A potential test system for detecting contaminations by bacterial lipoproteins     

Farhat K  Ulmer AJ  Jungi TW 《Veterinary immunology and immunopathology》2012,145(1-2):66-73

Biological specimens are often contaminated with bacteria-derived products such as LPS or lipoproteins (LP), which trigger unwanted inflammatory responses in hosts. Whereas a contamination by LPS can be determined by various test systems, a contamination by LP can as yet not be determined. TLR4 and TLR2 are key components of the LPS and the LP receptor complex, respectively. It was tested in this study whether HEK293 cell stably transfected with bovine TLR2 have the ability to react to low concentrations of diacylated and triacylated synthetic LP. The stable cell lines we present here recognize low concentrations of synthetic LP resembling LP of different bacteria. Therefore, these cells are suitable to detect low contaminations present in probes. For example, HEK293 cells stably transfected with bovine TLR2 recognized an egg albumin preparation as contaminated, as evidenced by copious production of IL-8. In contrast, these cells did not respond to a highly purified human serum albumin (HSA) preparation used in the clinic but responded to HSA containing small amounts of diacylated synthetic LP. The TLR4 ligand LPS is often said to activate TLR2. Here we present evidence that LP contaminations are responsible for TLR2 activity. HEK293 cells stably transfected with bovine TLR2 and TLR1 (e.g. clone 1) did not respond to ultra-pure Escherichia coli LPS preparations but acquired responsiveness when stimulated with differently purified commercial LPS. Thus, the described system involving HEK293 cells stably transfected with bovine TLR2 and TLR1 is the first test system described attempting to measure a contamination by LP.  相似文献   

Viewpoint: factors involved in type I interferon responses during porcine virus infections     

Summerfield A 《Veterinary immunology and immunopathology》2012,148(1-2):168-171

Since type I interferon (IFN-I) is considered a potent antiviral defence mechanism, it is not surprising that during evolution viruses have development of various mechanisms to counteract IFN-I induction or release. Despite this, certain virus infections are associated with very high levels of systemic IFN-I. One explanation for this observation is the presence of plasmacytoid dendritic cells (pDC), which are able to produce high levels of IFN-I despite the presence of viral IFN-I antagonists. Examples of virus infection in pigs including classical swine fever virus, influenza virus, foot-and-mouth disease virus, and porcine circo virus type 2 highlight factors involved in controlling such responses and illustrate potential negative and positive effects for the host. Based on published data, we propose that in addition to the ability to activate pDC, the ability to spread systemically, and the tropism for lymphoid tissue also represent important factors contributing to strong systemic IFN-I responses during certain virus infections.  相似文献   

Treatment with oral cyclosporin A of a case of vesicular cutaneous lupus erythematosus in a rough collie     

Font A  Bardagi M  Mascort J  Fondevila D 《Veterinary dermatology》2006,17(6):440-442

Cyclosporin A (CsA) is an immunosuppressive agent that can be used as alternative treatment to glucocorticoid therapy. This case report describes a case of vesicular cutaneous lupus erythematosus in a rough collie successfully controlled with oral CsA for more than 18 months.  相似文献   

Color properties of four cyanidin-pyruvic acid adducts     

Oliveira J  Fernandes V  Miranda C  Santos-Buelga C  Silva A  de Freitas V  Mateus N 《Journal of agricultural and food chemistry》2006,54(18):6894-6903

