Trichinella spiralis infection induces angiogenic factor thymosin β4 expression

Trichinella spiralis (T. spiralis) has been reported to up-regulate the expression of the angiogenic molecule vascular endothelial cell growth factor (VEGF) during nurse cell formation. In order to analyze the induction of angiogenesis by T. spiralis, the expression patterns of angiogenesis-related proteins were investigated by immunohistochemical analysis. VEGF expression was induced in the infected muscles at an early stage of infection (10 days after infection) and diminished after 3 weeks. Thymosin β4, a major factor which induces VEGF, showed increased expression in muscle fibers 10 days after infection, and the expression remained high in the nurse cells for 6 weeks, when the formation of the nurse cell complex was completed. The hypoxia inducible factor (HIF)-1α showed a diffuse expression pattern around the infected muscle fibers and was strongly expressed in inflammatory cells but was not related to the hypoxic condition caused by nurse cell formation. Localization of the hypoxic regions by the hypoxia marker, pimonidazole showed T. spiralis infection does not induce a hypoxic condition in nurse cells. These results suggest that the expression of VEGF and thymosin β4 induce angiogenesis and the expression of thymosin β4 remained elevated to potentially regulate nurse cell formation and maintenance.     

Influence of modifiable risk factors on the incidence of stillbirth/perinatal mortality in dairy cattle

Bovine perinatal mortality is defined as the death of a full-term calf before, during or up to 48 h after calving. Recent studies indicate that the prevalence of bovine perinatal mortality is increasing, particularly in Holstein primiparae. Factors leading to a greater incidence of dystocia are the most important modifiable variables influencing the risk of perinatal mortality. Modifiable predictors are largely (age at first calving, breeding method, sire, calving management, feto-maternal health status and gestational nutrition) or moderately (calf breed, sex, gestation length) under the control of the dairy farm manager. Unlike non-modifiable risk factors, such as primiparity and fetal plurality, these factors can be manipulated to reduce the incidence of perinatal mortality.     

An International Delphi Study of the Causes of Death and the Criteria Used to Assign Cause of Death in Bovine Perinatal Mortality

The objective of the present study was to elicit opinion from two groups of veterinarians [subject matter experts and non‐subject matter experts] about the causes of bovine perinatal mortality and the criteria used to assign such causes. The subject matter experts were selected on the basis of their scientific publications or experience of working in a veterinary diagnostic or research laboratory in the area of bovine perinatal mortality. The non‐subject matter experts were self‐selected as cattle veterinarians without particular expertise in bovine perinatology. A total of 74 veterinarians (46 subject matter experts and 28 non‐subject matter experts) from 23 countries responded. The study was conducted using Delphi methodology over seven rounds. Respondents were asked to agree the causes of bovine perinatal mortality and for each cause to agree the supporting diagnostic criteria. There was a close agreement between groups on 16 causes of death apart from intra‐uterine growth retardation (IUGR) and micronutrient imbalances which were accepted by fewer subject matter experts. There was inter‐group consensus on the criteria to diagnose accidents, congenital defects, dystocia, hyperthermia, infections, premature placental separation, prematurity and prolonged calving. There was inter‐group consensus on the criteria to diagnose anoxia, apart from gingival cyanosis; on haemorrhage, apart from haemorrhagic anaemia; on IUGR, apart from organ weights; and on iodine imbalance, apart from goitre and thyroid iodine content. The results from this study highlighted the current lack of standardization of the criteria used to define the cause of death for bovine perinatal mortality and the need for such standardization.     

Alteration of helper T-cell related cytokine production in splenocytes during Trichinella spiralis infection

Infection by Trichinella spiralis takes place in two distinct phases: one is the intestinal phase and the other is the muscle phase. To evaluate alterations in cytokine production during a T. spiralis infection, we periodically assessed the cytokine production of splenocytes in mice after infection (AI). The levels of Th2-related cytokines immediately increased after the initiation of T. spiralis larval intestinal invasion (1 week AI). These early elevations in the Th2 response might be associated with the innate immune responses of intestine epithelial cells against T. spiralis larval invasion. IL-4 and IL-13 levels reached a peak prior to the initiation of nurse cell formation (2 weeks AI). Additionally, all Th17-related cytokines, except for IL-17, increased slightly until 2 weeks AI. However, expression levels for all of the Th2 and Th17-related cytokines began to decrease after the initiation of nurse cell formation and reached basal levels at 4 weeks AI, except for IL-5. At the same time, the CD4(+)CD25(+)Foxp3(+) T (regulatory T, T(reg)) cell population increased significantly in the spleen. Additionally, the number of cells in the peripheral lymph nodes increased. In conclusion, T. spiralis larva intestinal invasion induced the production of Th2 and Th17 cell-related cytokines, and the cytokines decreased with T(reg) cell-related cytokine.     

