The enzyme creatine kinase (CK) plays a key role in energy homeostasis of cells with high and fluctuating energy requirements. As for spermatozoa, the activity of phosphocreatine shuttle, which directs energy from the mitochondria to sites of ATP consumption, is dependent on individual species. High activities of CK are observed in spermatozoa of nonvertebrate, fish, and birds, contrary to the low-level CK activity in mammalian spermatozoa. A new monoclonal antibody (MAb) to carp sperm creatine kinase was prepared. This antibody is applicable to large-scale immunochemical techniques. In this study, it was applied to the study of carp sperm motility, and the evaluation of the influence of CK on the quality and fertilization ability of carp spermatozoa.

• MeSH
• kapři imunologie   MeSH
• kreatinkinasa imunologie   MeSH
• lidé MeSH
• monoklonální protilátky fyziologie   MeSH
• prasata MeSH
• spermie enzymologie   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kreatinkinasa MeSH
• monoklonální protilátky MeSH

Background/Objectives: Dual-modality probes, combining positron emission tomography (PET) with fluorescence imaging (FI) capabilities in a single molecule, are of high relevance for the accurate staging and guided resection of tumours. We herein present a pair of candidates targeting the cholecystokinin-2 receptor (CCK2R), namely [68Ga]Ga-CyTMG and [68Ga]Ga-CyFMG. In these probes, the SulfoCy5.5 fluorophore and two units of a CCK2R-binding motif are coupled to the chelator acting as a core scaffold, triazacyclononane-phosphinic acid (TRAP), and Fusarinine C (FSC), respectively. Using this approach, we investigated the influence of these chelators on the final properties. Methods: The synthetic strategy to both precursors was based on the stoichiometric conjugation of the components via click chemistry. The characterization in vitro included the evaluation of the CCK2R affinity and internalization in A431-CCK2R cells. Ex vivo biodistribution as well as PET and FI studies were performed in xenografted mice. Results: 68Ga labelling was accomplished with high radiochemical yield and purity for both precursors. A CCK2R affinity in the subnanomolar range of the conjugates and a receptor-specific uptake of the radioligands in cells were observed. In A431-CCK2R/A431-mock xenografted mice, the investigated compounds showed specific accumulation in the tumours and reduced off-target uptake compared to a previously developed compound. Higher accumulation and prolonged retention in the kidneys were observed for [68Ga]Ga-CyTMG when compared to [68Ga]Ga-CyFMG. Conclusions: Despite the promising targeting properties observed, further probe optimization is required to achieve enhanced imaging contrast at early timepoints. Additionally, the results indicate a distinct influence of the chelators in terms of renal accumulation and retention.

• Klíčová slova
• FSC, PET, SulfoCy5.5, TRAP, cholecystokinin-2 receptor, dual-modality imaging agent, fluorescence guided surgery, gallium-68,
• Publikační typ
• časopisecké články MeSH

PURPOSE: Multimeric arginine-glycine-aspartic acid (RGD) peptides have advantages for imaging integrin αvβ3 expression. Here, we compared the in vitro and in vivo behavior of three different Ga-68-labeled multimeric Fusarinine C-RGD (FSC-RGD) conjugates, whereby RGD was coupled directly, via a succinic acid or PEG linker (FSC(RGDfE)3, FSC(succ-RGD)3, FSC(Mal-RGD)3). The positron emission tomography/X-ray computed tomography (PET/CT) imaging properties were further compared using [(68)Ga]FSC(succ-RGD)3 with the monomeric [(68)Ga]NODAGA-RGD in a murine tumor model. PROCEDURE: FSC-RGD conjugates were labeled with Ga-68, and stability properties were studied. For in vitro characterization, the partition coefficient, integrin αvβ3 binding affinity, and cell uptake were determined. To characterize the in vivo properties, biodistribution studies and microPET/CT were carried out using mice bearing either human M21/M21-L melanoma or human U87MG glioblastoma tumor xenografts. RESULTS: All FSC-RGD conjugates were quantitatively labeled with Ga-68 within 10 min at RT. The [(68)Ga]FSC-RGD conjugates exhibited high stability and hydrophilic character, with only minor differences between the different conjugates. In vitro and in vivo studies showed enhanced integrin αvβ3 binding affinity, receptor-selective tumor uptake, and rapid renal excretion resulting in good imaging properties. CONCLUSIONS: The type of linker between FSC and RGD had no pronounced effect on targeting properties of [(68)Ga]FSC-RGD trimers. In particular, [(68)Ga]FSC(succ-RGD)3 exhibited improved properties compared to [(68)Ga]NODAGA-RGD, making it an alternative for imaging integrin αvβ3 expression.