Four anthocyanin-pyruvic adducts were synthesized through the reaction of cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-sophoroside, and cyanidin 3-O-sambubioside with pyruvic acid, structurally characterized by liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR), and their chromatic properties were studied (pH and SO2 stability assays). Overall, these pigments were shown to have a higher resistance to discoloration toward pH variations and also in the presence of SO2, being that this resistance to discoloration was explained by a higher protection of the chromophore group against the water or bisulfite nucleophilic attack that gives rise to the colorless hemiacetal form. Only slight differences in the protection against the nucleophilic attack of water and bisulfite were found to occur between all of the cyanidin-pyruvic acid adducts studied. Indeed, anthocyanin-pyruvic acid adducts with glucose or sambubiose attached to the 3-O position of the flavylium moiety were shown to have smaller bleaching constants compared with similar pigments that possess a rutinosyl or sophorosyl moiety. The study of the pigments (A-D and cyanidin-3-O-glucoside) color parameters, namely, chroma (C), lightness (L), and the hue angle (h(a,b)), obtained from the CIELAB system, revealed that different patterns of sugars in the anthocyanin-pyruvic acid adduct moiety affected the referred three parameters of color. The loss of saturation (DeltaC < 0) and the increase of lightness (DeltaL > 0) presented by the cyanidin-pyruvic acid adduct solutions at acidic pH values (1.0 and 2.0) showed that they are much less colored than the cyanidin-3-O-glucoside. For higher pH values (5.0 and 7.0), the reverse trend was observed. This means that the cyanidin-pyruvic acid adducts A-D are much more colored than the anthocyanin at these pH values. The higher coloring capacity of these pigments at higher pH values may be an important feature, indicating a putative application of these compounds in food products.  相似文献   

Efficacy of a live attenuated vaccine in classical swine fever virus postnatally persistently infected pigs     

Sara Mu?oz-González  Marta Perez-Simó  Marta Mu?oz  José Alejandro Bohorquez  Rosa Rosell  Artur Summerfield  Mariano Domingo  Nicolas Ruggli  Llilianne Ganges 《Veterinary research》2015,46(1)

Classical swine fever (CSF) causes major losses in pig farming, with various degrees of disease severity. Efficient live attenuated vaccines against classical swine fever virus (CSFV) are used routinely in endemic countries. However, despite intensive vaccination programs in these areas for more than 20 years, CSF has not been eradicated. Molecular epidemiology studies in these regions suggests that the virus circulating in the field has evolved under the positive selection pressure exerted by the immune response to the vaccine, leading to new attenuated viral variants. Recent work by our group demonstrated that a high proportion of persistently infected piglets can be generated by early postnatal infection with low and moderately virulent CSFV strains. Here, we studied the immune response to a hog cholera lapinised virus vaccine (HCLV), C-strain, in six-week-old persistently infected pigs following post-natal infection. CSFV-negative pigs were vaccinated as controls. The humoral and interferon gamma responses as well as the CSFV RNA loads were monitored for 21 days post-vaccination. No vaccine viral RNA was detected in the serum samples and tonsils from CSFV postnatally persistently infected pigs for 21 days post-vaccination. Furthermore, no E2-specific antibody response or neutralising antibody titres were shown in CSFV persistently infected vaccinated animals. Likewise, no of IFN-gamma producing cell response against CSFV or PHA was observed. To our knowledge, this is the first report demonstrating the absence of a response to vaccination in CSFV persistently infected pigs.  相似文献   

Identification of anthocyanin-flavanol pigments in red wines by NMR and mass spectrometry     

Mateus N  Silva AM  Santos-Buelga C  Rivas-Gonzalo JC  de Freitas V 《Journal of agricultural and food chemistry》2002,50(7):2110-2116

Three newly formed Port wine pigments were isolated by Toyopearl HW-40(s) gel chromatography and semipreparative HPLC. Furthermore, the pigments were identified by mass spectrometry (LC/MS) and NMR techniques (1D and 2D). These anthocyanin-derived pigments showed UV-visible spectra different from those of the original grape anthocyanins. These pigments correspond to malvidin 3-glucoside linked through a vinyl bond to either (+)-catechin, (-)-epicatechin, or procyanidin dimer B3 [(+)-catechin-(+)-catechin]. NMR data of these pigments are reported for the first time.  相似文献   

The response of HEK293 cells transfected with bovine TLR2 to established pathogen-associated molecular patterns and to bacteria causing mastitis in cattle     

