In a previous study, we established a collection of appropriate porcine placental extracts using PBS at 80°C (PE-PBS80) as a food supplement to increase immune activities in a mice model. In this study, piglets were treated with 0.1%, 0.3%, and 0.5% PE-PBS80 for 3 wk after weaning. Experiments were performed at 2 separate farms using 2 different pig varieties. Composition of white blood cells, lymphocyte activation, and cytokine concentrations were analyzed to assess the immune modulation effect. In Exp. 1, the number of white blood cells increased significantly in the PE-PBS80 treatment and T- and B-cell activation increased as well (P < 0.01). Interestingly, piglets in all treatments in Exp. 2 were naturally infected by a rotavirus at the third day of the experiment but recovered after d 10. Increased lymphocyte activation was observed in the PE-PBS80 treatment (P < 0.01) regardless of viral infection. Additionally, unlike in Exp. 1, the percentage of granulocytes and concentrations of interferon-γ, IL-1β, and IgG increased in the PE-PBS80 treatment (P < 0.01) and were more active in the 0.3% PE-PBS80 treatment compared with the control and the other treatment. In conclusion, 0.3% PE-PBS80 treatment modulated immune activities in antigen-infected piglets. Therefore, the PE-PBS80 pig placental extract, particularly the 0.3% supplement to the normal diet, could be useful as an alternative feed supplement to modulate immune activity during the early piglet period.
To obtain insights into the cytoplasmic maturation status of cat oocytes recovered from cat ovaries following hormone treatment, we first examined microtubule and microfilament assembly in cat oocytes recovered from hormone-treated ovaries at various stages of maturation. Additionally, we determined the alteration of spindle and microfilament assembly, as well as mitogen-activated protein kinase (MAPK) activity, in cat oocytes at 0, 6, 12 and 18 h of further maturation in vitro. We then looked at pronuclear formation and cleavage of these oocytes following parthenogenetic activation. Similar to other species, microtubules are present in germinal vesicle (GV) stage cat oocytes, and following GV breakdown, microtubules encompassed condensed chromatin particles to form the meiotic metaphase spindle. Microfilaments were located in the cortex and around the GV. A microfilament-rich area, in which the chromatin is located, was observed in the oocytes during meiotic maturation. Maturation rates in aged oocytes (cultured for 18 h) were increased when compared with that in relatively fresh oocytes (<12 h culture), and the number of oocytes with abnormal spindle shapes was also increased in aged oocytes. Furthermore, in aged oocytes, the incidence of the metaphase plate observed outside the thick microfilament domain was higher compared with that of young oocytes, and this seemed to result in an increase in the number of oocytes with two pronuclei and one polar body following activation. Western blot analysis revealed a decrease in MAPK activity in aged cat oocytes. Taken collectively, these results suggest that the optimum time for improved cytoplasmic maturation is <12 h in cat oocytes recovered from hormone-treated ovaries.
Photoplethysmography (PPG) offers the clinically meaningful parameters, such as, heart rate, and respiratory rate. In this study, we presented three respiratory signal detection algorithms using photoplethysmography raw data generated from commercial PPG sensor: (1)Min-Max (2)Peak-to-Peak (3)Pulse Shape. As reference signal, nasal sensor signal was acquired simultaneously and compared and analyzed. We used two types of moving average filtering technique to process three PPG parameters. In laboratory experiment, 6 subjects' PPG signals were measured when they respire ten and fifteen, and arbitrary times per minute. From the results, following conclusions were drawn. Min-Max and Peak-to-Peak algorithms perform better than Pulse shape algorithm. They can be used to detect respiratory rate. But, Pulse Shape algorithm was accurate for subject 4 only. More experimental data is necessary to improve the accuracy and reliability.
