Synuclein Analysis in Adult Xenopus laevis

The α-, β- and γ-synucleins are small soluble proteins expressed in the nervous system of mammals and evolutionary conserved in vertebrates. After being discovered in the cartilaginous fish Torpedo californica, synucleins have been sequenced in all vertebrates, showing differences in the number of genes and splicing isoforms in different taxa. Although α-, β- and γ-synucleins share high homology in the N-terminal sequence, suggesting their evolution from a common ancestor, the three isoforms also differ in molecular characteristics, expression levels and tissue distribution. Moreover, their functions have yet to be fully understood. Great scientific interest on synucleins mainly derives from the involvement of α-synuclein in human neurodegenerative diseases, collectively named synucleinopathies, which involve the accumulation of amyloidogenic α-synuclein inclusions in neurons and glia cells. Studies on synucleinopathies can take advantage of the development of new vertebrate models other than mammals. Moreover, synuclein expression in non-mammalian vertebrates contribute to clarify the physiological role of these proteins in the evolutionary perspective. In this paper, gene expression levels of α-, β- and γ-synucleins have been analysed in the main organs of adult Xenopus laevis by qRT-PCR. Moreover, recombinant α-, β- and γ-synucleins were produced to test the specificity of commercial antibodies against α-synuclein used in Western blot and immunohistochemistry. Finally, the secondary structure of Xenopus synucleins was evaluated by circular dichroism analysis. Results indicate Xenopus as a good model for studying synucleinopathies, and provide a useful background for future studies on synuclein functions and their evolution in vertebrates.     

Electrochemical Study of Co-Based Alloys in Simulated Physiological Solution

The aim of the present work was to provide a better understanding of the biocompatibility of Co-based alloys in the physiological environment. Co-based alloys are widely used in orthopaedics for the manufacture of prosthetic devices. Two alloys, Co—Cr—Mo and Co—Ni—Cr—Mo, and their individual metal components were studied in simulated physiological solution at 37 °C, with and without the addition of two complexing agents, EDTA and citrate, in the concentration range from 0.1 to 50 mM. The processes were studied using electrochemical techniques: cyclic voltammetry, anodic potentiodynamic measurements, linear polarization and electrochemical impedance spectroscopy. The results are discussed in the context of the dependence of the passivation behaviour of the two alloys on their composition. Both alloys are shown to passivate very well in simulated physiological solution. The effect of complexing agents is more evident on Co—Ni—Cr—Mo than on Co—Cr—Mo alloy. This appears to be related to the presence of Ni.     

Lactones in the Synthesis of Prostaglandins and Prostaglandin Analogs

In the total stereo-controlled synthesis of natural prostaglandins (PGs) and their structural analogs, a vast class of compounds and drugs, known as the lactones, are encountered in a few key steps to build the final molecule, as: δ-lactones, γ-lactones, and 1,9-, 1,11-, and 1,15-macrolactones. After the synthesis of 1,9-PGF_2α and 1,15-PGF_2α lactones, many 1,15-lactones of E₂, E₃, F₂, F₃, A₂, and A₃ were found in the marine mollusc Tethys fimbria and the quest for understanding their biological role stimulated the research on their synthesis. Then 1,9-, 1,11-, and 1,15-PG lactones of the drugs were synthesized as an alternative to the corresponding esters, and the first part of the paper describes the methods used for their synthesis. The efficient Corey procedure for the synthesis of prostaglandins uses the key δ-lactone and γ-lactone intermediates with three or four stereocenters on the cyclopentane fragment to link the PG side chains. The paper describes the most used procedures for the synthesis of the milestone δ-Corey-lactones and γ-Corey-lactones, their improvements, and some new promising methods, such as interesting, new stereo-controlled and catalyzed enantioselective reactions, and methods based on the chemical/enzymatic resolution of the compounds in different steps of the sequences. The many uses of δ-lactones not only for the synthesis of γ-lactones, but also for obtaining 9β-halogen-PGs and halogen-substituted cyclopentane intermediates, as synthons for new 9β-PG analogs and future applications, are also discussed.     

