infection response arrays
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The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria.
- MeSH
- down regulace MeSH
- fibroblasty cytologie metabolismus mikrobiologie MeSH
- kultivované buňky MeSH
- kur domácí genetika metabolismus MeSH
- kuřecí embryo MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- makrofágy metabolismus mikrobiologie MeSH
- nemoci drůbeže metabolismus patologie MeSH
- Salmonella enteritidis fyziologie MeSH
- salmonelová infekce u zvířat metabolismus patofyziologie MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- transkriptom * MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Antiretroviral restriction factors may play an essential role in the safety of xenotransplantation. Therefore, the present study focused on investigation of the changes in the tripartite motif-containing family (TRIM) gene expression in normal human dermal fibroblasts with and without lipopolysaccharide stimulation in response to porcine endogenous retrovirus infection. Analysis of the expression profile of TRIMs was performed using oligonucleotide microarrays and QRT-PCR. Nine (TRIM1, TRIM2, TRIM5, TRIM14, TRIM16, TRIM18, TRIM22, TRIM27 and TRIM31) statistically significantly differentially expressed genes were found (P < 0.05, one-way ANOVA). In conclusion, comprehensive analysis of retroviral restriction factor gene expression in human dermal fibroblasts before and after porcine endogenous retrovirus infection with and without LPS stimulation may suggest association of the selected TRIMs with antiretroviral activity.
- MeSH
- aminokyselinové motivy MeSH
- endogenní retroviry fyziologie MeSH
- fibroblasty metabolismus virologie MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- reprodukovatelnost výsledků MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- škára patologie MeSH
- stanovení celkové genové exprese * MeSH
- Sus scrofa MeSH
- transportní proteiny genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- messenger RNA MeSH
- transportní proteiny MeSH
Polymeric biomaterials with antibacterial effects are requisite materials in the fight against hospital-acquired infections. An effective way for constructing a second generation of antibacterials is to exploit the synergic effect of (i) patterning of polymeric materials by a laser, and (ii) deposition of noble metals in their nanostructured forms. With this approach, we prepared highly-ordered periodic structures (ripples) on polyethylene naphthalate (PEN). Subsequent deposition of Ag under the glancing angle of 70° resulted in the formation of self-organized, fully separated Ag nanowire (Ag NW) arrays homogenously distributed on PEN surface. Surface properties of these samples were characterized by AFM and XPS. Vacuum evaporation of Ag at the glancing angle geometry of 70° caused that Ag NWs were formed predominantly from one side of the ripples, near to the top of the ridges. The release of Ag+ ions into physiological solution was studied by ICP-MS. The results of antibacterial tests predetermine these novel structures as promising materials able to fight against a broad spectrum of microorganisms, however, their observed cytotoxicity warns about their applications in the contact with living tissues.
- Klíčová slova
- Antibacterial effects, Laser patterning, Nanowires, Polyethylene naphthalate, Silver, Surface properties, Vacuum evaporation,
- MeSH
- antibakteriální látky chemie farmakologie MeSH
- Escherichia coli účinky léků MeSH
- fotoelektronová spektroskopie MeSH
- lasery * MeSH
- mikroskopie atomárních sil MeSH
- nanodráty chemie toxicita MeSH
- polyethylen chemie MeSH
- povrchové vlastnosti MeSH
- Staphylococcus epidermidis účinky léků MeSH
- stříbro chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- polyethylen MeSH
- stříbro MeSH
Tularaemia caused by inhalation of type A Francisella tularensis bacteria is one of the most aggressive infectious diseases known, but the reasons for the very rapid spread of the organism from the lungs to internal organs and the ensuing mortality are unknown. The present study used the mouse model to examine in detail the host immune response in the lung. After an aerosol challenge with 20 c.f.u. of the type A strain FSC033, all mice developed clinical signs of severe disease, showed weight loss by day 4 of infection and died the next day. Histopathological findings in the lung revealed acute inflammation and intense vasculitis and perivasculitis on day 4. Gene transcriptional changes in the mouse lung samples were examined on days 1, 2 and 4 of infection using a cDNA microarray with 20,600 mouse clones representing 18,500 genes. In total, 424 genes were found to be differentially expressed, some of which were both up- and downregulated at different time points, 192 of which were upregulated and 234 of which were downregulated for at least one time point. A high percentage of selected genes identified by the microarray analysis were confirmed to be differentially regulated by quantitative real-time PCR. Categorization of the differentially expressed genes showed that those preferentially involved in host immune responses were activated extensively on day 4 but hardly or not at all on days 1 and 2. Further analysis revealed that several of the genes upregulated on day 4 are known to depend on gamma interferon or tumour necrosis factor alpha for their regulation. In keeping with this finding, tumour necrosis factor alpha and gamma interferon levels were found to be increased significantly in bronchoalveolar lavage on day 4.
