Survival of hypoxic human mesenchymal stem cells is enhanced by a positive feedback loop involving miR-210 and hypoxia-inducible factor 1

The use of mesenchymal stem cells (MSCs) has emerged as a potential new treatment for myocardial infarction. However, the poor viability of MSCs after transplantation critically limits the efficacy of this new strategy. The expression of microRNA-210 (miR-210) is induced by hypoxia and is important for cell survival under hypoxic conditions. Hypoxia increases the levels of hypoxia inducible factor-1 (HIF-1) protein and miR-210 in human MSCs (hMSCs). miR-210 positively regulates HIF-1α activity. Furthermore, miR-210 expression is also induced by hypoxia through the regulation of HIF-1α. To investigate the effect of miR-210 on hMSC survival under hypoxic conditions, survival rates along with signaling related to cell survival were evaluated in hMSCs over-expressing miR-210 or ones that lacked HIF-1α expression. Elevated miR-210 expression increased survival rates along with Akt and ERK activity in hMSCs with hypoxia. These data demonstrated that a positive feedback loop involving miR-210 and HIF-1α was important for MSC survival under hypoxic conditions.     

GC/MS analysis of high-performance liquid chromatography fractions from Sophora flavescens and Torilis japonica extracts and their in vitro anti-neosporal effects on Neospora caninum

We analyzed alcoholic extracts of herbs possessing anti-neosporal activity against Neospora (N.) caninum. To identify the chemical components of Sophora (S.) flavescens and Torilis (T.) japonica associated with anti-neosporal activity, specific fractions were isolated by high-performance liquid chromatography (HPLC). In vitro activity of the fractions against N. caninum was then assessed. Gas chromatography/mass spectrometry (GC/MS) was used to identify and quantify specific anti-neosporal molecules in the herbal extracts. Almost all HPLC fractions of S. flavescens and T. japonica had higher levels of anti-neosporal activity compared to the not treated control. Active constituents of the extracts were sophoridane, furosardonin A, and tetraisopropylidene-cyclobutane in S. flavescens; 5,17-β-dihydroxy-de-A-estra-5,7,9,14-tetraene, furanodiene, and 9,12-octadecadienoic acid (Z,Z)-(CAS,1) in T. japonica.     

Rehabilitating Abandoned Pastures in Panama: Control of the Invasive Exotic Grass,Saccharum spontaneum L., Using Artificial Shade Treatments

ABSTRACT

The exotic grass, Saccharum spontaneum L., has invaded abandoned agricultural lands in the Panama Canal Watershed for decades. The grass aggressively competes with regenerating tree seedlings preventing natural forest regeneration. To estimate effectual light level for controlling the grass, the growth of S. spontaneum was measured under a range of artificial shading conditions. Five shade treatment subplots–light intensities of 100% (full sunlight), 50, 25, 15, and 5%–were established at each of eight plots in land adjacent to the Panama Canal Watershed. Each site was cleared of S. spontaneum and then the regrowth was harvested. The regrowth of the grass was harvested and measured four times every one and half months for six months. The biomass of S. spontaneum was significantly less in lower light conditions than in full sunlight. The results showed that comparing growth of the grass at each harvest date, except for the first harvest date, there were significant differences between full sunlight and light intensities of 5, 15 and 25%. When compared by different harvest dates for each light level, only full sunlight showed a significant difference in biomass of the grass. The study demonstrates that shading is an effective method for controlling S. spontaneum. The results can be applied to developing reforestation strategies for abandoned lands occupied by S. spontaneum.     

Classification of the conductance of moisture through wood cell components

This study aimed to clarify the conductance of moisture through wood cell components. Moisture diffusion coefficients were determined from three models (Stamm, Siau, and Kang et al.) and cell wall, pit, and ray dimensions were experimentally observed in a wood specimen. Fractions of moisture diffusing along each path in each of the models were analyzed. As moisture content decreased, the fraction of water diffusing as bound water through cell walls in tangential and longitudinal directions decreased while water vapor diffusion through lumens and pits became more dominant. Diffusion coefficients predicted by each model were compared with experimental values. Although predicted values differed from experimental values, predicted trends for diffusion rate dependence on moisture content were similar to the experimental results. In particular, the models of Stamm and Kang et al., which consider moisture transport through rays and pits, show a very consistent trend for transverse diffusion, which is always faster radially than tangentially. Input of more accurate dimensions of cell walls and cavities into the models should result in more reliable values, closer to the experimentally determined diffusion coefficients.     

Nasal immunization with major epitope-containing ApxIIA toxin fragment induces protective immunity against challenge infection with Actinobacillus pleuropneumoniae in a murine model

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439–801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.     

Generation of porcine induced pluripotent stem cells and evaluation of their major histocompatibility complex protein expression in vitro

Induced pluripotent stem cells (iPSCs) are thought to be highly beneficial in the field of regenerative medicine and are believed to overcome immunogenic barriers to cell transplantation. However, issues remain regarding their safety and efficiency for medical use. Furthermore, some recent reports have suggested that iPSCs could be targeted by the autologous immune system. To promote practical applications of iPSCs, in depth research using appropriate animal models is needed and porcine species appear to provide an ideal model. Recent studies have focused on the generation of porcine iPSC cells, but no investigations of their immunological properties have been conducted to date. In the present study, we generated putative iPSCs from porcine somatic cells and measured major histocompatibility complex (MHC) expression on the iPSCs and their derivatives. Compact colonies that expressed pluripotent markers appeared 11 days after viral infection. Embryonic bodies (EB) were produced and differentiated into three germ layers in vitro. Karyotyping and swine leukocyte antigen (SLA) typing showed that the iPSCs were identical to parental somatic cells. Porcine iPSCs expressed only low levels of MHC class I and moderately increased levels on their differentiated derivatives, whereas MHC class II was rarely expressed. In the presence of interferon-gamma (IFN-γ), the expression of MHC class I was elevated on differentiated iPSCs, and gradually decreased after withdrawal of the cytokine. Our data suggest that porcine iPSCs could be useful for preclinical studies of the efficiency and viability of iPSCs, and for devising strategies to rescue transplanted cells from the autologous immune system.     

