Two new Paramecium strains were isolated from an artificial pond on a university campus located in Tokyo, Japan. Morphological observations and molecular phylogenetic analyses based on partial sequences of heat shock protein 70 and cytochrome b gene sequences identified both strains as Paramecium pentaurelia , one of the sibling species within the Paramecium aurelia species complex. This represents the first confirmed record of P . pentaurelia in Japan. Although this species has rarely been reported on a worldwide basis, the phylogenetic analyses showed that the Japanese strains shared identical or nearly identical sequences with multiple P . pentaurelia strains from geographically distant regions, suggesting that this species may have a wider, possibly cosmopolitan, distribution than has currently been recognised. The limited number of records may reflect the infrequent occurrence of this species in environmental samples, which may hinder its detection.
Exogenous bacterial species, which mainly populate the intestines of Helix pomatia snails, also affect various biological processes that are crucial to the survival of the host. However, the microbiota undergoes changes during overwintering, as feeding is suspended, leading to serious health consequences for the host. This study aims to investigate whether the food supplementation of snails with a microbiological inoculation containing bacterial strains isolated from the intestines of overwintering snails could improve their cold tolerance in summer. The experiment was conducted on two groups: a control group without supplementation; and an experimental group given the microbiological inoculation. The supercooling point (SCP) of their body fluids was measured to assess the effects of supplementation on the cold tolerance of wild snails in the summer. The results showed that the SCP for the snails in the experimental group was statistically significantly lower than for the snails in the control group, with a mean SCP value of −1.73°C compared to −0.97°C (p < 0.05). These findings confirm that the microbiota has a significant impact on cold tolerance in snails, and the microbiological inoculation had a positive impact on lowering the supercooling temperature of H . pomatia in the summer. This sheds new light on the mechanism of this species’ adaptation to cold, which still remains a mystery.
Land-use changes are a major driver of biodiversity shifts in Southeast Asia, particularly within human modified landscapes. Understanding how migratory species use habitats during the non-breeding season is important for assessing their tolerance to environmental changes. We examined the non-breeding habitat associations of the Brown Shrike ( Lanius cristatus Linnaeus, 1758) in Laos by analysing 139 field-detected occurrence sites and comparing them with an equal number of randomly selected background points. Three landscape variables were most strongly associated with the detection of shrikes: road area (positive relationship), and the extent of built-up areas and forest cover (both negative relationships). Brown Shrikes were most frequently recorded in semi-open environments within agricultural mosaics, where structural elements such as perches and open foraging spaces were maintained. Our findings suggest that modified landscapes can provide suitable conditions for the species when structural habitat elements are retained. Our study provides the first fine-scale assessment of the non-breeding habitat associations of Brown Shrikes in mainland Southeast Asia and highlights the importance of maintaining mosaic agricultural landscapes for migratory insectivorous birds.
The Alps are recognised as a hotspot of biodiversity and have historically been extensively studied. However, despite the number of studies devoted to the biodiversity of the Alps, large knowledge gaps are still present. The Southwestern Alps are a biodiverse region known for hosting many endemic animals and plants. To increase the faunistic knowledge of the Southwestern Alps, we have produced the first survey of the tardigrades from Monviso. Notably, we sequenced the mitogenome of the rarely found Arctic-alpine Acanthechiniscus victor (Ehrenberg, 1853), which is an important addition to the existing knowledge as it is the second mitogenome to be sequenced for the tardigrade superfamily Echiniscoidea. The findings of this study highlight how continued explorations of the European mountain ecosystem are still needed to fill in the knowledge gaps regarding its biodiversity.
