Screening Collagenase Activity in Bacterial Lysate for Directed Enzyme Applications

Collagenases are essential enzymes capable of digesting triple-helical collagen under physiological conditions. These enzymes play a key role in diverse physiological and pathophysiological processes. Collagenases are used for diverse biotechnological applications, and it is thus of major interest to identify new enzyme variants with improved characteristics such as expression yield, stability, or activity. The engineering of new enzyme variants often relies on either rational protein design or directed enzyme evolution. The latter includes screening of a large randomized or semirational genetic library, both of which require an assay that enables the identification of improved variants. Moreover, the assay should be tailored for microplates to allow the screening of hundreds or thousands of clones. Herein, we repurposed the previously reported fluorogenic assay using 3,4-dihydroxyphenylacetic acid for the quantitation of collagen, and applied it in the detection of bacterial collagenase activity in bacterial lysates. This enabled the screening of hundreds of E. coli colonies expressing an error-prone library of collagenase G from C. histolyticum, in 96-well deep-well plates, by measuring activity directly in lysates with collagen. As a proof-of-concept, a single variant exhibiting higher activity than the starting-point enzyme was expressed, purified, and characterized biochemically and computationally. This showed the feasibility of this method to support medium-high throughput screening based on direct evaluation of collagenase activity.     

Durable CD8 T Cell Memory against SARS-CoV-2 by Prime/Boost and Multi-Dose Vaccination: Considerations on Inter-Dose Time Intervals

Facing the COVID-19 pandemic, anti-SARS-CoV-2 vaccines were developed at unprecedented pace, productively exploiting contemporary fundamental research and prior art. Large-scale use of anti-SARS-CoV-2 vaccines has greatly limited severe morbidity and mortality. Protection has been correlated with high serum titres of neutralizing antibodies capable of blocking the interaction between the viral surface protein spike and the host SARS-CoV-2 receptor, ACE-2. Yet, vaccine-induced protection subsides over time, and breakthrough infections are commonly observed, mostly reflecting the decay of neutralizing antibodies and the emergence of variant viruses with mutant spike proteins. Memory CD8 T cells are a potent weapon against viruses, as they are against tumour cells. Anti-SARS-CoV-2 memory CD8 T cells are induced by either natural infection or vaccination and can be potentially exploited against spike-mutated viruses. We offer here an overview of current research about the induction of anti-SARS-CoV-2 memory CD8 T cells by vaccination, in the context of prior knowledge on vaccines and on fundamental mechanisms of immunological memory. We focus particularly on how vaccination by two doses (prime/boost) or more (boosters) promotes differentiation of memory CD8 T cells, and on how the time-length of inter-dose intervals may influence the magnitude and persistence of CD8 T cell memory.     

Efficient solar photocatalyst based on cobalt oxide/iron oxide composite nanofibers for the detoxification of organic pollutants

A Co₃O₄/Fe₂O₃ composite nanofiber-based solar photocatalyst has been prepared, and its catalytic performance was evaluated by degrading acridine orange (AO) and brilliant cresyl blue (BCB) beneath solar light. The morphological and physiochemical structure of the synthesized solar photocatalyst was characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). FESEM indicates that the Co₃O₄/Fe₂O₃ composite has fiber-like nanostructures with an average diameter of approximately 20 nm. These nanofibers are made of aggregated nanoparticles having approximately 8.0 nm of average diameter. The optical properties were examined by UV-visible spectrophotometry, and the band gap of the solar photocatalyst was found to be 2.12 eV. The as-grown solar photocatalyst exhibited high catalytic degradation in a short time by applying to degrade AO and BCB. The pH had an effect on the catalytic performance of the as-grown solar photocatalyst, and it was found that the synthesized solar photocatalyst is more efficient at high pH. The kinetics study of both AO and BCB degradation indicates that the as-grown nanocatalyst would be a talented and efficient solar photocatalyst for the removal of hazardous and toxic organic materials.     

Differential effects of alcohol on working memory: Distinguishing multiple processes.

The authors assessed effects of alcohol consumption on different types of working memory (WM) tasks in an attempt to characterize the nature of alcohol effects on cognition. The WM tasks varied in 2 properties of materials to be retained in a 2-stimulus comparison procedure. Conditions included (a) spatial arrays of colors, (b) temporal sequences of colors, (c) spatial arrays of spoken digits, and (d) temporal sequences of spoken digits. Alcohol consumption impaired memory for auditory and visual sequences but not memory for simultaneous arrays of auditory or visual stimuli. These results suggest that processes needed to encode and maintain stimulus sequences, such as rehearsal, are more sensitive to alcohol intoxication than other WM mechanisms needed to maintain multiple concurrent items, such as focusing attention on them. These findings help to resolve disparate findings from prior research on alcohol's effect on WM and on divided attention. The results suggest that moderate doses of alcohol impair WM by affecting certain mnemonic strategies and executive processes rather than by shrinking the basic holding capacity of WM. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Fake ID ownership and heavy drinking in underage college students: Prospective findings.

The authors examined the ownership of false identification (fake ID) for the purpose of obtaining alcohol and the relation of fake ID ownership to heavy drinking in a longitudinal sample of college students under 21 years of age. A sample of 3,720 undergraduates was assessed the summer prior to college entrance and during the 4 semesters comprising freshman and sophomore years. Regression analyses were used to estimate bidirectional relations between consumption and fake ID ownership. Sex, Greek membership, and prior drinking were controlled. Results showed that fake ID ownership increased over time (12.5% precollege to 32.2% fourth semester) and that Greek members were more likely than others to own fake IDs. Fake ID ownership predicted concurrent and next-semester heavy drinking with increasing strength over time. Also, the acquisition (onset) of fake ID ownership at each time point was predicted by previous-semester consumption. When traditional, robust risk factors of consumption are controlled, fake ID ownership meaningfully relates to heavy drinking in college. It thus presents a significant public health problem, addressable through training for alcohol servers and retailers, punitive measures toward fake ID owners, and other possible interventions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Peristaltic pumping with double diffusive natural convective nanofluid in a lopsided channel with accounting thermophoresis and Brownian moment

In the current study, double-diffusive natural convective peristaltic flow of a nanofluid with induced magnetic field in an asymmetric porous channel is discussed. The five coupled equations are solved analytically using with new analytical techniques detail of which is given in solution section. The analytical solutions for pressure gradient and pressure rise, velocity profile, temperature, solutal concentration, and nanoparticle fraction are evaluated and presented through graphs.

     

An Empirical Study on the Reliability of Perceiving Correlation Indices using Scatterplots

Scatterplots have been in use for about two centuries, primarily for observing the relationship between two variables and commonly for supporting correlation analysis. In this paper, we report an empirical study that examines how humans’ perception of correlation using scatterplots relates to the Pearson's product‐moment correlation coefficient (PPMCC) – a commonly used statistical measure of correlation. In particular, we study human participants’ estimation of correlation under different conditions, e.g., different PPMCC values, different densities of data points, different levels of symmetry of data enclosures, and different patterns of data distribution. As the participants were instructed to estimate the PPMCC of each stimulus scatterplot, the difference between the estimated and actual PPMCC is referred to as an offset. The results of the study show that varying PPMCC values, symmetry of data enclosure, or data distribution does have an impact on the average offsets, while only large variations in density cause an impact that is statistically significant. This study indicates that humans’ perception of correlation using scatterplots does not correlate with computed PPMCC in a consistent manner. The magnitude of offsets may be affected not only by the difference between individuals, but also by geometric features of data enclosures. It suggests that visualizing scatterplots does not provide adequate support to the task of retrieving their corresponding PPMCC indicators, while the underlying model of humans’ perception of correlation using scatterplots ought to feature other variables in addition to PPMCC. The paper also includes a theoretical discussion on the cost‐benefit of using scatterplots.     

Cellulose acetate-iron oxide nanocomposites for trace detection of fluorene from water samples by solid-phase extraction technique

In this study, cellulose acetate (CA)/iron oxide nanocomposites (NC1 and NC2) were synthesized and utilized as extractors for the selective separation of fluorene from aqueous samples. The selectivity of nanocomposites to different organic compounds was assessed. Based on the results, fluorene was the most quantitatively adsorbed on NC2. Batch method was performed by varying the initial fluorene concentrations and contact time. Langmuir and Freundlich isotherms were used for modeling the experimental data and were best correlated by the Langmuir model. Adsorption data were also followed the pseudo-second-order kinetic model. Finally, validation of the developed method was achieved for fluorene determination in real water samples.