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A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model
2021
Kim, Yŏng-gyu | Lee, Ju Young | Hwang, Jeong Ho | Suh, Han Na
Because minipig skin is similar to human skin in anatomy and physiology, establishing an atopic dermatitis (AD) minipig model seems meaningful. We applied 1-fluoro-2,4-dinitrobenzene (DNFB) or ovalbumin onto the back skin of five Yucatan minipigs aged 8–10 months and 19 kg in median weight. Two minipigs with the same parameters served as controls. Both DNFB and ovalbumin mediated epithelial hyperplasia, spongiosis, and immune cell infiltration in the dermis, which is a typical histopathological feature of AD. Moreover, AD upregulated the Th1- and Th2-related cytokine expressions in DNFB- or in ovalbumin-treated skin. Notably, AD-induced minipigs exhibited greater cytokine serum concentrations. Histopathological finding and cytokine analysis revealed that DNFB or ovalbumin mediates AD. However, ovalbumin-treated minipig is a more reliable and precise AD model owing to the DNFB-induced severe skin damage. In summary, ovalbumin-treated skin shows similar AD as human in histopathological and molecular analysis.
Afficher plus [+] Moins [-]Evaluation of serum and urine neutrophil gelatinase-associated lipocalin and cystatin C as biomarkers of acute kidney injury in horses
2021
Siwińska, Natalia | Żak, Agnieszka | Pasławska, Urszula
Diagnosis of acute kidney injury (AKI) in horses is difficult at the subclinical stage, due to nonspecific clinical signs. The aim of this study was to evaluate the concentrations of selected serum and urinary biomarkers in healthy horses, horses at risk of AKI, and those with clinical AKI. Thirty healthy horses, 30 horses at risk of AKI and 11 horses with clinical AKI and azotaemia were included in the study. Serum and urinary neutrophil gelatinase-associated lipocalin (NGAL) and cystatin C were measured using commercially available enzyme immunoassay tests. The median and (in parentheses) first and third quartile concentrations of selected biomarkers in healthy horses, horses at risk of AKI and horses with AKI were respectively as follows: serum cystatin C – 0.25 (0.19–0.37), 0.23 (0.15–0.37) and 0.61 (0.37–1.13) mg/L; serum NGAL – 50.5 (38.8–58.8), 51.1 (40.4–66.9) and 98.1 (59.4–128.2) ng/mL; urinary NGAL – 20.7 (17.9–24.5), 32.3 (32.7–55.8) and 36.6 (26.8–89.9) ng/mL; and urinary cystatin C – 0.1 (0.07–0.13), 0.13 (0.1–0.2) and 0.34 (0.22–0.37) mg/L. There were significant differences in the concentration of all biomarkers between the healthy and AKI-affected horses. Horses with AKI all had biomarker concentrations higher than the healthy horses. None of the biomarkers made azotaemia recognisable in all affected horses. The obtained results indicate the need to create a serum and urinary biomarker panel to detect AKI.
Afficher plus [+] Moins [-]Phytoncides in the prevention and therapy of blackhead disease and their effect on the turkey immune system
2021
Tykałowski, Bartłomiej | Śmiałek, Marcin | Kowalczyk, Joanna | Dziewulska, Daria | Stenzel, Tomasz | Koncicki, Andrzej
Turkey histomonosis poses a serious threat to poultry production due to the ban on the use of effective drugs. The aim of the study was to evaluate the influence of a phytoncidal feed supplement on the course of histomonosis. The preparation was also analysed for immunomodulatory properties. Clinical observations and production monitoring were conducted in a flock of turkeys with histomonosis from their 11ᵗʰ to 56ᵗʰ weeks of life which were treated with the adiCoxSᴼᴸPF soluble supplement in a dose of 2.5 mL/L water. Later the preparation was used in a preventive dose (1 mL/L). The influence on the immune system was evaluated in broiler turkeys having been given adiCoxSᴼᴸPF for 3 days in doses of 1 or 3 mL/L. The T and B lymphocyte percentages in turkey blood and spleen tissue were analysed with flow cytometry. ELISA was implemented to evaluate antibody titres after Ornithobacterium rhinotracheale vaccination, and biochemical analyses were performed to evaluate the supplement’s safety. AdiCoxSᴼᴸPF was found effective in therapy and prevention of histomonosis. Additionally, adiCoxSᴼᴸPF stimulated both humoral and cell-mediated immune mechanisms, without impairing the functions of internal organs. The treated turkeys also yielded better production results (eggs/hen, fertility, and hatchability). AdiCoxSᴼᴸPF possesses immunomodulatory properties and it can be used successfully in the prevention and therapy of histomonosis in turkeys.
