Publications

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comprehensive analysis of formin localization in xenopus epithelial cells.reorganization of the actin cytoskeleton is crucial for cellular processes, including cytokinesis and cell-cell junction remodeling. formins are conserved processive actin-polymerizing machines that regulate actin dynamics by nucleating, elongating, and bundling linear actin filaments. because the formin family is large, with at least 15 members in vertebrates, there have not been any comprehensive studies examining formin localization and function within a common cell type. here, we characteriz ...201930379611
anticipation of food intake induces phosphorylation switch to regulate basolateral amino acid transporter lat4 (slc43a2) function.amino acid absorption requires luminal uptake into and subsequent basolateral efflux out of epithelial cells, with the latter step being critical to regulate the intracellular concentration of the amino acids. the basolateral essential neutral amino acid uniporter lat4 (slc43a2) has been suggested to drive the net efflux of non-essential and cationic amino acids via parallel amino acid antiporters by recycling some of their substrates; its deletion has been shown to cause defective postnatal gro ...201930379325
evaluating potential distribution of high-risk aquatic invasive species in the water garden and aquarium trade at a global scale based on current established populations.aquatic non-native invasive species are commonly traded in the worldwide water garden and aquarium markets, and some of these species pose major threats to the economy, the environment, and human health. understanding the potential suitable habitat for these species at a global scale and at regional scales can inform risk assessments and predict future potential establishment. typically, global habitat suitability models are fit for freshwater species with only climate variables, which provides ...201930428498
copper uptake mechanism of arabidopsis thaliana high-affinity copt transporters.copper (cu) is an essential plant micronutrient. under scarcity, cu2+ is reduced to cu+ and taken up through specific high-affinity transporters (copts). in arabidopsis, the copt family consists of six members, either located at the plasma membrane (copt1, copt2, and copt6) or in internal membranes (copt3 and copt5). cu uptake by copt proteins has been mainly assessed through complementation studies in corresponding yeast mutants, but the mechanism of this transport has not been elucidated. to t ...201930043153
following endocrine-disrupting effects on gene expression in xenopus laevis.endocrine-disrupting chemicals (edcs), found in all categories of chemicals, are suspected to be a cause of declining well-being and human health, both as single molecules and as mixtures. it is therefore necessary to develop high throughput methods to assess the endocrine-disrupting potential of multiple chemicals currently on the market that are as yet untested. an advantage of in vivo chemical screening is that it provides a full spectrum of physiological impacts exerted by a given chemical. ...201930042138
assessing antibody responses to pathogens or model antigens in xenopus by enzyme-linked immunosorbent assay (elisa).xenopus laevis-specific monoclonal antibodies recognize igm and igy antibodies not only from x. laevis but also x. tropicalis as well as a variety of amphibian species including ranidae, bufonidae, and even some salamanders. these reagents are very useful to assess antibody responses from the serum or other animal secretions (e.g., peritoneal fluid). we present here an enzyme-linked immunosorbent assay (elisa) optimized for amphibians that permits users to detect and titrate the presence of each ...201930042135
expression of the hormonal fgf co-receptor klotho beta in the xenopus laevis model.klotho beta (klb), a single-pass transmembrane protein, has been described as a co-receptor for endocrine fgfs, such as fgf15/19 and fgf21, to regulate critical metabolic processes in multiple organs and tissues in adult mice. however, its function during early embryonic development remains largely unknown. in this paper, we evaluated for the first time the expression of klb mrna during early development of xenopus laevis by rt-pcr and whole mount in situ hybridization. rt-pcr experiments showed ...201930259590
endoplasmic reticulum network formation with xenopus egg extracts.the endoplasmic reticulum (er) consists of morphologically distinct domains, including a polygonal network of tubules that is connected by three-way junctions. this network is found in all eukaryotic cells. extracts from xenopus laevis eggs contain stockpiles of components that allow the assembly of an er network in vitro. here we provide protocols for assembly of er networks in extracts that are arrested at different stages of the cell cycle. unfertilized xenopus laevis eggs contain a cytostati ...201929475993
studies of limb regeneration in larval xenopus.a basic protocol is given for animal maintenance and surgery in studies of hindlimb regeneration in larval xenopus laevis unlike urodele limbs, those of larval frogs typically show much more variation in the extent of regeneration after amputation. such variation can be reduced by optimizing the conditions of larval maintenance to regulate the rates of growth and development, by selecting only larvae with normal rates of growth and morphological development for experimental use, and by attention ...201929895566
investigating the cellular and molecular mechanisms of wound healing in xenopus oocytes and embryos.the african clawed frog xenopus has remarkable capacities to heal wounds rapidly and to regenerate complex tissues. because of its experimental tractability, studies using xenopus oocytes, embryos, and larvae have contributed extensively to our understanding of the molecular and cellular mechanisms underpinning wound healing and tissue regeneration. in this protocol, we describe wound-healing assays following mechanical or laser injuries of oocytes and multicellular epithelia in xenopus laevis e ...201929895564
ex vivo eye tissue culture methods for xenopus.lens regeneration can be studied in whole animals following removal of the original lens (lentectomy). however, culturing a whole animal can be impractical for assays involving small molecule inhibitors or proteins. ex vivo eye tissue culture is an alternative approach for examining lens regeneration. the ex vivo culture system offers certain advantages when compared to the in vivo regeneration assay, as the percentage of cases showing lens differentiation can exceed that seen in whole animals. ...201929895561
chromatin characterization in xenopus laevis cell-free egg extracts and embryos.xenopus laevis development is marked by accelerated cell division solely supported by the proteins maternally deposited in the egg. oocytes mature to eggs with concomitant transcriptional silencing. the unique maternal chromatin state contributing to this silencing and subsequent zygotic activation is likely established by histone posttranslational modifications and histone variants. therefore, tools for understanding the nature and function of maternal and embryonic histones are essential to de ...201929475998
