Effect of matrigel on the osteogenic potential of canine adipose tissue-derived mesenchymal stem cells

Adipose tissue-derived mesenchymal stem cells (Ad-MSCs) are a promising source of cells for bone tissue engineering. Matrigel is a basement membrane extract containing multiple extracellular components. This mixture may promote the osteogenic differentiation of MSCs and provide a more appropriate microenvironment for transplanted cells. Here, we investigated the effect of Matrigel on the osteogenic potential of Ad-MSCs. Canine Ad-MSCs were cultured in 2D and 3D matrices and implanted into subcutaneous pouches of dogs either with or without Matrigel. Culture mineralization, cell adhesion efficiency, cell proliferation, osteoid matrix production and alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase activities were quantified and compared. Ad-MSCs grown in 2D cultures with Matrigel showed higher levels of calcium deposition and ALP activity than those grown in the absence of Matrigel under osteogenic conditions. In 3D cultures, the cells cultivated with Matrigel showed greater attachment, proliferation and osteogenic differentiation than those grown without Matrigel. In vivo, Ad-MSCs implanted with Matrigel showed higher osteogenic potential than those without Matrigel. In conclusion, these data suggest that the use of Matrigel can increase the osteogenic potential of canine Ad-MSCs.     

Concurrent multicentric hemangiosarcoma and ovarian teratoma in an aged Père david's deer (Elaphurus davidianus).

A 20-yr-old female Père David's deer (Elaphurus davidianus) died following a 3-wk history of depression, anorexia, and progressive respiratory distress. At necropsy, numerous soft to firm, tan or blood-filled nodular structures, 1-6 cm in diameter, were noted in the lung and the visceral and parietal pleura and within the mediastinum. Similar nodules were also found in the liver, spleen, kidney, and lymph nodes. Histologically, the nodules were diagnosed as hemangiosarcoma. A benign teratoma was present in the left ovary.     

Monoclonal antibody-based immunohistochemical detection of porcine epidemic diarrhea virus antigen in formalin-fixed, paraffin-embedded intestinal tissues.

An immunohistochemistry technique was developed for the diagnosis of porcine epidemic diarrhea virus (PEDV). The technique was tested on formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV. Five different monoclonal antibodies (MAbs) were tested in this study. PEDV antigen was consistently detected in the PLP (4% paraformaldehyde, 100 mM L-lysine dihydrochloride, 10 mM sodium m-periodate in phosphate-buffered saline)-fixed PEDV-infected Vero cells or formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV. The C9-2-2 MAb gave the strongest reactivity and least background staining, detecting 10 of 10 infected pigs. The positive reaction was cytoplasmic. Positive enterocytes were distributed over the tip and along the sides of atrophied or fused villi in the jejunum and ileum. Positive-staining cells were not detected in the crypts. No staining was observed in cecum and colon. No positive cells were observed when the C9-2-2 MAb was reacted with the tissue sections from noninfected piglets or from transmissible gastroenteritus virus (TGEV)- and rotavirus-infected piglets. The selected anti-PEDV MAbs tested on formalin-fixed, paraffin-embedded tissue sections are useful for diagnosis when virus isolation is not available. This method would be of particular value in countries where both PEDV and TGEV are epizootic and would aid in differentiating between PEDV and TGEV infection.     

Sero-survey on Aino, Akabane, Chuzan, bovine ephemeral fever and Japanese encephalitis virus of cattle and swine in Korea

Vector-borne arboviruses produce mild to severe symptoms in domestic animals. Bovine ephemeral fever (BEF), Akabane, Aino, and Chuzan virus have been primarily attributed to reproductive disorders or febrile diseases in cattle, and Japanese encephalitis virus (JEV) is mainly associated with reproductive failures in swine. We investigated antibody titers from domestic swine against four bovine arboviruses (BEF, Akabane, Aino, and Chuzan virus) and from cattle against JEV in Korea. While the positive rates for Akabane and BEF were 37.4% and 15.7%, the positive incidence of Chuzan and Aino were relatively low, with positive rates of 3.04% and 0.4%, respectively, based on a virus neutralization assay. Antibody titers against more than one virus were also frequently detected in domestic swine. The incidence of JEV was 51.3% among domestic cattle. In addition, one positive case was detected in the thoracic fluids from 35 aborted calves, based on the hemagglutination inhibition test. Our results indicate that swine are susceptible hosts of bovine arboviruses without showing clinical symptoms in a natural environment. Moreover, we confirmed that JEV could be associated with reproductive failure in pregnant cattle, as were other vector-borne bovine arboviruses assessed in this study.     

In vitro starch digestibility and pasting properties of germinated brown rice after hydrothermal treatments

Germinated brown rice (GBR) recently has received renewed attention due to its enhanced nutritional value. Pasting properties and in vitro starch digestibility of GBR were examined before and after hydrothermal treatments. Steeping in water (30 °C, 24 h) raised the moisture content and germination percentage of brown rice. Pasting viscosity was substantially decreased but gelatinization temperatures and enthalpy were decreased only marginally by germination (30 °C, 48 h). However, annealing (50 °C, 24 h) and heat-moisture treatment (100 °C, 1 h at 30% moisture) after germination resulted in increased pasting viscosity and gelatinization temperatures. The hydrothermal treatments, however, induced browning reactions to darken the flour of GBR. The digestibility of starch in brown rice was increased by germination. The contents of rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS) in the cooked brown rice were 47.3%, 40.8%, and 11.9%, respectively, but changed to 57.7%, 39.1%, and 3.2%, respectively upon germination. The hydrothermal treatments, however, decreased the digestibility of starch in GBR. The heat-moisture treatment decreased the RDS content in GBR near to that of native brown rice. The digestibility and physical properties of brown rice can be controlled by germination and hydrothermal treatments.     

Exploration of Sugar Functionality in Sugar‐Snap and Wire‐Cut Cookie Baking: Implications for Potential Sucrose Replacement or Reduction

The sugar series—xylose, glucose, fructose, and sucrose—can be used diagnostically to explore the effects of sugar type on differential scanning calorimetry (DSC), Rapid Visco‐Analyser (RVA), and cookie baking performance because of the differences in glass‐forming abilities (related to plasticization) and solubility parameters (related to solvent preference) of different sugars. Sugar concentration (% S), total solvent (TS), and dough formulation defined a core experimental design for cookie baking with the four sugar types and two baking methods. Although wire‐cut cookie baking (66% S and 64 TS) showed the same trends as sugar‐snap cookie baking (73% S and 79 TS) for diameter, height, and moisture content, the wire‐cut formulation enabled greater discrimination among the effects of different sugar types on dough and cookie responses. Use of two different crystal sizes of sucrose confirmed the dominant impact of both gluten development during dough mixing and starch pasting during cookie baking on collapse: the greater rate of dissolution of smaller sucrose crystals resulted in greater surface crack for sugar‐snap cookies, and lower height for wire‐cut cookies. Because the historical definition of an “excellent quality cookie flour” is based on the performance of a flour in a cookie formulated with sucrose, the effect of sugar type on cookie making is to transform the apparent baking performance of a flour. Whereas formulation with sucrose optimizes the flour performance for cookie baking, formulation with xylose exaggerates the worst aspects of cookie flour functionality and makes even the best cookie flour look like a “poor quality cookie flour”. Use of solvent retention capacity (SRC), DSC, RVA, and wire‐cut cookie baking as predictive research tools demonstrated that identification of a flour with an optimized SRC pattern is the key to successful mitigation of the detrimental effects of sucrose replacement on cookie processing and product attributes.