Solutions Of Spermine Research Articles

Application of the "Plazmontest" Software and Hardware Complex for the Determination of Spermine Concentration

Introduction. One of the main areas of application of SPR sensors is biochemical analysis. The "Plasmontest" device, developed at the Institute of Cybernetics of the National Academy of Sciences of Ukraine, was used for rapid diagnosis of various concentrations of the tumor marker spermine. Colloidal gold nanoparticles stabilized by sodium citrate were used as a sensitive layer for the detection of biogenic polyamines as markers of prostate cancer development. Two methods of conducting an experiment on spermine detection using colloidal gold nanoparticles are proposed. The purpose of the paper is to show the possibility of detecting the tumor marker spermine in different concentrations using the "Plasmontest" device. To solve this problem it is necessary to provide a series of experiments on the application of colloidal gold solutions with different methods of deposition of reagents. The results. With the help of the "Plasmontest" software and technical complex, a series of biochemical experiments were carried out using solutions of spermine in the concentration range of 50 nM - 500 nM and colloidal gold in a concentration of 3·10-4 M. Two methods of applying reagents were proposed: spermine – colloidal gold and colloidal gold – spermine. Both techniques made it possible to observe a shift in the SPR angle, which means the adsorption of spermine on the surface of a gold film with colloidal gold nanoparticles. Conclusions. It is shown that the developed software and technical complex "Plasmontest" allows monitoring the course of biochemical reactions at low concentrations of the investigated reagents. Studies of the possibility of detecting spermine in different concentrations in the presence of gold nanoparticles using the PPR method open the prospect of creating a biosensor for spermine. Keywords: surface plasmon resonance, biosensor, nanoparticles, spermine.

Supramolecular Chemotherapy Based on the Host-Guest Complex of Lobaplatin-Cucurbit[7]uril.

This work aimed to develop a supramolecular chemotherapy-based strategy to reduce the cytotoxicity of the antitumor drug lobaplatin (LbPt) toward normal human intestinal cells and recover its antitumor bioactivity toward human intestinal tumor cells. Through host-guest interactions, cucurbit[7]uril (CB[7])-encapsulated LbPt decreased the cytotoxicity of LbPt toward normal human intestinal cells, and even at a concentration of 100.0 μM, LbPt-CB[7] exhibited 77.4% higher safety over a 24 h period compared with LbPt. At pH 6.0, the binding affinity constant (Ka) of CB[7] and spermine was (1.18 ± 0.12) × 106 M-1, which is an order of magnitude higher than that of CB[7] with LbPt [Ka = (2.09 ± 0.07) × 105 M-1]; thus, the encapsulated LbPt can be released from its host-guest complex of LbPt-CB[7] through competitive replacement with spermine solution. The LbPt-CB[7] complex exhibited good in vitro performance for spermine as a biomarker tumor, as demonstrated with human intestinal tumor cells. These results indicate that LbPt is released from its host-guest complex by spermine in the tumor microenvironment, and supramolecular strategies will likely be extended to other clinical antitumor drugs to decrease their severe side effects in normal tissues and recover their antitumor bioactivity in tumors, which will enrich the field of safe supramolecular chemotherapy.

Molecular ring toss of circular BAC DNA using micropillar array for single-molecule studies.

This paper reports a method for trapping circular DNA molecules and imaging the dynamics with high spatial resolution using a micropillar-array device. We successfully trapped circular bacterial artificial chromosome DNA molecules at a micropillar-based "ring toss" in the laminar flow of a microchannel under a fluorescence microscope and demonstrated the imaging of their extension by flow and condensation process induced by spermine solution. DNA molecules were visualized in an extended loop conformation, allowing high spatial resolution, and the results showed that the dynamics is induced by the microfluidic control of the surrounding chemical environment. The method is expected to lead to the elucidation of the physical characteristics and the dynamics of circular DNA molecules.

Detection of polyamines using immunobiosensor basedonzinc oxide nanoparticles.

To analyze the performance of biosensor based on nanoparticles of zinc oxide for the detection of spermine and spermidine in solution and in cell culture. Zinc oxide nanoparticles were used for preparing biosensor containing antibodies to spermine and spermidine. Polyamine concentration is solutions of spermine and spermidine as well as in lyophilisate of MCF-7cells was measured by luminescence of the samples excited by laser beam at 380nm. The minimum concentration for the detection of polyamines in model solutions is 10ng/ml, and maximum one is 100ng/ml. A higher level of luminescence intensity of nanoparticles was found during analysis the polyamines in MCF-7lyophilisate allowing for detecting polyamines at concentrations from 100cells/ml to 100,000cells/ml. The proposed biosensor system for determining the level of biogenic polyamines in cell lyophilisate using the optical properties of zinc oxide nanoparticles is promising for further improvement of the methodology and its implementation for detection and measurement of polyamines in biological systems.

