Production of fresh probiotic cheese with addition of transglutaminase

Svrha rada bila je istražiti utjecaj probiotičke kulture Lactobacillus acidophilus i enzima transglutaminaze (TG) na kvalitetu i senzorska svojstva autohtonog svježeg sira iz okolice Zagreba. Svježem, nepasteriziranom, obranom mlijeku dodan je enzim transglutaminaza (TG), a na različitim temperaturama i trajanju aktivacije (8 sati na 11 ºC i 4 sata na 25 ºC). Inaktivacija enzima provedena je postupkom pasterizacije (65 ºC/30 min). Nakon hlađenja na 25 ºC, mlijeko za proizvodnju svježeg sira inokulirano je mezofilnom kulturom bakterija mliječne kiseline MM101 i probiotičkom kulturom L. acidophilus LAC-1. Uz pokusne uzorke, s dodatkom TG i probiotičke kulture, proizvedeni su i kontrolni uzorci bez dodatka TG i probiotika te uzorci bez TG, ali uz dodatak probiotika. Uzorci svježeg sira proizvedeni uz dodatak TG, posebno oni gdje je TG djelovala 8 sati na 11 ºC, imali su veću masu od uzoraka proizvedenih bez dodatka enzima, pa je stoga i prinos sira bio veći, a pokazali su i manju sinerezu tijekom 10 dana čuvanja svježeg sira na 10 ºC negoli uzorci koji su proizvedeni bez dodatka enzima. Isti su uzorci dobili najviše ocjene senzorskih svojstava. Metabolička aktivnost mezofilne kulture MM101 i probiotičkog soja L. acidophilus LAC-1 utjecala je na bolji okus i miris svježeg sira, a broj živih stanica probiotičkog soja L. acidophilus LAC-1 u priređenim uzorcima bio je cca 5 x 106 cfu/g nakon 10 dana čuvanja na 10 ºC, što je više od minimalnog preporučenog broja u probiotičkim proizvodima. Dodatak transglutaminaze doprinosi boljoj konzistenciji i općem izgledu svježeg sira.The aim of this research was to examine the influence of probiotic culture Lactobacillus acidophilus and enzyme transglutaminase (TG) on quality and sensory properties of autochthonous fresh cheese from Zagreb region. Fresh, unpasteurized, skimmed milk was inoculated with TG at different temperatures and activation time (8 h at 11 ºC and 4 h at 25 ºC). Inactivation of the enzyme was carried out during the process of pasteurization (65ºC/30 min). The milk for fresh cheese production was further inoculated with mesophilic culture of lactic acid bacteria MM101 and probiotic strain Lactobacillus acidophilus LAC-1. Besides the trial samples with addition of TG and probiotic bacteria, control samples without addition of TG and probiotic were produced, as well as the samples without addition of TG but with probiotic bacteria addition. Samples of fresh cheese produced with addition of TG, especially in which TG was active at 11 ºC, had greater weight then samples produced without the enzyme addition. Therefore, their yield was also greater then yield of cheese produced without the addition of the enzyme. Furthermore, the samples of fresh cheese produced with addition of TG have shown lesser syneresis than other samples during 10 days of storage at 10 ºC. The same samples also had the best sensory properties. Metabolic activity of mesophilic culture MM101 and probiotic culture L. acidophilus LAC-1 has resulted in better taste and odour of fresh cheese. The viable cell number of probiotic strain L. acidophilus LAC-1 in prepared samples was around 5 x 106 cells/g after 10 days of storage at 10 ºC, which is higher than the minimal dose required for 27 probiotic products. Addition of transgultaminase contributed to better consistency and general appearance of produced fresh cheese

Production of fresh probiotic cheese with addition of transglutaminase

Svrha rada bila je istražiti utjecaj probiotičke kulture Lactobacillus acidophilus i enzima transglutaminaze (TG) na kvalitetu i senzorska svojstva autohtonog svježeg sira iz okolice Zagreba. Svježem, nepasteriziranom, obranom mlijeku dodan je enzim transglutaminaza (TG), a na različitim temperaturama i trajanju aktivacije (8 sati na 11 ºC i 4 sata na 25 ºC). Inaktivacija enzima provedena je postupkom pasterizacije (65 ºC/30 min). Nakon hlađenja na 25 ºC, mlijeko za proizvodnju svježeg sira inokulirano je mezofilnom kulturom bakterija mliječne kiseline MM101 i probiotičkom kulturom L. acidophilus LAC-1. Uz pokusne uzorke, s dodatkom TG i probiotičke kulture, proizvedeni su i kontrolni uzorci bez dodatka TG i probiotika te uzorci bez TG, ali uz dodatak probiotika. Uzorci svježeg sira proizvedeni uz dodatak TG, posebno oni gdje je TG djelovala 8 sati na 11 ºC, imali su veću masu od uzoraka proizvedenih bez dodatka enzima, pa je stoga i prinos sira bio veći, a pokazali su i manju sinerezu tijekom 10 dana čuvanja svježeg sira na 10 ºC negoli uzorci koji su proizvedeni bez dodatka enzima. Isti su uzorci dobili najviše ocjene senzorskih svojstava. Metabolička aktivnost mezofilne kulture MM101 i probiotičkog soja L. acidophilus LAC-1 utjecala je na bolji okus i miris svježeg sira, a broj živih stanica probiotičkog soja L. acidophilus LAC-1 u priređenim uzorcima bio je cca 5 x 106 cfu/g nakon 10 dana čuvanja na 10 ºC, što je više od minimalnog preporučenog broja u probiotičkim proizvodima. Dodatak transglutaminaze doprinosi boljoj konzistenciji i općem izgledu svježeg sira.The aim of this research was to examine the influence of probiotic culture Lactobacillus acidophilus and enzyme transglutaminase (TG) on quality and sensory properties of autochthonous fresh cheese from Zagreb region. Fresh, unpasteurized, skimmed milk was inoculated with TG at different temperatures and activation time (8 h at 11 ºC and 4 h at 25 ºC). Inactivation of the enzyme was carried out during the process of pasteurization (65ºC/30 min). The milk for fresh cheese production was further inoculated with mesophilic culture of lactic acid bacteria MM101 and probiotic strain Lactobacillus acidophilus LAC-1. Besides the trial samples with addition of TG and probiotic bacteria, control samples without addition of TG and probiotic were produced, as well as the samples without addition of TG but with probiotic bacteria addition. Samples of fresh cheese produced with addition of TG, especially in which TG was active at 11 ºC, had greater weight then samples produced without the enzyme addition. Therefore, their yield was also greater then yield of cheese produced without the addition of the enzyme. Furthermore, the samples of fresh cheese produced with addition of TG have shown lesser syneresis than other samples during 10 days of storage at 10 ºC. The same samples also had the best sensory properties. Metabolic activity of mesophilic culture MM101 and probiotic culture L. acidophilus LAC-1 has resulted in better taste and odour of fresh cheese. The viable cell number of probiotic strain L. acidophilus LAC-1 in prepared samples was around 5 x 106 cells/g after 10 days of storage at 10 ºC, which is higher than the minimal dose required for 27 probiotic products. Addition of transgultaminase contributed to better consistency and general appearance of produced fresh cheese

Matrix-M™ adjuvation broadens protection induced by seasonal trivalent virosomal influenza vaccine

Background: Influenza virus infections are responsible for significant morbidity worldwide and therefore it remains a high priority to develop more broadly protective vaccines. Adjuvation of current seasonal influenza vaccines has the potential to achieve this goal. Methods: To assess the immune potentiating properties of Matrix-M (TM), mice were immunized with virosomal trivalent seasonal vaccine adjuvated with Matrix-M (TM). Serum samples were isolated to determine the hemagglutination inhibiting (HAI) antibody titers against vaccine homologous and heterologous strains. Furthermore, we assess whether adjuvation with Matrix-M (TM) broadens the protective efficacy of the virosomal trivalent seasonal vaccine against vaccine homologous and heterologous influenza viruses. Results: Matrix-M (TM) adjuvation enhanced HAI antibody titers and protection against vaccine homologous strains. Interestingly, Matrix-M (TM) adjuvation also resulted in HAI antibody titers against heterologous influenza B strains, but not against the tested influenza A strains. Even though the protection against heterologous influenza A was induced by the adjuvated vaccine, in the absence of HAI titers the protection was accompanied by severe clinical scores and body weight loss. In contrast, in the presence of heterologous HAI titers full protection against the heterologous influenza B strain without any disease symptoms was obtained. Conclusion: The results of this study emphasize the promising potential of a Matrix-M (TM)-adjuvated seasonal trivalent virosomal influenza vaccine. Adjuvation of trivalent virosomal vaccine does not only enhance homologous protection, but in addition induces protection against heterologous strains and thus provides overall more potent and broad protective immunit

Recombinant low-seroprevalent adenoviral vectors Ad26 and Ad35 expressing the respiratory syncytial virus (RSV) fusion protein induce protective immunity against RSV infection in cotton rats

AbstractRSV is an important cause of lower respiratory tract infections in children, the elderly and in those with underlying medical conditions. Although the high disease burden indicates an urgent need for a vaccine against RSV, no licensed RSV vaccine is currently available. We developed an RSV vaccine candidate based on the low-seroprevalent human adenovirus serotypes 26 and 35 (Ad26 and Ad35) encoding the RSV fusion (F) gene. Single immunization of mice with either one of these vectors induced high titers of RSV neutralizing antibodies and high levels of F specific interferon-gamma-producing T cells. A Th1-type immune response was indicated by a high IgG2a/IgG1 ratio of RSV-specific antibodies, strong induction of RSV-specific interferon-gamma and tumor necrosis factor-alpha cytokine producing CD8 Tcells, and low RSV-specific CD4 T-cell induction. Both humoral and cellular responses were increased upon a boost with RSV-F expressing heterologous adenovirus vector (Ad35 boost after Ad26 prime or vice versa). Both single immunization and prime-boost immunization of cotton rats induced high and long-lasting RSV neutralizing antibody titers and protective immunity against lung and nasal RSV A2 virus load up to at least 30 weeks after immunization. Cotton rats were also completely protected against challenge with a RSV B strain (B15/97) after heterologous prime-boost immunization. Lungs from vaccinated animals showed minimal damage or inflammatory infiltrates post-challenge, in contrast to animals vaccinated with formalin-inactivated virus. Our results suggest that recombinant human adenoviral Ad26 and Ad35 vectors encoding the RSV F gene have the potential to provide broad and durable protection against RSV in humans, and appear safe to be investigated in infants

Anatomska i mikromorfološka građa listova Triticum aestivum L., Agropyrum repens (L) Beauv., Avena fatua L. i Lolium perenne L.

Poznato je da površinske karakteristike lista, kao i njegova unutrašnja struktura, mogu biti faktori od kojih zavisi usvajanje herbicida. Ovo istraživanje je bilo fokusirano na proučavanje listova pšenice (Triticum aestivum L.) i listova ekonomski štetnih uskolisnih korovskih vrsta, kao što su pirevina (Agropyrum repens (L.) Beauv.), divlji ovas (Avena fatua L.) i ljulj (Lolium perenne L.), kao osnove za bolje razumevanje značaja mikromorfoloških karakteristika, naročito broja stoma, kao i anatomskih karakteristika listova za prodiranje herbicida i posledične razlike u osetljivosti na herbicide. Uzorci pšenice kao i sve tri korovske vrste sakupljeni su u maju 2020. godine sa parcele pod usevom pšenice u selu Maovi (Šabac). Sve uzorkovane biljke su bile sa potpuno formiranim cvastima tj. neposredno pre cvetanja. Za morfoanatomsku analizu uzet je potpuno razvijen list koji se nalazi u čvoru ispod lista zastavičara. Uzeto je ukupno po deset listova od svake vrste, a iz središnjeg dela svakog lista uzet je uzorak veličine 2 cm i podeljen na dva dela. Jedna polovina korišćena je za analizu anatomske građe, pri čemu su, nakon procedure kalupljenja u parafin, sečenja i bojenja, dobijeni poprečni preseci sa kojih je merena debljina epidermisa i debljina mezofila. Druga polovina korišćena je za mikromorfološka ispitivanja tako što su lice i naličje lista tretirani providnim lakom za nokte, a otisci površine su preneti na mikroskopske pločice pomoću lepljive trake. Na osnovu ovih otisaka utvrđen je broj stoma po jedinici površine (gustina). Svi mikroskopski preparati su analizirani pomoću mikroskopa Leica DM2000 i snimljeni kamerom Leica DFC320. Merenja na digitalnim fotografijama su vršena u softverskom paketu Leica IM1000, a zatim je urađena statistička analiza dobijenih vrednosti. Utvrđeno je da su listovi svih ispitivanih vrsta amfistomatični. Najveći broj stoma na licu lista zabeležen je kod pirevine i ljulja, dok je kod pšenice i divljeg ovsa gustina stoma bila dvostruko manja. Na naličju lista pirevine, divljeg ovsa i pšenice broj stoma po jedinici površine lista je bio sličan, dok je kod ljulja bio značajno manji. Budući da se brojnost stoma može smatrati jednim od faktora koji mogu uticati na usvajanje herbicida, očekuje se da bi razlike u gustini (broj po jedinici površine) stoma između ovih vrsta mogle uticati na efikasnost usvajanja herbicida

Protection against H5N1 Influenza Virus Induced by Matrix-M Adjuvanted Seasonal Virosomal Vaccine in Mice Requires Both Antibodies and T Cells.

BACKGROUND:It remains important to develop the next generation of influenza vaccines that can provide protection against vaccine mismatched strains and to be prepared for potential pandemic outbreaks. To achieve this, the understanding of the immunological parameters that mediate such broad protection is crucial. METHOD:In the current study we assessed the contribution of humoral and cellular immune responses to heterosubtypic protection against H5N1 induced by a Matrix-M (MM) adjuvanted seasonal influenza vaccine by serum transfer and T-cell depletion studies. RESULTS:We demonstrate that the heterosubtypic protection against H5N1 induced by MM adjuvanted vaccine is partially mediated by antibodies. The serum contained both H5N1 cross-reactive hemagglutinin (HA)- and neuraminidase (NA)-specific antibodies but with limited virus neutralizing and no hemagglutination inhibiting activity. The cross-reactive antibodies induced antibody-dependent cellular cytotoxicity (ADCC) in vitro, suggesting a role for the Fc part of the antibodies in protection against H5N1. Besides H5N1 specific antibody responses, cross-reactive HA- and NA-specific T-cell responses were induced by the adjuvanted vaccine. T-cell depletion experiments demonstrated that both CD4+ and CD8+ T cells contribute to protection. CONCLUSION:Our study demonstrates that cross-protection against H5N1 induced by MM adjuvanted seasonal virosomal influenza vaccine requires both the humoral and cellular arm of the immune system

Maternal antibodies protect offspring from severe influenza infection and do not lead to detectable interference with subsequent offspring immunization

Abstract Background Various studies have shown that infants under the age of 6 months are especially vulnerable for complications due to influenza. Currently there are no vaccines licensed for use in this age group. Vaccination of pregnant women during the last trimester, recommended by the WHO as protective measure for this vulnerable female population, may provide protection of newborns at this early age. Although it has been observed that maternal vaccination can passively transfer protection, maternal antibodies could possibly also interfere with subsequent active vaccination of the offspring. Methods Using a mouse model, we evaluated in depth the ability of maternal influenza vaccination to protect offspring and the effect of maternal immunization on the subsequent influenza vaccination of the offspring. By varying the regimen of maternal immunization we explored the impact of different levels of maternal antibodies on the longevity of these antibodies in their progeny. We subsequently assessed to what extent maternal antibodies can mediate direct protection against influenza in their offspring, and whether these antibodies interfere with protection induced by active vaccination of the offspring. Results The number of immunizations of pregnant mice correlates to the level and longevity of maternal antibodies in the offspring. When these antibodies are present at time of influenza challenge they protect offspring against lethal influenza challenge, even in the absence of detectable HAI titers. Moreover, no detectable interference of passively-transferred maternal antibodies on the subsequent vaccination of the offspring was observed. Conclusion In the absence of a licensed influenza vaccine for young children, vaccination of pregnant women is a promising measure to provide protection of young infants against severe influenza infection

The Th1 Immune Response to Plasmodium falciparum Circumsporozoite Protein Is Boosted by Adenovirus Vectors 35 and 26 with a Homologous Insert ▿

The most advanced malaria vaccine, RTS,S, is comprised of an adjuvant portion of the Plasmodium falciparum circumsporozoite (CS) protein fused to and admixed with the hepatitis B virus surface antigen. This vaccine confers short-term protection against malaria infection, with an efficacy of about 50%, and induces particularly B-cell and CD4+ T-cell responses. In the present study, we tested by the hypothesis that the Th1 immune response to CS protein, in particular the CD8+ T-cell response, which is needed for strong and lasting malaria immunity, is boosted to sustainable levels vectors adenovirus and 26 with an homologous insert 35 (Ad35.CS/Ad26.CS). In this study, we evaluated immune responses induced with vaccination regimens based on an adjuvant-containing, yeast-produced complete CS protein followed by two recombinant low-seroprevalence adenoviruses expressing P. falciparum CS antigen, Ad35.CS (subgroup B) and Ad26.CS (subgroup D). Our results show that (i) the yeast (Hansenula polymorpha)produced, adjuvanted full-length CS protein is highly potent in inducing high CS-specific humoral responses in mice but produces poor T-cell responses, (ii) the Ad35.CS vector boosts the gamma interferon-positive (IFN-γ+) CD8+ T-cell response induced by the CS protein immunization and shifts the immune response toward the Th1 type, and (iii) a three-component heterologous vaccination comprised of a CS protein prime followed by boosts with Ad35.CS and Ad26.CS elicits an even more robust and sustainable IFN-γ+ CD8+ T-cell response than one- or two-component regimens. The Ad35.CS/Ad26.CS combination boosted particularly the IFN-γ+ and tumor necrosis factor alpha-positive (TNF-α+) T cells, confirming the shift of the immune response from the Th2 type to the Th1 type. These results support the notion of first immunizations of infants with an adjuvanted CS protein vaccine, followed by a booster Ad35.CS/Ad26.CS vaccine at a later age, to induce lasting protection against malaria for which the Th1 response and immune memory is required

Graphene-Based TiO2 Nanocomposite for Photocatalytic Degradation of Dyes in Aqueous Solution under Solar-Like Radiation

This study presents a novel method for the development of TiO2/reduced graphene oxide (rGO) nanocomposites for photocatalytic degradation of dyes in an aqueous solution. The synergistic integration of rGO and TiO2, through the formation of Ti–O–C bonds, offers an interesting opportunity to design photocatalyst nanocomposite materials with the maximum absorption shift to the visible region of the spectra, where photodegradation can be activated not only with UV but also with the visible part of natural solar irradiation. TiO2@rGO nanocomposites with different content of rGO have been self-assembled by the hydrothermal method followed by calcination treatment. The morphological and structural analysis of the synthesized photocatalysts was performed by FTIR, XRD, XPS, UV-Vis DRS, SEM/EDX, and Raman spectroscopy. The effectiveness of the synthesized nanocomposites as photocatalysts was examined through the photodegradation of methylene blue (MB) and rhodamine B (RhB) dye under artificial solar-like radiation. The influence of rGO concentration (5 and 15 wt.%) on TiO2 performance for photodegradation of the different dyes was monitored by UV-Vis spectroscopy. The obtained results showed that the synthesized TiO2@rGO nanocomposites significantly increased the decomposition of RhB and MB compared to the synthesized TiO2 photocatalyst. Furthermore, TiO2@rGO nanocomposite with high contents of rGO (15 wt.%) presented an improved performance in photodegradation of MB (98.1%) and RhB (99.8%) after 120 min of exposition to solar-like radiation. These results could be mainly attributed to the decrease of the bandgap of synthesized TiO2@rGO nanocomposites with the increased contents of rGO. Energy gap (Eg) values of nanocomposites are 2.71 eV and 3.03 eV, when pure TiO2 particles have 3.15 eV. These results show the potential of graphene-based TiO2 nanocomposite to be explored as a highly efficient solar light-driven photocatalyst for water purification

CD4⁺ and CD8⁺ T cells both contribute to H5N1 protection.

<p>Mice (n = 8-10/group) were immunized once with TVV+MM or PBS as negative control 4 weeks before challenge and (A) CD8⁺ or the combination of CD4⁺ and CD8⁺ T cells were depleted or (B) CD4⁺ T cells only were depleted with antibodies injected 4 days and 1 day before challenge. Mice were challenged with 12.5xLD₅₀ of wild type H5N1 A/Hong Kong/156/97 and monitored for 21 days for (A and B) survival and (C and D) bodyweight-loss. Graphs A and B represent the Kaplan-Meier survival curves and graphs C and D represent mean bodyweight change with 95% confidence interval.</p