Afficher plus [+] Moins [-]Identification of histamine in fish and fish products in Poland during 2014–2018
2021
Pawul-Gruba, Marzena | Osek, Jacek
Histamine is one of the most important and toxic biogenic amines which may be present in food and may cause food poisoning in humans when contained at a high level. It is produced during bacterial decarboxylation of histidine in fish muscles. The aim of the study was to investigate the presence of histamine in fish and fish products available in Poland during 2014–2018. A total of 421 samples of raw (248), smoked (107), canned (50), and marinated fish (16) were analysed by high-performance liquid chromatography with diode array detection. Histamine was detected in 14.1% samples of raw fish, 29% of smoked fish, 22% of canned fish and 93.8% of marinated fish in concentrations ranging from 3.4 to 156.4 mg/kg. Content of this amine above 100 mg/kg was found in four samples: raw Atlantic salmon, smoked European sprat and two samples of marinated Atlantic herring. The study showed that fish and fish products on the Polish market generally meet the food safety criteria for histamine and are safe for consumers.
Afficher plus [+] Moins [-]Monitored therapy of sporadic mycobacteriosis caused by Mycobacterium genavense in Atlantic canaries (Serinus canaria) and Bengalese finch (Lonchura striata)
2021
Ledwoń, Aleksandra | Dolka, Izabella | Adamczyk, Krzysztof | Szeleszczuk, Piotr
Mycobacteriosis is a significant disease of companion and wild birds which causes emaciation and widely distributed lesions, as well as being a potential zoonosis. Its primary aetiological agents in birds are Mycobacterium avium subsp. avium and the fastidious Mycobacterium genavense. This study monitored the therapy of birds naturally infected with Mycobacterium genavense to gain understanding of its effectiveness and the interrelation of co-infections with the disease course and pharmacotherapy. Five Atlantic canaries (Serinus canaria) and one Bengalese finch (Lonchura striata) with tentative diagnoses of mycobacteriosis resulting from M. genavense infection were treated twice daily with clarithromycin at 40 mg/kg, ethambutol at 30 mg/kg, and moxifloxacin at 10 mg/kg for 6 months. Two canaries were also found to be carriers of Cryptosporidium galli. Mycobacteria in faecal samples of all birds were investigated by bacterioscopy and quantitative PCR. Molecular tests yielded positive results for up to four months after treatment initiation for M. genavense and Cryptosporidium, but microscopy failed to detect the latter after four weeks in specimens from one canary. Co-infections with polyomavirus (in all birds) and circovirus and bornavirus (in canaries) were diagnosed. Two birds died during treatment and one was euthanised because of other disease, 1 month after treatment completion. Three canaries were in relatively good health a year after treatment. Canary circovirus and polyomavirus co-infection may suppress the immune system and this may facilitate the development of mycobacteriosis. The set of drugs used led to the complete cure of mycobacteriosis in three canaries. In one bird the disease returned. Clarithromycin was the active drug against C. galli. Molecular methods serve well to monitor mycobacteriosis therapy and identify M. genavense and C. galli carriage.
Afficher plus [+] Moins [-]Local and systemic influence of toxic levels of airborne ozone on the inflammatory response in rats
2021
Chmielewska-Krzesińska, Małgorzata | Wąsowicz, Krzysztof
Ozone is not harmful itself; however, it directly oxidises biomolecules and produces radical-dependent cytotoxicity. Exposure to ozone is by inhalation and therefore the lungs develop the main anti-inflammatory response, while ozone has an indirect impact on the other organs. This study investigated the local and systemic effects of the ozone-associated inflammatory response. Three groups each of 5 Wistar Han rats aged 6 months were exposed for 2h to airborne ozone at 0.5 ppm and a fourth identical group were unexposed controls. Sacrifice was at 3h after exposure for control rats and one experimental group and at 24 h and 48 h for the others. Lung and liver samples were evaluated for changes in expression of transforming growth factor beta 1, anti-inflammatory interleukin 10, pro-inflammatory tumour necrosis factor alpha and interleukin 1 beta and two nuclear factor kappa-light-chain-enhancer of B cells subunit genes. Total RNA was isolated from the samples in spin columns and cDNA was synthesised in an RT-PCR. Expression levels were compared to those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and analysed statistically. All variables changed non-linearly over time comparing experimental groups to the control. Conspicuous expression changes in the subunit genes and cytokines were observed in both evaluated organs. Locally and systemically, inflammation responses to ozone inhalation include regulation of certain genes’ expression. The mechanisms are unalike in lungs and liver but ozone exerts a similar effect in both organs. A broader range of variables influential on ozone response should be studied in the future.
Afficher plus [+] Moins [-]Modification of a haematoxylin, eosin, and natural saffron staining method for the detection of connective tissue
2021
Ceccopieri, Cassandra | Skonieczna, Joanna | Madej, Jan P.
The aim of our study was to optimise an existing staining procedure: haematoxylin-eosin saffron (HES). The method follows the classical haematoxylin and eosin protocol with the addition of a staining step using natural saffron to better identify the collagen fibres. The saffron solution was obtained by dissolving ground saffron stigmas in absolute alcohol. In order to test the HES method for its staining ability on four main types of collagen (I, II, III, and IV), specific tissues (skin, tooth, cartilage, aorta, spleen, and penis) were chosen. The procedure showed a sharp differentiation between muscle, stained red or pink, and connective tissue, stained bright yellow or orange. HES allows the diagnosis of reticulin fibrosis undetected in HE and in previous saffron staining procedures. HES represents an advantageous alternative to HE staining giving highly reproducible results with high diagnostic value.