Effect of maturity stages on shelf life of papaya (Carica papaya L.)

An experiment using cv. Taiwan was 786 laid out in factorial completely randomized design with thirty treatments and three replications was, carried out at Laboratory of Horticulture Section, College of Agriculture, Dhule (M.S.) during 2017. The experiment comprised of three factors viz. two maturity stages (25% mature and 50% mature), dipping them in aqueous solutions of spermine (1.0 mM and 2.0 mM) and calcium lactate (1% and 2%) for 5 minutes and drying for 30 minutes at room temperature, three packaging materials (newspaper, shrink film and polyethylene bags) were used. Results indicated a significant impact of interaction among maturity stages, spermine and calcium lactate and packaging material on all parameters included in the study. When considered interaction, twenty five percent mature fruits dipped in (2.0 mM) spermine with shrink packaging had the minimum physiological loss in weight. This treatment also recorded the maximum peel to pulp ratio, fruit firmness. The maximum shelf life of 16 days without sacrificing organoleptic quality by manipulating the time of harvesting, packaging and application of spermine.

Effect of oral polyamine supplementation pre-weaning on piglet growth and intestinal characteristics

A high proportion of piglets fail to adapt to the changing composition of their diet at weaning, resulting in weight loss and increased susceptibility to pathogens. Polyamines are present in sow milk and promote neonatal maturation of the gut. We hypothesised that oral spermine and spermidine supplementation before weaning would increase piglet growth and promote gastrointestinal development at weaning. In Experiment One, one pair of liveweight (LW)-matched piglets per litter from first and third lactation sows received 2 ml of a 0 (Control) or 463 nmol/ml spermine solution at 14, 16, 18, 20 and 22 days of age (n=6 piglets/treatment per parity). Villus height and crypt depth in the duodenum and jejunum were measured at weaning (day 23 postpartum). In Experiment Two, piglets suckling 18 first and 18 third lactation sows were used. Within each litter, piglets received 2 ml of either water (Control), 463 nmol/ml spermine solution or 2013 nmol/ml spermidine solution at 14, 16, 18, 22 and 24 days of age (n=54 piglets/treatment per sow parity). Piglets were weighed individually at 14, 18, 24 (weaning) and 61 days of age. In Experiment One, oral spermine supplementation resulted in a 41% increase in villus height, a 21% decrease in crypt depth and 79% decrease in the villus height : crypt depth ratio compared with control piglets (P<0.01). In Experiment Two, spermine and spermidine-supplemented piglets suckling first lactation sows grew faster (P<0.05) between days 14 and 18 postpartum than control piglets: 0.230±0.011 and 0.227±0.012 v. 0.183±0.012 kg/day, respectively. Spermine supplementation tended (P<0.1) to increase piglet LW gain from weaning to day 37 post-weaning compared with control piglets (0.373±0.009 v. 0.341±0.010 kg/day). In conclusion, spermine supplementation increased villus height at weaning, and appears to have the potential to improve the pre- and post-weaning growth of conventionally weaned piglets.

Determining the chemical exchange saturation transfer (CEST) behavior of citrate and spermine under in vivo conditions.

PurposeTo estimate the exchange rates of labile 1H in citrate and spermine, metabolites present in prostatic secretions, to predict the size of the citrate and spermine CEST effects in vivo.MethodsCEST z‐spectra were acquired at high‐field [11.7 Tesla (T)] from citrate and spermine solutions at physiological pH (6.5) using saturation power 6 μT. CEST was performed at different temperatures to determine exchange regimes (slow, intermediate or fast). For low pH solutions of spermine, exchange rates were estimated from resonance line width, fitting z‐spectra using the Bloch equations incorporating exchange, and using quantifying exchange using saturation time experiments (QUEST). These rates were extrapolated to physiological pH.ResultsCitrate showed little CEST effect at pH 6.5 and temperature (T) = 310 K (maximum 0.001% mM‐1), indicating fast exchange, whereas spermine showed greater CEST effects (maximum 0.2% mM‐1) indicating intermediate‐to‐fast exchange. Extrapolating data acquired from low pH spermine solutions predicts exchange rates at pH 6.5 and T of 310 K of at least 2 × 104s‐1.ConclusionCitrate and spermine show minimal CEST effects at 11.7T even using high saturation power. These effects would be much less than 2% at clinical field‐strengths due to relatively faster exchange and would be masked by CEST from proteins. Magn Reson Med 76:742–746, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Application of natural and synthetic polyamines as growth regulators to improve the freezing tolerance of winter wheat (Triticum aestivum L.)