• Klíčová slova
• Fusarinine C, Gallium-68, Integrins, Positron emission tomography (PET), RGD peptides,
• MeSH
• acetáty chemie   MeSH
• endocytóza MeSH
• heterocyklické sloučeniny monocyklické chemie   MeSH
• kyseliny hydroxamové chemie   MeSH
• lidé MeSH
• melanom diagnostické zobrazování   patologie   MeSH
• myši inbrední BALB C MeSH
• myši nahé MeSH
• nádorové buněčné linie MeSH
• oligopeptidy chemie   MeSH
• PET/CT * MeSH
• radiofarmaka chemie   MeSH
• radioizotopy galia MeSH
• tkáňová distribuce MeSH
• xenogenní modely - testy antitumorózní aktivity * MeSH
• železité sloučeniny chemie   MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• 1-(1,3-carboxypropyl)-4,7-carboxymethyl-1,4,7-triazacyclononane MeSH Prohlížeč
• acetáty MeSH
• arginyl-glycyl-aspartic acid MeSH Prohlížeč
• fusigen MeSH Prohlížeč
• heterocyklické sloučeniny monocyklické MeSH
• kyseliny hydroxamové MeSH
• oligopeptidy MeSH
• radiofarmaka MeSH
• radioizotopy galia MeSH
• železité sloučeniny MeSH

Flexible supercapacitors (FSCs) have received a lot of interest as portable power sources for wearable electronics. The biocompatibility of electrodes and electrolytes in wearable FSCs is important to consider although research into these topics is still in its early stages. In this work, we developed a wearable FSC that uses MXene Ti3C2 nanosheets and polypyrrole-carboxymethylcellulose nanospheres composite (Ti3C2@PPy-CMC) as the active electrode material and sweat as the electrolyte. The electrochemical performances of Ti3C2@PPy-CMC FSC were analyzed using an artificial sweat solution and exhibited excellent specific capacitance, power density, cycling stability, and bending stability. To demonstrate a real application of Ti3C2@PPy-CMC FSC, a sweat-chargeable FSC patch has been developed that can be applied directly to human clothing and skin to power a portable electronic gadget when the wearer is exercising. A comprehensive electrochemical study of the FSC patch was also conducted in various sweat secretion body regions such as the finger, foot sole, and wrist. Ti3C2@PPy-CMC composite's outstanding electrochemical performance indicates its potential capabilities and biocompatibility in wearable energy storage devices.

• Klíčová slova
• 2D materials, Biofluid, Flexible supercapacitor, Healthcare, Portable power source, Wearable bioelectronics,
• MeSH
• běh * MeSH
• biosenzitivní techniky * MeSH
• lidé MeSH
• nositelná elektronika * MeSH
• polymery MeSH
• pot MeSH
• pyrroly MeSH
• titan MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• polymery MeSH
• pyrroly MeSH
• titan MeSH

Filipin, a membrane beta-hydroxysterol probe, and polymyxin B (PXB), a probe for anionic lipids, were used to study human sperm plasma membrane (PM) with particular reference to changes induced by capacitation in vitro. In washed but noncapacitated spermatozoa the density of filipin/sterol complexes (FSC) was uniformly high in the PM overlying the acrosome, without any differences between its anterior and equatorial regions. The postacrosomal region was poor in FSC. The exclusion of FSC from small areas of the PM covering the two acrosomal regions and reduction of their number in the postacrosomal region were the main changes induced by sperm capacitation. Unlike filipin, PXB produced a capacitation-independent reaction pattern characterized by a high reactivity of the PM covering the anterior portion of the acrosome, probably conferring to this region its potential fusigenicity temporarily antagonized by the elevated sterol content. Only an exceptional and slight PXB binding was seen in the nonfusigenic equatorial and postacrosomal regions.

• MeSH
• buněčná membrána fyziologie   ultrastruktura   MeSH
• elektronová mikroskopie MeSH
• kapacitace spermií * MeSH
• lidé MeSH
• membránové lipidy analýza   MeSH
• motilita spermií MeSH
• mrazové lámání MeSH
• spermie fyziologie   ultrastruktura   MeSH
• steroly analýza   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• membránové lipidy MeSH
• steroly MeSH

The coexistence of lipid domains with different degrees of lipid packing in the plasma membrane of mammalian cells has been postulated, but direct evidence has so far been challenging to obtain because of the small size and short lifetime of these domains in live cells. Here, we use fluorescence spectral correlation spectroscopy in conjunction with a probe sensitive to the membrane environment to quantify spectral fluctuations associated with dynamics of membrane domains in live cells. With this method, we show that membrane domains are present in live COS-7 cells and have a lifetime lower bound of 5.90 and 14.69 ms for the ordered and disordered phases, respectively. Comparisons to simulations indicate that the underlying mechanism of these fluctuations is complex but qualitatively described by a combination of dye diffusion between membrane domains as well as the motion of domains within the membrane.

