Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-D-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite, N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other His.

• MeSH
• acetylglukosamin chemie   metabolismus   MeSH
• chromatografie afinitní MeSH
• endotoxiny biosyntéza   izolace a purifikace   MeSH
• Escherichia coli metabolismus   MeSH
• glutaminfruktosa-6-fosfáttransaminasa (izomerizující) biosyntéza   izolace a purifikace   MeSH
• kultivační média chemie   MeSH
• myši MeSH
• proteiny tepelného šoku HSP70 biosyntéza   izolace a purifikace   MeSH
• průmyslová mikrobiologie metody   MeSH
• rekombinantní proteiny biosyntéza   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

The Candida albicans heat shock protein 90 kDa (hsp90-CA) is an important target for protective antibodies in disseminated candidiasis of experimental mice and humans. Hsp90-CA is present in the cell wall of Candida pseudohyphae or hyphae--typical pathogenic morphotypes in both mucosal and systemic Candida infections. However, the potential protective effects of hsp90-CA-specific antibodies in vaginal candidiasis has not yet been reported. In the present study we used various vaccine formulations (recombinant hsp90-CA protein and hsp90-CA-encoding DNA vaccine) and routes of administration (intradermal, intranasal, and intravenous) to induce both hsp90-CA-specific systemic and vaginal mucosa immune responses in experimental BALB/c mice. The results showed that intradermal recombinant hsp90-CA protein priming, followed by intranasal or intradermal recombinant hsp90-CA protein boosting induced significant increases in both serum and vaginal hsp90-CA-specific IgG and IgA antibodies compared to the control group, as well as enhanced hsp90-CA-specific splenocyte responses in vitro. In the intradermally boosted group, subsequent experimental vaginal Candida infection induced additional increases in the hsp90-CA specific IgG isotype, suggesting that Candida has the ability to induce a local hsp90-specific antibody (IgG) response during vulvovaginal candidiasis. Further work is required to elucidate the importance of immunity to highly conserved antigens during infection of the human female reproductive tract where a balance between immunity to and tolerance for commonly antigens such as hsp90 is necessary for the maintenance of fertility.

• MeSH
• aplikace intranazální MeSH
• Candida albicans imunologie   MeSH
• DNA vakcíny aplikace a dávkování   imunologie   MeSH
• financování organizované MeSH
• fungální proteiny imunologie   MeSH
• fungální vakcíny aplikace a dávkování   imunologie   MeSH
• injekce intradermální MeSH
• injekce intravenózní MeSH
• kandidóza vulvovaginální imunologie   MeSH
• lymfocyty imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• proliferace buněk MeSH
• proteiny tepelného šoku HSP90 imunologie   MeSH
• protilátky fungální analýza   krev   MeSH
• sekundární imunizace MeSH
• slezina imunologie   MeSH
• syntetické vakcíny aplikace a dávkování   imunologie   MeSH
• tvorba protilátek MeSH
• vagina imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH

George L. Engel navrhl v roce 1977 (in Caplan, Englehart, McCartney, 1981, pp. 589) biopsycho- sociální model zdraví jako alternativu modelu výhradně biomedicínskému. Model tří determinant zdraví převzala WHO a podle něj definovala, že zdraví je stav úplné tělesné, duševní a sociální pohody, nejen nepřítomnost nemoci. Takto pojatá definice zdraví změnila zásadně přístup kjeho determinantám. V souladu s koncepcí WHO se v této práci se zaměřujeme na životní styl jako na jednu z nejvýznamnějších determinant zdravotního stavu, a to zkoumáním fyzických, psychických i sociálních komponent zdravého životního stylu. Cílem je sledování jejich vzájemných vztahů a také jejich vztahu ke zdravotní anamnéze. Dalším cílem je porovnání interaktivních složek jednotlivých komponent zdravého životního stylu (tj. poznání (K), postoje (A), jednání (B».

• Klíčová slova
• determinanty zdraví,
• MeSH
• lidé MeSH
• mladý dospělý MeSH
• ošetřovatelství MeSH
• postoj ke zdraví MeSH
• průzkumy a dotazníky využití   MeSH
• sociologie MeSH
• studenti ošetřovatelství MeSH
• zdravé chování MeSH
• zdraví - znalosti, postoje, praxe MeSH
• zdraví MeSH
• životní styl MeSH
• Check Tag
• lidé MeSH
• mladý dospělý MeSH

Vaccination is historically one of the most successful strategies for the prevention of infectious diseases. For safety reasons, modern vaccinology tends toward the usage of inactivated or attenuated microorganisms and uses predominantly subunit vaccines. The antigens need to be clearly defined, pure, stable, appropriately composed, and properly presented to the immune system of the host. Differing ratios of various proportions between specific CD4+ and CD8+ T cell responses are essential for conferring the required protection in the case of individual vaccines. To stimulate both CD4+ and CD8+ T cells, the antigens must be processed and presented to both antigen-presentation pathways, MHC I and MHC II. Protein antigens delivered by vaccination are processed as extracellular antigens. However, extracellularly delivered antigen can be directed towards intracellular presentation pathways in conjugation with molecules involved in antigen cross-presentation, e.g. heat shock proteins, or by genomic-DNA vaccination. In this overview, current knowledge of the host immune response to DNA vaccines is summarized in the introduction. The subsequent sections discuss techniques for enhancing DNA vaccine efficacy, such as DNA delivery to specific tissues, delivery of DNA to the cell cytoplasm or nucleus, and enhancement of the immune response using molecular adjuvants. Finally, the prospects of DNA vaccination and ongoing clinical trials with various DNA vaccines are discussed.

