The use of ultraviolet radiation as a non-thermal treatment for the inactivation of alicyclobacillus acidoterrestris spores in water, wash water from a fruit processing plant and grape juice concentrate

Alicyclobacillus acidoterrestris is a non-pathogenic, spore-forming bacterium that can survive the commercial pasteurisation processes commonly used during fruit juice production. Surviving bacterial endospores germinate, grow and cause spoilage of high acid food products. Fruit juices can be treated using ultraviolet light (UV-C) with a wavelength of 254 nm, which has a germicidal effect against micro-organisms. In this study, A. acidoterrestris was inoculated into water, used wash water from a fruit processing plant and grape juice concentrate. Ultraviolet dosage levels (J L−1) of 0, 61, 122, 183, 244, 305 and 367 J L−1 were applied using a novel UV-C turbulent flow system. The UV treatment method was shown to reliably achieve in excess of a 4 log10 reduction (99.99%) per 0.5 kJ L-1 of UV-C dosage in all the liquids inoculated with A. acidoterrestris. The applied novel UV technology could serve as an alternative to thermal treatments of fruit juices for the inactivation of Alicyclobacillus spores as well as in the treatment of contaminated wash water used in fruit processing.Department of HE and Training approved lis

Procjena probiotičkih svojstava bakterije Enterococcus mundtii, njezino preživljavanje u bozi i proizvodnja bakteriocina in situ

Boza is a low-pH and low-alcohol cereal-based beverage produced in the Balkan Peninsula. Barley was cooked and prepared according to a traditional recipe and inoculated with Enterococcus mundtii ST4V (a potential probiotic and bacteriocin-producing strain), commercially produced boza, Saccharomyces cerevisiae, and a combination of strain E. mundtii ST4V and Saccharomyces cerevisiae. Fermentation was carried out at 37 °C for 3 h. The organoleptic properties of fermented products were evaluated by a qualified taste panel. No significant differences in rheological properties were observed, suggesting that E. mundtii ST4V had no effect on the quality of the final product. Microbial cell numbers remained relatively unchanged during one week of storage. The preservative properties of bacteriocin ST4V were evaluated by contaminating boza with Lactobacillus sakei DSM 20017. Changes in microbial populations were monitored by using classical microbiological methods, PCR with species-specific primers and denaturing gradient gel electrophoresis (DGGE). Adsorption of bacteriocin ST4V to target cells is pH-dependent, with the highest adsorption (88 %) recorded at pH=8.0 and pH=10.0. Maximum adsorption of bacteriocin ST4V (75 %) to Enterococcus faecalis and Listeria innocua was recorded at 25 to 37 °C. Growth of E. mundtii ST4V was inhibited only by a few antibiotics and anti-inflammatory medicaments, suggesting that the strain may be used as a probiotic by individuals receiving medical treatment.Boza je napitak od žitarica, male pH-vrijednosti i neznatnog udjela alkohola, koji se priprema na Balkanu. U istraživanju je upotrijebljena boza pripremljena kuhanjem ječma prema tradicionalnom receptu, inokulirana bakterijskim sojem Enterococcus mundtii ST4V (koji proizvodi bakteriocin i ima probiotička svojstva), bozom proizvedenom na komercijalan način, kvascem Saccharomyces cerevisiae i kombinacijom bakterijskog soja i kvasca. Tim stručnjaka ispitao je organoleptička svojstva proizvoda dobivenih nakon 3 sata fermentacije pri 37 ºC. Nije utvrđena značajna razlika u reološkim svojstvima dobivenih proizvoda, što upućuje na zaključak da inokulacija sojem E. mundtii ST4V nije utjecala na kakvoću proizvoda. Čak ni nakon tjedan dana skladištenja nije se promijenio broj stanica mikroorganizama u proizvodima. Da bi se procijenio zaštitni učinak bakteriocina ST4V, ispitana je boza inokulirana sojem Lactobacillus sakei DSM 20017, pa je mikrobna populacija praćena klasičnim mikrobiološkim metodama, reakcijom polimeraze (PCR) sa specifičnim početnicama i elektroforezom u gradijentu denaturirajućega gela (DGGE). Adsorpcija bakteriocina ST4V na ciljane stanice ovisi o pH-vrijednosti, s najvećom adsorpcijom (88 %) zabilježenom pri pH=8 i pH=10. Maksimalna adsorpcija bakteriocina (75 %) na stanice Enterococcus faecalis i Listeria innocua zabilježena je u rasponu temperature od 25 do 37 ºC. Rast bakterije E. mundtii ST4V inhibirali su samo neki antibiotici i protuupalni lijekovi, što upućuje na to da taj soj ima probiotički učinak i na pacijente koji primaju određenu terapiju

The DACAPO-PESO campaign: Dynamics, Aerosol, Cloud and Precipitation Observations in the Pristine Environment of the Southern Ocean: An overview