Farhat K  Sauter KS  Brcic M  Frey J  Ulmer AJ  Jungi TW 《Veterinary immunology and immunopathology》2008,125(3-4):326-336

Toll-like receptors (TLRs) are key sensors of pathogen-associated molecular patterns (PAMPs). Their role in immunity is difficult to examine in species of veterinary interest, due to restricted access to the knockout technology and TLR-specific antibodies. An alternative approach is to generate cell lines transfected with various TLRs and to examine the recognition of PAMPs or relevant bacteria. In this report, we examined whether recognition of various PAMPs and mastitis-causing bacteria is achieved by transfection of recombinant bovine TLR2 (boTLR2). Therefore, human embryonic kidney (HEK) 293 cells were transfected by whole boTLR2. A clonal analysis of stably transfected cells disclosed variable recognition of several putative TLR2 agonists although expressing similar amounts of the transgene and endogenous TLR6. One clone (clone 25) reacted by copious interleukin-8 (IL-8) production to several stimulants of TLR2 such as di-palmitoylated cysteyl-seryl-lysyl-lysyl-lysyl-lysine (Pam2), a biochemical preparation of lipoteichoic acid from Staphylococcus aureus, a commercial preparation of peptidoglycan from S. aureus, and heat-killed Listeria monocytogenes (HKLM). TLR2-dependent induction of IL-8 release was stronger in medium containing human serum albumin than in medium containing fetal calf serum. Clone 25 cells responded to high concentrations of S. aureus and to Escherichia coli causing mastitis, but not to Streptococcus uberis and to Streptococcus agalactiae which also cause mastitis. Stimulation by S. aureus was relatively weak when compared (i) with stimulation of the same cells by HKLM and PAMPs derived from S. aureus, (ii) with a clone stably transfected with TLR4 and MD-2 and stimulated by E. coli causing mastitis, and (iii) with interferon-gamma-costimulated bovine macrophages stimulated by S. aureus and S. agalactiae. Thus, clone 25 is suitable for studying the interaction of putative TLR2 agonists with bovine TLR2-transfected cells, provides a cell to search for TLR2-specific antibodies, and is a tool for studying the interaction of TLR2 with bacteria causing disease, e.g. mastitis, in cattle.  相似文献   

Standardization of a set of microsatellite markers for use in cultivar identification studies in olive (Olea europaea L.)     

Silvia Doveri  Fabíola Sabino Gil  Aurora Díaz  Silvia Reale  Matteo Busconi  Artur da Cmara Machado  Antonio Martín  Corrado Fogher  Paolo Donini  David Lee 《Scientia Horticulturae》2008,116(4):367-373

The comparability of eight olive microsatellite profiles in 17 cultivars generated by four laboratories using different DNA genotyping platforms was tested. In total, 54 alleles were identified, from a minimum of 3 alleles (DCA15) to a maximum of 12 (DCA9), averaging 6.75 alleles per marker. A representative sample of the olive genetic variability can be obtained by selecting a relatively low number of sufficiently different cultivars. Initial comparison of the data generated, revealed the presence of a few discrepancies between the laboratories, most of them due to easily identifiable rounding errors. However, 94.9% of the genotypes were in agreement between at least two laboratories after the harmonisation of the results and only in seven cases it was not possible to determine the genotype. No discrepancies between the four laboratories were observed at all in 106 genotypes (77.9%), while 18 (13.2%) showed discrepancies at one allele and 12 (8.8%) at two alleles. Most of the differences (73.8%) were due to results obtained by only one different laboratory each time. Markers DCA3, DCA8, DCA11, DCA13, DCA14 and DCA15 showed the highest concordance percentage between datapoints scored from all partners, while DCA4 and DCA9 produced less concordant results. Forty-three percent of the discrepancies were due to heterozygous/homozygous misreadings, that is often related to the presence of stutter peaks. The determining factors for obtaining reproducible results seem to be the utilization of unique sources of plant material, the employment of the same reference cultivars by all the laboratories, the standardization of PCR conditions and the selection of the markers with the most robust amplification pattern.  相似文献   

Endoscopic findings of the stomach in pleasure horses in Poland     

Artur Nied?wied?  Krzysztof Kubiak  Józef Nicpoń 《Acta veterinaria Scandinavica》2013,55(1):45

Background

This study was performed to determine the prevalence of ulcers in the gastric squamous and glandular mucosa in Polish pleasure horses.