- MeSH
- aerosoly MeSH
- Francisella tularensis patogenita MeSH
- genetická transkripce * MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- organismy bez specifických patogenů MeSH
- plíce imunologie mikrobiologie patologie MeSH
- proteiny genetika metabolismus MeSH
- regulace genové exprese MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- stanovení celkové genové exprese * MeSH
- tularemie genetika imunologie mikrobiologie patologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- aerosoly MeSH
- proteiny MeSH
Burkholderia cenocepacia causes severe pulmonary infections in cystic fibrosis (CF) patients. Since the bacterium is virtually untreatable by antibiotics, chronic infections persist for years and might develop into fatal septic pneumonia (cepacia syndrome, CS). To devise new strategies to combat chronic B. cenocepacia infections, it is essential to obtain comprehensive knowledge about their pathogenesis. We conducted a comparative genomic analysis of 32 Czech isolates of epidemic clone B. cenocepacia ST32 isolated from various stages of chronic infection in 8 CF patients. High numbers of large-scale deletions were found to occur during chronic infection, affecting preferentially genomic islands and nonessential replicons. Recombination between insertion sequences (IS) was inferred as the mechanism behind deletion formation; the most numerous IS group was specific for the ST32 clone and has undergone transposition burst since its divergence. Genes functionally related to transition metal metabolism were identified as hotspots for deletions and IS insertions. This functional category was also represented among genes where nonsynonymous point mutations and indels occurred parallelly among patients. Another category exhibiting parallel mutations was oxidative stress protection; mutations in catalase KatG resulted in impaired detoxification of hydrogen peroxide. Deep sequencing revealed substantial polymorphism in genes of both categories within the sputum B. cenocepacia ST32 populations, indicating extensive adaptive evolution. Neither oxidative stress response nor transition metal metabolism genes were previously reported to undergo parallel evolution during chronic CF infection. Mutations in katG and copper metabolism genes were overrepresented in patients where chronic infection developed into CS. Among professional phagocytes, macrophages use both hydrogen peroxide and copper for their bactericidal activity; our results thus tentatively point to macrophages as suspects in pathogenesis towards the fatal CS.
- MeSH
- Burkholderia cenocepacia genetika MeSH
- chronická nemoc MeSH
- cystická fibróza komplikace mikrobiologie MeSH
- infekce bakteriemi rodu Burkholderia genetika MeSH
- infekce dýchací soustavy mikrobiologie MeSH
- lidé MeSH
- srovnávací genomová hybridizace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: More than 90% of cases of anal cancers are caused by high-risk human papillomavirus (HR HPV) infection and a history of cervical intraepithelial neoplasia (CIN) is established as possible risk factor. OBJECTIVES: To demonstrate relationship between anal and cervical HPV infection in women with different grades of CIN and microinvasive cervical cancer. STUDY DESIGN: A total of 272 women were enrolled in the study. The study group included 172 women who underwent conization for high-grade CIN or microinvasive cervical cancer. The control group consisted of 100 women with non-neoplastic gynecologic diseases or biopsy-confirmed CIN 1. All participants completed a questionnaire detailing their medical history and sexual risk factors and were subjected to anal and cervical HPV genotyping using Cobas and Lynear array HPV test. RESULTS: Cervical, anal, and concurrent cervical and anal HPV infections were detected in 82.6%, 48.3% and 42.4% of women in the study group, and in 28.0%, 26.0% and 8.0% of women in the control group, respectively. The prevalence of the HR HPV genotypes was higher in the study group and significantly increased with the severity of cervical lesion. Concurrent infections of the cervix and anus occurred 5.3-fold more often in the study group than in the control group. Any contact with the anus was the only significant risk factor for development of concurrent HPV infection. CONCLUSIONS: Concurrent anal and cervical HR HPV infection was found in nearly half of women with CIN 2+. The dominant genotype found in both anatomical locations was HPV 16. Any frequency and any type of contact with the anus were shown as the most important risk factor for concurrent HPV infection.
- Klíčová slova
- AIN, AIS, ASC-H, Anal cancer, Anal infection, CIN, CIN 2+, COC, Cervical infection, DNA, HIV, HPV, HR, HRT, HSIL, IUD, LR, OR, PCR, adenocarcinoma in situ, anal intraepithelial neoplasia, atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion, cervical intraepithelial neoplasia, combined oral contraception, deoxyribonucleic acid, high-grade squamous intraepithelial lesion, high-gradeCIN or microinvasive cervical cancer, high-risk, hormone replacement therapy, human immunodeficiency virus, human papillomavirus, intrauterine device, low-risk, odd ratio, polymerase chain reaction,
- MeSH
- anální kanál virologie MeSH
- cervix uteri virologie MeSH
- dospělí MeSH
- dysplazie děložního hrdla virologie MeSH
- genotyp MeSH
- infekce papilomavirem komplikace epidemiologie přenos virologie MeSH
- kohortové studie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- molekulární epidemiologie MeSH
- Papillomaviridae klasifikace genetika izolace a purifikace MeSH
- průzkumy a dotazníky MeSH
- sexuální chování MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: The aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections. METHODS: For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 10⁹ CFU pro toto). RESULTS: Subcutaneous inoculation of a single dose with 2.6 × 10⁹ colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation. CONCLUSIONS: Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.
- MeSH
- časové faktory MeSH
- Francisella tularensis * MeSH
- látky reagující s kyselinou thiobarbiturovou MeSH
- oxidační stres * MeSH
- sérový albumin metabolismus MeSH
- tularemie metabolismus patologie veterinární MeSH
- zajíci * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- látky reagující s kyselinou thiobarbiturovou MeSH
- sérový albumin MeSH
The feeding success of sand flies (Diptera: Phlebotominae) is linked to the vast array of pharmacological substances in their saliva, which interferes with the host haemostasis and immune response. Modification of feeding site plays also an important role in Leishmania transmission. In naive hosts, co-inoculation of saliva and Leishmania parasites increases the chance of successful transmission. Disease exacerbation seems to be associated with enhanced production of type 2 cytokines and selective inhibition of some macrophage functions including the production of NO and H202. On the other hand, hosts repeatedly exposed to sand fly bites develop anti-saliva immune response that results in a protection against Leishmania infection. This led to a new interesting approach to anti-Leishmania vaccine--using salivary components to block parasite transmission. The review is therefore focused on the interactions that run between immunomodulatory molecules in sand fly saliva and host immune response, with the impact on Leishmania infection development. Recent studies revealed that saliva-based vaccine for leishmaniasis might be effective and feasible, however, several questions still require to be solved. The knowledge based on experimental mouse model cannot be fully extrapolated to dogs or humans and due to differences in salivary antigens between sand fly species the protective effect is species-specific. On the other hand, the specificity of salivary antigens enables the use of anti-saliva antibodies for monitoring the exposure of hosts to sand fly bites and might be used as a marker of risks for Leishmania transmission in endemic areas.
- MeSH
- hmyz - vektory imunologie parazitologie MeSH
- Leishmania imunologie MeSH
- leishmanióza imunologie parazitologie přenos MeSH
- přenos infekční nemoci * MeSH
- Psychodidae imunologie parazitologie MeSH
- sliny imunologie parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
An array of 12 20 x 20 m quadrats in a mixed forest near Poteply, Central Bohemia, Czech Republic, was investigated for the abundance and spatial distribution of host-questing Ixodes ricinus nymphs and their infection with borreliae. Tick densities were estimated by flagging and their borrelia infection status was determined by direct immunofluorescence. While the tick population appeared to be continuous and homogeneous based on quadrat counts, their infection with borreliae evinced a mosaic pattern. The nymphal infection rates ranged between 1.6 and 10.5% (mean = 6.0%) with significant differences between adjacent quadrats. Home ranging of small rodents (Apodemus flavicollis and Clethrionomys glareolus) inhabiting the forest seems to be responsible for the spatial pattern of borrelial infection.
- MeSH
- Borrelia burgdorferi komplex imunologie izolace a purifikace MeSH
- fluorescenční protilátková technika přímá MeSH
- hlodavci MeSH
- infestace klíšťaty parazitologie veterinární MeSH
- klíště růst a vývoj mikrobiologie MeSH
- lymeská nemoc prevence a kontrola MeSH
- nemoci hlodavců parazitologie MeSH
- nymfa mikrobiologie MeSH
- protilátky bakteriální analýza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- protilátky bakteriální MeSH
Tick-borne infections are the most common vector-borne diseases in the USA. Ticks harbor and transmit several infections with Lyme disease being the most common tickborne infection in the US and Europe. Lack of awareness about tick populations, specific diagnostic tests, and overlapping signs and symptoms of tick-borne infections can often lead to misdiagnosis affecting treatment and the prevalence data reported especially for non-Lyme tick-borne infections. The diagnostic tests currently available for tick-borne diseases are severely limited in their ability to provide accurate results and cannot detect multiple pathogens in a single run. The multiplex protein microarray developed at Vibrant was designed to detect multiple serological antibodies thereby detecting exposure to multiple pathogens simultaneously. Our microarray in its present form can accommodate 400 antigens (molecules that can bind to specific antibodies) and can multiplex across antigen types, whole cell lysates, recombinant proteins, and peptides. A designed array containing multiple antigens of several microbes including Borrelia burgdorferi, the Lyme disease spirochete, was manufactured and evaluated. The immunoglobulin M (IgM) and G (IgG) responses against several tick-borne microbes and other infectious agents were analyzed for analytical and clinical performance. The microarray improved IgM and IgG sensitivities and specificities of individual microbes when compared with the respective gold standards. The testing was also performed in a single run in comparison to multiple runs needed for comparable testing standards. In summary, our study presents a flexible multiplex microarray platform that can provide quick results with high sensitivity and specificity for evaluating exposure to varied infectious agents especially tick-borne pathogens.
- Klíčová slova
- Co-infections, Immunoglobulin, Infectious disease, Lyme disease, Microarray, Multiplex, Tick bite, Tick-borne infections,
- MeSH
- Borrelia burgdorferi imunologie MeSH
- čipová analýza proteinů * metody MeSH
- imunoglobulin G imunologie krev MeSH
- imunoglobulin M imunologie krev MeSH
- klíšťata MeSH
- lidé MeSH
- lymeská nemoc diagnóza imunologie MeSH
- nemoci přenášené klíšťaty * diagnóza imunologie MeSH
- protilátky bakteriální * imunologie krev MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- imunoglobulin G MeSH
- imunoglobulin M MeSH
- protilátky bakteriální * MeSH