This article gives an overview of the DACAPO-PESO field experiment, which has taken place in Punta Arenas, Chile, from November 2018 to November 2021, and showcases first exciting research results that have already emerged from it.In diesem Artikel wird ein Überblick über das DACAPO-PESO Experiment gegeben, welches von November 2018 bis November 2021 in Punta Arenas, Chile, stattgefunden hat. Außerdem werden erste spannende Forschungsergebnisse vorgestellt, die bereits daraus gewonnen wurden

Erythropoietin Receptor Signaling Is Membrane Raft Dependent

Upon erythropoietin (Epo) engagement, Epo-receptor (R) homodimerizes to activate JAK2 and Lyn, which phosphorylate STAT5. Although recent investigations have identified key negative regulators of Epo-R signaling, little is known about the role of membrane localization in controlling receptor signal fidelity. Here we show a critical role for membrane raft (MR) microdomains in creation of discrete signaling platforms essential for Epo-R signaling. Treatment of UT7 cells with Epo induced MR assembly and coalescence. Confocal microscopy showed that raft aggregates significantly increased after Epo stimulation (mean, 4.3±1.4(SE) vs. 25.6±3.2 aggregates/cell; p≤0.001), accompanied by a >3-fold increase in cluster size (p≤0.001). Raft fraction immunoblotting showed Epo-R translocation to MR after Epo stimulation and was confirmed by fluorescence microscopy in Epo stimulated UT7 cells and primary erythroid bursts. Receptor recruitment into MR was accompanied by incorporation of JAK2, Lyn, and STAT5 and their activated forms. Raft disruption by cholesterol depletion extinguished Epo induced Jak2, STAT5, Akt and MAPK phosphorylation in UT7 cells and erythroid progenitors. Furthermore, inhibition of the Rho GTPases Rac1 or RhoA blocked receptor recruitment into raft fractions, indicating a role for these GTPases in receptor trafficking. These data establish a critical role for MR in recruitment and assembly of Epo-R and signal intermediates into discrete membrane signaling units

Local erythropoietin and endothelial progenitor cells improve regional cardiac function in acute myocardial infarction

<p>Abstract</p> <p>Background</p> <p>Expanded endothelial progenitor cells (eEPC) improve global left ventricular function in experimental myocardial infarction (MI). Erythropoietin beta (EPO) applied together with eEPC may improve regional myocardial function even further by anti-apoptotic and cardioprotective effects. Aim of this study was to evaluate intramyocardial application of eEPCs and EPO as compared to eEPCs or EPO alone in experimental MI.</p> <p>Methods and Results</p> <p>In vitro experiments revealed that EPO dosed-dependently decreased eEPC and leukocyte apoptosis. Moreover, in the presence of EPO mRNA expression in eEPC of proangiogenic and proinflammatory mediators measured by TaqMan PCR was enhanced. Experimental MI was induced by ligation and reperfusion of the left anterior descending coronary artery of nude rats (n = 8-9). After myocardial transplantation of eEPC and EPO CD68+ leukocyte count and vessel density were enhanced in the border zone of the infarct area. Moreover, apoptosis of transplanted CD31 + TUNEL + eEPC was decreased as compared to transplantation of eEPCs alone. Regional wall motion of the left ventricle was measured using Magnetic Resonance Imaging. After injection of eEPC in the presence of EPO regional wall motion significantly improved as compared to injection of eEPCs or EPO alone.</p> <p>Conclusion</p> <p>Intramyocardial transplantation of eEPC in the presence of EPO during experimental MI improves regional wall motion. This was associated with an increased local inflammation, vasculogenesis and survival of the transplanted cells. Local application of EPO in addition to cell therapy may prove beneficial in myocardial remodeling.</p

The Janus kinases (Jaks)

The Janus kinase (Jak) family is one of ten recognized families of non-receptor tyrosine kinases. Mammals have four members of this family, Jak1, Jak2, Jak3 and Tyrosine kinase 2 (Tyk2). Birds, fish and insects also have Jaks. Each protein has a kinase domain and a catalytically inactive pseudo-kinase domain, and they each bind cytokine receptors through amino-terminal FERM (Band-4.1, ezrin, radixin, moesin) domains. Upon binding of cytokines to their receptors, Jaks are activated and phosphorylate the receptors, creating docking sites for signaling molecules, especially members of the signal transducer and activator of transcription (Stat) family. Mutations of the Drosophila Jak (Hopscotch) have revealed developmental defects, and constitutive activation of Jaks in flies and humans is associated with leukemia-like syndromes. Through the generation of Jak-deficient cell lines and gene-targeted mice, the essential, nonredundant functions of Jaks in cytokine signaling have been established. Importantly, deficiency of Jak3 is the basis of human autosomal recessive severe combined immunodeficiency (SCID); accordingly, a selective Jak3 inhibitor has been developed, forming a new class of immunosuppressive drugs