Application of Adjoint-based Forecast Sensitivities to Asian Dust Transport Events in Korea

Phytoremediation is an attractive, economic alternative to soil removal and burial methods to remediate contaminated soil. However, it is also a slow process. The effect of humic acid in enhancing B and Pb phytoextraction from contaminated soils was studied (pot experiment) using transplanted vetiver grass (Vetiveria zizanioides (L.) Nash). Boron was applied at 0, 45, 90 and 180 kg B ha^?1 soil (as H₃BO₃) in 16 replicates. Of the 64 pots, four pots each were treated with 0, 100, 200 and 400 kg ha^?1 humic acid (HA) solution. In a separate experiment, Pb was applied (as Pb(NO₃)₂) at 0, 45, 90 and 180 kg Pb ha^?1 prior to addition of HA solutions at levels identical to the B experiment. Experiments were conducted using a randomized complete block design with four replicates. Vetiver grass was harvested 90 days after planting. Lead addition beyond 45 kg Pb ha^?1 decreased Pb uptake mostly due to a yield decline. Humic acid application increased Pb availability in soil and enhanced Pb uptake while maintaining or enhancing yield. An application of 200 kg HA ha^?1 was optimal for maintaining yield at elevated Pb levels. Boron application did not impact yield but greatly increased B content of roots and shoot. Boron uptake was greatest upon addition of 400 kg HA ha^?1. We conclude that HA addition to vetiver grass can be an effective way to enhance phytoremediation of B and Pb but optimum rates differ depending on soil B and Pb contamination levels.     

Oxygenation of arachidonoyl lysophospholipids by lipoxygenases from soybean, porcine leukocyte, or rabbit reticulocyte

Oxygenation of arachidonoyl lysophosphatidylcholine (lysoPC) or arachidonoyl lysophosphatidic acid (lysoPA) by lipoxygenase (LOX) was examined. The oxidized products were identified by HPLC/UV spectrophotometry/mass spectrometry analyses. Straight-phase and chiral-phase HPLC analyses indicated that soybean LOX-1 and rabbit reticulocyte LOX oxygenated arachidonoyl lysophospholipids mainly at C-15 with the S form as major enantiomer, whereas porcine leukocyte LOX oxygenated at C-12 with the S form. Next, the sequential exposure of arachidonoyl-lysoPC to soybean LOX-1 and porcine leukocyte LOX afforded two major isomers of dihydroxy derivatives with conjugated triene structure, suggesting that 15(S)-hydroperoxyeicosatetraenoyl derivatives were converted to 8,15(S)-dihydroxyeicosatetraenoyl derivatives. Separately, arachidonoyl-lysoPA, but not arachidonoyl-lysoPC, was found to be susceptible to double oxygenation by soybean LOX-1 to generate a dihydroperoxyeicosatetraenoyl derivative. Overall, arachidonoyl lysophospholipids were more efficient than arachidonic acid as LOX substrate. Moreover, the catalytic efficiency of arachidonoyl-lysoPC as substrate of three lipoxygenases was much greater than that of arachidonoyl-lysoPA or arachidonic acid. Taken together, it is proposed that arachidonoyl-lysoPC or arachidonoyl-lysoPA is efficiently oxygenated by plant or animal lipoxygenases, C12- or C15-specific, to generate oxidized products with conjugated diene or triene structure.     

Regulation of lipoxygenase activity by polyunsaturated lysophosphatidylcholines or their oxygenation derivatives