The alpine bullhead (Alpinocottus poecilopus) is a cold-water benthic fish with a disjunct distribution across Europe, including a recently discovered population in northern Poland's Pomeranian region. Prior to now, the taxonomic identity of this enclave population has not been confirmed using molecular methods. This study presents the first DNA barcoding analysis of the A. poecilopus from Pomerania. Sequences of the mitochondrial coI gene and phylogenetic reconstruction confirmed the species identity and revealed no, or a very low genetic divergence between the Pomeranian and the Scandinavian and Czech Republic A. poecilopus. The results also support the recent reclassification of A. poecilopus into the genus Alpinocottus, alongside A. volki and A. szanaga, forming a distinct clade with C. kolymensis, which may also belong to this genus. These findings confirm that the Pomeranian alpine bullhead represents a genetically valid relict population, which was likely preserved due to postglacial isolation and specific cold-water habitat conditions. Additionally, unverified reports of an alpine bullhead occurrence in the Drweca River basin (Vistula system) were clarified, as specimens from this site were identified as the European bullhead (Cottus gobio).
Obtaining population demography metrics in a non-lethal way is essential for the evidence-based management of reptiles that are declining in increasingly degraded European landscapes. Here, we quantified life-history traits of the sand lizard Lacerta agilis argus in Western Poland and tested whether they varied by sex and locality. Using skeletochronology, we aged 177 individuals from three populations inhabiting different habitats. The lizards reached sexual maturity after their second hibernation; and the mean adult age was 4.6 f 1.9 years (range 2-8). The annual survival rate was 0.80 and the adult life expectancy was 5.25 years. The Sexual Dimorphism Index (SDI) was female-biased but small (SDI = 0.04). Linear modelling showed that the snout-vent length increased with age, yet the age-size slope differed between sexes and among populations: females and lizards from the two warmer, drier sites grew faster and attained a larger body size than males or lizards from the cooler forest site. Sex had no main effect on the size once age was accounted for. These results demonstrate that even over short geographic distances, habitat thermal conditions modulate the growth trajectories in this ecologically flexible species. Our study highlights the value of skeletochronology for rapid, non-lethal assessments of the population demography in temperate lizards.
Repeat surveys of the shell colour and banding polymorphism in populations of the land snail Cepaea nemoralis in the cities of Sheffield (UK) and Wroclaw (Poland) and in its surrounding settlements were made after a 13-18 years interval. In all three cases, a substantial proportion of sites had been destroyed by development (18-30%) and a similar proportion no longer held the species, despite being apparently suitable; up to 50% of populations had disappeared. Overall, the pattern was one of stasis in morph-frequencies, and of little change in the range of variation as estimated by a modified version of Fst. There were, however, substantial changes in some populations, not consistent in direction, but larger than expected from sampling error. While there was tenuous evidence for selection against yellow shells, the large changes are attributed to bottlenecks, founder effects, and possible local extinction and recolonisation. Urban populations are often transitory and the results are contrasted with those from an area of long-term habitat stability and population connectivity. Newly established and sometimes transitory populations offer opportunities to study underestimated aspects of evolutionary processes.
With a dismal prognosis, cholangiocarcinoma (CCA) is a highly invasive cancer and its global incidence is increasing. Non-coding RNAs, particularly circRNAs, are increasingly recognized as important regulators in tumorigenesis, yet the mechanistic details of hsa_circ_0000977-mediated miR-338-3p regulation in CCA remain incompletely understood. This study aimed to investigate the mechanism of the hsa_circ_0000977/miR-338-3p/ETS1 axis in CCA progression, evaluate its prognostic significance and investigate its functional role in CCA cells. Real-time quantitative PCR (RT-qPCR) was performed to quantify hsa_circ_0000977, miR-338-3p and ETS1 mRNA expression levels. Pearson correlation was used to assess their association. To confirm the molecular interactions between these molecules, dual-luciferase reporter assays were employed. Cell migration capacity was evaluated via Transwell migration assays, while CCK-8 tests analysed proliferation. Prognostic value was assessed through survival analysis and multi-variate regression. The study revealed significant up-regulation of hsa_circ_0000977 in CCA tumour tissues, with elevated expression levels associated with poorer 5-year survival outcomes. Multivariate analysis confirmed that hsa_circ_0000977 over-expression is an independent predictor of survival. Functional assays indicated that hsa_circ_0000977 negatively regulated miR-338-3p, which was down-regulated in CCA and exhibited tumour-suppressive effects. CCA cell proliferation and migration were suppressed following hsa_circ_0000977 knockdown, effects that were partially reversed by miR-338-3p inhibition. Further investigation demonstrated that miR-338-3p exerts its tumour-suppressive effects by directly targeting ETS1, and the hsa_circ_0000977/miR-338-3p/ETS1 axis regulates the proliferation and migration of CCA cells. We have concluded that hsa_circ_0000977 drives CCA progression by sponging miR-338-3p and modulating its target ETS1, suggesting its clinical value for predicting outcomes and developing targeted therapies.
