Novel acid-degradable and thermo-sensitive poly(N-isopropylacrylamide) hydrogels cross-linked by α,α-trehalose diacetals

New trehalose-containing hydrogels and the first hydrogels cross-linked by mono- or disaccharide acetals that are degradable in acidic medium were designed and synthesized. The cross-linking agent was obtained in acetalization reaction of trehalose with x-allyloxybenzaldehyde in which diallyl compounds were formed and used afterward in free radical copolymerization with N-isopropylacrylamide (NIPAAm) to provide thermo-sensitive hydrogels. Depending on the trehalose diacetal isomer and solvent system used in copolymerization, as well the degree of network cross-linking, these hydrogels differed in morphology, swelling and deswelling behavior and time of degradation. Compared with conventional PNIPAAm hydrogels they are degradable what makes them good materials for biomedical applications.     

Studies on dimerisation of tocopherols under the influence of methyl linoleate peroxides

The stability of plant oils is related to the level of polyunsaturated fatty acids and the presence of native antioxidants--especially tocopherols. During storage, lipids or the fat products undergo oxidation and tocopherol dimers and trimers are formed. These compounds possess reducing and antioxidant properties and participate in oxidation clearly inhibiting this process. In the present study, the correlation between levels of peroxides formed during autoxidation of methyl linoleate and simultaneous decomposition of tocopherols was examined. The peroxide value was investigated. Quantities of decomposed tocopherols and formation of their dimers were determined by high-performance liquid chromatography (HPLC). Mass spectrum analysis confirmed thatthe analysed compounds were dimes. Dimerisation of gamma-T begins at the smaller quantity of the methyl linoleate peroxides than dimerisation of delta-T. At the beginning of methyl linoleate autoxidation dimerisation of gamma-T in relation to its loss was smaller. The quantity of gamma-T dimers with ether bonds in total dimers pointed to faster binding of phenoxy radicals than transformation into the phenyl ones. delta-T dimers with phenyl bonds constitute about 65% of the total. The quantity of peroxides in methyl linoleate, necessary for quantitative and qualitative changes of homologous tocopherols, decreased from delta-T to alpha-T.     

Genetic variation in the extract viscosity of rye (Secale cereale L.) bread made from endosperm and wholemeal flour: impact of high-molecular-weight arabinoxylan, starch and protein

BACKGROUND: To verify the viscous potential of rye bread made from population and hybrid cultivars, the overall extract viscosities (EVs) of endosperm and wholemeal breads (EBs and WMBs respectively) were assessed using extractants of different pH. Also, arabinoxylan‐dependent EV (AX‐EV) was determined after combined action of starch‐ and protein‐degrading enzymes. RESULTS: The synergistic effect of a combination of α‐amylase, amyloglucosidase and protease on EV was almost two times higher than the effects observed after single addition of each of them. However, water‐extractable arabinoxylans (WE AXs) were the major contributors to EV. Unexpectedly, a decrease in the mean WE AX contents observed in the following order, WMB (31.7 g kg^?1) and EB (29.3 g kg^?1) of hybrid rye and WMB (28.8 g kg^?1) and EB (23.8 g kg^?1) of population rye, was accompanied by an increase in their AX‐EVs (12, 21, 21 and 29 mPa s respectively). CONCLUSION: In some cases the structure of WE AXs is a more important determinant of AX‐EV than their content in rye bread. Nevertheless, both factors influence AX‐EV, which may reflect the viscosity of the human small intestine after rye bread consumption, related to some beneficial metabolic effects. Therefore AX‐EV can be a preliminary parameter for selection towards high‐viscosity bread. Copyright © 2010 Society of Chemical Industry     

Ultrasonic in vivo measurement of ocular surface expansion

Our aim was to ascertain whether the ultrasonic measurement of longitudinal corneal apex displacements carried out in a proper headrest is a credible method of ocular pulse (OP) detection. To distinguish between longitudinal movements of the eye globe treated as a rigid body and ocular surface expansion caused by the variations of the eye-globe volume, two ultrasound distance sensors were applied to noninvasively measure displacements of cornea and sclera. The same sensors were used to examine the influence of the anterio-posterior movements of a fixed head on the registration of corneal apex pulsation. In both experiments, ECG signals were synchronically recorded. Time, spectral, and coherence analyses obtained for four healthy subjects showed that the ocular surface expansion due to pulsatile ocular blood flow (POBF) is the main component of longitudinal corneal displacement. Ocular surface pulsation is always affected by the head movement. However, there exist some unique properties of signals, which help to distinguish between head and eye movements. A rigid headrest and a bite bar are required to stabilize the head during OP measurement. Ultrasonic technique enables noninvasive and accurate in vivo measurement of corneal pulsation, which could be of interest for indirectly estimating intraocular pressure propagation and POBF component.     

Maternal Separation Alters Ethanol Drinking and Reversal Learning Processes in Adolescent Rats: The Impact of Sex and Glycine Transporter Type 1 (GlyT1) Inhibitor

Adverse early life experiences are associated with an enhanced risk for mental and physical health problems, including substance abuse. Despite clinical evidence, the mechanisms underlying these relationships are not fully understood. Maternal separation (MS) is a commonly used animal model of early neglect. The aim of the current study is to determine whether the N-methyl-D-aspartate receptor (NMDAR)/glycine sites are involved in vulnerability to alcohol consumption (two-bottle choice paradigm) and reversal learning deficits (Barnes maze task) in adolescent rats subjected to the MS procedure and whether these effects are sex dependent. By using ELISA, we evaluated MS-induced changes in the NMDAR subunits (GluN1, GluN2A, GluN2B) expression, especially in the glycine-binding subunit, GluN1, in the prefrontal cortex (PFC) and ventral striatum (vSTR) of male/female rats. Next, we investigated whether Org 24598, a glycine transporter 1 (GlyT1) inhibitor, was able to modify ethanol drinking in adolescent and adult male/female rats with prior MS experience and reversal learning in the Barnes maze task. Our findings revealed that adolescent MS female rats consumed more alcohol which may be associated with a substantial increase in GluN1 subunit of NMDAR in the PFC and vSTR. Org 24598 decreased ethanol intake in both sexes with a more pronounced decrease in ethanol consumption in adolescent female rats. Furthermore, MS showed deficits in reversal learning in both sexes. Org 24598 ameliorated reversal learning deficits, and this effect was reversed by the NMDAR/glycine site inhibitor, L-701,324. Collectively, our results suggest that NMDAR/glycine sites might be targeted in the treatment of alcohol abuse in adolescents with early MS, especially females.     

Quantification of the compositional information provided by immonium ions on a quadrupole-time-of-flight mass spectrometer

Immonium ions have been largely overlooked during the rapid expansion of mass spectrometry-based proteomics largely due to the dominance of ion trap instruments in the field. However, immonium ions are visible in hybrid quadrupole-time-of-flight (QTOF) mass spectrometers, which are now widely available. We have created the largest database to date of high-confidence sequence assignments to characterize the appearance of immonium ions in CID spectra using a QTOF instrument under "typical" operating conditions. With these data, we are able to demonstrate excellent correlation between immonium ion peak intensity and the likelihood of the appearance of the expected amino acid in the assigned sequence for phenylalanine, tyrosine, tryptophan, proline, histidine, valine, and the indistinguishable leucine and isoleucine residues. In addition, we have clearly demonstrated a positional effect whereby the proximity of the amino acid generating the immonium ion to the amino terminal of the peptide correlates with the strength of the immonium ion peak. This compositional information provided by the immonium ion peaks could substantially improve algorithms used for spectral assignment in mass spectrometry analysis using QTOF platforms.     

Structural and Biofunctional Insights into the Cyclo(Pro-Pro-Phe-Phe-) Scaffold from Experimental and In Silico Studies: Melanoma and Beyond

Short peptides have great potential as safe and effective anticancer drug leads. Herein, the influence of short cyclic peptides containing the Pro-Pro-Phe-Phe sequence on patient-derived melanoma cells was investigated. Cyclic peptides such as cyclo(Leu-Ile-Ile-Leu-Val-Pro-Pro-Phe-Phe-), called CLA, and cyclo(Pro-homoPro-β³homoPhe-Phe-), called P11, exert the cytotoxic and the cytostatic effects in melanoma cells, respectively. CLA was the most active peptide as it reduced the viability of melanoma cells to 50% of control at about 10 µM, whereas P11 at about 40 µM after 48 h incubation. Interestingly, a linear derivative of P11 did not induce any effect in melanoma cells confirming previous studies showing that cyclic peptides exert better biological activity compared to their linear counterparts. According to in silico predictions, cyclic tetrapeptides show a better pharmacokinetic and toxic profile to humans than CLA. Notably, the spatial structure of those peptides containing synthetic amino acids has not been explored yet. In the Cambridge Structural Database, there is only one such cyclic tetrapeptide, cyclo((R)-β²homoPhe-D-Pro-Lys-Phe-), while in the Protein Data Bank—none. Therefore, we report the first crystal structure of cyclo(Pro-Pro-β³homoPhe-Phe-), denoted as 4B8M, a close analog of P11, which is crucial for drug discovery. Comparative molecular and supramolecular analysis of both structures was performed. The DFT findings revealed that 4B8M is well interpreted in the water solution. The results of complex Hirshfeld surface investigations on the cooperativity of interatomic contacts in terms of electrostatic and energetic features are provided. In short, the enrichment ratio revealed O^…H/H^…O and C^…H/H^…C as privileged intercontacts in the crystals in relation to basic and large supramolecular H-bonding synthon patterns. Furthermore, the ability of self-assemble 4B8M leading to a nanotubular structure is also discussed.     

Leydig Cells in Immunocastrated Polish Landrace Pig Testis: Differentiation Status and Steroid Enzyme Expression Status

Porker immunocastration against gonadoliberin (GnRH) secretion has been utilized since 2009; however, consumers are still skeptical of it. This is due to not having full information available on the problem of a boar taint, as well as a lack of research on morphological and molecular changes that may occur in the animal reproductive system and other body systems. The present study aimed to explore the functional status of steroidogenic Leydig cells of the testicular interstitial tissue in immunocastrated Polish Landrace pigs. Analyses were performed using Western blot, immunohistochemistry for relaxin (RLN), insulin-like 3 protein (INSL3), pelleted growth factor receptor α (PDGFRα), cytochrome P450scc, 3β- and 17β-hydroxysteroid dehydrogenases (3β-HSD, 17β-HSD), cytochrome P450arom, and 5α-reductase (5α-RED). Immunoassay ELISA was used to measure the androstenone, testosterone, and estradiol levels in the testis and serum of immunocastrates. We revealed disturbances in the distribution and expression of (i) RLN, indicating an inflammatory reaction in the interstitial tissue; (ii) INSL3 and PDGFRα, indicating alterations in the differentiation and function of fetal, perinatal, or adult Leydig cell populations; (iii) P450scc, 3β-HSD, 17β-HSD, P450arom, and 5α-RED, indicating disturbances in the sex steroid hormone production and disturbed functional status of Leydig cells; as well as (iv) decreased levels of androstenone, testosterone, and estradiol in testicular tissue and serum, indicating the dedicated action of Improvac to reduce boar taint at both the hypothalamic–hypophysis–gonadal axis and local level (Leydig cells). In summary, our study provides a significant portion of knowledge on the function of Leydig cells after immunocastration, which is also important for the diagnosis and therapy of testis dysfunction due to GnRH action failure and/or Leydig cell differentiational–functional alterations.     

Thermal stability of hydrostatically extruded EUROFER 97 steel

EUROFER 97 steel is a candidate structural material for the future fusion power reactors, as well as for the European Test Blanket Modules (TBMs) to be tested in ITER. In the reported study, the microstructure of EUROFER 97 was modified by hydrostatic extrusion (HE) which reduced the grain size from 400 to 86 nm and that of the carbide particles from 111 to 75 nm. The changes in the microstructure significantly improved the strength of the extruded samples. However, it is important that the enhanced properties of nanostructured materials are stable over the required range of intended service temperature. The thermal stability of the nanostructured EUROFER steel was evaluated by subjecting the hydrostatically extruded samples to annealing at temperatures ranging from 473 to 1073 K (200–800 °C) for 1 h. Tensile tests and microhardness measurements with a 200 g load were carried out on the annealed samples to determine the effect of the heat treatment. The results show that the highest microhardness (403 HV_0.2) was achieved for samples annealed at 673 K. However, the tensile and yield strength decreased at the higher temperature of 873 K and the total elongation increased to 15%, compared to only 3% for as-extruded samples. The changes in the mechanical properties were rationalized by the examination of the microstructural changes. During heating the initial grain size remains virtually unchanged below a temperature of 873 K. However, above 873 K the grain size increased and it is very likely that growth will be very rapid at higher temperatures.