• MeSH
• benzoxaziny chemie   MeSH
• buněčná membrána chemie   MeSH
• Cercopithecus aethiops MeSH
• COS buňky MeSH
• fluorescenční spektrometrie MeSH
• kvartérní amoniové sloučeniny chemie   MeSH
• membránové lipidy chemie   MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• benzoxaziny MeSH
• kvartérní amoniové sloučeniny MeSH
• membránové lipidy MeSH
• NR12S compound MeSH Prohlížeč

OBJECTIVE: Confocal laser scanning endomicroscopy (CLSE) is a diagnostic technology that produces virtual histology of the mucosal layer using fluorescence technique. Fluorescein (FSC) is the most commonly used fluorescence agent. Fluorescence light coming from a horizontal special focal plane is detected during confocal laser endomicroscopy of the gastrointestinal tract. FSC causes intensive yellowish discoloration of tissues, including skin and mucous membranes. This pre-clinical study was aimed to evaluate the tissue distribution and pharmacokinetics of FSC after its intravenous administration. METHODS: The study was performed in an adult experimental pig. A reversed-phase high-performance liquid chromatographic method with fluorescence detection was used for the determination of fluorescein in blood plasma and tissue samples. RESULTS AND CONCLUSION: The pharmacokinetic study of fluorescein determined the optimum time interval for diagnostic scanning (5-10 min.) The biodistribution study of fluorescein (aimed on the potential organ accumulation) proved the high concentration in the renal system followed by levels in bile > lung > adipose tissue > all other organs (including gastrointestinal wall) and these were relatively similar to each other. Fluorescein has a significantly low distribution in the brain (contrast with the level in adipose tissue indicates the low ability to penetrate the blood-brain barrier).

• MeSH
• fluorescein aplikace a dávkování   farmakokinetika   MeSH
• fluorescenční barviva aplikace a dávkování   farmakokinetika   MeSH
• gastrointestinální trakt cytologie   metabolismus   MeSH
• hematoencefalická bariéra MeSH
• injekce intravenózní MeSH
• konfokální mikroskopie * MeSH
• ledviny cytologie   metabolismus   MeSH
• modely u zvířat MeSH
• mozek cytologie   metabolismus   MeSH
• plíce cytologie   metabolismus   MeSH
• prasata MeSH
• tuková tkáň cytologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fluorescein MeSH
• fluorescenční barviva MeSH

Within the last years (89)Zr has attracted considerable attention as long-lived radionuclide for positron emission tomography (PET) applications. So far desferrioxamine B (DFO) has been mainly used as bifunctional chelating system. Fusarinine C (FSC), having complexing properties comparable to DFO, was expected to be an alternative with potentially higher stability due to its cyclic structure. In this study, as proof of principle, various FSC-RGD conjugates targeting αvß3 integrins were synthesized using different conjugation strategies and labeled with (89)Zr. In vitro stability, biodistribution, and microPET/CT imaging were evaluated using [(89)Zr]FSC-RGD conjugates or [(89)Zr]triacetylfusarinine C (TAFC). Quantitative (89)Zr labeling was achieved within 90 min at room temperature. The distribution coefficients of the different radioligands indicate hydrophilic character. Compared to [(89)Zr]DFO, [(89)Zr]FSC derivatives showed excellent in vitro stability and resistance against transchelation in phosphate buffered saline (PBS), ethylenediaminetetraacetic acid solution (EDTA), and human serum for up to 7 days. Cell binding studies and biodistribution as well as microPET/CT imaging experiments showed efficient receptor-specific targeting of [(89)Zr]FSC-RGD conjugates. No bone uptake was observed analyzing PET images indicating high in vivo stability. These findings indicate that FSC is a highly promising chelator for the development of (89)Zr-based PET imaging agents.