Long noncoding RNAs (lncRNAs) are known to play critical roles in the progression of osteosarcoma. Despite their recognized importance, the specific biological functions of lncRNAs in osteosarcoma remain unclear. In this context, prostate cancer-associated transcript 7 (PCAT7) has been identified as a bone metastasis-related lncRNA through the analysis of The Cancer Genome Atlas dataset. In this study, we investigated the expression of PCAT7 in osteosarcoma cells, particularly those exhibiting resistance to doxorubicin, a widely used chemotherapeutic agent in clinic. Functional assays including cell growth, invasion and apoptosis were conducted to elucidate the impact of PCAT7 inhibition on osteosarcoma cells, focusing on sensitivity to doxorubicin treatment. To understand the underlying molecular mechanisms, the interaction between PCAT7, miR-324-5p, and the TGF-β/SMAD signalling pathway was further explored. The study revealed that PCAT7 is up-regulated in osteosarcoma cells with doxorubicin resistance. Inhibition of PCAT7 could enhance the sensitivity to doxorubicin treatment by reducing cell growth, suppressing cell invasion and increasing cell apoptosis. Mechanistically, PCAT7 was shown to activate the TGF-β/SMAD signalling pathway by up-regulating the expression of TGFBR1 through sponging miR-324-5p. These findings unveil a novel mechanism contributing to the constitutive activation of TGF-β signalling in osteosarcoma. Targeting PCAT7 may offer a promising avenue for therapeutic interventions in osteosarcoma by disrupting the aberrant TGF-β signalling, thus presenting a potential strategy to improve treatment outcomes in this challenging cancer.
Nerol (NRL), a monoterpene in essential oils of various plants, exhibits anticancer activity, albeit with contrasting effects on normal cells. Because anaemia is induced by anticancer drugs, this study was initiated to profile the cytotoxic mechanisms of NRL in human erythrocytes. Cells were treated with NRL (0.01-0.1 %) for 24 h at 37 °C in Ringer buffers. Lytic cell death was measured photometrically and eryptosis was examined by flow cytometry using forward scatter, annexin V-affinity assay, H2DCFDA, and Fluo4/AM to evaluate cell volume, phosphatidylserine (PS) externalization, oxidative stress, and Ca2+, respectively. Small molecule inhibitors were used to probe the molecular mechanisms governing NRL-induced cytotoxicity. NRL led to eryptotic volume loss associa-ted with increased PS externalization and lysis through oxidative stress and Ca2+ nucleation. Ca2+ deprivation and K+ gradient dissipation, along with glucose, guanosine and sucrose, significantly blunted NRL-induced eryptosis and lytic death. Notably, both forms of cell death were inhibited by staurosporine, necrostatin 2 and myriocin, whereas urea, Z-VAD-FMK, SB205830 and D4476 only attenuated eryptosis. Furthermore, while lytic death was inhibited by polyethylene glycol, it was potentiated by Ca2+ deprivation and heparin, whereas eryptosis was aggravated by uric acid. In conclusion, NRL triggers cation channel- and redox-mediated eryptosis and lytic death through energy deprivation and activation of protein kinase C, receptor-interacting protein 1, serine palmitoyltransferase, caspase, p38 MAPK and casein kinase 1α. Altogether, these findings underscore the differential mechanisms by which NRL modulates divergent erythrocyte injury pathways and collectively advance the current knowledge of the extent of its cellular effects.