the role of ambient temperature and body mass on body temperature, standard metabolic rate and evaporative water loss in southern african anurans of different habitat specialisation.temperature and water availability are two of the most important variables affecting all aspects of an anuran's key physiological processes such as body temperature (tb), evaporative water loss (ewl) and standard metabolic rate (smr). since anurans display pronounced sexual dimorphism, evidence suggests that these processes are further influenced by other factors such as vapour pressure deficit (vpd), sex and body mass (mb). however, a limited number of studies have tested the generality of thes ...201931660269
environmental oxygen exposure allows for the evolution of interdigital cell death in limb patterning.amphibians form fingers without webbing by differential growth between digital and interdigital regions. amniotes, however, employ interdigital cell death (icd), an additional mechanism that contributes to a greater variation of limb shapes. here, we investigate the role of environmental oxygen in the evolution of icd in tetrapods. while cell death is restricted to the limb margin in amphibians with aquatic tadpoles, eleutherodactylus coqui, a frog with terrestrial-direct-developing eggs, has ce ...201931204171
transcriptome profiling reveals male- and female-specific gene expression pattern and novel gene candidates for the control of sex determination and gonad development in xenopus laevis.xenopus laevis is an amphibian (frog) species widely used in developmental biology and genetics. to unravel the molecular machinery regulating sex differentiation of xenopus gonads, we analyzed for the first time the transcriptome of developing amphibian gonads covering sex determination period. we applied microarray at four developmental stages: (i) nf50 (undifferentiated gonad during sex determination), (ii) nf53 (the onset of sexual differentiation of the gonads), (iii) nf56 (sexual different ...201930972573
expression of tcr genes in adult and larval xenopus laevis.in order to better understand the development and function of γδ t cells in xenopus frogs, it is necessary to determine where and when γδ t cells are found in xenopus tissues. this study examined the expression of tcr genes, focused primarily on tcr γ, in tissues of adult and larval xenopus laevis and provide new data about the expression pattern of these different tcr genes in this anuran amphibian. tcr gene expression was detected by rt-pcr in adult frog tissues including the thymus, spleen, s ...201930738793
preventing ethanol-induced brain and eye morphology defects using optogenetics.background: embryonic exposure to the teratogen ethanol leads to dysmorphias, including eye and brain morphology defects associated with fetal alcohol spectrum disorder (fasd). exposure of xenopus laevis embryos to ethanol leads to similar developmental defects, including brain and eye dysmorphism, confirming our work and the work of others showing xenopus as a useful system for studies of the brain and eye birth defects associated with fasd. several targets of ethanol action have been hypothesi ...201932685918
loc496300 is expressed in the endoderm of developing xenopus laevis embryos. 201932550462
using hdr and a template to introduce an in-frame ha tag on the 3' end of the xenopus laevis gata2.l open reading frame. 201932550411
a novel calcium-activated potassium channel controls membrane potential and intracellular ph in trypanosoma cruzi.trypanosoma cruzi develops in environments where nutrient availability, osmolarity, ionic concentrations, and ph undergo significant changes. the ability to adapt and respond to such conditions determines the survival and successful transmission of t. cruzi. ion channels play fundamental roles in controlling physiological parameters that ensure cell homeostasis by rapidly triggering compensatory mechanisms. combining molecular, cellular and electrophysiological approaches we have identified and ...201932010643
myelopoiesis of the amphibian xenopus laevis is segregated to the bone marrow, away from their hematopoietic peripheral liver.across vertebrates, hematopoiesis takes place within designated tissues, wherein committed myeloid progenitors further differentiate toward cells with megakaryocyte/erythroid potential (mep) or those with granulocyte/macrophage potential (gmp). while the liver periphery (lp) of the xenopus laevis amphibian functions as a principal site of hematopoiesis and contains meps, cells with gmp potential are instead segregated to the bone marrow (bm) of this animal. presently, using gene expression and w ...201932038608
antiarrhythmic properties of ranolazine: inhibition of atrial fibrillation associated task-1 potassium channels.background: atrial fibrillation (af) is the most common sustained cardiac arrhythmia and one of the major causes of cardiovascular morbidity and mortality. despite good progress within the past years, safe and effective treatment of af remains an unmet clinical need. the anti-anginal agent ranolazine has been shown to exhibit antiarrhythmic properties via mainly late ina and ikr blockade. this results in prolongation of the atrial action potential duration (apd) and effective refractory period ( ...201932038227
lissamphibian limbs and the origins of tetrapod hox domains.the expression and function of hox genes have played a key role in the debate on the evolution of limbs from fins. as an early branching tetrapod lineage, lissamphibians may provide information on the origin of the limb's hox domains and particularly how the plesiomorphic tetrapod pattern compares to the hox pattern present in fish fins. here, we comparatively investigated the expression of hox genes in the developing limbs of axolotl and xenopus laevis as well as in the fins of the direct devel ...201931445923
broad applicability of a streamlined ethyl cinnamate-based clearing procedure.turbidity and opaqueness are inherent properties of tissues that limit the capacity to acquire microscopic images through large tissues. creating a uniform refractive index, known as tissue clearing, overcomes most of these issues. these methods have enabled researchers to image large and complex 3d structures with unprecedented depth and resolution. however, tissue clearing has been adopted to a limited extent due to a combination of cost, time, complexity of existing methods and potential nega ...201930665888
the peptide transporter 1a of the zebrafish danio rerio, an emerging model in nutrigenomics and nutrition research: molecular characterization, functional properties, and expression analysis.peptide transporter 1 (pept1, alias slc15a1) mediates the uptake of dietary di/tripeptides in all vertebrates. however, in teleost fish, more than one pept1-type transporter might function, due to specific whole genome duplication event(s) that occurred during their evolution leading to a more complex paralogue gene repertoire than in higher vertebrates (tetrapods).201931890051
heterologous expression of aedes aegypti cation chloride cotransporter 2 (aeccc2) in xenopus laevis oocytes induces an enigmatic na⁺/li⁺ conductance.the yellow fever mosquito aedes aegypti possesses three genes encoding putative na⁺-coupled cation chloride cotransporters (cccs): aenkcc1, aeccc2, and aeccc3. to date, none of the aecccs have been functionally characterized. here we expressed aeccc2 heterologously in xenopus oocytes and measured the uptake of li⁺ (a tracer for na⁺) and rb⁺ (a tracer for k⁺). compared to control (h₂o-injected) oocytes, the aeccc2-expressing oocytes exhibited significantly greater uptake of li⁺, but not rb⁺. howe ...201930875796
the effects of insecticides on two splice variants of the glutamate-gated chloride channel receptor of the major malaria vector, anopheles gambiae.between half to 1 million people die annually from malaria. anopheles gambiae mosquitoes are major malaria vectors. unfortunately, resistance has emerged to the agents currently used to control a. gambiae, creating a demand for novel control measures. the pentameric glutamate-gated chloride channel (glucl) expressed in the muscle and nerve cells of these organisms are a potentially important biological target for malaria control. the pharmacological properties of anophiline glucl receptors are, ...201931479507
in vitro inhibition of human nucleoside transporters and uptake of azacitidine by an isocitrate dehydrogenase-2 inhibitor enasidenib and its metabolite agi-16903.1. the present study investigated inhibitory effects of enasidenib and its metabolite agi-16903 on (a) recombinant human nucleoside transporters (hnts) in hnt-producing xenopus laevis oocytes, and (b) azacitidine uptake in a normal b-lymphoblast peripheral blood cell line (pbc) and acute myeloid leukemia (aml) cell lines. 2. enasidenib inhibited hent1, hent2, hent3, and hent4 in oocytes with ic50 values of 7, 63, 27, and 76 μm, respectively, but exhibited little inhibition of hcnt1-3. agi-16903 ...201930394160
paracetamol-induced liver injury modelled in xenopus laevis embryos.failure to predict drug-induced liver injury (dili) remains a major contributing factor to lead compound drop-out during drug development. xenopus embryos are amenable for early stage medium throughput small molecule screens and so have the potential to be used in pre-clinical screens. to begin to assess the usefulness and limitations of xenopus embryos for safety assessment in the early phases of drug development, paracetamol was used as a model hepatotoxin. paracetamol overdose is associated w ...201930282005
conservation and divergence of protein pathways in the vertebrate heart.heart disease is the leading cause of death in the western world. attaining a mechanistic understanding of human heart development and homeostasis and the molecular basis of associated disease states relies on the use of animal models. here, we present the cardiac proteomes of 4 model vertebrates with dual circulatory systems: the pig (sus scrofa), the mouse (mus musculus), and 2 frogs (xenopus laevis and xenopus tropicalis). determination of which proteins and protein pathways are conserved and ...201931490923
discovery of the arp2 protein as a determining molecule in tumor cell death.cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ...201932091029
descubrimiento de la proteína reguladora de apoptosis 2 como determinante en la muerte de células tumorales.cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ...201931695224
a flipr assay for discovery of gabaa receptor modulators of natural origin.a fluorometric imaging plate reader (flipr) assay utilizing chinese hamster ovary (cho) cells stably transfected with gabaa receptors of α1β2γ2 subunit composition was evaluated and validated for rapid screening of plant extract libraries and efficient localization of active compounds in extracts. validation was performed with pure compounds and extracts known to contain allosteric gabaa receptor modulators. plants extracts that had been previously reported as active in an assay using xenopus la ...201931127604
[exploring the mysteries of megakaryocytopoiesis and thrombopoiesis through comparative hematology].in modern hematology, research on hematopoiesis and blood cells in vertebrates, such as birds, reptiles, amphibians, and fish, is lagging. this is because there are many experimental constraints when selecting subjects other than humans and mice as research subjects. currently, the availability of flow cytometry to count classified nucleated blood cells and utilization of whole genome information have led to novel findings. for example, in case of amphibian hematopoiesis studies, megakaryocytes ...201931597828
distinct host-mycobacterial pathogen interactions between resistant adult and tolerant tadpole life stages of xenopus laevis.mycobacterium marinum is a promiscuous pathogen infecting many vertebrates, including humans, whose persistent infections are problematic for aquaculture and public health. among unsettled aspects of host-pathogen interactions, the respective roles of conventional and innate-like t (it) cells in host defenses against m. marinum remain unclear. in this study, we developed an infection model system in the amphibian xenopus laevis to study host responses to m. marinum at two distinct life stages, t ...201931591148
different pharmacological properties of glut9a and glut9b: potential implications in preeclampsia.glucose transporter 9 (glut9/slc2a9) is the major regulator of uric acid homeostasis in humans. hyperuricemia due to impaired regulation by glut9 in pregnancy is closely associated with preeclampsia. while glut9 is expressed in two alternative splice variants, glut9a and glut9b, with different subcellular localizations, no functional differences of the two splice variants are known to date. the aim of this study was to investigate the function of both glut9 isoforms.201931502429
heterologous hyaluronic acid production in kluyveromyces lactis.hyaluronic acid (ha) is a biopolymer composed by the monomers glucuronic acid (glcua) and n-acetyl glucosamine (glcnac). it has a broad range of applications in the field of medicine, being marketed between usd 1000-5000/kg. its primary sources include extraction of animal tissue and fermentation using pathogenic bacteria. however, in both cases, extensive purification protocols are required to prevent toxin contamination. in this study, aiming at creating a safe ha producing microorganism, the ...201931466214
advancing genetic and genomic technologies deepen the pool for discovery in xenopus tropicalis.xenopus laevis and xenopus tropicalis have long been used to drive discovery in developmental, cell, and molecular biology. these dual frog species boast experimental strengths for embryology including large egg sizes that develop externally, well-defined fate maps, and cell-intrinsic sources of nutrients that allow explanted tissues to grow in culture. development of the xenopus cell extract system has been used to study cell cycle and dna replication. xenopus tadpole tail and limb regeneration ...201931254427
adaptive correction of craniofacial defects in pre-metamorphic xenopus laevis tadpoles involves thyroid hormone-independent tissue remodeling.although it is well established that some organisms can regenerate lost structures, the ability to remodel existing malformed structures has been less well studied. therefore, in this study we examined the ability of pre-metamorphic xenopus laevis tadpoles to self-correct malformed craniofacial tissues. we found that tadpoles can adaptively improve and normalize abnormal craniofacial morphology caused by numerous developmental perturbations. we then investigated the tissue-level and molecular me ...201931253636
the amphibian (xenopus laevis) colony-stimulating factor-1 and interleukin-34-derived macrophages possess disparate pathogen recognition capacities.pathogens such as the frog virus 3 (fv3) ranavirus are contributing to the worldwide amphibian declines. while amphibian macrophages (mϕs) are central to the immune defenses against these viruses, the pathogen recognition capacities of disparate amphibian mϕ subsets remain unexplored. in turn, mϕ differentiation and functionality are interdependent on the colony-stimulating factor-1 receptor (csf-1r), which is ligated by colony-stimulating factor-1 (csf-1) and the unrelated interleukin-34 (il-34 ...201931029710
the voltage sensing phosphatase (vsp) localizes to the apical membrane of kidney tubule epithelial cells.voltage-sensing phosphatases (vsps) are transmembrane proteins that couple changes in membrane potential to hydrolysis of inositol signaling lipids. vsps catalyze the dephosphorylation of phosphatidylinositol phosphates (pips) that regulate diverse aspects of cell membrane physiology including cell division, growth and migration. vsps are highly conserved among chordates, and their rna transcripts have been detected in the adult and embryonic stages of frogs, fish, chickens, mice and humans. how ...201930964862
developmental exposure to chemicals associated with unconventional oil and gas extraction alters immune homeostasis and viral immunity of the amphibian xenopus.although aquatic vertebrates and humans are increasingly exposed to water pollutants associated with unconventional oil and gas extraction (uog), the long-term effects of these pollutants on immunity remains unclear. we have established the amphibian xenopus laevis and the ranavirus frog virus 3 (fv3) as a reliable and sensitive model for evaluating the effects of waterborne pollutants. x. laevis tadpoles were exposed to a mixture of equimass amount of uog chemicals with endocrine disrupting act ...201930939317
developmental regulation of wnt signaling by nagk and the udp-glcnac salvage pathway.in a screen for human kinases that regulate xenopus laevis embryogenesis, we identified nagk and other components of the udp-glcnac glycosylation salvage pathway as regulators of anteroposterior patterning and wnt signaling. we find that the salvage pathway does not affect other major embryonic signaling pathways (fgf, tgfβ, notch, or shh), thereby demonstrating specificity for wnt signaling. we show that the role of the salvage pathway in wnt signaling is evolutionarily conserved in zebrafish a ...201930904594
stage-dependent cardiac regeneration in xenopus is regulated by thyroid hormone availability.despite therapeutic advances, heart failure is the major cause of morbidity and mortality worldwide, but why cardiac regenerative capacity is lost in adult humans remains an enigma. cardiac regenerative capacity widely varies across vertebrates. zebrafish and newt hearts regenerate throughout life. in mice, this ability is lost in the first postnatal week, a period physiologically similar to thyroid hormone (th)-regulated metamorphosis in anuran amphibians. we thus assessed heart regeneration in ...201930755533
ketone body modulation of ligand-gated ion channels.ketogenesis is a metabolic process wherein ketone bodies are produced from the breakdown of fatty acids. in humans, fatty acid catabolism results in the production of acetyl-coa which can then be used to synthesize three ketone bodies: acetoacetate, acetone, and β-hydroxybutyrate. ketogenesis occurs at a higher rate in situations of low blood glucose, such as during fasting, heavy alcohol consumption, and in situations of low insulin, as well as in individuals who follow a 'ketogenic diet' consi ...201930562540
evaluation of a multiplexed, multispecies nuclear receptor assay for chemical hazard assessment.sensitivity to potential endocrine disrupting chemicals in the environment varies across species and is influenced by sequence conservation of their nuclear receptor targets. here, we evaluated a multiplexed, in vitro assay testing receptors relevant to endocrine and metabolic disruption from five species. the trans-factorial™ system of human nuclear receptors was modified to include additional species: mouse (mus musculus), frog (xenopus laevis), zebrafish (danio rerio), chicken (gallus gallus) ...202033049310
organelle size scaling over embryonic development.cell division without growth results in progressive cell size reductions during early embryonic development. how do the sizes of intracellular structures and organelles scale with cell size and what are the functional implications of such scaling relationships? model organisms, in particular caenorhabditis elegans worms, drosophila melanogaster flies, xenopus laevis frogs, and mus musculus mice, have provided insights into developmental size scaling of the nucleus, mitotic spindle, and chromosom ...202032003549
polyunsaturated fatty acid-derived iks channel activators shorten the qt interval ex-vivo and in-vivo.we aimed to assess the ability of natural and modified polyunsaturated fatty acids (pufas) to shorten qt interval in ex-vivo and in-vivo guinea pig hearts.202032223014
periodic albinism of a widely used albino mutant of xenopus laevis caused by deletion of two exons in the hermansky-pudlak syndrome type 4 gene.the periodic albino mutant of xenopus laevis is a recessive mutant, in which reduced amounts of melanin appear in the retinal pigment epithelium (rpe) and in melanophores at the late embryonic stage, after which both rpe and melanophores gradually depigment. three types of pigment cells (melanophores, iridophores, and xanthophores) have been reported to be affected in this albino. however, the causative gene of the periodic albinism remains unknown. hermansky-pudlak syndrome (hps) is an autosoma ...202033147376
disease-associated hcn4 v759i variant is not sufficient to impair cardiac pacemaking.the hyperpolarization-activated cation current if is a key determinant for cardiac pacemaker activity. it is conducted by subunits of the hyperpolarization-activated cyclic nucleotide-gated (hcn) channel family, of which hcn4 is predominant in mammalian heart. both loss-of-function and gain-of-function mutations of the hcn4 gene are associated with sinus node dysfunction in humans; however, their functional impact is not fully understood yet. here, we sought to characterize a hcn4 v759i variant ...202033095298
using an aquatic model, xenopus laevis, to uncover the role of chromodomain 1 in craniofacial disorders.the chromodomain family member chromodomain 1 (chd1) has been shown to have numerous critical molecular functions including transcriptional regulation, splicing, and dna repair. complete loss of function of this gene is not compatible with life. on the other hand, missense and copy number variants of chd1 can result in intellectual disabilities and craniofacial malformations in human patients including cleft palate and pilarowski-bjornsson syndrome. we have used the aquatic developmental model o ...202032918369
acute consequences of a unilateral viiith nerve transection on vestibulo-ocular and optokinetic reflexes in xenopus laevis tadpoles.loss of peripheral vestibular function provokes severe impairments of gaze and posture stabilization in humans and animals. however, relatively little is known about the extent of the instantaneous deficits. this is mostly due to the fact that in humans a spontaneous loss often goes unnoticed initially and targeted lesions in animals are performed under deep anesthesia, which prevents immediate evaluation of behavioral deficits. here, we use isolated preparations of xenopus laevis tadpoles with ...202032915311