Tumor-Shed PGE2 Impairs IL2Rγc-Signaling to Inhibit CD4+ T Cell Survival: Regulation by Theaflavins

BACKGROUND:Many tumors are associated with decreased cellular immunity and elevated levels of prostaglandin E2 (PGE2), a known inhibitor of CD4+ T cell activation and inducer of type-2 cytokine bias. However, the role of this immunomodulator in the survival of T helper cells remained unclear. Since CD4+ T cells play critical roles in cell-mediated immunity, detail knowledge of the effect tumor-derived PGE2 might have on CD4+ T cell survival and the underlying mechanism may, therefore, help to overcome the overall immune deviation in cancer. METHODOLOGY/PRINCIPAL FINDINGS:By culturing purified human peripheral CD4+ T cells or Jurkat cells with spent media of theaflavin- or celecoxib-pre-treated MCF-7 cells, we show that tumor-shed PGE2 severely impairs interleukin 2 receptor gammac (IL2Rgammac)-mediated survival signaling in CD4+ T cells. Indeed, tumor-shed PGE2 down-regulates IL2Rgammac expression, reduces phosphorylation as well as activation of Janus kinase 3 (Jak-3)/signal transducer and activator of transcription 5 (Stat-5) and decreases Bcl-2/Bax ratio thereby leading to activation of intrinsic apoptotic pathway. Constitutively active Stat-5A (Stat-5A1 6) over-expression efficiently elevates Bcl-2 levels in CD4+ T cells and protects them from tumor-induced death while dominant-negative Stat-5A over-expression fails to do so, indicating the importance of Stat-5A-signaling in CD4+ T cell survival. Further support towards the involvement of PGE2 comes from the results that (a) purified synthetic PGE2 induces CD4+ T cell apoptosis, and (b) when knocked out by small interfering RNA, cyclooxygenase-2 (Cox-2)-defective tumor cells fail to initiate death. Interestingly, the entire phenomena could be reverted back by theaflavins that restore cytokine-dependent IL2Rgammac/Jak-3/Stat-5A signaling in CD4+ T cells thereby protecting them from tumor-shed PGE2-induced apoptosis. CONCLUSIONS/SIGNIFICANCE:These data strongly suggest that tumor-shed PGE2 is an important factor leading to CD4+ T cell apoptosis during cancer and raise the possibility that theaflavins may have the potential as an effective immunorestorer in cancer-bearer

Electroanalysis may be used in the Vanillin Biotechnological Production

This study shows that electroanalysis may be used in vanillin biotechnological production. As a matter of fact, vanillin and some molecules implicated in the process like eugenol, ferulic acid, and vanillic acid may be oxidized on electrodes made of different materials (gold, platinum, glassy carbon). By a judicious choice of the electrochemical method and the experimental conditions the current intensity is directly proportional to the molecule concentrations in a range suitable for the biotechnological process. So, it is possible to imagine some analytical strategies to control some steps in the vanillin biotechnological production: by sampling in the batch reactor during the process, it is possible to determine out of line the concentration of vanillin, eugenol, ferulic acid, and vanillic acid with a gold rotating disk electrode, and low concentration of vanillin with addition of hydrazine at an amalgamated electrode. Two other possibilities consist in the introduction of electrodes directly in the batch during the process; the first one with a gold rotating disk electrode using linear sweep voltammetry and the second one requires three gold rotating disk electrodes held at different potentials for chronoamperometry. The last proposal is the use of ultramicroelectrodes in the case when stirring is not possible

PCR and DGGE detection limits for wine spoilage microbes

The original publication is available at http://www.sasev.org/.In this study the culture-independent technique, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE), was investigated for the early detection and identification of possible spoilage microbes in wine. PCR and DGGE conditions were successfully optimised with the universal primers HDA1GC and HDA2, the bacteria-specific primers WBAC1GC-WBAC2, and the yeast-specific primers NL1GC and LS2. PCR and DGGE detection limits were determined for Lactobacillus plantarum, Pediococcus pentosaceus, Acetobacter pasteurianus and Brettanomyces bruxellensis when inoculated into sterile saline solution (SSS) and white wine at 106 cfu/mL respectively. PCR detection limits were more sensitive (101 to 102 cfu/mL) than DGGE detection limits (101 to 104 cfu/ mL), with the exception of B. bruxellensis, which had higher PCR and DGGE detection limits than the other reference microbes. PCR-DGGE analysis was also used successfully to detect and identify Lb. plantarum, A. pasteurianus and B. bruxellensis at a concentration of 108 cfu/mL as part of mixed populations in SSS and white wine. PCR detection limits of 101 cfu/mL were determined for all three reference microbes in mixed populations. The DGGE detection limits were higher for mixed populations when compared to single strains.Publishers' versio