• MeSH
• aktivace lymfocytů MeSH
• DNA vakcíny aplikace a dávkování   imunologie   škodlivé účinky   MeSH
• financování organizované MeSH
• genetické vektory MeSH
• klinické zkoušky jako téma MeSH
• lidé MeSH
• předpověď MeSH
• prezentace antigenu MeSH
• systémy cílené aplikace léků MeSH
• tvorba protilátek MeSH
• vakcinace trendy   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: Borrelia burgdorferi sensu lato is a group of at least twelve closely related species some of which are responsible for Lyme disease, the most frequent zoonosis in Europe and the USA. Many of the biological features of Borrelia are unique in prokaryotes and very interesting not only from the medical viewpoint but also from the view of molecular biology. METHODS: Relevant recent articles were searched using PubMed and Google search tools. RESULTS AND CONCLUSION: This is a review of the biological, genetic and physiological features of the spirochete species group, Borrelia burgdorferi sensu lato. In spite of a lot of recent articles focused on B. burgdorferi sensu lato, many features of Borrelia biology remain obscure. It is one of the main reasons for persisting problems with prevention, diagnosis and therapy of Lyme disease. The aim of the review is to summarize ongoing current knowledge into a lucid and comprehensible form.

• MeSH
• Borrelia burgdorferi komplex fyziologie   MeSH
• lidé MeSH
• lymeská nemoc mikrobiologie   přenos   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Verification of the efficacy of Biocan B inj. ad us. vet. (Bioveta, a.s.) was done by challenge testing. Ticks collected in the nature were used as natural vectors of the infection. Six beagles and two control ones were used in the test. Formation of outer surface protein A specific antibodies (OspA antibodies) and borrelia specific immonoglobulins (IgG) was measured by Western blot and EIA in the sera samples. The tissue samples were used for detection of borreliae by cultivation method and dark field microscopy (DFM). Formation of IgG antibodies and OspA antibodies after vaccination was observed. The maximum titer level of antibodies was reached between 21. and 49. day after vaccination and then slowly decreased. Presence of borreliae was detected only in skin biopsies of non-vaccinated dogs. The post mortem tissue samples showed presence of borreliae in all of the samples of the non-vaccinated dogs. The tissues of the vaccinated dogs were not infected with borreliae, except for two samples of dog with low titer levels of OspA antibodies. The development of the new vaccine is based on preparation of recombinant outer surface proteins (e.g. rOspA and rOspC) of B. afzelii, B. burgdorferi and B. garinii origin. Chosen recombinant proteins were successfully expressed in E. coli. The obtained purified proteins are currently being tested on laboratory BALB/c mice. Formation of specific antibodies against some recombinant proteins has been confirmed. These proteins are suitable candidates for preparation of a vaccine prototype and they will be subsequently used in challenge tests.

• MeSH
• antigeny povrchové imunologie   MeSH
• bakteriální vakcíny imunologie   MeSH
• Borrelia burgdorferi komplex imunologie   MeSH
• infekce bakteriemi rodu Borrelia prevence a kontrola   veterinární   MeSH
• lipoproteiny imunologie   MeSH
• nemoci psů imunologie   prevence a kontrola   MeSH
• proteiny vnější bakteriální membrány imunologie   MeSH
• protilátky bakteriální krev   MeSH
• psi MeSH
• syntetické vakcíny imunologie   MeSH
• vakcína proti lymeské nemoci imunologie   MeSH
• vakcinace MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this work was isolation and purification of the major immunodominant protein, Outer surface protein C (OspC) of three members of the species group Borrelia burgdorferi, the causative agent of Lyme disease. Our aim was to obtain this protein in a quantity and purity sufficient for immunization of experimental animals. For optimalization of protein purification's yield we used immobilized metal ion affinity chromatography (IMAC) under different conditions. The greatest efficiency was achieved by using of HiTrap Chelating Column under native conditions.

• MeSH
• antigeny bakteriální biosyntéza   izolace a purifikace   MeSH
• DNA vakcíny biosyntéza   izolace a purifikace   MeSH
• Escherichia coli MeSH
• genetické vektory MeSH
• proteiny vnější bakteriální membrány biosyntéza   izolace a purifikace   MeSH
• vakcína proti lymeské nemoci biosyntéza   izolace a purifikace   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The recombinant Outer surface protein A (rOspA) from Borrelia burgdorferi is a possible immunogen for protection of infected humans and animals against development of Lyme borreliosis (Lyme disease), a chronic tick-borne disease characterised by diverse dermatologic, neurologic, rheumatic, and cardiac manifestations. For several years, research and development have been directed towards a vaccine for the prevention of this debilitating disease. Numerous animal studies demonstrate that pre-existing antibodies against the outer surface proteins of B. burgdorferi can prevent infection and disease caused by this organism. In this communication, using recombinant DNA technology, genes from B. burgdorferi sensu stricto and B. afzelii were inserted into E. coli-expression vectors and the rOspA were produced. Our aim was to obtain rOspA protein in a purity and quantity desirable for immunization of experimental animals. rOspA is currently the most developed, molecularly-defined vaccine candidate for the prevention of Lyme borreliosis.

• MeSH
• antigeny povrchové biosyntéza   izolace a purifikace   MeSH
• bakteriální vakcíny biosyntéza   izolace a purifikace   MeSH
• Borrelia burgdorferi komplex imunologie   MeSH
• Borrelia burgdorferi imunologie   MeSH
• DNA vakcíny biosyntéza   izolace a purifikace   MeSH
• Escherichia coli MeSH
• genetické vektory MeSH
• lipoproteiny biosyntéza   izolace a purifikace   MeSH
• proteiny vnější bakteriální membrány biosyntéza   izolace a purifikace   MeSH
• vakcína proti lymeské nemoci biosyntéza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH