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1.
Mol Psychiatry ; 2023 Oct 05.
Article in English | MEDLINE | ID: mdl-37798419

ABSTRACT

The Wnt/ß-catenin pathway contains multiple high-confidence risk genes that are linked to neurodevelopmental disorders, including autism spectrum disorder. However, its ubiquitous roles across brain cell types and developmental stages have made it challenging to define its impact on neural circuit development and behavior. Here, we show that TCF7L2, which is a key transcriptional effector of the Wnt/ß-catenin pathway, plays a cell-autonomous role in postnatal astrocyte maturation and impacts adult social behavior. TCF7L2 was the dominant Wnt effector that was expressed in both mouse and human astrocytes, with a peak during astrocyte maturation. The conditional knockout of Tcf7l2 in postnatal astrocytes led to an enlargement of astrocytes with defective tiling and gap junction coupling. These mice also exhibited an increase in the number of cortical excitatory and inhibitory synapses and a marked increase in social interaction by adulthood. These data reveal an astrocytic role for developmental Wnt/ß-catenin signaling in restricting excitatory synapse numbers and regulating adult social behavior.

2.
J Inorg Biochem ; 248: 112364, 2023 11.
Article in English | MEDLINE | ID: mdl-37689037

ABSTRACT

Hepcidin is an iron regulatory hormone that does not bind iron directly. Instead, its mature 25-peptide form (H25) contains a binding site for other metals, the so-called ATCUN/NTS (amino-terminal Cu/Ni binding site). The Cu(II)-hepcidin complex was previously studied, but due to poor solubility and difficult handling of the peptide the definitive account on the binding equilibrium was not obtained reliably. In this study we performed a series of fluorescence competition experiments between H25 and its model peptides containing the same ATCUN/NTS site and determined the Cu(II) conditional binding constant of the CuH25 complex at pH 7.4, CK7.4 = 4 ± 2 × 1014 M-1. This complex was found to be very inert in exchange reactions and poorly reactive in the ascorbate consumption test. The consequences of these findings for the putative role of Cu(II) interactions with H25 are discussed.


Subject(s)
Hepcidins , Iron , Humans , Binding Sites , Fluorescence , Protein Domains
3.
Acta Histochem ; 125(1): 151975, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36455340

ABSTRACT

BACKGROUND: The motor ability of cancer cells to cross the basement membrane contributes to their implantation in a new location. Metastasis is a significant factor that worsens the prognosis of cancer patients. Thus, reducing cell invasiveness is an important aspect of anticancer therapy, also in bladder cancer treatment. MATERIAL: The study material was the T24 cell line of human urinary bladder cancer. The migratory potential of the cells and the effect of the treatment with individually doses and synergistic combination of doxorubicin and metformin in the 500:1 ratio for 24 h were analyzed. RESULTS: The results obtained show a compound-initiated decrease in the motor abilities of bladder cancer cells compared to controls. A decrease in the rate of colony formation was observed, as well as inhibition of migration through inserts. The visualized reorganization of the vimentin and actin networks confirms the drug-initiated limitation of the metastatic potential of T24 cells. CONCLUSION: According to our knowledge, we are the first to show, that combination of doxorubicin and metformin also worth considering in the treatment of bladder cancer. We showed that simultaneous administration of these cytostatic enhances the antiproliferative effect of drugs, but also limits cells' migratory potential.


Subject(s)
Metformin , Urinary Bladder Neoplasms , Humans , Metformin/pharmacology , Doxorubicin/pharmacology , Urinary Bladder Neoplasms/drug therapy , Cell Line, Tumor , Cell Movement , Cell Proliferation
4.
Int J Mol Sci ; 23(24)2022 Dec 07.
Article in English | MEDLINE | ID: mdl-36555126

ABSTRACT

Hepcidin (DTHFPICIFCCGCCHRSKCGMCCKT), an iron-regulatory hormone, is a 25-amino-acid peptide with four intramolecular disulfide bonds circulating in blood. Its hormonal activity is indirect and consists of marking ferroportin-1 (an iron exporter) for degradation. Hepcidin biosynthesis involves the N-terminally extended precursors prepro-hepcidin and pro-hepcidin, processed by peptidases to the final 25-peptide form. A sequence-specific formation of disulfide bonds and export of the oxidized peptide to the bloodstream follows. In this study we considered the fact that prior to export, reduced hepcidin may function as an octathiol ligand bearing some resemblance to the N-terminal part of the α-domain of metallothioneins. Consequently, we studied its ability to bind Zn(II) and Cd(II) ions using the original peptide and a model for prohepcidin extended N-terminally with a stretch of five arginine residues (5R-hepcidin). We found that both form equivalent mononuclear complexes with two Zn(II) or Cd(II) ions saturating all eight Cys residues. The average affinity at pH 7.4, determined from pH-metric spectroscopic titrations, is 1010.1 M-1 for Zn(II) ions; Cd(II) ions bind with affinities of 1015.2 M-1 and 1014.1 M-1. Using mass spectrometry and 5R-hepcidin we demonstrated that hepcidin can compete for Cd(II) ions with metallothionein-2, a cellular cadmium target. This study enabled us to conclude that hepcidin binds Zn(II) and Cd(II) sufficiently strongly to participate in zinc physiology and cadmium toxicity under intracellular conditions.


Subject(s)
Cadmium , Hepcidins , Cadmium/metabolism , Peptides , Iron , Disulfides , Metallothionein/metabolism
5.
Cells ; 11(19)2022 09 20.
Article in English | MEDLINE | ID: mdl-36230905

ABSTRACT

HAX1 is a human protein with no known homologues or structural domains. Mutations in the HAX1 gene cause severe congenital neutropenia through mechanisms that are poorly understood. Previous studies reported the RNA-binding capacity of HAX1, but the role of this binding in physiology and pathology remains unexplained. Here, we report the transcriptome-wide characterization of HAX1 RNA targets using RIP-seq and CRAC, indicating that HAX1 binds transcripts involved in translation, ribosome biogenesis, and rRNA processing. Using CRISPR knockouts, we find that HAX1 RNA targets partially overlap with transcripts downregulated in HAX1 KO, implying a role in mRNA stabilization. Gene ontology analysis demonstrated that genes differentially expressed in HAX1 KO (including genes involved in ribosome biogenesis and translation) are also enriched in a subset of genes whose expression correlates with HAX1 expression in four analyzed neoplasms. The functional connection to ribosome biogenesis was also demonstrated by gradient sedimentation ribosome profiles, which revealed differences in the small subunit:monosome ratio in HAX1 WT/KO. We speculate that changes in HAX1 expression may be important for the etiology of HAX1-linked diseases through dysregulation of translation.


Subject(s)
Proteins , Ribosomes , Adaptor Proteins, Signal Transducing/metabolism , Humans , Mutation , Proteins/metabolism , RNA/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Ribosomes/genetics , Ribosomes/metabolism
6.
Sensors (Basel) ; 22(5)2022 Feb 25.
Article in English | MEDLINE | ID: mdl-35270990

ABSTRACT

Hydrographic surveys enable the acquisition and processing of bathymetric data, which after being plotted onto nautical charts, can help to ensure safety of navigation, monitor changes in the coastal zone, and assess hydro-engineering structure conditions. This study involves the measurement of waterbody depth, identification of the seabed shape and geomorphology, the coastline course, and the location of underwater obstacles. Hydroacoustic systems mounted on vessels are commonly used in bathymetric measurements. However, there is also an increasing use of Unmanned Aerial Vehicles (UAV) that can employ sensors such as LiDAR (Light Detection And Ranging) or cameras previously not applied in hydrography. Current systems based on photogrammetric and remote sensing methods enable the determination of shallow waterbody depth with no human intervention and, thus, significantly reduce the duration of measurements, especially when surveying large waterbodies. The aim of this publication is to present and compare methods for determining shallow waterbody depths based on an analysis of images taken by UAVs. The perspective demonstrates that photogrammetric techniques based on the SfM (Structure-from-Motion) and MVS (Multi-View Stereo) method allow high accuracies of depth measurements to be obtained. Errors due to the phenomenon of water-wave refraction remain the main limitation of these techniques. It was also proven that image processing based on the SfM-MVS method can be effectively combined with other measurement methods that enable the experimental determination of the parameters of signal propagation in water. The publication also points out that the Lyzenga, Satellite-Derived Bathymetry (SDB), and Stumpf methods allow satisfactory depth measurement results to be obtained. However, they require further testing, as do methods using the optical wave propagation properties.


Subject(s)
Image Processing, Computer-Assisted , Unmanned Aerial Devices , Photogrammetry , Water
7.
Front Neuroanat ; 16: 838567, 2022.
Article in English | MEDLINE | ID: mdl-35356436

ABSTRACT

The pretectum has a distinct nuclear arrangement and complex neurochemical anatomy. While previous genoarchitectural studies have described rostrocaudal and dorsoventral progenitor domains and subdomains in different species, the relationship between these early partitions and its later derivatives in the mature anatomy is less understood. The signals and transcription factors that control the establishment of pretectal anatomy are practically unknown. We investigated the possibility that some aspects of the development of pretectal divisions are controlled by Wnt signaling, focusing on the transitional stage between neurogenesis and histogenesis in zebrafish. Using several molecular markers and following the prosomeric model, we identified derivatives from each rostrocaudal pretectal progenitor domain and described the localization of gad1b-positive GABAergic and vglut2.2-positive glutamatergic cell clusters. We also attempted to relate these clusters to pretectal nuclei in the mature brain. Then, we examined the influence of Wnt signaling on the size of neurochemically distinctive pretectal areas, using a chemical inhibitor of the Wnt pathway and the CRISPR/Cas9 approach to knock out genes that encode the Wnt pathway mediators, Lef1 and Tcf7l2. The downregulation of the Wnt pathway led to a decrease in two GABAergic clusters and an expansion of a glutamatergic subregion in the maturing pretectum. This revealed an instructive role of the Wnt signal in the development of the pretectum during neurogenesis. The molecular anatomy presented here improves our understanding of pretectal development during early postmitotic stages and support the hypothesis that Wnt signaling is involved in shaping the neurochemical organization of the pretectum.

8.
Cells ; 11(1)2022 01 01.
Article in English | MEDLINE | ID: mdl-35011697

ABSTRACT

In the adult brain, new neurons are constitutively derived from postnatal neural stem cells/progenitors located in two neurogenic regions: the subventricular zone (SVZ) of the lateral ventricles (migrating and differentiating into different subtypes of the inhibitory interneurons of the olfactory bulbs), and the subgranular layer of the hippocampal dentate gyrus. Cyclin D2 knockout (cD2-KO) mice exhibit reduced numbers of new hippocampal neurons; however, the proliferation deficiency and the dysregulation of adult neurogenesis in the SVZ required further investigation. In this report, we characterized the differentiation potential of each subpopulation of the SVZ neural precursors in cD2-KO mice. The number of newly generated cells in the SVZs was significantly decreased in cD2-KO mice compared to wild type mice (WT), and was not accompanied by elevated levels of apoptosis. Although the number of B1-type quiescent precursors (B1q) and the overall B1-type activated precursors (B1a) were not affected in the SVZ neurogenic niche, the number of transit-amplifying progenitors (TaPs) was significantly reduced. Additionally, the subpopulations of calbindin D28k and calretinin interneurons were diminished in the olfactory bulbs of cD2-KO mice. Our results suggest that cyclin D2 might be critical for the proliferation of neural precursors and progenitors in the SVZ-the transition of B1a into TaPs and, thereafter, the production of newly generated interneurons in the olfactory bulbs. Untangling regulators that functionally modulate adult neurogenesis provides a basis for the development of regenerative therapies for injuries and neurodegenerative diseases.


Subject(s)
Cyclin D2/metabolism , Lateral Ventricles/metabolism , Animals , Cell Differentiation , Cell Proliferation , Genotype , Mice , Mice, Knockout
9.
Sensors (Basel) ; 21(23)2021 Nov 25.
Article in English | MEDLINE | ID: mdl-34883835

ABSTRACT

The integration of geospatial data in hydrography, performed using different measurement systems, involves combining several study results to provide a comprehensive analysis. Each of the hydroacoustic and optoelectronic systems is characterised by a different spatial reference system and the method for technical implementation of the measurement. Therefore, the integration of hydrographic data requires that problems in selected fields of electronics, geodesy and physics (acoustics and optics) be solved. The aim of this review is to present selected fusion methods applying the data derived from Global Navigation Satellite System (GNSS), Real Time Kinematic (RTK) measurements, hydrographic surveys, a photogrammetric pass using unmanned vehicles and Terrestrial Laser Scanning (TLS) and compare their accuracy. An additional goal is the evalution of data integration methods according to the International Hydrographic Organization (IHO) S-44 standard. The publication is supplemented by implementation examples of the integration of geospatial data in the Geographic Information System (GIS). The methods described indicate the lack of a uniform methodology for data fusion due to differences in both the spatial reference systems and the techniques used. However, the integration of hydroacoustic and optoelectronic data allows for high accuracy geospatial data to be obtained. This is confirmed by the methods cited, in which the accuracy of integrated geospatial data was in the order of several centimetres.

10.
Int J Mol Sci ; 22(23)2021 Nov 26.
Article in English | MEDLINE | ID: mdl-34884588

ABSTRACT

Metastasis is one of the most urgent issues in breast cancer patients. One of the factors necessary in the migration process is the remodeling of the extracellular matrix (ECM). Metalloproteinases (MMPs) can break down the elements of the ECM, which facilitates cell movement. Many highly aggressive tumors are characterized by high levels of MMPs. In the case of breast cancer, the association between MMP-9 and the migration potential and invasiveness of cells has been demonstrated. In addition, reports indicating increased migration of breast cancer cells after the administration of the commonly used cytostatic cyclophosphamide (CP) are particularly disturbing. Hence, our research aimed to assess the effect of CP treatment on MDA-MB-231 and MCF-7 cells and how this response is influenced by the downregulation of the MMP-9 level. The obtained results suggest that CP causes a decrease in the survival of breast cancer cells of various invasiveness, and the downregulation of MMP-9 enhances this effect, mainly by inducing apoptosis. Moreover, in the group of MMP-9 siRNA-transfected CP-treated cells, a more severe reduction in invasion and migration of cells of both lines was observed, as indicated by the migration and invasion transwell assays and Wound healing assay. Hence, we suggest that CP alone may not result in satisfactory therapeutic effects. On the other hand, the use of combination therapy targeting MMP-9, together with the CP, could improve the effectiveness of the treatment. Additionally, we confirmed a relationship between the levels of MMP-9 and cytokeratin 19 (CK19).


Subject(s)
Breast Neoplasms/pathology , Cell Movement , Cyclophosphamide/pharmacology , Gene Expression Regulation, Neoplastic/drug effects , Matrix Metalloproteinase 9/chemistry , Antineoplastic Agents, Alkylating/pharmacology , Apoptosis , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Cycle , Cell Proliferation , Female , Humans , Keratin-19/genetics , Keratin-19/metabolism , Prognosis , Tumor Cells, Cultured
11.
Sensors (Basel) ; 21(18)2021 Sep 15.
Article in English | MEDLINE | ID: mdl-34577395

ABSTRACT

Mineral excavation is a common process throughout the world. The open pits remaining after the closure of a mine require well-considered and meticulous reclamation activities aimed at restoring the environmental properties of a given area. The inspections carried out in Poland indicate numerous irregularities in implementing the reclamation process. The research in this study was conducted in six measurement series and includes both chemical and olfactometry determinations by devices: multisensor portable gas detector and field olfactometer. Statistical analysis of the results obtained show high concentrations in ambient air of both chemical compounds (NH3, VOCs, H2S, CH3SH) and odour, excluding the possibility of occurrence in the pit of only waste types contained in the administrative decision on reclamation. In addition to the unpleasant odour, the listed compounds can have dangerous effects on the health and life of living organisms. This paper presents a suitable method of control and detection of irregularities in the conducted processes. The main advantage is the relatively low cost of purchasing sensors and field olfactometers compared to other devices, and the possibility to test the polluted air in situ, without the risk of chemical processes occurring during transport of gas samples to the laboratory.


Subject(s)
Air Pollutants , Odorants , Air Pollutants/analysis , Odorants/analysis , Olfactometry , Poland
12.
Int J Mol Sci ; 22(11)2021 Jun 04.
Article in English | MEDLINE | ID: mdl-34200045

ABSTRACT

Ischemic stroke is a disturbance in cerebral blood flow caused by brain tissue ischemia and hypoxia. We optimized a multifactorial in vitro model of acute ischemic stroke using rat primary neural cultures. This model was exploited to investigate the pro-viable activity of cell-penetrating peptides: arginine-rich Tat(49-57)-NH2 (R49KKRRQRRR57-amide) and its less basic analogue, PTD4 (Y47ARAAARQARA57-amide). Our model included glucose deprivation, oxidative stress, lactic acidosis, and excitotoxicity. Neurotoxicity of these peptides was excluded below a concentration of 50 µm, and PTD4-induced pro-survival was more pronounced. Circular dichroism spectroscopy and molecular dynamics (MD) calculations proved potential contribution of the peptide conformational properties to neuroprotection: in MD, Tat(49-57)-NH2 adopted a random coil and polyproline type II helical structure, whereas PTD4 adopted a helical structure. In an aqueous environment, the peptides mostly adopted a random coil conformation (PTD4) or a polyproline type II helical (Tat(49-57)-NH2) structure. In 30% TFE, PTD4 showed a tendency to adopt a helical structure. Overall, the pro-viable activity of PTD4 was not correlated with the arginine content but rather with the peptide's ability to adopt a helical structure in the membrane-mimicking environment, which enhances its cell membrane permeability. PTD4 may act as a leader sequence in novel drugs for the treatment of acute ischemic stroke.


Subject(s)
Brain Ischemia/prevention & control , Cell-Penetrating Peptides/pharmacology , Disease Models, Animal , Ischemic Stroke/prevention & control , Neurons/drug effects , Neuroprotective Agents/pharmacology , Animals , Brain Ischemia/etiology , Brain Ischemia/pathology , Cell Membrane Permeability , Female , Ischemic Stroke/etiology , Ischemic Stroke/pathology , Rats , Rats, Wistar
13.
Cancer Manag Res ; 13: 91-106, 2021.
Article in English | MEDLINE | ID: mdl-33447082

ABSTRACT

PURPOSE: Metastasis remains a serious clinical problem in which epithelial-to-mesenchymal transition is strictly involved. The change of cell phenotype is closely related to the dynamics of the cytoskeleton. Regarding the great interest in microfilaments, the manipulation of ABPs (actin-binding proteins) appears to be an interesting treatment strategy. MATERIAL: The research material was the highly aggressive A549 cells with FHOD1 (F FH1/FH2 domain-containing protein 1) downregulation. The metastatic potential of the cells and the sensitivity to treatment with alkaloids (piperlongumine, sanguinarine) were analyzed. RESULTS: In comparison to A549 cells with naïve expression of FHOD1, those after manipulation were characterized by a reduced migratory potential. The obtained results were associated with microfilaments and vimentin reorganization induced by the manipulation of FHOD1 together with alkaloids treatment. The result was also an increase in the percentage of late apoptotic cells. CONCLUSION: Downregulation of FHOD1 induced reorganization of microfilament network followed by the reduction in the metastatic potential of the A549 cells, as well as their sensitization to selected compounds. The presented results and the analysis of clinical data indicate the possibility of transferring research from the basic level to in vivo models in the context of manipulation of ABPs as a new therapeutic target in oncology.

14.
Molecules ; 25(24)2020 Dec 18.
Article in English | MEDLINE | ID: mdl-33352986

ABSTRACT

Benzoxaboroles emerged recently as molecules of high medicinal potential with Kerydin® (Tavaborole) and Eucrisa® (Crisaborole) currently in clinical practice as antifungal and anti-inflammatory drugs, respectively. Over a dozen of 3-amino benzoxaboroles, including Tavaborole's derivatives, have been synthetized and characterized in terms of their activity against Candida albicans as a model pathogenic fungus. The studied compounds broaden considerably the structural diversity of reported benzoxaboroles, enabling determination of the influence of the introduction of a heterocyclic amine, a fluorine substituent as well as the formyl group on antifungal activity of those compounds. The determined zones of the growth inhibition of examined microorganism indicate high diffusion of majority of the studied compounds within the applied media as well as their reasonable activity. The Minimum Inhibitory Concentration (MIC) values show that the introduction of an amine substituent in position "3" of the benzoxaborole heterocyclic ring results in a considerable drop in activity in comparison with Tavaborole (AN2690) as well as unsubstituted benzoxaborole (AN2679). In all studied cases the presence of a fluorine substituent at position para to the boron atom results in lower MIC values (higher activity). Interestingly, introduction of a fluorine substituent in the more distant piperazine phenyl ring does not influence MIC values. As determined by X-ray studies, introduction of a formyl group in proximity of the boron atom results in a considerable change of the boronic group geometry. The presence of a formyl group next to the benzoxaborole unit is also detrimental for activity against Candida albicans.


Subject(s)
Antifungal Agents/pharmacology , Candida albicans/drug effects , Antifungal Agents/chemical synthesis , Antifungal Agents/chemistry , Microbial Sensitivity Tests , Molecular Structure
15.
Acta Histochem ; 122(7): 151625, 2020 Oct.
Article in English | MEDLINE | ID: mdl-33066845

ABSTRACT

The aim of this investigation was to determine the effect of doxorubicin on F-actin rearrangement and ß-catenin and cofilin-1 in a rat glioma C6 cell line in combination with changes in their morphology and ultrastructure. The experimental material constituted rat glioma C6 cell line. The cells were incubated with sublethal doses of doxorubicin in the concentration of 50, 100 and 200 nM. The blue trypan dye method was used to determine the number of dead cells. Morphological and ultrastructural changes in the cells were evaluated using light and transmission electron microscope, respectively. In order to determine the rearrangements and level of expression of F-actin, ß-catenin and cofilin-1 they were analyzed using a fluorecence microscope. In turn, cell death and cell cycle were evaluated by Guava 6HT-2 L Cytometer. The performed experiments showed a dose-dependent decrease in the survival of C6 cells after treatment with doxorubicin. The analysis of cell death showed a dose-dependent increase in the population of apoptotic and necrotic cells. These results were confirmed by microscopy observation. The changes in morphology, ultrastructure, and rearrangements of F-actin, ß-catenin and cofilin-1 were also observed. The results obtained in the study showed that sublethal concentrations of doxorubicin influenced the structure of F-actin and other proteins involved in cell-cell interactions. Moreover, mitotic catastrophe may preceding apoptosis, what suggest the cytotoxic effect of low dose of doxorubicin. Furthermore, our results confirmed the multi-dimensional mechanism of DOX action in tumor cells.


Subject(s)
Cell Cycle/drug effects , Cell Death/drug effects , Doxorubicin/pharmacology , Glioma/drug therapy , Actins/metabolism , Animals , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Glioma/ultrastructure
16.
Cells ; 9(10)2020 10 06.
Article in English | MEDLINE | ID: mdl-33036298

ABSTRACT

The actin cytoskeleton plays a crucial role in many cellular processes while its reorganization is important in maintaining cell homeostasis. However, in the case of cancer cells, actin and ABPs (actin-binding proteins) are involved in all stages of carcinogenesis. Literature has reported that ABPs such as SATB1 (special AT-rich binding protein 1), WASP (Wiskott-Aldrich syndrome protein), nesprin, and villin take part in the initial step of carcinogenesis by regulating oncogene expression. Additionally, changes in actin localization promote cell proliferation by inhibiting apoptosis (SATB1). In turn, migration and invasion of cancer cells are based on the formation of actin-rich protrusions (Arp2/3 complex, filamin A, fascin, α-actinin, and cofilin). Importantly, more and more scientists suggest that microfilaments together with the associated proteins mediate tumor vascularization. Hence, the presented article aims to summarize literature reports in the context of the potential role of actin and ABPs in all steps of carcinogenesis.


Subject(s)
Actins/metabolism , Carcinogenesis/metabolism , Microfilament Proteins/metabolism , Cell Movement , Humans
17.
Int J Mol Sci ; 21(17)2020 Aug 27.
Article in English | MEDLINE | ID: mdl-32867146

ABSTRACT

The tripeptide NH2-Gly-His-Lys-COOH (GHK), cis-urocanic acid (cis-UCA) and Cu(II) ions are physiological constituents of the human body and they co-occur (e.g., in the skin and the plasma). While GHK is known as Cu(II)-binding molecule, we found that urocanic acid also coordinates Cu(II) ions. Furthermore, both ligands create ternary Cu(II) complex being probably physiologically functional species. Regarding the natural concentrations of the studied molecules in some human tissues, together with the affinities reported here, we conclude that the ternary complex [GHK][Cu(II)][cis-urocanic acid] may be partly responsible for biological effects of GHK and urocanic acid described in the literature.


Subject(s)
Copper/chemistry , Oligopeptides/chemistry , Urocanic Acid/chemistry , Chelating Agents/chemistry , Chelating Agents/pharmacology , Circular Dichroism , Electron Spin Resonance Spectroscopy , Humans , Imidazoles/chemistry , Oligopeptides/pharmacology , Protein Multimerization , Serum/chemistry , Urocanic Acid/pharmacology
18.
Development ; 147(16)2020 08 25.
Article in English | MEDLINE | ID: mdl-32675279

ABSTRACT

Neuronal phenotypes are controlled by terminal selector transcription factors in invertebrates, but only a few examples of such regulators have been provided in vertebrates. We hypothesised that TCF7L2 regulates different stages of postmitotic differentiation in the thalamus, and functions as a thalamic terminal selector. To investigate this hypothesis, we used complete and conditional knockouts of Tcf7l2 in mice. The connectivity and clustering of neurons were disrupted in the thalamo-habenular region in Tcf7l2-/- embryos. The expression of subregional thalamic and habenular transcription factors was lost and region-specific cell migration and axon guidance genes were downregulated. In mice with a postnatal Tcf7l2 knockout, the induction of genes that confer thalamic terminal electrophysiological features was impaired. Many of these genes proved to be direct targets of TCF7L2. The role of TCF7L2 in terminal selection was functionally confirmed by impaired firing modes in thalamic neurons in the mutant mice. These data corroborate the existence of master regulators in the vertebrate brain that control stage-specific genetic programmes and regional subroutines, maintain regional transcriptional network during embryonic development, and induce terminal selection postnatally.