Cardiovascular diseases due to atherosclerosis remain a dominant medical problem. Macro-phages play a crucial role in both atherosclerosis progression and recycling of body iron. Epidemio-logical data point to elevation of body iron stores as one of non-classical cardiovascular risk factors, and we know that iron must be contained within macrophages to be atherogenic. Presumably, iron already contained within circulating monocytes turns cells to a more pro-inflammatory and hence atherogenic phenotype, but experimental evidence for such relationship remains limited. In this study, human monocytic THP-1 cells were loaded with micromolar iron in the forms of transferrin, ferric-ammonium citrate (FAC) and haemin for 2 and 24 hours. Only haemin was cytotoxic. All kinds of iron elevated the labile iron pool at 2 hours, as well as ferritin expression at 24 hours. Expression of scavenger receptors A and B, pro-inflammatory as well as anti-inflammatory cytokines, haem oxygenase and haptoglobin receptor were measured by quantitative PCR. Very few changes, none pro-inflammatory, were observed in response to transferrin or FAC. Haemin suppressed the expression of scavenger receptors, increased the expression of pro-inflammatory, and variably, anti-inflammatory cytokines, and dramatically induced haem oxygenase. The effects of haemin were not prevented by apotransferrin. We conclude that iron must be in the haemin form to act as a pro-inflammatory stimulus in THP-1 monocytes. Non-haem iron might require the presence of other factors to be atherogenic. Haemin treatment of THP-1 cells may represent a convenient experimental model to study the pro-inflammatory effects of haem that are observed in late stages of atherosclerosis.
Aphids of the genus Stomaphis are among the largest aphids, living on tree trunks in the Palaearctic, always with an obligate mutualistic relationship with ants. Some species are very common and abundant, but a few of them are very rare and known only from single sites. Stomaphis radicicola was described in 1947 and since its description, it has been found only three times prior to 2024, when an abundant population was discovered in Germany, close to the UNESCO biosphere reserve Upper Lusatia Heath and Pond Landscape. Detailed studies on the species' biology and modelling of its potential past, current and future distribution indicate its relatively narrow habitat specialisation towards riparian forests, with Alnus glutinosa as a predominant host plant. With such a level of specialisation, its distribution during the Last Glacial Maximum was very limited, and the current rarity across Europe may result from a bottleneck effect. The species' future range may shift eastwards under climate change, but it will depend on finding riparian habitats and host ant species.
The biodiversity of protists, which are key players in many ecosystems, remains understudied, particularly in tropical regions. The ciliate Paramecium sexaurelia, a cryptic species within the Paramecium aurelia complex, is typically restricted to warm climates. In this study, we examine the genetic variability of P. sexaurelia populations collected over three years (2016-2018) from water bodies in the palm houses of the Jagiellonian University Botanical Garden in Krakow. These artificial palm-house environments, which contain tropical plants, may serve as reservoirs for microbial eukaryotes native to warm climates, thereby providing a unique opportunity to study protist diversity outside their native regions. Our molecular analysis revealed a considerable amount of genetic diversity within these populations, as we detected 13 distinct COI haplotypes (Pa6COI_02, 07, 14-24). While two haplotypes (Pa6COI_02 and 07) matched previously known sequences, the remaining eleven haplotypes (Pa6COI_14-24) were novel to this study, demonstrating the unexplored genetic richness of P. sexaurelia, even in artificial habitats. Given the high genetic diversity and widespread distribution of the species, these results provide valuable insights into its population structure in controlled environments. The presence of P. sexaurelia in atemperate-climate palm house suggests possible plant-mediated introductions, raising intriguing questions about the dispersal and persistence of tropical protists beyond their native ranges. These findings highlight the often-overlooked role of botanical gardens in preserving microbial and eukaryotic diversity, while underscoring the value of such artificial habitats as natural laboratories for studying the biodiversity of tropical protists in non-native settings.