Pathogenesis of type 1 (European genotype) porcine reproductive and respiratory syndrome virus in male gonads of infected boar

The objective of this study was to determine the pathogenesis of experimental infection with a type 1 porcine reproductive and respiratory syndrome virus (PRRSV) by defining the sites of viral replication and apoptosis in male gonads from infected boars for a period of 21 days after intranasal inoculation. Microscopically, hypospermatogenesis and abundant germ cell depletion and death were observed in the testes. Such germ cell death occurs by apoptosis, as determined by a characteristic histological patterns and evidence of massive DNA fragment detected in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) reaction. PRRSV was detected in the testicular tissue of infected boars only. Viral nucleic acid was localized in spermatogonia, spermatocytes and spermatids but not in the vesicular and bulbourethral gland. In serial sections, PRRSV-positive cells did not co-localized with apoptotic cells. TUNEL-positive apoptotic cells were more numerous than PRRSV-positive cells in testicular sections. The present study demonstrated that type 1 PRRSV infects the spermatogonia and their progeny, and induces apoptosis in these germ cells.     

Thromboelastography platelet mapping in healthy dogs using 1 analyzer versus 2 analyzers

The objective of this study was to describe the results of thromboelastography platelet mapping (TEG-PM) carried out using 2 techniques in 20 healthy dogs. Maximum amplitudes (MA) generated by thrombin (MA_thrombin), fibrin (MA_fibrin), adenosine diphosphate (ADP) receptor activity (MA_ADP), and thromboxane A₂ (TxA₂) receptor activity (stimulated by arachidonic acid, MA_AA) were recorded. Thromboelastography platelet mapping was carried out according to the manufacturer’s guidelines (2-analyzer technique) and using a variation of this method employing only 1 analyzer (1-analyzer technique) on 2 separate blood samples obtained from each dog. Mean [± standard deviation (SD)] MA values for the 1-analyzer/2-analyzer techniques were: MA_thrombin = 51.9 mm (± 7.1)/52.5 mm (± 8.0); MA_fibrin = 20.7 mm (± 21.8)/23.0 mm (± 26.1); MA_ADP = 44.5 mm (± 15.6)/45.6 mm (± 17.0); and MA_AA = 45.7 mm (± 11.6)/45.0 mm (± 15.4). Mean (± SD) percentage aggregation due to ADP receptor activity was 70.4% (± 32.8)/67.6% (± 33.7). Mean percentage aggregation due to TxA₂ receptor activity was 77.3% (± 31.6)/78.1% (± 50.2). Results of TEG-PM were not significantly different for the 1-analyzer and 2-analyzer methods. High correlation was found between the 2 methods for MA_fibrin [concordance correlation coefficient (r) = 0.930]; moderate correlation was found for MA_thrombin (r = 0.70) and MA_ADP (r = 0.57); correlation between the 2 methods for MA_AA was lower (r = 0.32). Thromboelastography platelet mapping (TEG-PM) should be further investigated to determine if it is a suitable method for measuring platelet dysfunction in dogs with thrombopathy.     

Efficacy of a piglet-specific commercial inactivated vaccine against Porcine circovirus type 2 in clinical field trials

The efficacy of a piglet-specific inactivated Porcine circovirus type 2 (PCV2) vaccine was evaluated with clinical field trials, as recommended by the Republic of Korea’s Animal, Plant & Fisheries Quarantine & Inspection Agency. Three farms were selected on the basis of their history of postweaning multisystemic wasting syndrome. On each farm 60, 1-week-old pigs were randomly allocated to 1 of 2 treatment groups: vaccination at 1 and 3 wk of age or no vaccination. The 2-dose schedule of vaccination with inactivated PCV2 vaccine improved the average daily weight gain from birth to 16 wk of age, the PCV2 load in the blood, and the frequency and severity of lymph node lesions. Inactivated PCV2 vaccine seems to be very effective in controlling PCV2 infection under field conditions.     

Program of vaccination and antibiotic treatment to control polyserositis caused by Haemophilus parasuis under field conditions

The present study investigated the effects of vaccinating sows and piglets or piglets alone against Haemophilus parasuis on the prevalence of H. parasuis in nasal swabs, on the humoral and cellular immune responses, and on the production parameters of piglets at 3 Korean farms with a clinical history of polyserositis caused by H. parasuis. Piglets born to vaccinated or non-vaccinated sows were subdivided into 3 groups: vaccinated sows and vaccinated pigs (VS-VP), non-vaccinated sows and vaccinated pigs (NVS-VP), and non-vaccinated sows and non-vaccinated pigs (NVS-NVP). The proportion of piglets with positive nasal swabs was significantly lower (P < 0.05) in the vaccinated animals (VS-VP and NVS-VP groups) than in the non-vaccinated animals (NVS-NVP group) at 35 and 60 d of age at the 3 farms. The overall growth performance (from 7 to 60 d of age) of the vaccinated piglets was significantly better (P < 0.05) than that of the non-vaccinated piglets at the 3 farms. Piglets in the VS-VP group had significantly higher levels (P < 0.05) of H. parasuis-specific IgG antibodies, lymphocyte proliferation, and interferon-γ-secreting cells than piglets in the NVS-VP and NVS-NVP groups on days 1, 7, 21, 35, and 60 after birth at the 3 farms.