l-glutamic acid monosodium salt reduces the harmful effect of lithium on the development of xenopus laevis embryos.many xenobiotics in the environment affect the human body in various ways. among those xenobiotics, lithium chloride (li, licl) and monosodium glutamate (l-glutamic acid monosodium salt, msg) compounds affect the crucial processes of stem cell differentiation, cell proliferation, developmental gene expression, and overall development in animals. in this study, we aimed to examine the developmental effects of exposure to flavor enhancer msg and lici medicament on xenopus embryos using the frog em ...202032705564
uncoupling sodium channel dimers restores the phenotype of a pain-linked nav 1.7 channel mutation.the voltage-gated sodium channel nav 1.7 is essential for adequate perception of painful stimuli. mutations in the encoding gene, scn9a, cause various pain syndromes in humans. the hnav 1.7/a1632e channel mutant causes symptoms of erythromelalgia and paroxysmal extreme pain disorder (pepd), and its main gating change is a strongly enhanced persistent current. on the basis of recently published 3d structures of voltage-gated sodium channels, we investigated how the inactivation particle binds to ...202032663327
trpm6 and trpm7: novel players in cell intercalation during vertebrate embryonic development.a common theme in organogenesis is how the final structure of organs emerge from epithelial tube structures, with the formation of the neural tube being one of the best examples. two types of cell movements co-occur during neural tube closure involving the migration of cells toward the midline of the embryo (mediolateral intercalation or convergent extension) as well as the deep movement of cells from inside the embryo to the outside of the lateral side of the neural plate (radial intercalation) ...202032315468
effects of bisphenol a and its alternative bisphenol f on notch signaling and intestinal development: a novel signaling by which bisphenols disrupt vertebrate development.we previously found bisphenol a (bpa) alternative, bisphenol f (bpf) upregulated notch-related gene expression in intestines of the african clawed frog xenopus laevis, suggesting an agonistic action on notch signaling, a crucial signaling in multiple biological processes during development. here, we aimed to confirm the actions of bpa and bpf on notch signaling and to reveal their effects on intestinal development. using x. laevis, an excellent model for developmental biology, we found that 10-1 ...202032311622
modeling bainbridge-ropers syndrome in xenopus laevis embryos.the additional sex combs-like (asxl1-3) genes are linked to human neurodevelopmental disorders. the de novo truncating variants in asxl1-3 proteins serve as the genetic basis for severe neurodevelopmental diseases such as bohring-opitz, shashi-pena, and bainbridge-ropers syndromes, respectively. the phenotypes of these syndromes are similar but not identical, and include dramatic craniofacial defects, microcephaly, developmental delay, and severe intellectual disability, with a loss of speech an ...202032132929
hplc reveals novel features of nucleoside and nucleobase homeostasis, nucleoside metabolism and nucleoside transport.humans possess three members of the cation-coupled concentrative nucleoside transporter cnt (slc 28) family, hcnt1-3: hcnt1 is selective for pyrimidine nucleosides but also transports adenosine, hcnt2 transports purine nucleosides and uridine, and hcnt3 transports both pyrimidine and purine nucleosides. hcnt1/2 transport nucleosides using the transmembrane na+ electrochemical gradient, while hcnt3 is both na+- and h+-coupled. by producing recombinant hcnt3 in xenopus laevis oocytes, we have used ...202032126230
evaluation of the effects of low concentrations of bisphenol af on gonadal development using the xenopus laevis model: a finding of testicular differentiation inhibition coupled with feminization.developmental exposures to estrogenic chemicals possibly cause structural and functional abnormalities of reproductive organs in vertebrates. bisphenol af (bpaf), a bisphenol a (bpa) analogue, has been shown to have higher estrogenic activity than bpa, but little is known about the effects of bpaf on gonadal development, particularly gonadal differentiation. we aimed to determine whether low concentrations of bpaf could disrupt gonadal differentiation and subsequent development using xenopus lae ...202031991354
cfap43 modulates ciliary beating in mouse and xenopus.malfunctions of motile cilia cause a variety of developmental defects and diseases in humans and animal model organisms. defects include impaired mucociliary clearance of the airways, sperm immotility, hydrocephalus and organ laterality. here, we characterize the evolutionary conserved cfap43 gene by loss-of-function experiments in the mouse and the frog xenopus laevis. cfap43 is expressed in tissues carrying motile cilia and acts as a target gene of the transcription factor foxj1, which is esse ...202031884020
insights regarding skin regeneration in non-amniote vertebrates: skin regeneration without scar formation and potential step-up to a higher level of regeneration.skin wounds are among the most common injuries in animals and humans. vertebrate skin is composed of an epidermis and dermis. after a deep skin injury in mammals, the wound heals, but the dermis cannot regenerate. instead, collagenous scar tissue forms to fill the gap in the dermis, but the scar does not function like the dermis and often causes disfiguration. in contrast, in non-amniote vertebrates, including fish and amphibians, the dermis and skin derivatives are regenerated after a deep skin ...202031831357
impact of copper in xenopus laevis liver: histological damages and atp7b downregulation.copper is an essential micronutrient but its excess in the dietary can be toxic. both copper deficiency and abundance can occur in natural conditions and can lead to pathological dysfunctions. many of the toxic effects of copper, such as increased lipid peroxidation in cell membranes and dna damage, are due to its role in the generation of oxygen free radicals. copper is released into the environment by both natural sources and human activities and it can damage organisms and ecosystems. in the ...202031757510
effects of copper on the neuromasts of xenopus laevis.fish and aquatic amphibians possess neuromasts on the surface of their body that constitute the lateral line, a sensory system used to detect water displacement. copper is known to inactivate the neuromast organs of this system. copper-induced neuromast loss in african clawed frogs, xenopus laevis, was examined by exposing nieuwkoop-faber stage 54-55 larvae to copper concentrations of 0, 100, 200, 300, and 400 µg/l for 96 h, followed by an examination of neuromast counts, staining intensity, and ...202033180147
ecotoxicological assessment of commercial boron nitride nanotubes toward xenopus laevis tadpoles and host-associated gut microbiota.despite the growing interest for boron nitride nanotubes (bnnt) due to their unique properties, data on the evaluation of the environmental risk potential of this emerging engineered nanomaterial are currently lacking. therefore, the ecotoxicity of a commercial form of bnnt (containing tubes, hexagonal-boron nitride, and boron) was assessed in vivo toward larvae of the amphibian xenopus laevis. following the exposure, multiple endpoints were measured in the tadpoles as well as in bacterial commu ...202033171057
