1,720,968 research outputs found
Evaluation of the immunoremodulatory effect of the chimeric protein Bet v 1- BanLec and its mutants on cells of the innate immune system
No full textKao jedna od najčešćih hroničnih bolesti, alergija predstavlja ozbiljan zdravstveni problem,koji osim toga utiče i na socioekonomski aspekt obolele osobe. Za razliku od farmakoterpijekoja omogućava samo kratkoročno uklanjanje simptoma, alergen- specifična imunoterapija(AIT) je jedino dugoročno rešenje za problem alergija. U cilju poboljšanja AIT, istraživanja seusredsređuju na inovativne strategije koje obuhvataju zamenu konvencionalnih alergenihekstrakata dobijenih iz prirodnih izvora. Ovaj pristup uključuje korišćenje rekombinantnoproizvedenih alergena, alergenih peptida, ili hipoalergenih izoformi alergena, kao i primenuodgovarajućih adjuvanasa radi smanjenja potrebne doze alergena. Odabir idealnog adjuvansau kombinaciji sa alergenom je ključan, te se teži pronalaženju adjuvanasa koji će efikasnoaktivirati ćelije urođenog imunskog sistema, što bi dalje modulisalo adaptivni imunskiodgovor. Na ovaj način bi se unapredila efikasnost i sigurnost AIT. Lektin banane (rBLwt) jeprotein koji specifično prepoznaje manozu i moduliše imuni odgovor mišijih makrofaga,najverovatnije kroz vezivanje za oligosaharidne komponente na površini TLR2 i CD14. U ovojdoktorskoj disertaciji je ispitivan imunomodulatorni efekat lektina banane kao potencijalnogadjuvansa na modelu glavnog alergena polena breze (rBet v 1 a; rBv1a). Rekombinantnehimere koju čine rBLwt i rBv1a (rBv1a-BLwt; Cwt), kao i hipoalergena izoforma Bet v 1(Bv1l) i mutant BLH84T (rBv1l-BLH84T (C1) i rBLH84T-Bv1l (C2)) su uspešno dizajniranain silico nakon čega su eksprimirane u Escherichia coli ekspresionom sistemu. Nakonprečišćavanja, biohemijske i imunološke karakterizacije testiran je imunomodulatorni efekatproteina na peritonealnim mišjim makrofagima kao i na humanim ćelijskim linijamauključujući i ćelijske kokulture. Među testiranim himerama, Cwt je pokazalaimunomodulatorni potencijal kroz stimulaciju sekrecije IL-10 i povećanjem odnosa izlučenihIFN-γ/IL-4. Ova studija ukazuje da Cwt himera ima najveći potencijal među analiziranimproteinima za primenu u sublingvalnoj terapiji pacijenata alergičnih na polen breze.As one of the most common chronic diseases, allergy represents a serious health issue, whichfurther affects the socio-economic aspect of allergic individuals. Unlike pharmacotherapy,which provides short-term relief of symptoms, allergen-specific immunotherapy (AIT) iscurrently the only long-term solution to the problem of allergies. To advance AIT, novelresearch focuses on innovative strategies involving the replacement of conventional allergenicextracts obtained from natural sources. This approach includes the use of recombinantlyproduced allergens, allergenic peptides, or hypoallergenic isoforms of allergens, as well as theuse of appropriate adjuvants to reduce the therapeutic allergen dose. The selection of an idealadjuvant in combination with the allergen is crucial. The aim is to find adjuvants that effectivelyactivate cells of the innate immune system, thereby modulating the adaptive immune response.likely through binding to oligosaccharide components on the surface of TLR2 and CD14. Inthis doctoral dissertation, the immunomodulatory effect of banana lectin as a potential adjuvantwas investigated on the model of the major birch pollen allergen (rBet v 1a; rBv1a). Therecombinant chimeras consisting of rBLwt and rBv1a (rBv1a-BLwt; Cwt), as well as chimerascomposed of a hypoallergenic isoform of Bet v 1 (rBv1l) and mutant banana lectin (BLH84T)(rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2)), were successfully designed in silico wassuccessfully designed in