Wheat cultivars were grown as soil culture under normal growth conditions. Twoweek- old seedlings were exposed to 4°C for 6 h and then transferred to −12°C for 24 h in the dark. Twenty-four hours before freezing stress, some of the plants were sprayed with aqueous solutions of spermine, spermidine, putrescine, 1,3-diaminopropane (1,3-DAP) and diethylenetriamine (DETA). The data showed that freezing stress caused a decrease in the fresh weight, chlorophyll content and plant survival rate, accompanied by a simultaneous accumulation of free proline and the enhanced leakage of electrolytes. Preliminary treatment with polyamines caused a decline in electrolyte leakage and a considerable augmentation in proline quantity, indicating that the compounds are capable of preventing frost injury. Additionally, the foliar application of polyamines retarded the destruction of chlorophyll, and lessened fresh weight losses due to freezing stress. The synthetic triamine DETA was the most effective, having the most pronounced action in all the experiments, followed by the tetraamine spermine. The application of polyamines to wheat crops could be a promising approach for improving plant growth under unfavourable growth conditions, including freezing temperatures. The results demonstrate that treatment with polyamines could protect winter wheat by reducing the stress injuries caused by subzero temperatures.

A mucoadhesive nanoparticulate system for the simultaneous delivery of macromolecules and permeation enhancers to the intestinal mucosa

The feasibility of combining safe permeation enhancers in a mucoadhesive particulate system for the oral delivery of peptide drugs was investigated in this study. Polyelectrolyte complex nanoparticles (NPs) were prepared by ionic interaction of spermine (SPM) with polyacrylic acid (PAA) polymer. Cytotoxicity studies in Caco-2 monolayers revealed the safety of the delivery system in the concentration range used for permeation enhancement. The cellular transport of fluorescein isothiocyanate dextran (FD4) showed higher permeation enhancing profiles of SPM–PAA NPs, as compared to SPM solution or PAA NPs prepared by ionic gelation with MgCl 2 (Mg-PAA NPs). These permeation enhancing effects were associated with a reversible decrease in TEER values, suggesting a paracellular permeation pathway by reversible opening of the tight junctions. Furthermore, confocal microscopy results revealed strong association of the NPs prepared using fluorescence labeled PAA to Caco-2 cells. The permeation enhancing properties of SPM–PAA NPs were further evaluated in vivo after oral administration to rats, using FD4 and calcitonin as models of poorly permeating drugs. Confocal microscopy images of rats' small intestine confirmed previous findings in Caco-2 cells and revealed a strong and prolonged penetration of FD4 from the mucosal to the basolateral side of the intestinal wall. In addition, the proposed NPs were efficient in improving the oral absorption of calcitonin, as evidenced by the significant and prolonged reduction of the blood calcemia in rats.

Transient outward current carried by inwardly rectifying K+ channels in guinea pig ventricular myocytes dialyzed with low-K+ solution.

There have been periodic reports of nonclassic (4-aminopyridine insensitive) transient outward K+ current in guinea pig ventricular myocytes, with the most recent one describing a novel voltage-gated inwardly rectifying type. In the present study, we have investigated a transient outward current that overlaps inward Ca2+ current (I(Ca,L)) in myocytes dialyzed with 10 mM K+ solution and superfused with Tyrode's solution. Although depolarizations from holding potential (Vhp) -40 to 0 mV elicited relatively small inward I(Ca,L) in these myocytes, removal of external K+ or addition of 0.2 mM Ba2+ more than doubled the amplitude of the current. The basis of the enhancement of I(Ca,L) was the suppression of a large transient outward K+ current. Similar enhancement was observed when Vhp was moved to -80 mV and test depolarizations were preceded by short prepulses to -40 mV. Investigation of the time and voltage properties of the outward K+ transient indicated that it was inwardly rectifying and unlikely to be carried by voltage-gated channels. The outward transient was attenuated in myocytes dialyzed with high-Mg2+ solution, accelerated in myocytes dialyzed with 100 microM spermine solution, and abolished with time in myocytes dialyzed with ATP-free solution. These and other findings suggest that the outward transient is a component of classic "time-independent" inwardly rectifying K+ current.