Preferential expression of L-type amino acid transporter 1 in ameloblasts during rat tooth development

Certain amino acid transport systems play an important role in supplying organic nutrients to each cell and for cell proliferation during tooth development. However, the mechanisms responsible for such actions are unclear. This study demonstrated for the first time that LAT1 and 4F2hc are expressed during tooth development in prenatal and postnatal rats, and that the transporters show cell-specific expression in ameloblasts, which are the epithelium-derived dental cells. LAT1 and 4F2hc expression was not observed in other dental cells of the developing teeth such as odontoblasts and cementoblasts. Overall, these results suggest that LAT1 and 4F2hc might play an important role in enamel formation.     

Monoclonal antibody analysis of porcine reproductive and respiratory syndrome virus epitopes associated with antibody-dependent enhancement and neutralization of virus infection

Enhanced infection and replication of porcine reproductive and respiratory syndrome (PRRS) virus in the presence of specific antibody has been demonstrated in vitro and in vivo, a phenomenon known as antibody-dependent enhancement (ADE). ADE is considered to be a significant obstacle to developing effective vaccines for many viruses for which ADE has been reported, since virus-specific antibodies of maternal origin or those conferred by vaccination can facilitate the entry of the virus into target cells, sometimes resulting in increased severity of the disease. In this study, the role of specific PRRS viral epitopes in ADE and/or virus neutralization (VN) was assessed in vitro using 14 monoclonal antibodies (mAbs) to 4 PRRS viral proteins: nucleocapsid (N), matrix (M), glycoprotein (GP) 5, and GP3. Each mAb recognized a distinct epitope on one of these proteins. One-way ADE and VN assays were performed in vitro using homologous PRRS virus isolates in the presence or absence of each mAb. ADE activity was determined by detecting a significant increase of progeny virus yield in porcine alveolar macrophage cultures in the presence of individual mAbs. Neutralizing activity was determined by detecting a significant reduction or complete blocking of virus replication in MARC-145 cells in the presence of individual mAbs. mAbs could be categorized into 3 groups: enhancing, neutralizing and neither. Viral epitopes which are capable of inducing neutralizing antibodies appeared to reside on the M, GP3 and GP5 proteins, while epitopes that may induce ADE-mediating antibody were associated with the N and GP5 proteins. Identification of the viral proteins and antigens and epitopes responsible for ADE- and VN-mediating antibodies may provide the basis for developing efficacious second-generation vaccines for the control of PRRS virus; yet, further epitope mapping remains to be done.     

Impaired control of IRES-mediated translation in X-linked dyskeratosis congenita

The DKC1 gene encodes a pseudouridine synthase that modifies ribosomal RNA (rRNA). DKC1 is mutated in people with X-linked dyskeratosis congenita (X-DC), a disease characterized by bone marrow failure, skin abnormalities, and increased susceptibility to cancer. How alterations in ribosome modification might lead to cancer and other features of the disease remains unknown. Using an unbiased proteomics strategy, we discovered a specific defect in IRES (internal ribosome entry site)-dependent translation in Dkc1(m) mice and in cells from X-DC patients. This defect results in impaired translation of messenger RNAs containing IRES elements, including those encoding the tumor suppressor p27(Kip1) and the antiapoptotic factors Bcl-xL and XIAP (X-linked Inhibitor of Apoptosis Protein). Moreover, Dkc1(m) ribosomes were unable to direct translation from IRES elements present in viral messenger RNAs. These findings reveal a potential mechanism by which defective ribosome activity leads to disease and cancer.     

Comparative effects of organic and inorganic selenium on selenium transfer from sows to nursing pigs

To investigate the effects of supplemental Se on the transfer of Se to nursing pigs when sows are fed diets containing a Se level above the NRC recommendation (0.15 ppm), sows were fed diets containing no supplemental Se or supplemental (0.3 ppm) Se from sodium selenite or Se yeast. A nonSe-fortified corn-soybean meal basal diet with a high endogenous Se content served as the negative control (0.20 to 0.23 ppm Se). Fifty-two sows were fed diets from 60 d prepartum until 14 d of lactation. Six sows per treatment were bled at 60 and 30 d prepartum, at farrowing, and at 14 d postpartum to measure serum Se concentrations. Colostrum was collected within 12 h postpartum, and milk was collected at 14 d of lactation. Blood was obtained from 3 pigs each from 12 litters per treatment at birth and at weaning (d 14), and pooled serum was analyzed for Se and immunoglobulin G concentrations and glutathione peroxidase activity. Regardless of treatment, serum Se in sows declined throughout gestation and gradually increased during lactation. Sows fed Se yeast tended (P < 0.06) to have greater serum Se at farrowing than sows fed unsupplemented diets. Colostrum and milk (d 14) Se concentrations increased (P < 0.01) when sows were fed Se from yeast but not from sodium selenite. At birth, serum Se was increased (P < 0.01) for pigs whose dams were fed Se yeast compared with pigs from sows fed the basal diet. At 14 d of age, there was no difference in serum Se concentration of pigs from dams fed any of the treatments. Pig serum immunoglobulin G concentrations and glutathione peroxidase-1 activity were unaffected by dietary Se source. Supplementation of gestating and lactating sow diets with Se (0.3 ppm) from an organic or inorganic source reduced the number of stillbirths per litter. However, only pigs born to sows fed organic Se (Se yeast) had greater serum Se at birth. Organic Se increased Se concentration of colostrum and 14-d milk to a greater degree than inorganic Se.     

Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid

Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.     

Comparative analysis of pathogenesis-related protein 10 (<Emphasis Type="Italic">PR10</Emphasis>) genes between fungal resistant and susceptible peppers

To elucidate the functional roles of PR10 genes from two pepper species during plant-pathogen interactions, PR10 genes were isolated from fungal-resistant (Capsicum baccatum var. PBC80) and fungal-susceptible (C. annuum var. Yeoju) pepper fruits infected with anthracnose fungus (Colletotrichum acutatum). Despite strong nucleotide sequence identity, there were significant differences in the patterns of gene expression and protein accumulation between the genes from the two host species. Induced expression of the PR10 mRNA in PBC80 (bacPR10) was highly maintained from 24 h after infection (HAI) rather than that in Yeoju (annPR10). These mRNA expression patterns were correlated with the level of respective protein that was detected as two or three bands in each species. Substantial induction of bacPR10 proteins was confirmed by 2D-gel analysis followed by immunoblotting. Immunolocalization study showed that deposition of bacPR10 was exclusively observed in the pericarp of PBC80 fruits after fungal infection, suggesting functional significance in defence. Additionally, in vitro analysis of the enzymatic properties of PR10 proteins revealed that recombinant bacPR10 had higher ribonucleolytic activity and exhibited less sensitivity to proteinase treatment than did annPR10. Taken together, these results support the idea that relative abundance and prolonged longevity of bacPR10 in PBC80 fruits may contribute to their increased resistance in response to the anthracnose fungus, as compared with Yeoju fruit.     

Fucoidan regulate blood glucose homeostasis in C57BL/KSJ m+/+db and C57BL/KSJ db/db mice

Type 2 diabetes mellitus is a multisystem disease that is characterized by hyperglycemia and is associated with the dysfunction and failure of various organs. The control of postprandial hyperglycemia is important in the prevention and intervention of type 2 diabetes. Fucoidan has several biological activities in vitro and in vivo. However, the effect of fucoidan on hyperglycemia in non-diabetic and diabetic mice has not been investigated. This study was undertaken to study the effects of different molecular weight forms (5 kilodalton (k), 5-30 k and crude) of fucoidan on oral glucose tolerance tests in non-diabetic mice and on food intake, weight gain, fasting blood glucose and blood biochemistry of db/db mice. Treatment with 200 mg/mL 5 k, 5-30 k and crude fucoidan substantially prevented hyperglycemia according to oral glucose tolerance tests in non-diabetic mice. In addition, fucoidan fractions significantly reduced blood glucose levels in diabetic mice.     

Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects

Alternative sources of mesenchymal stem cells (MSCs) for replacing bone marrow (BM) have been extensively investigated in the field of bone tissue engineering. The purpose of this study was to compare the osteogenic potential of canine MSCs derived from adipose tissue (AT), BM, umbilical cord blood (UCB), and Wharton''s jelly (WJ) using in vitro culture techniques and in vivo orthotopic implantation assays. After canine MSCs were isolated from various tissues, the proliferation and osteogenic potential along with vascular endothelial growth factor (VEGF) production were measured and compared in vitro. For the in vivo assay, MSCs derived from each type of tissue were mixed with β-tricalcium phosphate and implanted into segmental bone defects in dogs. Among the different types of MSCs, AT-MSCs had a higher proliferation potential and BM-MSCs produced the most VEGF. AT-MSCs and UCB-MSCs showed greater in vitro osteogenic potential compared to the other cells. Radiographic and histological analyses showed that all tested MSCs had similar osteogenic capacities, and the level of new bone formation was much higher with implants containing MSCs than cell-free implants. These results indicate that AT-MSCs, UCB-MSCs, and WJ-MSCs can potentially be used in place of BM-MSCs for clinical bone engineering procedures.