Genome-Wide Identification and Analysis of CC-NBS-LRR Family in Response to Downy Mildew and Black Rot in Chinese Cabbage

The nucleotide-binding site–leucine-rich repeat (NBS–LRR) gene family is the largest group of plant disease resistance (R) genes widespread in response to viruses, bacteria, and fungi usually involved in effector triggered immunity (ETI). Forty members of the Chinese cabbage CC type NBS–LRR family were investigated in this study. Gene and protein characteristics, such as distributed locations on chromosomes and gene structures, were explored through comprehensive analysis. CC–NBS–LRR proteins were classified according to their conserved domains, and the phylogenetic relationships of CC–NBS–LRR proteins in Brassica rapa, Arabidopsis thaliana, and Oryza sativa were compared. Moreover, the roles of BrCC–NBS–LRR genes involved in pathogenesis-related defense were studied and analyzed. First, the expression profiles of BrCC–NBS–LRR genes were detected by inoculating with downy mildew and black rot pathogens. Second, sensitive and resistant Chinese cabbage inbred lines were screened by downy mildew and black rot. Finally, the differential expression levels of BrCC–NBS–LRR genes were monitored at 0, 1, 3, 6, 12 and 24 h for short and 0, 3, 5, 7, 10 and 14 days for long inoculation time. Our study provides information on BrCC–NBS–LRR genes for the investigation of the functions and mechanisms of CC-NBS-LRR genes in Chinese cabbage.     

Role of Retinal Amyloid-β in Neurodegenerative Diseases: Overlapping Mechanisms and Emerging Clinical Applications

Amyloid-β (Aβ) accumulations have been identified in the retina for neurodegeneration-associated disorders like Alzheimer’s disease (AD), glaucoma, and age-related macular degeneration (AMD). Elevated retinal Aβ levels were associated with progressive retinal neurodegeneration, elevated cerebral Aβ accumulation, and increased disease severity with a decline in cognition and vision. Retinal Aβ accumulation and its pathological effects were demonstrated to occur prior to irreversible neurodegeneration, which highlights its potential in early disease detection and intervention. Using the retina as a model of the brain, recent studies have focused on characterizing retinal Aβ to determine its applicability for population-based screening of AD, which warrants a further understanding of how Aβ manifests between these disorders. While current treatments directly targeting Aβ accumulations have had limited results, continued exploration of Aβ-associated pathological pathways may yield new therapeutic targets for preserving cognition and vision. Here, we provide a review on the role of retinal Aβ manifestations in these distinct neurodegeneration-associated disorders. We also discuss the recent applications of retinal Aβ for AD screening and current clinical trial outcomes for Aβ-associated treatment approaches. Lastly, we explore potential future therapeutic targets based on overlapping mechanisms of pathophysiology in AD, glaucoma, and AMD.     

Response to fertilization and nutrient deficiency diagnostics in peach palm in Central Amazonia

