Heterologous Processing and Export of the Bacteriocins Pediocin PA-1 and Lactococcin A in Lactococcus Lactis:A Study with Leader Exchange

The bacteriocins pediocin PA-1 and lactococcin A are synthesized as precursors carrying N-terminal extensions with a conserved cleavage site preceded by two glycine residues in positions -2 and -1. Each bacteriocin is translocated through the cytoplasmic membrane by an integral membrane protein of the ABC cassette superfamily which, in the case of pediocin PA-1, has been shown to possess peptidase activity responsible for proteolytic cleavage of the pre-bacteriocin. In each case, another integral membrane protein is essential for bacteriocin production. In this study, a two-step PCR approach was used to permutate the leaders of pediocin PA-1 and lactococcin A. Wild-type and chimeric pre-bacteriocins were assayed for maturation by the processing/export machinery of pediocin PA-1 and lactococcin A. The results show that pediocin PA-1 can be efficiently exported by the lactococcin machinery whether it carries the lactococcin or the pediocin leader. It can also compete with wild-type lactococcin A for the lactococcin machinery. Pediocin PA-1 carrying the lactococcin A leader or lactococcin A carrying that of pediocin PA-1 was poorly secreted when complemented with the pediocin PA-1 machinery, showing that the pediocin machinery is more specific for its bacteriocin substrate. Wild-type pre-pediocin and chimeric pre-pediocin were shown to be processed by the lactococcin machinery at or near the double-glycine cleavage site. These results show the potential of the lactococcin LcnC/LcnD machinery as a maturation system for peptides carrying double-glycine-type amino-terminal leaders

The 5 ' end of the 18S rRNA can be positioned from within the mature rRNA

In yeast, the 5' end of the mature 18S rRNA is generated by endonucleolytic cleavage at site

Vestigingssteun voor bedrijfsopvolgers in de agrarische sector : een analyse van drie mogelijke varianten

In dit rapport is voor drie varianten van vestigingssteun het effect op de financiële positie na de bedrijfsovername onderzocht. De financiële positie na bedrijfsovername is zeer verschillend tussen bedrijven. Deze uitgangssituatie na overname is van essentieel belang voor de levensvatbaarheid. Voor de analyse van de drie varianten is een rekenmodel gemaakt. Van de drie varianten komt de variant zoals die door het Nederlands Agrarisch Jongeren Kontakt is voorgesteld het beste naar voren: het 'Next Generation Fund'. De opvolger ontvangt bij bedrijfsovername 50.000 gulden, die na 10 jaar moet worden terugbetaald. Dit is met name gunstig voor de liquiditeitspositie van de opvolger. Het effect van vestigingssteun op de levensvatbaarheid valt in het niet vergeleken met de effecten van de verschillen in uitgangssituatie

Zorglandbouw op koers, maar blik op verdere professonalisering

Het aantal agrarische bedrijven met een zorgtak is de afgelopen jaren sterk toegenomen. Terwijl de opbrengsten vrij goed in beeld te brengen zijn, is er weinig zicht op de kostprijs van een zorgplaats. In een verdere professionaliseringsslag zou de sector zich, naast de nadere concretisering van de kostprijs, moeten richten op een verdieping van het ondernemerschap

Efficiency of time series homogenization: method comparison with 12 monthly temperature test datasets

The aim of time series homogenization is to remove nonclimatic effects, such as changes in station location, instrumentation, observation practices, and so on, from observed data. Statistical homogenization usually reduces the nonclimatic effects but does not remove them completely. In the Spanish ‘‘MULTITEST’’ project, the efficiencies of automatic homogenization methods were tested on large benchmark datasets of a wide range of statistical properties. In this study, test results for nine versions, based on five homogenization methods—the adapted Caussinus-Mestre algorithm for the homogenization of networks of climatic time series (ACMANT), ‘‘Climatol,’’ multiple analysis of series for homogenization (MASH), the pairwise homogenization algorithm (PHA), and ‘‘RHtests’’—are presented and evaluated. The tests were executed with 12 synthetic/surrogate monthly temperature test datasets containing 100–500 networks with 5–40 time series in each. Residual centered root-mean-square errors and residual trend biases were calculated both for individual station series and for network mean series. The results show that a larger fraction of the nonclimatic biases can be removed from station series than from network-mean series. The largest error reduction is found for the long-term linear trends of individual time series in datasets with a high signal-to-noise ratio (SNR), where the mean residual error is only 14%–36% of the raw data error. When the SNR is low, most of the results still indicate error reductions, although with smaller ratios than for large SNR. In general, ACMANT gave the most accurate homogenization results. In the accuracy of individual time series ACMANT is closely followed by Climatol, and for the accurate calculation of mean climatic trends over large geographical regions both PHA and ACMANT are recommended.This research was funded by the Spanish MULTITESTproject (Ministry of Economics and Competitiveness, CGL2014-52901-P)

