distribution ranges under two scenarios. In particular, we found that three critically endangered species-Three-striped roofed turtle (Batagur dhongoka), Durango mud turtle (Kinosternon durangoense), and Colombian mud turtle (Kinosternon dunni)-displayed extraction of their distribution ranges and faced extinction under global climate change. Additionally, our analysis revealed that the potential distribution ranges of some species might increase under future climate scenarios. However, these findings must be interpreted with caution as they do not account for other significant factors such as biological invasions, population structure, land-use change, anthropogenic disturbances, and inter-organism interrelationships. Future studies should incorporate these factors to provide a more comprehensive assessment of extinction risks. Our findings suggest that climate change, in conjunction with habitat degradation and human activities, must be considered when assessing the extinction risks of Testudoformes.

Adaptation of skin structures to environmental variations in anurans from southern and southwestern China.

Variations in skin structures can possibly reflect local adaptation to distinct environmental factors. As the primary interface with the surrounding environment, amphibian skin undergoes phenotypic innovations that play a key role in protection, water absorption, and respiration. However, the effects of environmental factors on skin structures have been examined in only a limited number of species. Here, we conducted a comparative analysis of the skin structures of 102 Chinese anuran species across varying geographical distributions and habitat types. Our results revealed that the total volume of granular glands and capillary density in the dorsal skin significantly increased with increasing latitude. We also found that the thickness of calcified layers in both dorsal and ventral skin was positively correlated with annual temperature and negatively correlated with humidity. Additionally, terrestrial species exhibited the largest dorsal granular gland, whereas arboreal species had the smallest one. Likewise, the largest dorsal mucous gland was observed in aquatic species, while the smallest was found in terrestrial species. These results highlighted the importance of understanding the relationship between skin phenotypes and environmental variables and thus providing conservation strategies based on the evolutionary adaptations in anurans. Our study can contribute to the broader knowledge of evolutionary biology in anurans by demonstrating how specific skin traits are linked to survival and fitness across various ecological contexts.

Assessing Reptile Conservation Status under Global Climate Change.

Global climate change drives variations in species distribution patterns and affects biodiversity, potentially increasing the risk of species extinction. Investigating the potential distribution range of species under future global climate change is crucial for biodiversity conservation and ecosystem management. In this study, we collected distributional data for 5282 reptile species to assess their conservation status based on distributional ranges using species distribution models. Our predictions indicate that the potential distribution ranges for over half of these species are projected to decrease under different scenarios. Under future scenarios with relatively low carbon emissions, the increase in the number of threatened reptiles is significantly lower, highlighting the importance of human efforts. Surprisingly, we identified some endangered species that are projected to expand their distribution ranges, underscoring the potential positive effects of climate change on some special species. Our findings emphasize the increased extinction risk faced by reptile species due to climate change and highlight the urgent need to mitigate the effects of habitat degradation and human activities on their potential distribution in the future.

Sexual Dimorphism in the Limb Bones of Asiatic Toad (Bufo gargarizans) in Relation to Sexual Selection.

Sexual dimorphism is often considered to be the result of differences in the intensity of sexual selection between sexes. From this point of view, the sexual dimorphism of the limb bones of the Bufo gargarizans in southwest China was studied. Results showed that the fore- and hindlimb skeletons of this species were sexually dimorphic in anatomy. The humerus, radioulna, and total lengths of the forelimb skeleton of males were substantially longer than those of females, but the hand length of males was smaller than that of females. Several other features of males, such as deltoid and medial crest areas and humerus and radioulnar weights, were also significantly larger than those of females. The femoris, tibiofibula, talus-calcaneus, and foot lengths; total hindlimb skeleton length; and femoral upper crest areas of males were significantly greater than those of females. However, no significant intersexual difference in femoris and tibiofibular weights was observed. These findings suggested that robust forelimb bones and long hindlimb bones could contribute to the mating success of males; if so, sexual selection promotes the evolution of sexual size and shape dimorphism in the limb bones of the B. gargarizans.

Geographic Variation in Organ Size in a Toad (Duttaphrynus melanostictus).