mechanisms activating latent functions of pip aquaporin water channels via the interaction between pip1 and pip2 proteins.plant plasma-membrane type pip aquaporins are classified into two groups, pip1s and pip2s. in this study, we focused on hvpip1; 2, a pip1 in barley (hordeum vulgare), to dissect the molecular mechanisms that evoke hvpip1-mediated water transport. no hvpip1; 2 protein was localized to the plasma membrane when expressed alone in xenopus laevis oocytes. in contrast, a chimeric hvpip1; 2 protein (hvpip1; 2_24nc), in which the n- and c-terminal regions were replaced with the corresponding regions fro ...202033169164
epigenetic regulation of replication origin assembly: a role for histone h1 and chromatin remodeling factors.during early embryonic development in several metazoans, accurate dna replication is ensured by high number of replication origins. this guarantees rapid genome duplication coordinated with fast cell divisions. in xenopus laevis embryos this program switches to one with a lower number of origins at a developmental stage known as mid-blastula transition (mbt) when cell cycle length increases and gene transcription starts. consistent with this regulation, somatic nuclei replicate poorly when trans ...202033165968
synthesis of glun2a-selective nmda receptor antagonists with an electron-rich aromatic b-ring.glutamatergic n-methyl-d-aspartate (nmda) receptors are heterotetrameric ion channels that can be comprised of different subunits. glun2a subunit-containing nmda receptors are associated with diseases like anxiety, depression, and schizophrenia. however, the exact contribution of these nmda receptor subtypes is still unclear. to understand better the role of the glun2a-containing receptors, novel ligands were designed. in co-crystallization with the isolated binding site, tcn-201 (1) and analogs ...202033162207
trim-away mediated knock down uncovers a new function for lbh during gastrulation of xenopus laevis.we previously identified the protein lbh as necessary for cranial neural crest (cnc) cell migration in xenopus through the use of morpholinos. however, lbh is a maternally deposited protein and morpholinos achieve knockdowns through prevention of translation. in order to investigate the role of lbh in earlier embryonic events, we employed the new technique "trim-away" to degrade this maternally deposited protein. trim-away utilizes the e3 ubiquitin ligase trim21 to degrade proteins targeted with ...202033159936
kunitz-type peptides from the sea anemone heteractis crispa demonstrate potassium channel blocking and anti-inflammatory activities.the kunitz/bpti peptide family includes unique representatives demonstrating various biological activities. electrophysiological screening of peptides hcrg1 and hcrg2 from the sea anemone heteractis crispa on six kv1.x channel isoforms and insect shaker ir channel expressed in xenopus laevis oocytes revealed their potassium channels blocking activity. hcrg1 and hcrg2 appear to be the first kunitz-type peptides from sea anemones blocking kv1.3 with ic50 of 40.7 and 29.7 nm, respectively. in addit ...202033158163
development of antimicrobial peptides from amphibians.since the discovery of magainins from the skin secretions of the african toad xenopus laevis by michael zasloff in 1987, an increasing number of antimicrobial peptides (amps) has been identified in different anuran species and studied in detail [...].202033158031
antimicrobial peptide induced-stress renders staphylococcus aureus susceptible to toxic nucleoside analogs.cationic antimicrobial peptides (amps) are active immune effectors of multicellular organisms and are also considered as new antimicrobial drug candidates. one of the problems encountered when developing amps as drugs is the difficulty of reaching sufficient killing concentrations under physiological conditions. here, using pexiganan, a cationic peptide derived from a host defense peptide of the african clawed frog and the first amp developed into an antibacterial drug, we studied whether sub-le ...202033133056
nicotinic acetylcholine receptors in the synganglion of the tick ixodes ricinus: functional characterization using membrane microtransplantation.nicotinic acetylcholine receptors are an important class of excitatory receptors in the central nervous system of arthropods. in the ticks ixodes ricinus, the functional and pharmacological properties of nicotinic receptors located in their neurons are still unknown. the objective of this study was to characterize the pharmacological properties of tick nicotinic receptors using membrane microtransplantation in xenopus laevis oocytes and two-electrodes voltage clamp method. the membranes microtra ...202033120248
observation of nucleic acids inside living human cells by in-cell nmr spectroscopy.the intracellular environment is highly crowded with biomacromolecules such as proteins and nucleic acids. under such conditions, the structural and biophysical features of nucleic acids have been thought to be different from those in vitro. to obtain high-resolution structural information on nucleic acids in living cells, the in-cell nmr method is a unique tool. following the first in-cell nmr measurement of nucleic acids in 2009, several interesting insights were obtained using xenopus laevis ...202033110737
high affinity na+ transport by wheat hkt1;5 is blocked by k.the wheat sodium transporters tmhkt1;5-a and tahkt1;5-d are encoded by genes underlying the major shoot na+ exclusion loci nax2 and kna1 from triticum monococcum (tm) and triticum aestivum (ta), respectively. in contrast to hkt2 transporters that have been shown to exhibit high affinity k+-dependent na+ transport, hkt1 proteins have, with one exception, only been shown to catalyze low affinity na+ transport and no k+ transport. here, using heterologous expression in xenopus laevis oocytes we unc ...202033103046
x-box-binding protein 1 is required for pancreatic development in xenopus laevis.x-box-binding protein 1 (xbp1) is a protein containing the basic leucine zipper structure. it belongs to the camp-response element binding protein (creb)/activating transcription factor transcription factor family. as the main transcription factor, spliced xbp1 (xbp1s) participates in many physiological and pathological processes and plays an important role in embryonic development. previous studies showed that xbp1-knockout mice died because of pancreatic exocrine function deficiency, indicatin ...202033098302
sclerostin inhibits wnt signaling through tandem interaction with two lrp6 ectodomains.low-density lipoprotein receptor-related protein 6 (lrp6) is a coreceptor of the β-catenin-dependent wnt signaling pathway. the lrp6 ectodomain binds wnt proteins, as well as wnt inhibitors such as sclerostin (sost), which negatively regulates wnt signaling in osteocytes. although lrp6 ectodomain 1 (e1) is known to interact with sost, several unresolved questions remain, such as the reason why sost binds to lrp6 e1e2 with higher affinity than to the e1 domain alone. here, we present the crystal ...202033097721