silico, expressed in Escherichia coli expression system. Afterpurification, biochemical, and immunological characterization the immunomodulatory abilityof chimeras was examined on peritoneal murine macrophages and on human cell linesincluding the cell co-culture. Among tested chimeras, Cwt showed immunomodulatorypotential through the stimulation of IL-10 secretion and increasment of IFN-γ/IL-4 ratio. Thisstudy suggests that among analysed chimeras, the Cwt chimera is the most suitable for furthertesting for the application in sublingual therapy for birch pollen allergic patients
Evaluation of the immunoremodulatory effect of the chimeric protein Bet v 1- BanLec and its mutants on cells of the innate immune system
No full textKao jedna od najčešćih hroničnih bolesti, alergija predstavlja ozbiljan zdravstveni problem,
koji osim toga utiče i na socioekonomski aspekt obolele osobe. Za razliku od farmakoterpije
koja omogućava samo kratkoročno uklanjanje simptoma, alergen- specifična imunoterapija
(AIT) je jedino dugoročno rešenje za problem alergija. U cilju poboljšanja AIT, istraživanja se
usredsređuju na inovativne strategije koje obuhvataju zamenu konvencionalnih alergenih
ekstrakata dobijenih iz prirodnih izvora. Ovaj pristup uključuje korišćenje rekombinantno
proizvedenih alergena, alergenih peptida, ili hipoalergenih izoformi alergena, kao i primenu
odgovarajućih adjuvanasa radi smanjenja potrebne doze alergena. Odabir idealnog adjuvansa
u kombinaciji sa alergenom je ključan, te se teži pronalaženju adjuvanasa koji će efikasno
aktivirati ćelije urođenog imunskog sistema, što bi dalje modulisalo adaptivni imunski
odgovor. Na ovaj način bi se unapredila efikasnost i sigurnost AIT. Lektin banane (rBLwt) je
protein koji specifično prepoznaje manozu i moduliše imuni odgovor mišijih makrofaga,
najverovatnije kroz vezivanje za oligosaharidne komponente na površini TLR2 i CD14. U ovoj
doktorskoj disertaciji je ispitivan imunomodulatorni efekat lektina banane kao potencijalnog
adjuvansa na modelu glavnog alergena polena breze (rBet v 1 a; rBv1a). Rekombinantne
himere koju čine rBLwt i rBv1a (rBv1a-BLwt; Cwt), kao i hipoalergena izoforma Bet v 1
(Bv1l) i mutant BLH84T (rBv1l-BLH84T (C1) i rBLH84T-Bv1l (C2)) su uspešno dizajnirana
in silico nakon čega su eksprimirane u Escherichia coli ekspresionom sistemu. Nakon
prečišćavanja, biohemijske i imunološke karakterizacije testiran je imunomodulatorni efekat
proteina na peritonealnim mišjim makrofagima kao i na humanim ćelijskim linijama
uključujući i ćelijske kokulture. Među testiranim himerama, Cwt je pokazala
imunomodulatorni potencijal kroz stimulaciju sekrecije IL-10 i povećanjem odnosa izlučenih
IFN-γ/IL-4. Ova studija ukazuje da Cwt himera ima najveći potencijal među analiziranim
proteinima za primenu u sublingvalnoj terapiji pacijenata alergičnih na polen breze.As one of the most common chronic diseases, allergy represents a serious health issue, which
further affects the socio-economic aspect of allergic individuals. Unlike pharmacotherapy,
which provides short-term relief of symptoms, allergen-specific immunotherapy (AIT) is
currently the only long-term solution to the problem of allergies. To advance AIT, novel
research focuses on innovative strategies involving the replacement of conventional allergenic
extracts obtained from natural sources. This approach includes the use of recombinantly
produced allergens, allergenic peptides, or hypoallergenic isoforms of allergens, as well as the
use of appropriate adjuvants to reduce the therapeutic allergen dose. The selection of an ideal
adjuvant in combination with the allergen is crucial. The aim is to find adjuvants that effectively
activate cells of the innate immune system, thereby modulating the adaptive immune response.