Lysophosphatidylcholines (lysoPCs) have been known to play a role as lipid mediators in various cellular responses. In this study, we examined whether lysoPC containing linoleoyl, arachidonoyl, or docosahexaenoyl groups or their peroxy derivatives affect lipoxygenase (LOX)-catalyzed oxygenation of native substrates. First, arachidonoyl lysoPC and docosahexaenoyl lysoPC were found to inhibit potato 5-LOX-catalyzed oxygenation of linoleic acid (LA) in a noncompetitive type with Ki values of 0.38 and 1.90 microM, respectively. Likewise, arachidonoyl lysoPC and docosahexaenoyl lysoPC also inhibited 5-LOX activity from rat basophilic leukemia cells-2H3 (RBL-2H3) with IC50 values (50% inhibitory concentration) of 18.5 +/- 3.06 and 30.6 +/- 1.06 microM, respectively. Additionally, arachidonoyl lysoPC and docosahexaenoyl lysoPC also inhibited 15-LOX activity from RBL-2H3 with IC50 values of 16.6 +/- 1.3 and 24.1 +/- 2.4 microM, respectively. In a separate experiment, where lysoPC peroxides were tested for the effect on soybean LOX-1, 15(S)-hydroperoxy-5,8,11,13-eicosatetraenoyl lysoPC and 17(S)-hydroperoxy-4,7,10,13,15,19-docosahexaenoyl lysoPC potently inhibited soybean LOX-1 activity with Ki values of 6.8 and of 1.54 microM, respectively. In contrast, 13(S)-hydroperoxy-9,11-octadecadienoyl lysoPC was observed to stimulate soybean LOX-1-catalyzed oxygenation of LA markedly with AC50 value (50% activatory concentration) of 1.5 microM. Taken together, it is proposed that lysoPCs containing polyunsaturated acyl groups or their peroxy derivatives may participate in the regulation of LOX activity in biological systems.     

Effect of restricting silage feeding prepartum on time of calving,dystocia and stillbirth in Holstein-Friesian cows

A study was carried out to investigate the effect of restricting silage feeding on time of calving and calving performance in Holstein-Friesian cows. In the treatment group (n = 1,248 cows, 12 herds) silage feeding commenced in the evening (17:00 to 20:00 h), after a period of restricted access (2 to 10 h) while in the control group ad-libitum access to silage was provided over the 24 h period (n = 1,193 cows, 12 herds). Daytime and nighttime calvings were defined as calvings occurring between the hours of 06:30 and 00:29 and between 00:30 and 06:29, respectively. Restricting access to silage resulted in less calvings at night compared to cows with ad-libitum access to silage (18 vs 22%, P < 0.05). Cows with restricted access to silage had a higher percentage of difficult calvings (11 vs 7%, P < 0.001) and stillbirths (7 vs 5%, P < 0.05) compared to cows in the control group. The percentage of calvings at night was lower (13%) when access to silage was restricted for 10 h compared to 2, 4 or 6 h (22, 18, 25%, respectively) (P < 0.001). Calf sire breed, calf gender or cow parity did not influence time of calving. In conclusion, offering silage to pregnant Holstein-Friesian cows in the evening, after a period of restricted access, reduced the incidence of nighttime calvings, but increased the incidence of dystocia and stillbirth.     

Inhibition of lysophospholipase D activity by unsaturated lysophosphatidic acids or seed extracts containing 1-linoleoyl and 1-oleoyl lysophosphatidic acid

Lysophospholipase D (lysoPLD), generating lipid mediator lysophosphatidic acid (LPA) from lysophosphatidyclcholine (LPC), is known to be inhibited by lysophosphatidic acids. Meanwhile, some plant lipids are known to contain lysophospholipids as minor components. Therefore, it is interesting to test whether edible seed samples, rich in phospholipids, may contain lysophospholipids, which express a strong inhibition of lysoPLD activity. First, the structural importance of fatty acyl group in LPAs was examined by determining the inhibitory effect of various LPAs on bovine lysoPLD activity. The most potent in the inhibition of lysoPLD activity was linoleoyl-LPA ( K i, 0.21 microM), followed by arachidonoyl-LPA ( K i, 0.55 microM), oleoyl-LPA ( K i, 1.2 microM), and palmitoyl-LPA ( K i, 1.4 microM), based on the fluoresecent assay. The same order of inhibitory potency among LPA analogs with different acyl chains was also found in the spectrophotometric assay. Subsequently, the extracts of 12 edible seeds were screened for the inhibition of lysoPLD activity using both spectrophotometric and fluorescent assays. Among seed extracts tested, the extract from soybean seed, sesame seed, or sunflower seed (30 mg seed weight/mL) was found to exhibit a potent inhibition (>80%) of lysoPLD activity. In further study employing ESI-MS/MS analysis, major LPA components in seed extracts were identified to be 1-linoleoyl LPA, 1-oleoyl LPA, and 1-palmitoyl LPA with 1-linoleoyl LPA being more predominant. Thus, the potent inhibition of lysoPLD activity by seed extracts might be ascribed to the presence of LPA with linoleoyl group rather than other acyl chains.