Peach palm (Bactris gasipaes Kunth) is increasingly grown in the tropics for its heart-of-palm and fruit. Determining fertilization response and diagnosing nutrient status in peach palm may require methods that consider the particularities in nutrient acquisition and recycling of perennial crops. Responses to nutrient additions, and the diagnostic value of soil and foliar analyses were examined in three field experiments with three-year old peach palm stands on Oxisols in Central Amazonia. To diagnose P-deficiency levels in soils, samples from 0–5 cm and 5–20 cm depth were analyzed for available P by different methods (Mehlich-1, Mehlich-3 and Modified Olsen). The second and fifth leaves were analyzed to assess N, P and K deficiencies. Field experiments involved several combinations of N (from 0 to 225 kg ha^–1 yr^–1), K (from 0 to 225 kg ha^–1 yr^–1) and P (from 0 to 59 kg ha^–1 yr^–1). Palms on control plots (unfertilized) and those receiving 225 kg ha^–1 yr^–1 N and 2 Mg ha^–1 of lime yielded between 4 and 19% of the maximum growth which was obtained with N, P and K applications. In one of the experiments, yield of heart-of-palm was positively related to N additions at the lowest levels of P (8.6 kg ha^–1 yr^–1) and K (60 kg ha^–1 yr^–1) additions. In one experiment, critical leaf N level was 2.5% for the second leaf and 2.2% for the fifth leaf. Some growth responses to P additions at constant N and K levels were observed (e.g., 797 kg ha^–1 yr^–1 of heart-of-palm with 39.3 kg ha^–1 yr^–1 of applied P, and 632 kg ha^–1 yr^–1 of heart-of-palm with 10.9 kg ha^–1 yr^–1 of applied P in one experiment, and 2334 kg ha^–1 yr^–1 of heart-of-palm with 39.3 kg ha^–1 yr^–1 of P and 1257 kg ha^–1 yr^–1 of heart-of-palm with 19.7 kg ha^–1 yr^–1 of P in another trial). In the experiment for fruit production from peach palm, total plant height did not respond to P additions between 19.7 and 59 kg ha^–1 yr^–1 and K additions between 75 and 225 kg ha^–1 yr^–1. Leaf P levels were found to be above the proposed critical levels of 0.23% for the third leaf and 0.16% for the fifth leaf. Plants in this experiment, however, showed evident symptoms of Mg deficiency, which was associated with a steep gradient of increasing Mg concentration from the fifth leaf to the second leaf. Standard leaf diagnostic methods in most cases proved less useful to show plant N and P status and growth responses to N and P additions. Soil P determined by common extractions was in general too variable for prediction of growth.     

Melatonin as a Signaling Molecule for Metabolism Regulation in Response to Hypoxia in the Crab Neohelice granulata

Melatonin has been identified in a variety of crustacean species, but its function is not as well understood as in vertebrates. The present study investigates whether melatonin has an effect on crustacean hyperglycemic hormone (CHH) gene expression, oxygen consumption (VO₂) and circulating glucose and lactate levels, in response to different dissolved-oxygen concentrations, in the crab Neohelice granulata, as well as whether these possible effects are eyestalk- or receptor-dependent. Melatonin decreased CHH expression in crabs exposed for 45 min to 6 (2, 200 or 20,000 pmol·crab⁻¹) or 2 mgO₂·L⁻¹ (200 pmol·crab⁻¹). Since luzindole (200 nmol·crab⁻¹) did not significantly (p > 0.05) alter the melatonin effect, its action does not seem to be mediated by vertebrate-typical MT1 and MT2 receptors. Melatonin (200 pmol·crab⁻¹) increased the levels of glucose and lactate in crabs exposed to 6 mgO₂·L⁻¹, and luzindole (200 nmol·crab⁻¹) decreased this effect, indicating that melatonin receptors are involved in hyperglycemia and lactemia. Melatonin showed no effect on VO₂. Interestingly, in vitro incubation of eyestalk ganglia for 45 min at 0.7 mgO₂·L⁻¹ significantly (p < 0.05) increased melatonin production in this organ. In addition, injections of melatonin significantly increased the levels of circulating melatonin in crabs exposed for 45 min to 6 (200 or 20,000 pmol·crab⁻¹), 2 (200 and 20,000 pmol·crab⁻¹) and 0.7 (200 or 20,000 pmol·crab⁻¹) mgO₂·L⁻¹. Therefore, melatonin seems to have an effect on the metabolism of N. granulata. This molecule inhibited the gene expression of CHH and caused an eyestalk- and receptor-dependent hyperglycemia, which suggests that melatonin may have a signaling role in metabolic regulation in this crab.     