A worldwide flu pandemic occurred in 2009, resulting in many victims and high social damages. In the A (H1N1) virus spreading process, the pig is the intermediate host, and this virus is amplified and genetically changed through recombination in pigs. The objective of this study was to develop influenza-resistant pigs. In interferon-α and interferon-γ treated cells, the porcine Mx2 protein has been observed near the nuclear envelope, which consequently has been linked with inhibition of influenza virus proliferation. Therefore, we attempted to produce transgenic (TG) pigs overexpressing the Mx2 gene by somatic cell nuclear transfer (SCNT). Porcine fetal fibroblasts were transfected with the cytomegalovirus vector, which includes the porcine Mx2 gene. The established transgenic cell was injected into the enucleated ooplasm to produce Mx2-TG cloned embryos. In total, 511 female TG porcine embryos were transferred to 5 surrogates. Two recipients were diagnosed pregnant (pregnancy rate, 40%) on Day 25. On Day 114, 6 fetuses and 4 mummies were collected. The PCR analysis concluded that there was no integration of the Mx2 gene. Then, a male Mx2-TG cell line was established to use as donor cell of SCNT. In total, 547 male TG-SCNT embryos were produced. Of these, 38 embryos were cultured in vitro to confirm the developmental capacity of the embryos. Among these porcine SCNT-TG embryos, 26 embryos (68.4%) cleaved and 5 (13.2%) developed to the blastocyst stage. The PCR analysis confirmed that all male TG-SCNT blastocysts were for integration of the Mx2 gene. The remaining 509 male embryos were transferred to 5 surrogates. Two recipients (pregnancy rate, 40%) were diagnosed pregnant at Day 25. To date, 1 of the surrogate has maintained pregnancy and another recipient gave birth to 9 piglets. Two days after birth, 2 piglets died and the remaining piglets remain healthy. Verification analysis of gene targeting and resistance to influenza is in progress. This study has presented new possibilities of production of influenza virus resistant pig by SCNT for translational research. This work was supported by a grant from Next-Generation BioGreen 21 program (# PJ008121012011), Rural Development Administration, Republic of Korea.
The putative mouse homologue of cytochrome P-450 4F16 (Cyp4f16) is induced by interleukin-1 (Il-1), interleukin-6 (Il-6) and tumour necrosis factor-α (Tnf-α) and repressed by interleukin-10 (Il-10) and lipopolysaccharide (LPS). The Cyp4F16 is a subfamily of Cyp4F and it is also related to eicosanoids that are important mediators in the inflammatory cascade (Cui et al. 2001). To investigate the role of Cyp4F16, in the present study, we report the production of Cyp4f16 gene knock-down mice in 2 strains of mice, namely A/J and C57BL/6. The A/J is susceptible to infection and it is associated with Cyp4F16, whereas C57BL/6 is relatively resistant to infection. An shRNA-Cyp4F16 expression vector was microinjected into pronuclei of fertilized mouse oocytes and the embryos were transferred into pseudopregnant recipients. As a result, 25 and 50 mice were produced in the A/J and C57BL, respectively. Two mice in the A/J strain and 6 in the C57BL strain were confirmed by PCR as transgenic. Organs were collected in each of the lines produced by inbreeding and screened with real-time PCR for Cyp4f16 transcripts. The Cyp4f16 gene was expressed in a tissue-specific manner with high expression in the pancreas, spleen and lung and a lower level of transcription in the heart, muscle, thymus, kidney, testis and liver. In the spleen of transgenic Cyp4f16 knock-down mice, Cyp4f16 mRNA and protein expression levels were significantly lower than those of wild-type mice. The A/J Cyp4f16 knock-down mice suffered an inflammatory skin disease and tumours, but wild-type A/J mice and knock-down C57BL mice did not. Taken together, these results suggest that Cyp4f16 may play a regulatory role in the immune system and point to the use of the Cyp4f16 knock-down mouse as an experimental animal model for the study of the inflammatory process. This work was supported by a grant PJ0070762010 from BioGreen 21 Program, Rural Development Administration, Republic of Korea.
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