Changes in the composition of the extracellular matrix accumulated by mesenchymal stem cells during in vitro expansion

One of the approaches to preserve the properties of mesenchymal stem cells (MSCs) during in vitro expansion is to use cell culture substrates. MSCs are known to generate the extracellular matrix (ECM) proper to preserve their proliferative capacity in vitro, but extensive expansion is considered to deprive MSCs of the capacity to prepare such ECM. In order to examine the features of ECM proper that is required to preserve the proliferative capacity of MSCs, we analyzed the changes in the composition of ECM accumulated by MSCs during in vitro expansion. Biochemical and immunological analysis showed that collagen and laminin content decreased during expansion. Immunofluorescence and ultrastructural analyses showed that the ECM structure changed from a dynamic fibrous, porous and steric structure to a static, crammed, and planar one. The results of Western blotting analysis suggested loose intermolecular association in ECM molecules accumulated by extensively proliferated MSCs. The ECM prepared by extensively proliferated MSCs was less effective to recover their proliferative capacity than that prepared by less proliferated cells. Our results suggest that a cell culture substrate to expand MSCs requires abundance in collagen and basement membrane components, and steric, porous and fibrous structure in which ECM molecules are tightly associated.     

Regolith migration and sorting on asteroid Itokawa

High-resolution images of the surface of asteroid Itokawa from the Hayabusa mission reveal it to be covered with unconsolidated millimeter-sized and larger gravels. Locations and morphologic characteristics of this gravel indicate that Itokawa has experienced considerable vibrations, which have triggered global-scale granular processes in its dry, vacuum, microgravity environment. These processes likely include granular convection, landslide-like granular migrations, and particle sorting, resulting in the segregation of the fine gravels into areas of potential lows. Granular processes become major resurfacing processes because of Itokawa's small size, implying that they can occur on other small asteroids should those have regolith.     

Improvement of short straws for sperm cryopreservation: installing an air-permeable filter facilitates handling

Saving space for sperm cryopreservation would aid mouse genetics research. We previously developed the ST (sperm freezing in ShorT STraw to reduce STorage space) method for cryopreserving mouse sperm in a smaller storage space than conventional methods. However, our ST method has two drawbacks: difficulties during freeze-thaw procedures and the potential risk of sperm loss during storage. Here, we refine ST, terming the new method improved ST (iST). In iST, the straw has an air-permeable filter and the straw container (2-ml cryotube) is endowed with air vents. As in our ST method, iST frozen-thawed sperm showed good performance upon in vitro fertilization. Moreover, up to nine straws can be stored in one cryotube, occupying less storage space than conventional methods. This method provides an easy and space-saving cryopreservation method for mouse sperm, and thus will be valuable for mouse genetics researchers.     

Influence of Dietary Metformin on the Growth Performance and Plasma Concentrations of Amino Acids and Advanced Glycation End Products in Two Types of Chickens

Glycation is a non-enzymatic reaction inducing the bonding of glucose to amino acids and proteins. Glycated amino acids are not useful for protein synthesis, suggesting that glycation reduces the utilization of amino acids. Metformin (MF) is well known as a therapeutic drug for type II diabetes that inhibits glycation. It is possible that treatment with MF raises the utilization of amino acids by the inhibition of glycation, thereby improving the growth performance of chickens. In the present study, therefore, we investigated the influence of dietary MF on the growth performance, and plasma concentrations of free amino acids and N^ε-(Carboxymethyl)lysine (CML), which is an advanced glycation end product, in layer (Experiment 1) and broiler (Experiment 2) chickens. From 7 d of age, chicks were allowed free access to one of the experimental diets containing MF at 3 supplementation levels (0, 150, and 300 mg/kg diet) for 14 days. Body weight and feed intake were measured every week. At the end of the experiments, blood and breast muscle (M. pectoralis major) were collected for further analysis. Dietary MF did not affect weight gain, feed intake, or feed efficiency in both layer and broiler chickens. Dietary MF at the level of 150 mg/kg diet increased breast muscle weight in both layer and broiler chickens. Dietary MF increased plasma concentrations of branched chain amino acids and decreased concentrations of CML in layer chickens, although it did not affect plasma concentrations of glucose. The present study suggested that dietary MF might have the potency to increase breast muscle weight of layer chickens with an increment in plasma concentrations of branched-chain amino acids.     

Hydrogen bonding in alcoholic beverages (distilled spirits) and water-ethanol mixtures

The hydrogen-bonding properties of water-ethanol of alcoholic beverages and water-ethanol mixtures of the corresponding ethanol contents were examined on the basis of OH proton NMR chemical shifts and the Raman OH stretching spectra of water and ethanol. Japanese shochu, an unaged distilled spirit of 25% (v/v) alcoholic content made from various grains, was provided for the samples; it is a high-purity spirit as it contains only small amounts of dissolved components, like typical vodka, gin, and white rum. The hydrogen-bonding structure in shochu containing some acids was found to be different from that of the water-ethanol mixture with corresponding ethanol content. It was concluded that, by the presence of small amounts of organic acids, the water-ethanol hydrogen-bonding structure was strengthened, at the same time, the proton exchange between water and ethanol molecules was promoted in shochu, compared with the water-ethanol mixture. The NMR chemical shifts of fruit cocktail drinks suggested that the hydrogen bonding of water-ethanol in the solution was developed by organic acids and (poly)phenols from fruit juices.     