Metformin Preserves β-Cell Compensation in Insulin Secretion and Mass Expansion in Prediabetic Nile Rats

Prediabetes is a high-risk condition for type 2 diabetes (T2D). Pancreatic β-cells adapt to impaired glucose regulation in prediabetes by increasing insulin secretion and β-cell mass expansion. In people with prediabetes, metformin has been shown to prevent prediabetes conversion to diabetes. However, emerging evidence indicates that metformin has negative effects on β-cell function and survival. Our previous study established the Nile rat (NR) as a model for prediabetes, recapitulating characteristics of human β-cell compensation in function and mass expansion. In this study, we investigated the action of metformin on β-cells in vivo and in vitro. A 7-week metformin treatment improved glucose tolerance by reducing hepatic glucose output and enhancing insulin secretion. Although high-dose metformin inhibited β-cell glucose-stimulated insulin secretion in vitro, stimulation of β-cell insulin secretion was preserved in metformin-treated NRs via an indirect mechanism. Moreover, β-cells in NRs receiving metformin exhibited increased endoplasmic reticulum (ER) chaperones and alleviated apoptotic unfold protein response (UPR) without changes in the expression of cell identity genes. Additionally, metformin did not suppress β-cell mass compensation or proliferation. Taken together, despite the conflicting role indicated by in vitro studies, administration of metformin does not exert a negative effect on β-cell function or cell mass and, instead, early metformin treatment may help protect β-cells from exhaustion and decompensation.     

DNA stretching on the wall surfaces in curved microchannels with different radii

DNA molecule conformation dynamics and stretching were made on semi-circular surfaces with different radii (500 to 5,000 μm) in microchannels measuring 200 μm × 200 μm in cross section. Five different buffer solutions - 1× Tris-acetate-EDTA (TAE), 1× Tris-borate-EDTA (TBE), 1× Tris-EDTA (TE), 1× Tris-phosphate-EDTA (TPE), and 1× Tris-buffered saline (TBS) solutions - were used with a variety of viscosity such as 40, 60, and 80 cP, with resultant 10⁻⁴ ≤ Re ≤ 10⁻³ and the corresponding 5 ≤ Wi ≤ 12. The test fluids were seeded with JOJO-1 tracer particles for flow visualization and driven through the test channels via a piezoelectric (PZT) micropump. Micro particle image velocimetry (μPIV) measuring technique was applied for the centered-plane velocity distribution measurements. It is found that the radius effect on the stretch ratio of DNA dependence is significant. The stretch ratio becomes larger as the radius becomes small due to the larger centrifugal force. Consequently, the maximum stretch was found at the center of the channel with a radius of 500 μm.     

Interaction between Galectin-3 and Integrins Mediates Cell-Matrix Adhesion in Endothelial Cells and Mesenchymal Stem Cells

Galectin-3 (Gal-3) is a β-galactoside-binding protein that influences various cell functions, including cell adhesion. We focused on the role of Gal-3 as an extracellular ligand mediating cell-matrix adhesion. We used human adipose tissue-derived stem cells and human umbilical vein endothelial cells that are promising for vascular tissue engineering. We found that these cells naturally contained Gal-3 on their surface and inside the cells. Moreover, they were able to associate with exogenous Gal-3 added to the culture medium. This association was reduced with a β-galactoside LacdiNAc (GalNAcβ1,4GlcNAc), a selective ligand of Gal-3, which binds to the carbohydrate recognition domain (CRD) in the Gal-3 molecule. This ligand was also able to detach Gal-3 newly associated with cells but not Gal-3 naturally present on cells. In addition, Gal-3 preadsorbed on plastic surfaces acted as an adhesion ligand for both cell types, and the cell adhesion was resistant to blocking with LacdiNAc. This result suggests that the adhesion was mediated by a binding site different from the CRD. The blocking of integrin adhesion receptors on cells with specific antibodies revealed that the cell adhesion to the preadsorbed Gal-3 was mediated, at least partially, by β1 and αV integrins—namely α5β1, αVβ3, and αVβ1 integrins.     