Study design

Medical records from gastroscopic examinations of 108 pleasure horses of different breeds were reviewed. The study population consisted of two groups; group I (n = 48) with horses that expressed mild clinical signs of gastric ulcer syndrome (EGUS) including poor appetite, slight weight loss or poor body condition, and group II (n = 60) with horses that had no signs of gastrointestinal problems. The age range was 4–10 years, including 5 males, 34 castrated males (geldings) and 69 mares. The prevalence, distribution and severity of gastric ulcers were recorded. Lesions involving the squamous mucosa and the glandular mucosa of the antrum and pylorus were graded and compared between groups.

Results

Significant difference was found in the presence and severity of gastric ulcers between the two groups of horses. The overall prevalence of gastric ulcers in the first group of horses (n = 48) was 59% while in the group of clinically healthy horses (n = 60) the prevalence of gastric lesion was 40% (P = 0.004). Almost 19% of horses from group I had between 6–10 lesions (EGUS score III) and nearly 19% had either >10 localized lesions or very large diffuse lesions (EGUS number score IV). The number of ulcerations in affected horses were significantly lower in group II compared to group I (P = 0.016) as 10% of horses had 6–10 lesions (EGUS number score III) and nearly 14% had either >10 localized lesions or very large diffuse lesions (EGUS number score IV). Gastroscopy revealed that nearly 32% of horses from the second group had an ulceration EGUS score ≥ II.

Discussion and conclusions

This study confirms that gastric ulcerations can be prevalent in apparently clinically normal pleasure horses and a complete gastroscopic examination including the examination of the pylorus is advisable to evaluate this syndrome.  相似文献   

Porcine B-cell activating factor promotes anti-FMDV antibodies in vitro but not in vivo after DNA vaccination of pigs     

Bergamin F  Saurer L  Neuhaus V  McCullough KC  Summerfield A 《Veterinary immunology and immunopathology》2007,120(3-4):115-123

‘B-cell activating factor belonging to the TNF family’ (BAFF) represents a cytokine produced by antigen presenting cells promoting B-cell maturation, activation and immunoglobulin class switching. In the present study, we demonstrate expression of BAFF on cultured monocyte-derived dendritic cells, which is further enhanced by interferon- or interferon-γ treatment. From these cells, porcine BAFF was cloned and the recombinant protein was expressed in mammalian cells with and without a FLAG tag at the carboxyl terminus. Only the protein without the FLAG tag was bioactive in vitro, and promoted B-cell survival and the differentiation of foot-and-mouth disease virus (FMDV)-specific memory B cells into antibody producing cells. Based on this result it was tested whether BAFF can enhance FMDV antibody responses in the context of a DNA vaccination. To this end, pigs were immunised with the anti-FMDV DNA vaccine plasmid pcDNA3.1/P1-2A3C3D and a pCI plasmid expressing porcine BAFF. Using a needle-free transdermal application method, also referred to as ‘jet injection’, pigs were vaccinated three times and their humoral response quantified by ELISA and a virus neutralisation test. After the third vaccination, three out of six animals vaccinated with the pcDNA3.1/P1-2A3C3D alone but none of the animals that also received the BAFF expressing plasmid had seroconverted. These data suggest that BAFF is not appropriate as a genetic adjuvant when applied as a simple co-injection with the antigen-encoding plasmid.  相似文献