Surviving the storm:manual vs. mechanical chest compressions onboard a lifeboat during bad weather conditions

Objective: It is challenging for rescuers to perform cardiopulmonary resuscitation (CPR) onboard lifeboats, particularly during rough weather. A mechanical chest compression device (MCD) may provide better quality chest compressions. The aim of this study was to compare the quality of chest compressions performed by lifeboat-crewmembers with those of a MCD during rough-sea conditions.Methods: Lifeboat-crewmembers were scheduled to provide compression-onlyCPR on a resuscitation-mannequin during two sets of five 6-min epochs on alifeboat at sea in two different weather-conditions. Simultaneously a MCD wasused for compression-only CPR on another mannequin onboard the lifeboat. Ona third occasion compressions by MCD only were measured due to COVID-19restrictions. The primary outcome variable was the quality of chest compression,evaluated using published variables and standards (mean compression depthand compression frequency, percentage correct compression depth, percentageof not leaning on the thorax, percentage of correct hand placement on thethorax, hands-off-time).Results: Six male lifeboat-crewmembers (mean age 35 years) performed CPRduring two different weather conditions. In weather-conditions one (wind∼6–7 Beaufort/wave-height: 100–150 cm) quality of manual compressions wassignificantly worse than mechanical compressions for mean compression depth(p < 0.05) and compression frequency (p < 0.05), percentage correct compression depth (p < 0.05), percentage of not leaning on the thorax (p < 0.05), and hands off time (p < 0.05). Crewmembers could only perform CPR for a limited time-period (sea-conditions/seasickness) and after one set of five epochs measurements were halted. In weather-condition two (wind ∼9 Beaufort/wave-height ∼200 cm) similar results were found during two epochs, after which measurements were halted (sea-conditions/seasickness). In weather-condition three (wind ∼7 Beaufort/wave-height ∼300–400 cm) MCD compressions were according to resuscitation-guidelines except for three epochs during which the MCD was displaced.Conclusion: Crewmembers were only able to perform chest-compressions for alimited time because of the weather-conditions. The MCD was able to providegood quality chest compressions during all but three epochs during the studyperiod. More research is needed to determine whether MCD-use in real-lifecircumstances improves outcome. Inclusion of data on use of a MCD on lifeboatsshould be considered in future revisions of the USFD and resuscitation guidelines

Understanding human gut diseases at single-cell resolution

Our understanding of gut functioning and pathophysiology has grown considerably in the past decades, and advancing technologies enable us to deepen this understanding. Single-cell RNA sequencing (scRNA-seq) has opened a new realm of cellular diversity and transcriptional variation in the human gut at a high, single-cell resolution. ScRNA-seq has pushed the science of the digestive system forward by characterizing the function of distinct cell types within complex intestinal cellular environments, by illuminating the heterogeneity within specific cell populations, and by identifying novel cell types in the human gut that could contribute to a variety of intestinal diseases. In this review, we highlight recent discoveries made with scRNA-seq that significantly advance our understanding of the human gut both in health and across the spectrum of gut diseases, including inflammatory bowel disease, colorectal carcinoma and celiac disease

Bioconversion of polyphenols and organic acids by gut microbiota of predigested Hibiscus sabdariffa L. calyces and Agave (A. tequilana Weber) fructans assessed in a dynamic in vitro model (TIM-2) of the human colon

The present work aimed at understanding gut microbiota bioconversion of phenolic compounds (PC) and organic acids in predigested Hibiscus sabdariffa (Hb) calyces and the mixture of Hb and Agave (Agave tequilana Weber) fructans (AF). With this purpose, dried Hb and Hb/AF were predigested with enzymatic treatment, and then fermented in a dynamic in vitro model of the human colon (TIM-2). After HPLC-ESI-QToF-MS analysis of samples taken at 0, 24, 48 and 72 h of fermentation, it was observed that hydroxycinnamic acids, flavanols, flavonols, and anthocyanins were mainly transformed into derivatives of hydroxyphenylpropionic, hydroxyphenylacetic and hydroxybenzoic acids. Moreover, organic acids, such as hydroxycitric and hibiscus acids, were formed along with unidentified lactones and reduced compounds. Interestingly, no differences were observed between microbial-derived metabolites formed after the fermentation of Hb and Hb/AF. In conclusion, colonic fermentation of polyphenol-rich Hb yields a wide range of microbial phenolic metabolites with potential effects on health.RMB and SGSA acknowledge to Science and Technology for Development Program (CYTED) through the Ibero-American Network of Underutilized Autonomous Foods (ALSUB-CYTED, 118RT0543). SGSA acknowledges CONACYT-Mexico for the sabbatical grant 260935. The Spanish Ministry of Science, Innovation and Universities (Project AGL2015-69986-R) is also acknowledged. This research has been made possible with the support of the Dutch Province of Limburg.Peer reviewe