Carbon sequestration in soil aggregates under different crop rotations and nitrogen fertilization in an inceptisol in southeastern Norway

Effects of crop rotation and fertilization (nitrogen and manure) on concentrations of soil organic carbon (SOC) and total soil nitrogen (TSN) in bulk soil and in soil aggregates were investigated in a long-term field experiment established in 1953 at Ås, Norway. The effect of these management practices on SOC sequestration was estimated. The experiment had three six-course rotations: (I) continuous spring grain, (II) spring grain for 3 years followed by root crops for 3 years, and (III) spring grain for 2 years followed by meadow for 4 years. Three fertilizer treatments compared were: (A) 30–40 kg N ha^–1; (B) 80–120 kg N ha^–1; and (C) a combination of B and 60 Mg farmyard manure (FYM) ha^–1. All plots received a basal rate of PK fertilizer. Soil samples from these treatments were collected in autumn 2001 and analyzed for aggregate size, SOC and TSN concentrations. There were significant increases in 0.6–2 mm and < 0.6 mm aggregate size fractions, and reduction in the 6–20 mm and the > 20 mm sizes for rotation III only. There were also significant differences among rotations with regard to water stable aggregation. The order of increase in stability was II < I < III. Fertilizer treatment had no effect on aggregation or aggregate size distribution, but there was a slight tendency of increased stability with the application of FYM. Aggregate stability increased with increasing concentration of SOC (r2 = 0.53). The SOC and TSN concentrations in bulk soil were significantly higher in rotation III than in rotations II and I. Application of FYM increased SOC and TSN concentrations significantly in the 0–10 cm soil depth, but there were few significant differences between fertility treatments A and B. There was a trend of increase in concentration of SOC and TSN with decreasing aggregate size, but significant differences in these parameters in different aggregate size fractions were found only in few cases. The SOC and TSN concentrations were higher in >0.25 mm than in < 0.25 mm aggregates. The SOC sequestration rate was 77–167 kg SOC ha^–1 yr^–1 by increasing the N rate and 40–162 kg SOC ha^–1 yr^–1 by applying FYM. The SOC sequestration rate by judicious use of inorganic fertilizer was the greatest in the grain–meadow rotation, while that by application of FYM was the greatest in the all grain rotation.     

Organization of contemporary systems for transport-warehouse servicing in the manufacture of man-made fibres

Conclusions -- It is necessary to begin the development of contemporary transport—warehouse systems promptly. Delay in development threatens in the future large losses and potential possibilities for new technological equipment.-- Work should be carried out in two directions: development of a complex of automated equipment for transport—warehouse systems; and performing investigative work on developing a complex of program—technical media and a mathematical—statistical model of transport—servicing systems.Translated from Khimicheskie Volokna, No. 5, pp. 50–51, September–October, 1990.     

Transition metal-based cathodes for hydrogen evolution in alkaline solution: Electrocatalysis on nickel-based ternary electrolytic codeposits

Nickel-molybdenum-iron, nickel-molybdenum-copper, nickel-molybdenum-zinc, nickel-molybdenum-cobalt, nickel-molybdenum-tungsten and nickel-molybdenum-chromium ternary codeposits, obtained through electrodeposition on mild steel strips have been characterized with the objective of qualitatively comparing and assessing their electrocatalytic activities as hydrogen electrodes in alkaline solution. It has been concluded that their electrocatalytic effects for the hydrogen evolution reaction (h.e.r.) rank in the following order: Ni–Mo–Fe>Ni–Mo–Cu>Ni–Mo–Zn>Ni–Mo–Co Ni–Mo–W>Ni–Mo–Cr>Ni-plated steel. Further investigations on these electrocatalysts have revealed that the cathodic overpotential contribution to the electrolysis voltage can be brought down by 0.3 V when compared with conventional steel cathodes. The best and most stable hydrogen evolving cathode of these, namely Ni–Mo–Fe, exhibited an overpotential of about 0.187 V for over 1500 h of continuous electrolysis in 6 M KOH at 300 mA cm^–2 and 353 K. The salient features of the codeposits, such as physical characteristics, chemical composition, current-potential behaviour and the varying effects of the catalytic activation method were analysed with a view to correlating the micro-structural characteristics of the coatings with the hydrogen adsorption process. The stability under open circuit conditions, the tolerance to electrochemical corrosion and the long term stability of Ni–Mo–Fe codeposit cathodes were very encouraging. An attempt to identify the pathway for the h.e.r. on these codeposit cathodes was made, in view of the electrochemical parameters obtained experimentally.     

Resistivity dependence of magnetoresistance in Co/ZnO films

We report the dependence of magnetoresistance effect on resistivity (ρ) in Co/ZnO films deposited by magnetron sputtering at different sputtering pressures with different ZnO contents. The magnitude of the resistivity reflects different carrier transport regimes ranging from metallic to hopping behaviors. Large room-temperature magnetoresistance greater than 8% is obtained in the resistivity range from 0.08 to 0.5 Ω · cm. The magnetoresistance value decreases markedly when the resistivity of the films is less than 0.08 Ω · cm or greater than 0.5 Ω · cm. When 0.08 Ω · cm < ρ < 0.5 Ω · cm, the conduction contains two channels: the spin-dependent tunneling channel and the spin-independent second-order hopping (N = 2). The former gives rise to a high room-temperature magnetoresistance effect. When ρ > 0.5 Ω · cm, the spin-independent higher-order hopping (N > 2) comes into play and decreases the tunneling magnetoresistance value. For the samples with ρ < 0.08 Ω · cm, reduced magnetoresistance is mainly ascribed to the formation of percolation paths through interconnected elongated metallic Co particles. This observation is significant for the improvement of room-temperature magnetoresistance value for future spintronic devices.     

Association of IgG1 Antibody Clearance with FcγRIIA Polymorphism and Platelet Count in Infliximab-Treated Patients

The FcγRIIA/CD32A is mainly expressed on platelets, myeloid and several endothelial cells. Its affinity is considered insufficient for allowing significant binding of monomeric IgG, while its H131R polymorphism (histidine > arginine at position 131) influences affinity for multimeric IgG2. Platelet FcγRIIA has been reported to contribute to IgG-containing immune-complexe clearance. Given our finding that platelet FcγRIIA actually binds monomeric IgG, we investigated the role of platelets and FcγRIIA in IgG antibody elimination. We used pharmacokinetics analysis of infliximab (IgG1) in individuals with controlled Crohn’s disease. The influence of platelet count and FcγRIIA polymorphism was quantified by multivariate linear modelling. The infliximab half-life increased with R allele number (13.2, 14.4 and 15.6 days for HH, HR and RR patients, respectively). It decreased with increasing platelet count in R carriers: from ≈20 days (RR) and ≈17 days (HR) at 150 × 10⁹/L, respectively, to ≈13 days (both HR and RR) at 350 × 10⁹/L. Moreover, a flow cytometry assay showed that infliximab and monomeric IgG1 bound efficiently to platelet FcγRIIA H and R allotypes, whereas panitumumab and IgG2 bound poorly to the latter. We propose that infliximab (and presumably any IgG1 antibody) elimination is partly due to an unappreciated mechanism dependent on binding to platelet FcγRIIA, which is probably tuned by its affinity for IgG2.     

Morphological effect of oscillating magnetic nanoparticles in killing tumor cells

Forced oscillation of spherical and rod-shaped iron oxide magnetic nanoparticles (MNPs) via low-power and low-frequency alternating magnetic field (AMF) was firstly used to kill cancer cells in vitro. After being loaded by human cervical cancer cells line (HeLa) and then exposed to a 35-kHz AMF, MNPs mechanically damaged cell membranes and cytoplasm, decreasing the cell viability. It was found that the concentration and morphology of the MNPs significantly influenced the cell-killing efficiency of oscillating MNPs. In this preliminary study, when HeLa cells were pre-incubated with 100 μg/mL rod-shaped MNPs (rMNP, length of 200 ± 50 nm and diameter of 50 to 120 nm) for 20 h, MTT assay proved that the cell viability decreased by 30.9% after being exposed to AMF for 2 h, while the cell viability decreased by 11.7% if spherical MNPs (sMNP, diameter of 200 ± 50 nm) were used for investigation. Furthermore, the morphological effect of MNPs on cell viability was confirmed by trypan blue assay: 39.5% rMNP-loaded cells and 15.1% sMNP-loaded cells were stained after being exposed to AMF for 2 h. It was also interesting to find that killing tumor cells at either higher (500 μg/mL) or lower (20 μg/mL) concentration of MNPs was less efficient than that achieved at 100 μg/mL concentration. In conclusion, the relatively asymmetric morphological rod-shaped MNPs can kill cancer cells more effectively than spherical MNPs when being exposed to AMF by virtue of their mechanical oscillations.     