likely through binding to oligosaccharide components on the surface of TLR2 and CD14. In
this doctoral dissertation, the immunomodulatory effect of banana lectin as a potential adjuvant
was investigated on the model of the major birch pollen allergen (rBet v 1a; rBv1a). The
recombinant chimeras consisting of rBLwt and rBv1a (rBv1a-BLwt; Cwt), as well as chimeras
composed of a hypoallergenic isoform of Bet v 1 (rBv1l) and mutant banana lectin (BLH84T)
(rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2)), were successfully designed in silico was
successfully designed in silico, expressed in Escherichia coli expression system. After
purification, biochemical, and immunological characterization the immunomodulatory ability
of chimeras was examined on peritoneal murine macrophages and on human cell lines
including the cell co-culture. Among tested chimeras, Cwt showed immunomodulatory
potential through the stimulation of IL-10 secretion and increasment of IFN-γ/IL-4 ratio. This
study suggests that among analysed chimeras, the Cwt chimera is the most suitable for further
testing for the application in sublingual therapy for birch pollen allergic patients
Evaluation of the immunoremodulatory effect of the chimeric protein Bet v 1- BanLec and its mutants on cells of the innate immune system
No full textKao jedna od najčešćih hroničnih bolesti, alergija predstavlja ozbiljan zdravstveni problem,
koji osim toga utiče i na socioekonomski aspekt obolele osobe. Za razliku od farmakoterpije
koja omogućava samo kratkoročno uklanjanje simptoma, alergen- specifična imunoterapija
(AIT) je jedino dugoročno rešenje za problem alergija. U cilju poboljšanja AIT, istraživanja se
usredsređuju na inovativne strategije koje obuhvataju zamenu konvencionalnih alergenih
ekstrakata dobijenih iz prirodnih izvora. Ovaj pristup uključuje korišćenje rekombinantno
proizvedenih alergena, alergenih peptida, ili hipoalergenih izoformi alergena, kao i primenu
odgovarajućih adjuvanasa radi smanjenja potrebne doze alergena. Odabir idealnog adjuvansa
u kombinaciji sa alergenom je ključan, te se teži pronalaženju adjuvanasa koji će efikasno
aktivirati ćelije urođenog imunskog sistema, što bi dalje modulisalo adaptivni imunski
odgovor. Na ovaj način bi se unapredila efikasnost i sigurnost AIT. Lektin banane (rBLwt) je
protein koji specifično prepoznaje manozu i moduliše imuni odgovor mišijih makrofaga,
najverovatnije kroz vezivanje za oligosaharidne komponente na površini TLR2 i CD14. U ovoj
doktorskoj disertaciji je ispitivan imunomodulatorni efekat lektina banane kao potencijalnog
adjuvansa na modelu glavnog alergena polena breze (rBet v 1 a; rBv1a). Rekombinantne
himere koju čine rBLwt i rBv1a (rBv1a-BLwt; Cwt), kao i hipoalergena izoforma Bet v 1
(Bv1l) i mutant BLH84T (rBv1l-BLH84T (C1) i rBLH84T-Bv1l (C2)) su uspešno dizajnirana
in silico nakon čega su eksprimirane u Escherichia coli ekspresionom sistemu. Nakon
prečišćavanja, biohemijske i imunološke karakterizacije testiran je imunomodulatorni efekat
proteina na peritonealnim mišjim makrofagima kao i na humanim ćelijskim linijama
uključujući i ćelijske kokulture. Među testiranim himerama, Cwt je pokazala
imunomodulatorni potencijal kroz stimulaciju sekrecije IL-10 i povećanjem odnosa izlučenih
IFN-γ/IL-4. Ova studija ukazuje da Cwt himera ima najveći potencijal među analiziranim
proteinima za primenu u sublingvalnoj terapiji pacijenata alergičnih na polen breze.As one of the most common chronic diseases, allergy represents a serious health issue, which
further affects the socio-economic aspect of allergic individuals. Unlike pharmacotherapy,
which provides short-term relief of symptoms, allergen-specific immunotherapy (AIT) is
currently the only long-term solution to the problem of allergies. To advance AIT, novel
research focuses on innovative strategies involving the replacement of conventional allergenic
extracts obtained from natural sources. This approach includes the use of recombinantly
produced allergens, allergenic peptides, or hypoallergenic isoforms of allergens, as well as the
use of appropriate adjuvants to reduce the therapeutic allergen dose. The selection of an ideal
adjuvant in combination with the allergen is crucial. The aim is to find adjuvants that effectively
activate cells of the innate immune system, thereby modulating the adaptive immune response.
likely through binding to oligosaccharide components on the surface of TLR2 and CD14. In
this doctoral dissertation, the immunomodulatory effect of banana lectin as a potential adjuvant
was investigated on the model of the major birch pollen allergen (rBet v 1a; rBv1a). The
recombinant chimeras consisting of rBLwt and rBv1a (rBv1a-BLwt; Cwt), as well as chimeras
composed of a hypoallergenic isoform of Bet v 1 (rBv1l) and mutant banana lectin (BLH84T)
(rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2)), were successfully designed in silico was
successfully designed in silico, expressed in Escherichia coli expression system. After
purification, biochemical, and immunological characterization the immunomodulatory ability
of chimeras was examined on peritoneal murine macrophages and on human cell lines
including the cell co-culture. Among tested chimeras, Cwt showed immunomodulatory
potential through the stimulation of IL-10 secretion and increasment of IFN-γ/IL-4 ratio. This
study suggests that among analysed chimeras, the Cwt chimera is the most suitable for further
testing for the application in sublingual therapy for birch pollen allergic patients
Production, purification and structural characterisation of recombinant BanLec-Bet v 1
Full text linkThe sublingual route of allergens administration in allergen-specific immunotherapy (ASIT) is proven to be a successful way to treat patients with respiratory allergy. The trend of replacing natural extracts with purified recombinant allergens is growing. Although the purified allergens themselves are not good immunogens, the combined vector systems and adjuvans can improve their immunogenicity 1. Cell surfaces are decorated by different glycan structures, so the lectins specific for these glycans can be used to deliver particular therapeutic to target specific tissue 2. Banana lectin (BanLec) is mannose-specific protein which belongs to the subfamily of Jacalin related lectins 3. Apart from its characteristic to bind glycans, BanLec also modulates immune cells in vitro 4. On the other hand, Bet v 1 (Betula verrucosa) is the major birch pollen allergen. T-cell epitops are distributed over almost entire protein structure 5. In the study the recombinant BanLec-Bet v 1 construct is designed, produced by the recombinant DNA technology, purified and characterized by classical biochemical methods for the application in the ASIT of birch pollen allergy. The expression of newly designed BanLec-Bet v 1 was performed in E. coli BL21 (DE3). After expression the protein was found in the inclusion bodies from which it was extracted with 4 M urea solution. After renaturation, affinity chromatography (Sephadex G-75 superfine) was used for protein purification. Biochemical characterization of the chimera was performed by: SDS PAGE electrophoreses, CD spectroscopy and mass spectrometry. Biological activity of the construct was confirmed by binding of BanLecBet v 1 to a horseradish peroxidase glycoprotein in ELISA. Purified BanLec-Bet v 1 showed molecular mass of 32 kDa. CD spectra of the recombinant construct revealed well defined secondary structures with predominant beta sheets (41.2%). By mass spectrometry 51.8% of the BanLec-Bet v 1 primary structure was confirmed. Biologicaly active recombinant BanLec-Bet v 1 was produced by the recombinant DNA technology. Further in vitro and in vivo studies will evaluate immunomodulatory potential of BanLec-Bet v 1 for application in ASIT
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Full text linkWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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