Gene transfer for Japanese flounder fertilized eggs by particle gun bombardment

ABSTRACT:   Fish transgenesis has progressed considerably. However, the technique of gene transfer for most marine aquaculture species has not been established. A method to introduce foreign genes into fertilized eggs of Japanese flounder Paralichthys olivaceus by particle gun bombardment was developed by the authors. A recombinant plasmid which contains the Japanese flounder keratin gene promoter linked to the green fluorescence protein (GFP) gene was introduced into fertilized eggs by particle gun bombardments twice at 250 psi. In each experimental group, 25 000–35 000 eggs were treated. However, the survival rate was 12.8% which is lower than that of the control groups (56.8%). Of 2606–5205 the embryos survived for 24 h, 43–61 GFP positive embryos were obtained in one experiment, giving a final gene transfer efficiency of 1.4%. All GFP-positive embryos developed and hatched normally. GFP expression was observed in epithelial tissue throughout the developmental stages. At 3 months after gene transfer, foreign DNA was detected by genome polymerase chain reaction analysis in 37 of 69 fry (53.6%). These results suggest that the particle gun method is an effective method to use with fertilized Japanese flounder eggs.     

Relationship of glomerular filtration rate based on serum iodixanol clearance to IRIS staging in cats with chronic kidney disease

We examined the correlation between the glomerular filtration rate (GFR) estimated from an equation based on the serum iodixanol clearance technique and International Renal Interest Society (IRIS) stages of chronic kidney disease (CKD) in cats. The equation included the injection dose, sampling time, serum concentration and estimated volume of distribution (Vd) of the isotonic, nonionic, contrast medium iodixanol as a test tracer. The percent changes in the median basal GFR values calculated from the equation in CKD cats resembled those of IRIS stages 1–3. These data validate the association between the GFR derived from the simplified equation and IRIS stages based on the serum creatinine concentration in cats with CKD. They describe the GFR ranges determined using single-sample iodixanol clearance for healthy cats and cats with various IRIS stages of CKD.     

Arrhythmogenic right ventricular cardiomyopathy coincided with the cardiac fibrosis in the inner muscle layer of the left ventricular wall in a boxer dog

A 7-year-old female boxer dog died suddenly without any clinical signs. It was suspected that the dog had arrhythmogenic right ventricular cardiomyopathy (ARVC) due to ventricular premature complexes and ventricular tachycardia at 3 years of age. The final diagnosis of ARVC was confirmed by histological characteristics, such as loss of cardiocytes and fibrofatty replacement, occurring in the right and left ventricular walls. In the cardiocytes, non-lipid vacuoles were observed. Cardiac fibrosis and intimal thickening of the small arteries occurred without fatty replacement in the inner muscle layer including the papillary muscles of the left ventricular wall. This paper describes the pathomorphological details of an ARVC case with coincidental cardiac fibrosis in the inner muscle layer of the left ventricular wall.     

Mixed poly (ethylene glycol) and oligo (ethylene glycol) layers on gold as nonfouling surfaces created by backfilling

Backfilling a self-assembled monolayer (SAM) of long poly (ethylene glycol) (PEG) with short PEG is a well-known strategy to improve its potential to resist fouling. Here it is shown, using x-ray photoelectron spectroscopy, contact angle, and atomic force microscopy, that backfilling PEG thiol with oligo (ethylene glycol) (OEG) terminated alkane thiol molecules results in underbrush formation. The authors also confirm the absence of phase separated arrangement, which is commonly observed with backfilling experiments involving SAMs of short chain alkane thiol with long chain alkane thiol. Furthermore, it was found that OEG addition caused less PEG desorption when compared to alkane thiol. The ability of surface to resist fouling was tested through serum adsorption and bacterial adhesion studies. The authors demonstrate that the mixed monolayer with PEG and OEG is better than PEG at resisting protein adsorption and bacterial adhesion, and conclude that backfilling PEG with OEG resulting in the underbrush formation enhances the ability of PEG to resist fouling.     

Mild drying of sandy soil can physically limit the uptake of phosphorus by rainfed lowland rice in northeast Thailand

ABSTRACT

Poor response of rice to phosphorus (P) fertilization and low phytoavailability of soil P have been reported in sandy rainfed fields in northeast Thailand. In order to evaluate the effects of mild soil drying on the uptake of P by rainfed lowland rice, we carried out nutrient omission trials for nitrogen (N) and P at Ubon Ratchathani Rice Research Center under rainfed and flooded conditions. The surface soil was classified as sandy loam. To avoid severe soil drying and drought stress in the rainfed field, soil water potential at a depth of 20 cm was maintained at the field capacity (> ?20 kPa) by flush irrigation. The effects of flooding and drying on the soil properties were also evaluated in the laboratory using soils with diverse textures in and around the center. In the field experiments, the above-ground biomass of rice plants (RD6) did not respond significantly to P fertilization in the rainfed field, although it responded positively to N fertilization. Root length in the surface 10 cm under the rainfed condition was significantly smaller than that under the flooded condition due partly to the increased soil hardness upon drying, but this could not quantitatively explain the large discrepancy of P uptake observed between the rainfed and flooded conditions. Under the rainfed condition, the P uptake did not increase significantly, even when the concentration of soil Bray P was tripled by transferring the surface soil from the flooded to the rainfed field. From the laboratory experiments, it was further suggested that soil P was supplied mainly by diffusion and that the effective diffusion coefficient for P can become less than one-tenth of the value in the flooded field when the sandy soil with clay at around 10% dried to ?100 kPa. Our results suggest that the uptake of P by the rainfed lowland rice grown in sandy soil can be limited physically by mild soil drying that reduces the supply of P to roots by diffusion rather than the chemical extractability of soil P.