Afficher plus [+] Moins [-]Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli
2021
Yu, Hai-Yang | Gao, Dong-Mei | Zhou, Wei | Xia, Bing-Bing | He, Zhi-Yuan | Wu, Bo | Jiang, Min-Zhi | Wang, Mingli | Zhao, Jun
Ovine interferon-tau (oIFN-τ) is a newly discovered type I interferon. This study used biochemical techniques to transform the oIFN-τ gene into Escherichia coli to obtain the mass and soluble expression of the recombinant protein. First, total RNA was extracted from fresh sheep embryonic tissues with TRIzol reagent and then used as a template to reverse transcribe and amplify the mature oIFN-τ gene with RT-PCR. The amplified product was next digested with the HindIII and XhoI restriction enzymes and inserted into the pET-32a(+) vector to construct the prokaryotic expression plasmid. The corrected in-frame recombinant plasmid, pET-32a(+)-oIFN-τ, was transformed into E. coli Rosetta (DE3) competent cells. After induction with isopropyl-beta-D-thiogalactopyranoside (IPTG), the recombinant protein was detected in bacteria. Finally, the bacteria were lysed by sonication, and the recombinant protein was purified by nickel affinity chromatography and DEAE anion exchange chromatography. The protein was confirmed to be oIFN-τ, which mainly existed in the soluble lysate fraction, as proven by SDS-PAGE and Western blot assays. Purified IFN-τ exists mostly in a soluble form, and its anti-vesicular stomatitis virus (VSV) activity reached 7.08×10(6)IU/mL.
Afficher plus [+] Moins [-]Mink SARS-CoV-2 infection in Poland – short communication
2021
Domańska-Blicharz, Katarzyna | Orłowska, Anna | Smreczak, Marcin | Niemczuk, Krzysztof | Iwan, Ewelina | Bomba, Arkadiusz | Lisowska, Anna | Opolska, Justyna | Trębas, Paweł | Potyrało, Patrycja | Kawiak-Sadurska, Magdalena | Rola, Jerzy
Since April 2020, when the first SARS-CoV-2 infection was reported in mink and subsequently in mink farm workers in the Netherlands, it has been confirmed that human-to-mink and mink-to-human transmission can occur. Later, SARS-CoV-2 infections in mink were reported in many European and North American countries. Samples from 590 mink from a total of 28 farms were tested by real-time RT-PCR. Whole genome sequences from one positive farm were generated and genetic relatedness was established. SARS-CoV-2 RNA was detected on a breeder farm with stock of 5,850 mink. Active viraemia was confirmed in individually tested samples with Ct values respectively between 19.4 and 29.6 for E and N gene fragments. Further testing of samples from culled animals revealed 70% positivity in throat swabs and 30% seropositivity in blood samples. Phylogenetic analysis of full-length nucleotide sequences of two SARS-CoV-2 isolates revealed that they belong to the 20B Nextstrain clade. Several nucleotide mutations were found in analysed samples compared to the reference Wuhan HU-1 strain and some of them were nonsynonymous. We report the infection of mink with SARS-CoV-2 on one farm in Poland and the results of subsequent analysis of virus sequences from two isolates. These data can be useful for assessment of the epidemiological situation of SARS-CoV-2 in Poland and how it endangers public health.
Afficher plus [+] Moins [-]Luteolin attenuates cognitive dysfunction induced by chronic cerebral hypoperfusion through the modulation of the PI3K/Akt pathway in rats
2021
He, Haitao | Chen, Xi
In our study, we evaluated the beneficial effect of luteolin in the treatment of cognitive dysfunction in rat models induced by cerebral hypoperfusion by two-vessel occlusion (2-VO). Seventy-five male Sprague Dawley rats were subjected to 2-VO surgery, in all but 15 (the sham group, group I) the ligation being permanent to impair cognitive abilities. The sham group rats received saline instead of a drug; 15 2-VO rats were not injected at all (the model group, group II); 15 2-VO rats were administered luteolin at 50 mg/kg b.w. (the lut 50 group, group III); to a further 15 luteolin was given at 100 mg/kg b.w. (the lut 100 group, group IV); and the final 15 received nimodipine at 16 mg/kg b.w. as positive controls (the nimodipine group, group V). Object recognition and Morris water maze tests were performed to investigate memory ability. A Western blot test was also conducted to assess expression of phosphatidylinositol 3-kinase (PI3K), its downstream target protein kinase B (Akt), and the phosphorylated form (P-Akt) in cerebral cortex and hippocampus tissue samples. Significant variations in the discrimination index in the object recognition test, the escape latencies in the Morris water maze test, and expression levels of PI3K-p110α and PI3K-p85 were observed three months after 2-VO surgery in both lut groups, with a significant change in the nimodipine group compared to the model group. P-Akt and Akt were expressed significantly higher in both lut groups and the nimodipine group than in the model group. Luteolin treatment of rats cognitively dysfunctional after experimental cerebral hypo perfusion was neuroprotective by activating the PI3K/Akt signals which inhibit neuronal death in the cerebral cortex and hippocampal region.
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