Dynamics of Heating of Fluoride Stock in Manufacture of Optical Fibres

Heating of fluoride stock was analyzed. A module for drawing fluoride stock into optical fibres was developed. The transient responses and transmitting functions of the object of control in “current strength—temperature” and “gas flow—temperature” channels were obtained. The structure of a system for controlling heating of fluoride stock was proposed based on the results of the study.__________Translated from Khimicheskie Volokna, No. 2, pp. 16–18, March–April, 2005.     

Long-Term Vitamin D Deficiency Results in the Inhibition of Cell Proliferation and Alteration of Multiple Gastric Epithelial Cell Lineages in Mice

Over one billion people globally are vitamin D (VD) deficient. Studies on the biological roles of VD are numerous but very little on the stomach. This project aims to understand how gastric homeostasis is affected by VD deficiency caused by prolonged exposure to darkness alone or combined with VD deficient diet. Three groups of C57/BL6 mice were subjected to different light exposure conditions and diets for 12 months (n = 8–12/group): control—12 h/12 h light/dark SDL (Standard Diet/Light), 24 h dark SDD (Standard Diet/Dark), and 24 h dark VDD (VD deficient diet/Dark). Stomach samples were collected for different multi-label lectin-/immuno-histochemical and qRT-PCR analyses, and the serum for LC-MS-MS. We found that the membrane VD receptor is expressed widely in the stomach when compared to nuclear VD receptors. Compared to SDL, VDD mice developed mucous cell expansion with increased mucins-mRNA (3.27 ± 2.73 (p < 0.05)) increased apoptotic cells, 15 ± 7 (p ≤ 0.001)); decreased cell proliferation, 4 ± 4 (p < 0.05)) and decreased acid secretion 33 ± 2 μEq/kg (p ≤ 0.0001)). Interestingly, mice exposed to full darkness developed mild VD deficiency with higher VD epimer levels: 11.9 ± 2.08 ng/mL (p ≤ 0.0001)), expansion in zymogenic cell number (16 ± 3 (p ≤ 0.01)), and a reduction in acid secretion (18 ± 2 μEq/kg (p ≤ 0.0001)). In conclusion, changes in light exposure or VD levels have serious physiological effects on the gastric mucosa, which should be considered during the management of gastric disorders.     

Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars

The low-molecular weight glutenin subunit (LMW-GS) composition of wheat (Triticum aestivum) flour has important effects on end-use quality. However, assessing the contributions of each LMW-GS to flour quality remains challenging because of the complex LMW-GS composition and allelic variation among wheat cultivars. Therefore, accurate and reliable determination of LMW-GS alleles in germplasm remains an important challenge for wheat breeding. In this study, we used an optimized reversed-phase HPLC method and proteomics approach comprising 2-D gels coupled with liquid chromatography–tandem mass spectrometry (MS/MS) to discriminate individual LMW-GSs corresponding to alleles encoded by the Glu-A3, Glu-B3, and Glu-D3 loci in the ‘Aroona’ cultivar and 12 ‘Aroona’ near-isogenic lines (ARILs), which contain unique LMW-GS alleles in the same genetic background. The LMW-GS separation patterns for ‘Aroona’ and ARILs on chromatograms and 2-D gels were consistent with those from a set of 10 standard wheat cultivars for Glu-3. Furthermore, 12 previously uncharacterized spots in ‘Aroona’ and ARILs were excised from 2-D gels, digested with chymotrypsin, and subjected to MS/MS. We identified their gene haplotypes and created a 2-D gel map of LMW-GS alleles in the germplasm for breeding and screening for desirable LMW-GS alleles for wheat quality improvement.