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Characterisation of the exopolysaccharide (EPS)-producing Lactobacillus paraplantarum BGCG11 and its non-EPS producing derivative strains as potential probiotics
No full textTraditional fermented foods are the best source for the isolation of strains with specific traits to act as functional starters and to keep the biodiversity of the culture collections. Besides, these strains could be used in the formulation of foods claimed to promote health benefits, i.e. those containing probiotic microorganisms. For the rational selection of strains acting as probiotics, several in vitro tests have been proposed. In the current study, we have characterized the probiotic potential of the strain Lactobacillus paraplanta rum BGCG11, isolated from a Serbian soft, white, homemade cheese, which is able to produce a "ropy" exopolysaccharide (EPS). Three novobiocin derivative strains, which have lost the ropy phenotype, were characterized as well in order to determine the putative role of the EPS in the probiotic potential. Under chemically gastrointestinal conditions, all strains were able to survive around 1-2% (10(6)-10(7) cfu/ml cultivable bacteria) only when they were included in a food matrix (1% skimmed milk). The strains were more resistant to acid conditions than to bile salts and gastric or pancreatic enzymes, which could be due to a pre-adaptation of the parental strain to acidic conditions in the cheese habitat. The ropy EPS did not improve the survival of the producing strain. On the contrary, the presence of an EPS layer surrounding the strain BGCG11 hindered its adhesion to the three epithelial intestinal cell lines tested, since the adhesion of the three non-ropy derivatives was higher than the parental one and also than that of the reference strain Lactobacillus rhamnosus CC. Aiming to propose a potential target application of these strains as probiotics, the cytokine production of peripheral blood mononuclear cells (PBMC) was analyzed. The EPS-producing L paraplantarum BGCG11 strain showed an anti-inflammatory or immunosuppressor profile whereas the non-ropy derivative strains induced higher pro-inflammatory response. In addition, when PBMC were stimulated with increasing concentrations of the purified ropy EPS (1, 10 and 100 mu g/ml) the cytokine profile was similar to that obtained with the EPS-producing lactobacilli, therefore pointing to a putative role of this biopolymer in its immune response
Novel vaccine strategy for ocular surface infections using conjunctiva-associated lymphoid tissue as a route of immunization: local and systemic responses to model antigen
No full textTwo seasons' experience with pandemic A H1N1 influenza infection in neonates
No full textThere are only a few reports on influenza A H1N1 infection in neonates. In this paper, we present our additional experience on the clinical characteristics, epidemiology and treatment of influenza A H1N1 (2009) infection in 10 newborn infants (aged 9-24 days). Influenza A H1N1 infection was confirmed by real-time reverse transcription-polymerase chain reaction of the nasopharyngeal swab specimens. The majority of neonates presented with fever, respiratory symptoms and lethargy. The respiratory illness ranged from mild symptoms to severe pneumonia requiring mechanical ventilation. Antiviral treatment with oseltamivir was started in five patients (50%). One lethal outcome was observed, while nine patients (90%) had complete recovery. To our knowledge, this is the largest presented series of neonatal cases with different clinical symptoms. We discuss the necessity of initiation of oseltamivir in infants with different clinical features
Optimizacija heterologe proizvodnje glukanaze iz banane u E. coli
Get PDFFor the heterologous production of a banana glucanase in Escherichia coli, its gene (GenBank GQ268963) was cloned into a pG EX-4T expression vector as a fusion protein with glutathione-S-transferase (GST). BL21 cells transformed with the GST-Mus a 5 con struct were employed for production of the protein induced by 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG). The conditions for protein expression were optimized by varying the temperature (25, 30 and 37°C) and duration of protein expression (3, 6 and 12 h). The level of protein production was analyzed by densitometry of the sodium dodecyl sulfate-polyacrylamide gel (SDS-PAG) after electrophoretic resolution of the respective cell lysates. The optimal protein expression for downstream processing was obtained after 12 h of cell growth at 25°C upon addition of IPTG. Recombinant GST-Mus a 5 purified by glutathione affinity chromatography revealed a molecular mass of a bout 60 kDa. The IgE and IgG reactivity of the rGST-Mus a 5 was confirmed by dot blot an analysis with sera of individual patients from subjects with banana allergy and polyclonal rabbit antibodies against banana extract, respectively. The purified recombinant glucanase is a potential candidate for banana allergy diagnosis.Za potrebe proizvodnje u Escherichia coli gen glukanaze iz banane (GenBank GQ268963) je ukloniran u ekspresioni vektor pGEX-4T sa glutation-S-transferazom (GST). Proizvodnja ovog proteina u ćelijama je indukovana 1 mM izopropil-β-D-tiogalaktopiranozidom (IPTG). Uslovi za ekspresiju proteina su optimizovani variranjem temperature (25, 30 i 37°C) i dužine trajanja proteinske sinteze (3, 6 i 12 h). Nivo proizvodnje proteina je analiziran denzitometrijom SDS-PA gela nakon elektroforetskog razdvajanja ćelijskih lizata. Optimalna proizvodnja proteina za njegovo dalje procesovanje je dobijena gajenjem ćelija nakon dodatka IPTG na 25°C tokom 12 h. Rekombinantni GST-Mus a 5 prečišćen afinitetnom hromatografijom sa glutationom pokazuje molekulsku masu od 60 kDa. IgE i IgG reaktivnost izolovane glukanaze potvrđena je u 'dot blot' sa pojedinačnim serumima osoba alergičnih na bananu, i sa poliklonskim zečijim antitelima na ekstrakt banane, redom. Prečišćena rekombinantna glukanaza je potencijalan kandidat za dijagnozu alergije na bananu.Link to the corrected article: [https://hdl.handle.net/21.15107/rcub_intor_364
Role of gonadal hormones in programming developmental changes in thymopoietic efficiency and sexual diergism in thymopoiesis
No full textThere is a growing body of evidence indicating the important role of the neonatal steroid milieu in programming sexually diergic changes in thymopoietic efficiency, which in rodents occur around puberty and lead to a substantial phenotypic and functional remodeling of the peripheral T-cell compartment. This in turn leads to an alteration in the susceptibility to infection and various immunologically mediated pathologies. Our laboratory has explored interdependence in the programming and development of the hypothalamo-pituitary-gonadal axis and thymus using experimental model of neonatal androgenization. We have outlined critical points in the complex process of T-cell development depending on neonatal androgen imprinting and the peripheral outcome of these changes and have pointed to underlying mechanisms. Our research has particularly contributed to an understanding of the putative role of changes in catecholamine-mediated communications in the thymopoietic alterations in adult neonatally androgenized rats
Uticaj starenja na fenotipske i funkcijske karakteristike dendritskih ćelija slezine pacova
No full textStarenje dovodi do promena skoro svnh komponenti urođenog i stečenog imuniteta. Brojna istraživanja kod čoveka i miša su pokazala promene u fenotipu i funkciji dendritskih ćelija (DĆ) takom starenja i ukazala na značajne specijes specifične razlike. Budući da nema podataka, a uticaju starenja na DĆ pacova, ispitivane su kanvencionalne OH62+ DĆ (kDĆ) izolovane iz slezina mladih (3 meseca) i starih (26 meseci) AO pacova. Određivana je zastupljenost glavnih subpopulacija ovih ćelija, ekspresija MHC II i kostimulatornih molekula i sposobnost preuzimanja antigena u suspenzijama sveže izolovanih ćelija, kao i njihove fenotiiske i funkcijske karakteristike nakon in vitro stimulacije lipopalisaharidom (LPS). U suspenziji sveže izolovanih slezinskih kDĆ starih nađena je veća zastupljenost CD11b+CD4- ćelija, manja površinska gustina CD80 i CD86 molekula (što ukazuje na manji stepen zrelosti) i pokazano da su one efikasnije u endocitoznoj aktivnosti posredovanoj manoznim receptorom. Ovi nalazi se mogu povezati sa većom količinom iRNK za IL-10 u frakciji ćelija slezine niske gustine starih pacova. Izraženija „spontana" aktivacija/maturacija kDĆ starih životinja u kulturi sugergiše da starenjem uslovljene promene u sazrevanju kDĆ in vivo najverovatnije nastaju usled supresivnog delovanja IL-10, koga produkuju ćelije iz njihovog okruženja. Nakon in vitro stimulacije LPS-om, kDĆ starih živatinja su ispoljile zreliji fenotipski profil u odnosu na kDĆ mladih životinja i pokazale izmenjenu sposobnost stimulacije alogenih CD4+ T ćelija u mešanoj leukocitnoj reakciji. Osim taga, nakon stimulacije LPS-om, u kDĆ starih životinja je nađena povećana ekspresija iRNK za TNF-alfa (što bi, makar
delimično, moglo da o6jasni opisane fenotipske promene) i iRENK za IL-12/IL-23, p 40 i IL-23 p19, dok je ekspresija iRNK za IL-10 i IL-12 p35 bila smanjena. U zaključku, rezultati pokazuju da tokom ctapenja dolazi do promene u odnosu subpopulacija kDĆ u slezini, da se menja njihova sposobnost sazrevanja, alostimulatorni kapacitet i citokinski profil
Neuropeptide Y: The story, the players, the outcomes
No full textAdvances in the held of neuroimmunomodulation have shown both anatomical and functional link between the brain and the
immune system. Aside from being the most abundant peptide in the central and peripheral nervous system, Neuropeptide У (NPY)
is also distinguished by its pleiotropic functions in many other physiological systems, including the immune system, Namely, NPY
is synthesised in postganglionic nerves innervating the lymphoid organs and released upon neuronal stimulation that, together
with expression of specific У receptors by mmune cells, enables a direct effect of nerve-derived NPY on immune cell functions. On
the other hand, immune stimulation can change the NPY content in the nervous system. Therefore, the role for МРУ in bidirectional
communication between the brain and immune system has been acknowledged. immune cells are capable of producing NPY upon
appropriate stimulation, providing г basis for autocrine/paracrine regulation of immune cell functions by NPY. Moreover, there has
been increasing evidence that NPY 6 involved in the pathogenesis of various immune-mediated disorders including autoimmune
diseases and cancer. However, further investigations are needed to elucidate the role of NPY in immunity and NPY-signaling pathways
involved in modulating immune homeostasis and autoinmune pathogenesis In that sense, the future growth in the field of
NPY research will open an avenue for potential new pharmacological treatments
Ongoing mumps outbreak in Novi Sad, the autonomous province of Vojvodina, Serbia, January to April 2012
No full textFrom 16 January to 30 April 2012, a total of 119 cases of mumps were notified in Novi Sad, Serbia. Of these cases, 89 (75%), were among students. The average age of cases was 22 years-old ( range 3-37). The outbreak is still ongoing in Novi Sad and is spreading to other parts of the Vojvodina province. As of 30 April, 209 cases have been notified in the province among those 119 from Novi Sad
NPY suppressed development of experimental autoimmune encephalomyelitis in Dark Agouti rats by disrupting costimulatory molecule interactions
No full textNeuropeptide Y (NPY) suppressed clinical experimental autoimmune encephalomyelitis (EAE) and reduced numbers of CD28+, CD11b+ and CD80+ cells among spinal cord infiltrating cells at the peak of disease in Dark Agouti rat strain. Suppression of EAE was accompanied by the reduced expression of costimulatory CD80 and CD86 molecules on ED1+ macrophages and OX62+ dendritic cells in draining lymph nodes during the inductive phase of EAE. An inhibitor of dipeptidyl peptidase 4, an enzyme which terminates the action of NPY on 11 receptor subtype, did not sustain the suppressive effect of NPY on the EAE development, suggesting involvement of Y2 and Y5 receptors. (C) 2012 Elsevier B.V. All rights reserved
Evaluation of IgE reactivity of active and thermally inactivated actinidin, a biomarker of kiwifruit allergy
No full textActinidin, an abundant cysteine protease from kiwifruit, is a specific biomarker of isolated allergy to kiwifruit. This study evaluates the IgE-binding properties of biologically active and thermally inactivated actinidin. Employing two different activity assays (caseinolytic assay and zymogram with gelatin) we showed that actinidin obtained from kiwifruit extract under native conditions represents a mixture of inactive and active enzyme. The structural integrity of actinidin was confirmed by SDS-PAGE. Edman degradation, mass fingerprint and Western blot with polyclonal antibodies. Although it was capable of inducing positive skin prick test reactions, we failed to detect IgE reactivity of active actinidin in Western blot with patient sera. Thermally inactivated actinidin exhibited IgE reactivity both in vivo and in vitro, indicating that heat processed kiwifruit products may induce clinical reactivity. These findings imply that apart from the allergenic epitopes on its surface, actinidin also contains hidden epitopes inside the protein which become accessible to IgE upon thermal treatment. (C) 2011 Elsevier Ltd. All rights reserved