The aim of the study was to assess the effectiveness of a microsatellite DNA analysis to genetically identify dog breeds and crossbreeds, using reference populations of purebred individuals and Bayesian clustering methods. The study was conducted based on 21 microsatellite markers (STR) that are recommended by the International Society for Animal Genetics (ISAG) for routine canine pedigree verification. The genetic diversity and population structure were assessed for 4 selected breeds: Golden Retriever, Bernese Mountain Dog, Poodle and Chinese Crested Dog. These reference populations were then used to analyse two case studies involving breed verification. Key genetic parameters were calculated, including the observed (H-O) and expected heterozygosity (H-E), degree of inbreeding (F-IS) and the polymorphism information content (PIC). The probability of exclusion was estimated in cases of knowing the genotype of one of the parents (CPE1) and both parents (CPE2). A relatively high level of genetic diversity among the studied breeds, constituting reference populations, was found to be above 50% for H-O and PIC, with no inbreeding. The probability of CPE1 and CPE2 was obtained at the level of 99% and 99.99%, respectively, which allowed the use of these markers for a parentage analysis. Bayesian clustering (STRUCTURE) and a Principal Coordinates Analysis (PCoA) were applied to assess the genetic structure and to identify the admixture in genotypes of the crossbred individuals. In both cases, the analyses successfully confirmed the breed's pedigree or detected mixed ancestry, which was further supported by the parentage testing. The results confirm that the applied STR marker panel is suitable for breed identification and the detection of hybrid ancestries in dogs. While further research is needed to validate the method across a wider range of breeds, the study provides a valuable foundation for the development of DNA-based tests to support breed verification and the assessment of breed compositions in designer and hybrid dogs.
The aim of this study was to evaluate the seasonal diet composition and feeding strategies of northern pike (Esox lucius) located in S & imath;dd & imath;kl & imath; K & uuml;& ccedil;& uuml;kbo & gbreve;az Dam Lake (K & imath;r & scedil;ehir, T & uuml;rkiye). Monthly sampling from September 2015 to August 2016 resulted in the collection of 133 northern pike individuals, with their total lengths ranging from 23.4 to 86.6 cm (mean length 53.84 +/- 10.51 cm). Stomach content analyses and statistical tests, including a Kruskal-Wallis analysis, were employed to assess the seasonal variation in feeding intensity. Although numerical differences in the feeding intensity were observed, with the highest values recorded in summer (FI = 1.59) and the lowest in winter (FI = 0.51), these differences were not statistically significant (K-W test, p < 0.05). Among the physicochemical parameters of the surface water, temperature emerged as the primary environmental factor influencing the feeding patterns, based on the results of a Spearman's rank correlation. The northern pike exhibited an exclusively piscivorous diet, primarily consuming Tinca tinca (44.83% by number and 56.16% by frequency of occurrence) as well as Atherina boyeri. Despite the availability of multiple prey species within the habitat, the northern pike demonstrated a strong prey specialisation toward these two fish species throughout the year. The observed consistency in this dietary preference underscores the species' role as a specialised top predator and provides critical ecological insights for the management and conservation of similar freshwater reservoir ecosystems.
In recent years, G protein-coupled oestrogen receptor 1 (GPER1) has been revealed as an important mediator of oestrogen's action on the reproductive processes in mammals; however, its role in the avian reproductive system is not known. In the present study, it was examined whether GPER1 is present in the hen's reproductive system and if so, its mRNA and protein distribution in the ovary and oviduct. Immunolocalisation of GPER1 in the ovarian follicles and all oviductal parts (i.e.infundibulum, magnum, isthmus, shell gland and vagina) was also investigated. The GPER1 mRNA transcript and protein were detected in all of the examined tissues by real-time PCR and Western Blot analyses, respectively. The relative expression of GPER1 depended on the follicle development and the follicular wall layer (p < 0.001). It was highest in the white and yellowish follicles and lowest in the granulosa layer of F3-F1 follicles. Immunoreactivity for the GPER1 protein was most prominent in the ovarian stroma with primordial and primary follicles, white follicles and yellowish follicles. Within the oviduct, the highest level of the GPER1 transcript was in the infundibulum, while the lowest was in the magnum (p < 0.001). Moreover, protein abundance was higher (p < 0.05) in the shell gland than in the vagina. The strongest immunopositive reaction was observed in the shell gland and vagina, especially in the luminal epithelium. The obtained preliminary results indicate that the oestrogen action in the reproductive systems of female birds also involves GPER1 signalling.
