Enzymatic Disruption of Biofilms During Cheese Manufacturing: A Mini Review

Bacteria are capable of colonizing industrial processing surfaces creating biofilms on them which may adversely affect the quality and safety of products. Traditional cleaning-in-place (CIP) treatments using caustic and nitric acid solutions have been known to exhibit variable efficiency in eliminating biofilm bacteria. Here, we introduce enzymes as an alternative to traditional CIP treatments and discuss their mechanism of action against bacterial biofilms in cheese manufacturing. In addition, we discuss research gaps namely thermal stability, substrate specificity and residual activity of enzymes that may play a vital role in the selection of enzymes with optimal effectiveness against multi species biofilms. The outcome of this mini review will aid in the development of a novel and sustainable enzyme-based CIP treatment during cheese manufacturing in the future.

Inhibition of selective autophagy by members of the herpesvirus ubiquitin-deconjugase family

Autophagy is an important component of the innate immune response that restricts infection by different types of pathogens. Viruses have developed multiple strategies to avoid autophagy to complete their replication cycle and promote spreading to new hosts. Here, we report that the ubiquitin deconjugases encoded in the N-terminal domain of the large tegument proteins of Epstein–Barr virus (EBV), Kaposi Sarcoma herpesvirus (KSHV) and human cytomegalovirus (HCMV), but not herpes simplex virus-1 (HSV-1), regulate selective autophagy by inhibiting the activity of the autophagy receptor SQSTM1/p62. We found that all the homologs bind to and deubiquitinate SQSTM1/p62 but with variable efficiency, which correlates with their capacity to prevent the colocalization of light chain 3 (LC3) with SQSTM1/p62 aggregates and promote the accumulation of a model autophagy substrate. The findings highlight important differences in the strategies by which herpesviruses interfere with selective autophagy.

Inhibition of selective autophagy by members of the herpesvirus ubiquitin-deconjugase family

Autophagy is an important component of the innate immune response that restricts infection by different types of pathogens. Viruses have developed multiple strategies to avoid autophagy to complete their replication cycle and promote spreading to new hosts. Here we report that the ubiquitin deconjugases encoded in the N-terminal domain of the large tegument proteins of Epstein-Barr virus (EBV), Kaposi Sarcoma herpesvirus (KSHV) and human cytomegalovirus (HCMV), but not herpes simplex virus-1 (HSV-1), regulate selective autophagy by inhibiting the activity of the autophagy receptor SQSTM1/p62. We found that all the homologs bind to and deubiquitinate SQSTM1/p62 but with variable efficiency, which correlates with their capacity to prevent the colocalization of LC3 with SQSTM1/p62 aggregates and promote the accumulation of a model autophagy substrate. The findings highlight important differences in the strategies by which herpesviruses interfere with selective autophagy.

Ocular Involvement of SARS-CoV-2 in a Polish Cohort of COVID-19-Positive Patients

The coronavirus SARS-CoV-2 responsible for the current human COVID-19 pandemic has shown tropism toward different organs with variable efficiency, eyes included. The purpose of this study has been to investigate the presence of detectable SARS-CoV-2 infection in ocular swabs in patients affected by COVID-19. A consecutive series of 74 COVID-19-positive patients (age 21–89) were enrolled at two Polish COVID-19 hospitals for 4 months and were characterized by PCR for the presence of the SARS-CoV-2 genetic material in nasopharyngeal (NP) and ocular swabs, while their respiratory and ocular symptoms were noted. Almost 50% of them presented with severe/critical respiratory involvement, and some degree of eye disease. No tight correlation was observed between the presence of ocular and respiratory symptoms. Three male patients presenting with severe/critical lung disease tested positive in ocular swab, however with mild/moderate ocular symptoms. In conclusion, our study lends further support to the view that overt ocular infection by the SARS-CoV-2 virus is not such a frequent occurrence.