• Klíčová slova
• 89Zr, RGD peptide, fusarinine C, positron emission tomography (PET), triacetylfusarinine C,
• MeSH
• chelátory chemie   MeSH
• kyseliny hydroxamové chemie   MeSH
• lidé MeSH
• oligopeptidy chemie   MeSH
• pozitronová emisní tomografie MeSH
• radionuklidy chemie   MeSH
• železité sloučeniny chemie   MeSH
• zirkonium chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• arginyl-glycyl-aspartic acid MeSH Prohlížeč
• chelátory MeSH
• fusigen MeSH Prohlížeč
• kyseliny hydroxamové MeSH
• N,N',N''-triacetylfusarinine C MeSH Prohlížeč
• oligopeptidy MeSH
• radionuklidy MeSH
• železité sloučeniny MeSH
• zirkonium MeSH

Flexible energy storage devices are becoming significantly important to power wearable and portable devices that monitor physiological parameters for many biomedical applications. Many hybrid nanomaterials based on 2D materials are used in order to improve the performance of flexible energy storage devices. Here, a hybrid nanocomposite is synthesized through in situ polymerization of aniline in the presence of black phosphorus (BP) nanoflakes. This nanocomposite, polyaniline (PANI)@BP, is employed to fabricate flexible supercapacitor (FSC) electrodes. PANI@BP FSCs can provide a power source for biometric devices. The generated signal can be transmitted to a smartphone in real time via wireless communication. Such a compact and lightweight integrated device has been used to track a human heart beat while powered by PANI@BP FSC. These findings are providing a promising example of a flexible energy storage device that can be integrated with different real-time health monitoring devices.

• Klíčová slova
• 2D materials, conductive polymers, flexible energy storage devices, health monitoring devices, portable electronics,
• MeSH
• aniliny MeSH
• biologický monitoring MeSH
• fosfor * MeSH
• lidé MeSH
• nanokompozity * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aniliny MeSH
• fosfor * MeSH
• polyaniline MeSH Prohlížeč

PURPOSE: Aspergillus fumigatus produces the siderophore triacetylfusarinine C (TAFC) for iron acquisition which is essential for its virulence. Therefore, TAFC is a specific marker for invasive aspergillosis. We have shown previously that positron emission tomography (PET) imaging with [68Ga]TAFC exhibited excellent targeting properties in an A. fumigatus rat infection model. In this study, we aimed to prepare TAFC analogs modifying fusarinine C (FSC) by acylation with different carbon chain lengths as well as with charged substituents and investigated the influence of introduced substituents on preservation of TAFC characteristics in vitro and in vivo. PROCEDURES: Fifteen TAFC derivatives were prepared and labeled with gallium-68. In vitro uptake assays were carried out in A. fumigatus under iron-replete as well as iron-depleted conditions and distribution coefficient was determined. Based on these assays, three compounds, [68Ga]tripropanoyl(FSC) ([68Ga]TPFC), [68Ga]diacetylbutanoyl(FSC) ([68Ga]DABuFC), and [68Ga]trisuccinyl(FSC) ([68Ga]FSC(suc)3), with high, medium, and low in vitro uptake in fungal cultures, were selected for further evaluation. Stability and protein binding were evaluated and in vivo imaging performed in the A. fumigatus rat infection model. RESULTS: In vitro uptake studies using A. fumigatus revealed specific uptake of mono- and trisubstituted TAFC derivatives at RT. Lipophilicities as expressed by logD were 0.34 to - 3.80. The selected compounds displayed low protein binding and were stable in PBS and serum. Biodistribution and image contrast in PET/X-ray computed tomography of [68Ga]TPFC and [68Ga]DABuFC were comparable to [68Ga]TAFC, whereas no uptake in the infected region was observed with [68Ga]FSC(suc)3. CONCLUSIONS: Our studies show the possibility to modify TAFC without losing its properties and specific recognition by A. fumigatus. This opens also new ways for multimodality imaging or theranostics of fungal infection by introducing functionalities such as fluorescent dyes or antifungal moieties.

• Klíčová slova
• Aspergillus fumigatus, Gallium-68, Infection imaging, PET, Siderophores, Triacetylfusarinine C,
• MeSH
• Aspergillus fumigatus fyziologie   MeSH
• krevní proteiny metabolismus   MeSH
• krysa rodu Rattus MeSH
• kyseliny hydroxamové chemie   MeSH
• lidé MeSH
• molekulární zobrazování * MeSH
• mutace genetika   MeSH
• mykózy diagnostické zobrazování   mikrobiologie   MeSH
• myši inbrední BALB C MeSH
• PET/CT MeSH
• plicní aspergilóza diagnostické zobrazování   mikrobiologie   MeSH
• radioizotopy galia chemie   MeSH
• siderofory chemická syntéza   chemie   MeSH
• tkáňová distribuce MeSH
• vazba proteinů MeSH
• železité sloučeniny chemie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Gallium-68 MeSH Prohlížeč
• krevní proteiny MeSH
• kyseliny hydroxamové MeSH
• N,N',N''-triacetylfusarinine C MeSH Prohlížeč
• radioizotopy galia MeSH
• siderofory MeSH
• železité sloučeniny MeSH