Blood parameters are widely used to evaluate the condition of birds. Typically, only a few parameters are used, such as the haemoglobin concentration, haematocrit, or the erythrocyte and leukocyte counts. The aim of this study was to present reference values of the haematological parameters of a free-living population of red-backed shrike ( Lanius collurio ) nestlings in east-central Poland. We assessed various blood parameters (haemoglobin concentration, erythrocyte and leukocyte, on blood smears for the erythrogram, leukogram and thrombocyte count), and examined their relationships with the brood size, biometrics conditions and hatching date. To reduce dimensionality and address the potential collinearity among variables, we conducted a Principal Component Analysis (PCA) and used the resulting components in Generalised Linear Mixed Models. None of the models yielded statistically significant effects. These results suggest a relatively stable physiological status among these nestlings, likely supported by a food-rich agricultural landscape. The haematological profiles presented here offer valuable baseline data for assessing the health of red-backed shrike populations and may support future comparative studies in wild birds.
A new genus Afrocranus (Delphacidae Leach, 1815, Stenocraninae Wagner, 1963) has been established for two species: Afrocranus ujotdensis Gębicki, Walczak & Świerczewski from continental Africa (Namibia); and Afrocranus mixtus Walczak, Gębicki & Świerczewski from Madagascar. A set of distinguishing characteristics is provided to differentiate this new taxon from the closely related Afrotropical genus Embolophora Stål, 1853, as well as the Neotropical species Tanycranus elongatus Bartlett, 2010 and other representatives of the subfamily Stenocraninae. Additionally, a DNA analysis was performed, and a phylogram illustrating the relationships among selected Stenocraninae planthopper species is presented.
The karyotypes of felids are considered to be very conservative. Comparative chromosome painting is a technique used to determine the homology of regions between the chromosomes of different species. In order to supplement the existing knowledge and to obtain information on large-scale genome structure variations within the Felidae family, the fluorescence in situ hybridisation technique was performed in this study. Molecular probes specific to the cat chromosome A1, B1, C1, X and Y were used. The probes were hybridised with chromosomes belonging to the Amur tiger (♀), African lion (♂) and the leopard (♀). No significant differences were observed in the morphology or the banding pattern of both autosomes and heterosomes, and the homology of the tested DNA of fragments was demonstrated.
The two land snail species Trochulus striolatus (C. Pfeiffer, 1828) and Xerocampylaea erjaveci (Brusina, 1870) are distantly-related within the Hygromiidae, and while geographically separated, have similar habitats. They are conchologically very similar in their shape. The growth, reproductive strategies and life histories of these species reared under constant laboratory conditions were studied in order to detect any similarities and differences between the species. In both species, isolated individuals did not reproduce; thus, uniparental reproduction appears not to occur. While morphological characteristics, such as the egg size and number of whorls at hatching and at sexual maturity, were relatively constant and similar in both species, there were differences in terms of the growth rate, age at first reproduction, output of eggs over the lifespan and in the pattern of mortality. While the range of variation within species was much greater in relation to these traits, it appears that X. erjaveci grows faster than T. striolatus : they become adults earlier; they have a shorter lifespan as adults, but less juvenile mortality; and individuals lay many more clutches, over a similar time span, laying more than five times as many eggs as T. striolatus over the course of the experiment. Under these admittedly artificial conditions, the individuals of both species (although more so in X. erjaveci ) can lay more than one clutch over a period of more than two years, in an environment where no winter conditions were imposed. These results, which can also be compared with similar data for Trochulus hispidus , a species closely-related to T. striolatus, emphasise the fact that life histories, even among species of a similar size and habitat, may differ considerably. They also illustrate the difficulties inherent in laboratory studies of life histories.