rad21 is involved in corneal stroma development by regulating neural crest migration.previously, we identified rad21r450c from a peripheral sclerocornea pedigree. injection of this rad21 variant mrna into xenopus laevis embryos disrupted the organization of corneal stroma fibrils. to understand the mechanisms of rad21-mediated corneal stroma defects, gene expression and chromosome conformation analysis were performed using cells from family members affected by peripheral sclerocornea. both gene expression and chromosome conformation of cell adhesion genes were affected in cells ...202033096935
phosphoproteomic analysis of xenopus laevis reveals expression and phosphorylation of hypoxia-inducible pfkfb3 during dehydration.xenopus laevis tolerate dehydration when their environments evaporate during summer months. protein phosphorylation has previously shown to regulate important adaptations in x. laevis, including the transition to anaerobic metabolism. we therefore performed phosphoproteomic analysis of x. laevis to further elucidate the cellular and metabolic responses to dehydration. phosphoproteins were enriched in cellular functions and pathways related to glycolysis/gluconeogenesis, the tca cycle, and protei ...202033083755
derivation of proliferative islet1-positive cells during metamorphosis and wound response in xenopus.in mammalian hearts, cardiomyocytes retain a transient capacity to proliferate and regenerate following injury before birth, whereas they lose proliferative capacity immediately after birth. it has also been known that cardiac progenitor cells including islet1-positive cells do not contribute to the cardiac repair and regeneration in mammals. in contrast, hearts of zebrafish, amphibians and reptiles maintain a regenerative ability throughout life. here, we analyzed proliferative capacity of card ...202033070205
non-junctional role of cadherin3 in cell migration and contact inhibition of locomotion via domain-dependent, opposing regulation of rac1.classical cadherins are well-known adhesion molecules responsible for physically connecting neighboring cells and signaling this cell-cell contact. recent studies have suggested novel signaling roles for "non-junctional" cadherins (njcads); however, the function of cadherin signaling independent of cell-cell contacts remains unknown. in this study, mesendodermal cells and tissues from gastrula stage xenopus laevis embryos demonstrate that deletion of extracellular domains of cadherin3 (cdh3; for ...202033060598
scorpion toxin meunatxα-1 sensitizes primary nociceptors by selective modulation of voltage-gated sodium channels.venoms are a rich source of highly specific toxins, which allow the identification of novel therapeutic targets. we have now applied high content screening (hcs) microscopy to identify toxins that modulate pain sensitization signaling in primary sensory neurons of rat and elucidated the underlying mechanism. a set of venoms and fractions thereof were analyzed for their ability to activate type ii protein kinase a (pka-ii) and extracellular signal-regulated kinases (erk1/2). we identified meunatx ...202033051988
glutamate, d-(-)-2-amino-5-phosphonopentanoic acid, and n-methyl-d-aspartate do not directly modulate glycine receptors.replication studies play an essential role in corroborating research findings and ensuring that subsequent experimental works are interpreted correctly. a previously published paper indicated that the neurotransmitter glutamate, along with the compounds n-methyl-d-aspartate (nmda) and d-(-)-2-amino-5-phosphonopentanoic acid (ap5), acts as positive allosteric modulators of inhibitory glycine receptors. the paper further suggested that this form of modulation would play a role in setting the spina ...202033051383
whole-brain calcium imaging in larval xenopus.sensory systems detect environmental stimuli and transform them into electrical activity patterns interpretable by the central nervous system. en route to higher brain centers, the initial sensory input is successively transformed by interposed secondary processing centers. mapping the neuronal activity patterns at all of those stages is essential to understand sensory information processing. larval xenopus laevis is very well-suited for whole-brain imaging of neuronal activity. this is mainly d ...202033037078
elucidating the framework for specification and determination of the embryonic retina.how cell determination is regulated remains a major unsolved problem in developmental biology. the early embryonic rudiments of many tissues and organs are difficult or impossible to identify, isolate and study at the time when determination occurs. we have examined the commitment process leading to retina formation in xenopus laevis, where presumptive eye tissue can be identified and studied to assay its biological properties during the events leading up to determination. we find that for the r ...202033031807
microtubule-dependent pushing forces contribute to long-distance aster movement and centration in xenopus laevis egg extracts.during interphase of the eukaryotic cell cycle, the microtubule (mt) cytoskeleton serves as both a supportive scaffold for organelles and an arborized system of tracks for intracellular transport. at the onset of mitosis, the position of the astral mt network, specifically its center, determines the eventual location of the spindle apparatus and ultimately the cytokinetic furrow. positioning of the mt aster often results in its movement to the center of a cell, even in large blastomeres hundreds ...202033026931
cross-talk between p2x and nmda receptors.purinergic p2x receptors (p2x) are atp-gated ion channels widely expressed in the cns. while the direct contribution of p2x to synaptic transmission is uncertain, p2x reportedly affect n-methyl-d-aspartate receptor (nmdar) activity, which has given rise to competing theories on the role of p2x in the modulation of synapses. however, p2x have also been shown to participate in receptor cross-talk: an interaction where one receptor (e.g., p2x2) directly influences the activity of another (e.g., nic ...202033003406
dna content contributes to nuclear size control in xenopus laevis.cells adapt to drastic changes in genome quantity during evolution and cell division by adjusting the nuclear size to exert genomic functions. however, the mechanism by which dna content within the nucleus contributes to controlling the nuclear size remains unclear. here, we experimentally evaluated the effects of dna content by utilizing cell-free xenopus egg extracts and imaging of in vivo embryos. upon manipulation of dna content while maintaining cytoplasmic effects constant, both plateau si ...202032997613
torc1 selectively regulates synaptic maturation and input convergence in the developing visual system.newly synthesized proteins support the development of functional neural circuits and previous work has suggested that dysregulated translation mediates certain forms of autism spectrum disorder (asd). here, we investigated the role of target of rapamycin complex 1 (torc1) in synaptic and dendritic development in vivo in the retinotectal system of xenopus laevis tadpoles. we found that torc1 signaling regulates dendritic growth and branching and that acute over-activation of torc1 by rheb overexp ...202032996262