Adaptive evolution is the process by which organisms change their morphological, physiological and biochemical characteristics to adapt to different environments during long-term natural selection. Especially, researching variation in organ size can provide important insights into morphological adaptation in amphibians. In this study, we comparatively studied differences in organ sizes (heart, lungs, liver, gallbladder, kidneys, spleen, digestive tract, testes and brain) among five geographical populations of the Asian common toad Duttaphrynus melanostictus. Our results revealed significant variations in the size of these nine specific organs among the populations. Notably, we observed a significant positive correlation between the relative size of the testes and latitude and/or altitude. However, no correlation was found between the relative size of the heart and the length of the digestive tract with altitude across populations, respectively, contradicting Hesse's rule and the digestion theory. These findings suggest that our study does not provide substantial theoretical support for the adaptive evolution of organ size in this particular toad species, but rather contributes to the understanding of the evolution and adaptations of species' different environmental conditions. Further research is warranted to delve deeper into the factors influencing organ size in amphibian populations.

Geographical Variation in Body Size in the Asian Common Toad (Duttaphrynus melanostictus).

The geographic variation in life-history traits of organisms and the mechanisms underlying adaptation are interesting ideas in evolutionary biology. This study investigated age and body size of the Asian common toad (Duttaphrynus melanostictus) among five populations along a geographical gradient. We found that geographical variation in age was non-significant among populations but there was a significant and positive correlation between mean age and body size. Although the body size values at 1043 m are quite different from other sites, after controlling for age effects, there was a significant positive correlation between altitude and body size. Our findings followed the predictions of Bergmann's rule, suggesting that the body size of D. melanostictus is potentially influenced by the low air temperatures at higher altitudes.

Predicting the Potential Distribution of the Szechwan Rat Snake (Euprepiophis perlacea) and Its Response to Climate Change in the Yingjing Area of the Giant Panda National Park.

Climate change is a significant driver of changes in the distribution patterns of species and poses a threat to biodiversity, potentially resulting in species extinctions. Investigating the potential distribution of rare and endangered species is crucial for understanding their responses to climate change and for the conservation of biodiversity and ecosystem management. The Szechwan rat snake (Euprepiophis perlacea) is an endemic and endangered species co-distributed with giant pandas, and studying its potential distribution contributes to a better understanding of the distribution pattern of endangered species. In this study, we confirmed seven presence points of this species in the Yingjing Area of the Giant Panda National Park, and selected eleven key factors to predict the potential distribution of E. perlacea under current and future scenarios using MaxEnt models. Our study consistently achieved AUC values exceeding 0.79, meeting the precision requirements of the models. The results indicated that the high potential distribution area of E. perlacea is mainly located near Yunwu mountain and the giant panda rewilding and reintroduction base, accounting for approximately 12% of the protected area. Moreover, we identified the primary environmental factors influencing the distribution of E. perlacea as the distance from streams and the slope degree, with their contribution rates exceeding 41% and 31%, respectively. In comparison to the current scenario, the potential habitat range for E. perlacea did not show an overall reduction in the context of future climate scenarios. To ensure the long-term preservation of E. perlacea, it is advisable to validate its actual distribution based on the models' results. Particular attention should be given to safeguarding its core distribution areas and raising awareness among residents within the potential distribution range about the conservation of E. perlacea.

Protein Inhibitor of Activated STAT Y (PIASy) Regulates Insulin Secretion by Interacting with LIM Homeodomain Transcription Factor Isl1.

It is known that the LIM homeodomain transcription factor Isl1 is highly expressed in all pancreatic endocrine cells and functions in regulating pancreatic development and insulin secretion. The Isl1 mutation has been found to be associated with type 2 diabetes, but the mechanism responsible for Isl1 regulation of insulin synthesis and secretion still needs to be elucidated. In the present study, the protein inhibitor of activated STAT Y (PIASy) was identified as a novel Isl1-interacting protein with a yeast two-hybrid system, and its interaction with Isl1 was further confirmed by a co-immunoprecipitation experiment. PIASy and Isl1 colocalize in human and mouse pancreas and NIT beta cells. Furthermore, PIASy and Isl1 upregulate insulin gene expression and insulin secretion in a dose-dependent manner by activating the insulin promoter. PIASy and Isl1 mRNA expression levels were also increased in type 2 diabetic db/db mice. In addition, our results demonstrate that PIASy and Isl1 cooperate to activate the insulin promoter through the Isl1 homeodomain and PIASy ring domain. These data suggest that that PIASy regulates insulin synthesis and secretion by interacting with Isl1 and provide new insight into insulin regulation, although the detailed molecular mechanism needs to be clarified in future studies.