CRISPR-finder: A high throughput and cost-effective method to identify successfully edited Arabidopsis thaliana individuals

Genome editing with the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR associated protein) system allows mutagenesis of a targeted region of the genome using a Cas endonuclease and an artificial guide RNA. Both because of variable efficiency with which such mutations arise and because the repair process produces a spectrum of mutations, one needs to ascertain the genome sequence at the targeted locus for many individuals that have been subjected to mutagenesis. We provide a complete protocol for the generation of amplicons up until the identification of the exact mutations in the targeted region. CRISPR-finder can be used to process thousands of individuals in a single sequencing run. We successfully identified an ISOCHORISMATE SYNTHASE 1 mutant line in which the production of salicylic acid was impaired compared to the wild type, as expected. These features establish CRISPR-finder as a high-throughput, cost-effective and efficient genotyping method of individuals whose genomes have been targeted using the CRISPR/Cas9 system.

Chemical evolution of the Milky Way: constraints on the formation of the thick and thin discs

ABSTRACT We study the evolution of Milky Way thick and thin discs in the light of the most recent observational data. In particular, we analyse abundance gradients of O, N, Fe, and Mg along the thin disc as well as the [Mg/Fe] versus [Fe/H] relations and the metallicity distribution functions at different Galactocentric distances. We run several models starting from the two-infall paradigm, assuming that the thick and thin discs formed by means of two different infall episodes, and we explore several physical parameters, such as radial gas flows, variable efficiency of star formation, different times for the maximum infall on to the disc, different distributions of the total surface mass density of the thick disc, and enriched gas infall. Our best model suggests that radial gas flows and variable efficiency of star formation should be acting together with the inside-out mechanism for the thin disc formation. The time-scale for maximum infall on to the thin disc, which determines the gap between the formation of the two discs, should be tmax ≃ 3.25 Gyr. The thick disc should have an exponential, small-scale length density profile and gas infall on the inner thin disc should be enriched. We also compute the evolution of Gaia–Enceladus system and study the effects of possible interactions with the thick and thin discs. We conclude that the gas lost by Enceladus or even part of it could have been responsible for the formation of the thick disc but not the thin disc.

CRISPR-finder: A high throughput and cost effective method for identifying successfully edited A. thaliana individuals

BackgroundGenome editing with the CRISPR/Cas9 system allows the user to mutate a targeted region of the genome using an endonuclease (Cas9) and an artificial single-guide RNA (sgRNA). Both because of variable efficiency with which such mutations arise and because the repair process produces a spectrum of mutations, one needs to ascertain the genome sequence at the targeted locus for many individuals that have been subjected to CRISPR/Cas9 mutagenesis. This process can be laborious, expensive and inefficient with conventional methods such as the T7E1 assay or Sanger sequencing. An alternative comprises methods for amplicon sequencing, but most available protocols do not include a facile way for high throughput generation of the samples for sequencing.ResultsIn this study we provide a full pipeline based on amplicon sequencing, CRISPR-finder. We provide a complete protocol for the generation of amplicons up until the identification of the exact mutations in the targeted region. CRISPR-finder can be used to process thousands of individuals in a single sequencing run. For example, we were able to analyze in one sequencing reaction over 900 Arabidopsis thaliana individuals whose genomes had been targeted with the CRISPR/Cas9 system.ConclusionsIn order to validate the potential of CRISPR-finder, we targeted the ISOCHORISMATE SYNTHASE 1 gene in A. thaliana using the CRISPR/Cas9 system. We successfully identified a mutant line in which the production of salicylic acid was impaired compared to the wild type, as expected. These features establish CRISPR-finder as a high-throughput, cost-effective and -efficient genotyping method of individuals whose genomes have been targeted using the CRISPR/Cas9 system.

EFISIENSI, PERSAINGAN, DAN RISIKO DEFAULT PERBANKAN INDONESIA SEBELUM DAN SETELAH PBI 14/26/2012

This study try to find out about the impact of PBI / 14/26/2012 for banks in Indonesia. This regulation limiting the activities of banks based on their core capital. The purpose of this regulation is to improve the efficiency, durability and banking competition. Efficiency is measured by using DEA, endurance proxy by Zscore and competititon proxy by HHI. The results showed that there was a significant difference before and after the regulations issued for variable efficiency and competition but not for durability. The next stage of this research is to know the effect of endurance, competition, regulations and numbers of Board of Directors (BOD) of the efficiency. Measurements were made by a panel regression and stated that the results of the BOD and year significantly affecting efficiency. Keywords: efficiency, competition, default risk, BOD, banking