Subject(s)
Cell Differentiation , Gene Expression Regulation, Developmental , Mitosis , Synaptic Transmission , Thalamus/embryology , Transcription Factor 4/metabolism , Animals , Mice , Mice, Knockout , Thalamus/cytology , Transcription Factor 4/genetics
19.
Molecules ; 25(13)2020 Jul 03.
Article in English | MEDLINE | ID: mdl-32635287

ABSTRACT

BACKGROUND: Cancers are one of the leading causes of deaths nowadays. The development of new treatment schemes for oncological diseases is an interesting direction in experimental medicine. Therefore, the evaluation of the influence of two alkaloids-piperlongumine (PL), sanguinarine (SAN) and their combination-on the basic life processes of the A549 cell line was considered reasonable. METHODS: The aim was achieved by analyzing the cytotoxic effects of PL and SAN and their combination in the ratio of 4:1 on the induction of cell death, changes in the distribution of cell cycle phases, reorganization of cytoskeleton and metastatic potential of A549 cells. The versatility of the applied concentration ratio was evaluated in terms of other cancer cell lines: MCF-7, H1299 and HepG2. RESULTS: The results obtained from the MTT assay indicated that the interaction between the alkaloids depends on the concentration and type of cells. Additionally, the compounds and their combination did not exhibit a cytotoxic effect against normal cells. The combined effects of PL and SAN increased apoptosis and favored metastasis inhibition. CONCLUSION: Selected alkaloids exhibit a cytotoxic effect on A549 cells. In turn, treatment with the combination of PL and SAN in a 4:1 ratio indicates a synergistic effect and is associated with an increase in the level of reactive oxygen species (ROS).


Subject(s)
Benzophenanthridines/pharmacology , Carcinoma, Non-Small-Cell Lung/drug therapy , Dioxolanes/pharmacology , Drug Synergism , Isoquinolines/pharmacology , Lung Neoplasms/drug therapy , Anti-Infective Agents/pharmacology , Apoptosis , Carcinoma, Non-Small-Cell Lung/pathology , Cell Cycle , Cell Movement , Cell Proliferation , Humans , Lung Neoplasms/pathology , Neoplasm Invasiveness , Reactive Oxygen Species/metabolism , Tumor Cells, Cultured
20.
Sci Total Environ ; 727: 138709, 2020 Jul 20.
Article in English | MEDLINE | ID: mdl-32334232

ABSTRACT

Wetland buffer zones (WBZs) are riparian areas that form a transition between terrestrial and aquatic environments and are well-known to remove agricultural water pollutants such as nitrogen (N) and phosphorus (P). This review attempts to merge and compare data on the nutrient load, nutrient loss and nutrient removal and/or retention from multiple studies of various WBZs termed as riparian mineral soil wetlands, groundwater-charged peatlands (i.e. fens) and floodplains. Two different soil types ('organic' and 'mineral'), four different main water sources ('groundwater', 'precipitation', 'surface runoff/drain discharge', and 'river inundation') and three different vegetation classes ('arboraceous', 'herbaceous' and 'aerenchymous') were considered separately for data analysis. The studied WBZs are situated within the temperate and continental climatic regions that are commonly found in northern-central Europe, northern USA and Canada. Surprisingly, only weak differences for the nutrient removal/retention capability were found if the three WBZ types were directly compared. The results of our study reveal that for example the nitrate retention efficiency of organic soils (53 ± 28%; mean ± sd) is only slightly higher than that of mineral soils (50 ± 32%). Variance in load had a stronger influence than soil type on the N retention in WBZs. However, organic soils in fens tend to be sources of dissolved organic N and soluble reactive P, particularly when the fens have become degraded due to drainage and past agricultural usage. The detailed consideration of water sources indicated that average nitrate removal efficiencies were highest for ground water (76 ± 25%) and lowest for river water (35 ± 24%). No significant pattern for P retention emerged; however, the highest absolute removal appeared if the P source was river water. The harvesting of vegetation will minimise potential P loss from rewetted WBZs and plant biomass yield may promote circular economy value chains and provide compensation to land owners for restored land now unsuitable for conventional farming.


Subject(s)
Phosphorus/analysis , Wetlands , Canada , Europe , Hydrology , Nitrogen/analysis , Soil
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