a survey of barley pip aquaporin ionic conductance reveals ca2+-sensitive hvpip2;8 na+ and k+ conductance.some plasma membrane intrinsic protein (pip) aquaporins can facilitate ion transport. here we report that one of the 12 barley pips (pip1 and pip2) tested, hvpip2;8, facilitated cation transport when expressed in xenopus laevis oocytes. hvpip2;8-associated ion currents were detected with na+ and k+, but not cs+, rb+, or li+, and was inhibited by ba2+, ca2+, and cd2+ and to a lesser extent mg2+, which also interacted with ca2+. currents were reduced in the presence of k+, cs+, rb+, or li+ relativ ...202032992595
inferring a complete genotype-phenotype map from a small number of measured phenotypes.understanding evolution requires detailed knowledge of genotype-phenotype maps; however, it can be a herculean task to measure every phenotype in a combinatorial map. we have developed a computational strategy to predict the missing phenotypes from an incomplete, combinatorial genotype-phenotype map. as a test case, we used an incomplete genotype-phenotype dataset previously generated for the malaria parasite's 'chloroquine resistance transporter' (pfcrt). wild-type pfcrt (pfcrt3d7) lacks signif ...202032991585
the mechanism of nmda receptor hyperexcitation in high pressure helium and hyperbaric oxygen.professional divers exposed to pressures greater than 1.1 mpa may suffer from the high pressure neurological syndrome (hpns). divers who use closed-circuit breathing apparatus face the risk of cns hyperbaric oxygen toxicity (hbotox). both syndromes are characterized by reversible cns hyperexcitability, accompanied by cognitive and motor deficits. previous studies have demonstrated that the hyperexcitability of hpns is induced mainly by nmda receptors (nmdars). in our recent studies, we demonstra ...202032982789
in-cell nmr spectroscopy of functional riboswitch aptamers in eukaryotic cells.we report here the in-cell nmr-spectroscopic observation of the binding of the cognate ligand 2'-deoxyguanosine to the aptamer domain of the bacterial 2'-deoxyguanosine-sensing riboswitch in eukaryotic cells, namely xenopus laevis oocytes and in human hela cells. the riboswitch is sufficiently stable in both cell types to allow for detection of binding of the ligand to the riboswitch. most importantly, we show that the binding mode established by in vitro characterization of this prokaryotic rib ...202032975353
design, synthesis, and in vitro evaluation of p2x7 antagonists.the p2x7 receptor is a promising target for the treatment of various diseases due to its significant role in inflammation and immune cell signaling. this work describes the design, synthesis, and in vitro evaluation of a series of novel derivatives bearing diverse scaffolds as potent p2x7 antagonists. our approach was based on structural modifications of reported (adamantan-1-yl)methylbenzamides able to inhibit the receptor activation. the adamantane moieties and the amide bond were replaced, an ...202032964578
endoparasites infecting exotic captive amphibian pet and zoo animals (anura, caudata) in germany.alongside exotic reptiles, amphibians, such as toads, frogs, salamanders, and newts, are nowadays considered popular pets worldwide. as reported for other exotic pet animals, amphibians are known to harbor numerous gastrointestinal parasites. nonetheless, very little data are available on captive amphibian parasitic diseases. in this study, we applied direct saline fecal smears (dsfs) to examine in total 161 stool samples from 41 different amphibian species belonging to the orders anura and caud ...202032960371
amiodarone bioconcentration and suppression of metamorphosis in xenopus.trace concentrations of a number of pharmaceutically active compounds have been detected in the aquatic environment in many countries, where they are thought to have the potential to exert adverse effects on non-target organisms. amiodarone (amd) is one such high-risk compound commonly used in general hospitals. amd is known to alter normal thyroid hormone (th) function, although little information is available regarding the specific mechanism by which this disruption occurs. anuran tadpole meta ...202032956954
aquaporin expression in the alimentary canal of the honey bee apis mellifera l. (hymenoptera: apidae) and functional characterization of am_eglp 1.aquaporins (aqp) are a family of plasma membrane proteins responsible for water transport through cell membranes. they are differentially expressed in different parts of the alimentary canal of insects where they regulate water transport. these proteins have been studied in detail in some insects, but few data are available for aquaporins of the honey bee, apis mellifera. we used quantitative pcr to study the expression of six putative aquaporin genes in forager honey bees. we found differential ...202032956406
do muscle contractile properties drive differences in locomotor performance in invasive populations of xenopus laevis in france?jumping and swimming are key locomotor traits in frogs intimately linked to survival and dispersal. french populations of the frog xenopus laevis from the invasion front are known to possess greater terrestrial locomotor endurance. here, we tested whether individuals from the invasion front show differences in their muscle physiology that may underlie the observed whole-organism performance differences. we measured muscle contractile properties of the isolated gastrocnemius muscle in vitro, incl ...202032955613
effect of 158 herbal remedies on human trpv1 and the two-pore domain potassium channels kcnk2, 3 and 9.herbal medicines are used to treat a broad number of maladies. however, the pharmacological profile of most remedies is poorly understood. we investigated the effect of herbal remedies from kampo, traditional chinese medicine (tcm) and other phytotherapies on human two-pore domain potassium channels (kcnk channels; trek-1, task-1 and task-3) as well as the human trpv1 channel. kcnk channels are responsible for the background potassium current of excitable cells, thus essential for the maintenanc ...202032953560
exposure impacts of environmentally relevant concentrations of a glufosinate ammonium herbicide formulation on larval development and thyroid histology of xenopus laevis.thyroid hormones play critical roles in body growth and development as well as reproduction. they also influence the activities of a wider variety of tissues and biological functions, such as osmoregulation, metabolism, and especially metamorphosis in organisms, such as frogs. these complex activities of thyroid hormones are prone to disruption by agricultural pesticides, often leading to modulation of growth and the reproductive system in particular. these substances include glufosinate ammoniu ...202032948887
an integrated approach to assess the sublethal effects of colloidal gold nanorods in tadpoles of xenopus laevis.gold nanorods (aunr) have been explored for many applications, including innovative nanomedicines, which also might contribute to its increase in the environment, namely due to inadequate disposable of wastes into aquatic environments. early-life stages of amphibians are usually aquatic and sensitive to chemical contamination. accordingly, this study aimed to determine the sublethal effects of ctab functionalized aunr on xenopus laevis tadpoles. as such, tadpoles were exposed to serial concentra ...202032947686
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