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Skern, Rasmus; Frost, Petter; Nilsen, Frank

doi:10.1186/1471-2199-6-10

Cited by 52 publications

(17 citation statements)

References 11 publications

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“…Gene-specific primers for the amplicons of zonula occludens (ZO)-1, occludin, and β-actin are listed in Table 1 . Relative transcript levels were quantified by the 2 −∆∆CT method as described previously [ 28 ].…”

Section: Methodsmentioning

confidence: 99%

Lysozyme improves gut performance and protects against enterotoxigenic Escherichia coli infection in neonatal piglets

Liu

et al. 2018

Vet Res

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Diarrhea remains one of the leading causes of morbidity and mortality globally, with enterotoxigenic Escherichia coli (ETEC) constituting a major causative pathogen. The development of alternative treatments for diarrhea that do not involve chemotherapeutic drugs or result in antibiotic resistance is critical. Considering that lysozyme is a naturally occurring antimicrobial peptide, in a previous study we developed a transgenic pig line that expresses recombinant human lysozyme (hLZ) in its milk. In the present study, we examined the protective effects of the consumption of this milk against ETEC infection in neonatal piglets. We found that consuming hLZ milk facilitated faster recovery from infection and decreased mortality and morbidity following an ETEC oral inoculation or infection acquired by contact-exposure. The protective effect of hLZ was associated with the enrichment of intestinal bacteria that improve gut health, such as Lactobacillus, and the enhancement of the mucosal IgA response to the ETEC-induced diarrhea. Our study revealed potential protective mechanisms underlying the antimicrobial activity of human lysozyme, validating the use of lysozyme as an effective preventive measure for diarrhea.

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Section: Methodsmentioning

confidence: 99%

Lysozyme improves gut performance and protects against enterotoxigenic Escherichia coli infection in neonatal piglets

Liu

et al. 2018

Vet Res

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“…For PCR amplifications, 1 µL of cDNA was added to a mixture containing 10 µL of 2× SYBR Green Master Mix (Takara, Dalian, China), 0.4 µL of ROX reference dye, and 0.4 µL of each primer (50 pmol/μL). Three transgenic and five wild-type pigs were used to analyze the relative mRNA levels between the two groups with the 2 −ΔΔCT method ( 42 ). The mean ΔCT of five wild-type samples was used as the control (set to 1).…”

Section: Methodsmentioning

confidence: 99%

Improved Cytotoxic T Lymphocyte Responses to Vaccination with Porcine Reproductive and Respiratory Syndrome Virus in 4-1BB Transgenic Pigs

et al. 2017

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Vaccination is the most reliable measure to prevent infectious diseases in domestic animals. Development of novel vaccines demands extensive studies with new technologies, such as using novel adjuvants and immunomodulatory molecules. The co-stimulatory molecule 4-1BB provides a key signal that directs the fate of T cells during activation, and thus is important to their function in immune protection. To determine whether host immune responses to viral infection could be promoted by enhancing 4-1BB co-stimulation, in this study, we produced transgenic pig clones expressing an extra copy of the 4-1BB gene by clustered regularly interspaced short palindromic repeats/CRISPR-associated gene 9-mediated homologous recombination at the Rosa26 locus. The immune responses of transgenic pigs to porcine reproductive and respiratory syndrome virus (PRRSV) vaccine were determined on day 14. We show that peripheral blood lymphocytes of transgenic pigs expressed around twice the level of 4-1BB mRNA than those of control pigs. We also found IL-2, TNF-α, and granzyme B mRNA levels as well as PRRSV-specific IFN-γ response were significantly upregulated in 4-1BB transgenic pigs, leading to more efficient cytotoxic T lymphocyte (CTL) killing, whereas the expressions of IL-4, IL-17, and Foxp3 were not affected. These results indicate that higher levels of 4-1BB expression involve in promoting Th1 differentiation and enhancing specific CTL responses to PRRSV, and provide a novel approach to increase the efficacy of current vaccines to control the infectious diseases.

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“…Standard curves were performed for each of the primers to verify their efficiency. The fold change for each gene was determined based on the ΔΔ C t method with the 18S reactions serving as the endogenous control (Skern et al . 2005) and DMSO treatment as the calibrator sample.…”

Section: Methodsmentioning

confidence: 99%

Comparative effects of estradiol, methyl-piperidino-pyrazole, raloxifene, and ICI 182 780 on gene expression in the murine uterus

Davis

Mao²,

Naz³

et al. 2008

Journal of Molecular Endocrinology

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Selective estrogen receptor modulators (SERMs) are potentially useful in treating various endometrial disorders, including endometrial cancer, as they block some of the detrimental effects of estrogen. It remains unclear whether each SERM regulates a unique subset of genes and, if so, whether the combination of a SERM and 17b-estradiol has an additive or synergistic effect on gene expression. We performed microarray analysis with Affymetrix Mouse Genome 430 2.0 short oligomer arrays to determine gene expression changes in uteri of ovariectomized mice treated with estradiol (low and high dose), methyl-piperidino-pyrazole (MPP), ICI 182 780, raloxifene, and combinations of high dose of estradiol with one of the SERM and dimethyl sulfoxide (DMSO) vehicle control. The nine treatments clustered into two groups, with MPP, raloxifene, and high dose of estradiol in one, and low dose of estradiol, ICI C estradiol, ICI, MPP C estradiol, and raloxifene C estradiol in the second group. Surprisingly, combining a high dose of estradiol with a SERM markedly increased (P!0 . 02) the number of regulated genes compared with each individual treatment. Analysis of expression for selected genes in uteri of estradiol and SERM-treated mice by quantitative (Q)RT-PCR generally supported the microarray results. For some cancer-associated genes, including Klk1, Ihh, Cdc45l, and Cdca8, administration of MPP or raloxifene with estradiol resulted in greater expression than estradiol alone (P!0 . 05). By contrast, ICI 182 780 suppressed more genes governing DNA replication compared with MPP and raloxifene treatments. Therefore, ICI 182 780 might be superior to MPP and raloxifene to treat estrogen-induced endometrial cancer in women.

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Abstract: Background: When estimating relative transcript abundances by quantitative real-time PCR (Q-PCR) we found that the results can vary dramatically depending on the method chosen for data analysis.

Cited by 52 publications

References 11 publications

Lysozyme improves gut performance and protects against enterotoxigenic Escherichia coli infection in neonatal piglets

Lysozyme improves gut performance and protects against enterotoxigenic Escherichia coli infection in neonatal piglets

Improved Cytotoxic T Lymphocyte Responses to Vaccination with Porcine Reproductive and Respiratory Syndrome Virus in 4-1BB Transgenic Pigs

Comparative effects of estradiol, methyl-piperidino-pyrazole, raloxifene, and ICI 182 780 on gene expression in the murine uterus

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