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Trichinella spiralis: impact on the expression of Toll-like receptor 4 (TLR4) gene during the intestinal phase of experimental trichinellosis
2018
Wojtkowiak-Giera, Agnieszka | Wandurska-Nowak, Elżbieta | Derda, Monika | Jankowska, Wiesława | Jagodziński, Paweł P. | Hadas, Edward
Introduction: Toll-like receptors (TLRs) play a key role in the rapid activation of the innate immune response to a variety of pathogens. The aim of this study was to evaluate the effect of Trichinella spiralis infection on the level of expression of the tlr4 gene in mouse intestines during the intestinal phase of experimental trichinellosis. Material and Methods: The experimental material consisted of the small and large intestines of BALB/c mice infected with Trichinella spiralis sampled at 4, 8, and 16 days post infection (dpi). Results: A statistically significant increase was demonstrated in the tlr4 mRNA level isolated from the infected mice jejunum at 4, 8, and 16 dpi over the uninfected control. Moreover, at 4, 8, and 16 dpi in the jejunum of infected mice, a strong positive reaction for the presence of TLR4 protein compared with that of uninfected mice was observed. Conclusion: Infection with T. spiralis changes the expression of the tlr4 gene in the small intestine of the mouse host.
Show more [+] Less [-]Antibody against synthetic peptide of prolactin-inducible protein homologue precursor (PIP-HP) of Bali Cattle (Bos javanicus) saliva as a potential biomarker for immunoassay development
2018
Djoko Kisworo | Sulaiman Ngongu Depamede
Objective: The present study aimed to produce and analyze antibody against a synthetic amino acid sequence fragment of PIP-HP of Bali cattle saliva.Materials and Methods: The synthetic amino acid sequence of the PIP-HP (VIRELGICPDDWAVIPIKANRF) was developed, conjugated to bovine serum albumin and was used to immunize Indonesian local rabbits. Positive sera that specific against the PIP-HP were pooled and purified sequentially by implying ammonium sulfate precipitation and protein A affinity methods. Purified antibody was then employed to analyze of PIP-HP in the ruminants saliva by means of westernblot assays.Results: A polyclonal antibody specific to asynthetic amino acid sequence fragment of PIP-HP of Bali cattle saliva was successfully produced. Our results show that the antibody potentially to be used to develop an immuno-diagnostic kit. Furthermore, the antibody was also able to inhibit the growth of both Escherichia coli and Staphylococcus aureus cultures significantly (P [J Adv Vet Anim Res 2018; 5(2.000): 182-187]
Show more [+] Less [-]Phylogeny and Differential Gene Expression Profile of IL-1R8 in Normal Dog Tissue
2022
Lauretta Turin | Laura Bianchessi | Joel Filipe | Alessia Inglesi | Federica Riva
Interleukin-1 Receptor 8 (IL-1R8) is a transmembrane protein of the IL-1 receptor family and represents an important regulator of the balance of innate and inflammatory responses. Depending on the immunological insult, IL1-R8 protects from the immunopathology or impairs the protective immune response against the insult. The expression pattern of IL-1R8 in dog tissues is unknown. Given the relevance of inflammatory diseases in dog, the aims of this study were to identify the sequence, analyze the phylogenesis and investigate the differential expression and distribution of IL-1R8 in a wide panel of non-pathologic tissues and organs by means of quantitative Real-Time PCR and uncover species-specific peculiarities. In Canis lupus familiaris, the IL1-R8 gene maps on chromosome 18, and includes ten exons. We first compared the coding sequence of dog IL-1R8 with sequences of other carnivors. Phylogenetic analysis revealed that IL-1R8 shares significantly high sequence homology with IL-1R8 of other canids particularly fox, sharing a common progenitor. Our study demonstrated that IL-1R8 is highly expressed in pancreas, considerably expressed in kidney, heart, liver, skeletal muscle, thymus, salivary gland, lymph node and lung. Interestingly, the expression pattern disclosed a unique profile for canine tissues when compared to tissues from other animal’s species. Imbalance of pro-inflammatory response leads to a vicious loop whither pro-inflammatory signaling and injury sustain each other and booster the disease. Therefore, it is crucial to investigate key regulator molecules such as IL-1R8, which functions both in homeostasis and disease and has potential to be a valid diagnostic, prognostic and therapeutic biomarker.
Show more [+] Less [-]Analytical validation of a point-of-care test and an automated immunoturbidimetric assay for the measurement of canine C-reactive protein in serum
2021
Covin, Marshal A. | Gomez, Robynne R. | Suchodolski, Jan S. | Steiner, Jörg M. | Lidbury, Jonathan A.
C-reactive protein (CRP) is an acute phase protein, which is used to evaluate and monitor the response of the innate immune system to a variety of inflammatory processes in the dog. The purpose of this study was to analytically validate a point-of-care assay (IDEXX Catalyst CRP Test) and an immunoturbidimetric assay (Gentian Canine CRP Immunoassay) for the measurement of serum CRP concentrations in dogs. These 2 assays (Catalyst, Gentian) were compared to a previously validated enzyme-linked immunosorbent assay (Tridelta Development EIA Canine CRP Assay). Linearity, precision, reproducibility, and accuracy were assessed using leftover serum samples. Agreement between assays was assessed using leftover serum samples and serum from clinically healthy dogs. Observed to expected ratios (O/E) for dilutional parallelism were 83.9 to 163.1% and 108.3 to 160.6% for the Catalyst and the Gentian assays, respectively. Coefficients of variation for intra-assay variability ranged from 6.4 to 9.5% for the Catalyst assay and 1.5 to 2.6% for the Gentian assay. Coefficients of variation for inter-assay variability ranged from 3.8 to 18.2% for the Catalyst assay and 4.5 to 5.8% for the Gentian assay. The mean O/E for recovery were 97.9% and 98.5% for the Catalyst and Gentian assays, respectively. Correlations between assays were as follows: Catalyst and Tridelta (R 2 = 0.76), Gentian and Tridelta (R 2 = 0.79), and Catalyst and Gentian (R (2) = 0.98). The Catalyst and Gentian assays are both acceptable for measuring CRP in dog serum, but their results are not directly comparable with the Tridelta assay.
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