Adaptation by Type V-A and V-B CRISPR-Cas Systems Demonstrates Conserved Protospacer Selection Mechanisms Between Diverse CRISPR-Cas Types.

Adaptation of clustered regularly interspaced short palindromic repeats (CRISPR) arrays is a crucial process responsible for the unique, adaptive nature of CRISPR-Cas immune systems. The acquisition of new CRISPR spacers from mobile genetic elements has previously been studied for several types of CRISPR-Cas systems. In this study, we used a high-throughput sequencing approach to characterize CRISPR adaptation of the type V-A system from and the type V-B system from .

Mechanism for Cas4-assisted directional spacer acquisition in CRISPR-Cas.

Prokaryotes adapt to challenges from mobile genetic elements by integrating spacers derived from foreign DNA in the CRISPR array. Spacer insertion is carried out by the Cas1-Cas2 integrase complex. A substantial fraction of CRISPR-Cas systems use a Fe-S cluster containing Cas4 nuclease to ensure that spacers are acquired from DNA flanked by a protospacer adjacent motif (PAM) and inserted into the CRISPR array unidirectionally, so that the transcribed CRISPR RNA can guide target searching in a PAM-dependent manner.

Cas4-Cas1 Is a Protospacer Adjacent Motif-Processing Factor Mediating Half-Site Spacer Integration During CRISPR Adaptation.

The immunization of bacteria and archaea against invading viruses via CRISPR adaptation is critically reliant on the efficient capture, accurate processing, and integration of CRISPR spacers into the host genome. The adaptation proteins Cas1 and Cas2 are sufficient for successful spacer acquisition in some CRISPR-Cas systems. However, many CRISPR-Cas systems additionally require the Cas4 protein for efficient adaptation.

Correction: Wachs, S., et al. Associations between Witnessing and Perpetrating Online Hate in Eight Countries: The Buffering Effects of Problem-Focused Coping. 2019, , 3992.

The authors wish to add the following corrections to their paper published in the International Journal of Environmental Research and Public Health [...

A Routine Activity Approach to Understand Cybergrooming Victimization Among Adolescents from Six Countries.

Little attention has been given academically to empirically tested theoretical frameworks that aim at measuring the risk of adolescents falling victim to cybergrooming. To this end, we have applied the routine activity theory (RAT) to investigate whether exposure to motivated offenders (PC/laptop ownership and Internet access in one's own bedroom), capable guardianship (parental mediation strategies of Internet use), and target suitability (adolescents' online disclosure of private information) might predict cybergrooming victimization among adolescents. Using data from a cross-sectional survey of 5,938 adolescents from Germany, India, South Korea, Spain, Thailand, and the United States, ranging in age from 12 to 18 ( = 14.

Direct Visualization of Native CRISPR Target Search in Live Bacteria Reveals Cascade DNA Surveillance Mechanism.

CRISPR-Cas systems encode RNA-guided surveillance complexes to find and cleave invading DNA elements. While it is thought that invaders are neutralized minutes after cell entry, the mechanism and kinetics of target search and its impact on CRISPR protection levels have remained unknown. Here, we visualize individual Cascade complexes in a native type I CRISPR-Cas system.

Are Sexist Attitudes and Gender Stereotypes Linked? A Critical Feminist Approach With a Spanish Sample.

This study aims to verify the psychometric properties of the Spanish versions of the Social Roles Questionnaire (SRQ; Baber and Tucker, 2006), Modern Sexism Scale (MS), and Old-Fashioned Sexism Scale (OFS; Swim et al., 1995; Swim and Cohen, 1997). Enough support was found to maintain the original factor structure of all instruments in their Spanish version.

Associations between Witnessing and Perpetrating Online Hate in Eight Countries: The Buffering Effects of Problem-Focused Coping.

Online hate is a topic that has received considerable interest lately, as online hate represents a risk to self-determination and peaceful coexistence in societies around the globe. However, not much is known about the explanations for adolescents posting or forwarding hateful online material or how adolescents cope with this newly emerging online risk. Thus, we sought to better understand the relationship between a bystander to and perpetrator of online hate, and the moderating effects of problem-focused coping strategies (e.

Development and Validation of a Measure of Emotional Distress in Survivors of Group Psychological Abuse.

In the context of the negative consequences of psychological abuse, a scale was developed to specifically assess the emotional disturbances in individuals who had experienced abusive behaviors over a period of time within a cultic group. The Emotional Distress Scale in Survivors of Abusive Groups (EDS-SAG) was administered, along with other relevant measures of group psychological abuse and psychopathological symptoms, to 706 Spanish-speaking former members of different groups, distributed into two samples according to whether they had experienced group psychological abuse (n = 413) or not (n = 293). Analyses supported a unidimensional structure of the 18 items on the EDS-SAG, explaining 50.

Conserved motifs in the CRISPR leader sequence control spacer acquisition levels in Type I-D CRISPR-Cas systems.

Integrating short DNA fragments at the correct leader-repeat junction is key to successful CRISPR-Cas memory formation. The Cas1-2 proteins are responsible to carry out this process. However, the CRISPR adaptation process additionally requires a DNA element adjacent to the CRISPR array, called leader, to facilitate efficient localization of the correct integration site.

Cas4-Cas1 fusions drive efficient PAM selection and control CRISPR adaptation.

Microbes have the unique ability to acquire immunological memories from mobile genetic invaders to protect themselves from predation. To confer CRISPR resistance, new spacers need to be compatible with a targeting requirement in the invader's DNA called the protospacer adjacent motif (PAM). Many CRISPR systems encode Cas4 proteins to ensure new spacers are integrated that meet this targeting prerequisite.

CRISPR-Cas Systems Reduced to a Minimum.

In two recent studies in Molecular Cell, Wright et al. (2019) report complete spacer integration by a Cas1 mini-integrase and Edraki et al. (2019) describe accurate genome editing by a small Cas9 ortholog with less stringent PAM requirements.

Using CAPTURE to detect spacer acquisition in native CRISPR arrays.

CRISPR-Cas systems are able to acquire immunological memories (spacers) from bacteriophages and plasmids in order to survive infection; however, this often occurs at low frequency within a population, which can make it difficult to detect. Here we describe CAPTURE (CRISPR adaptation PCR technique using reamplification and electrophoresis), a versatile and adaptable protocol to detect spacer-acquisition events by electrophoresis imaging with high-enough sensitivity to identify spacer acquisition in 1 in 10 cells. Our method harnesses two simple PCR steps, separated by automated electrophoresis and extraction of size-selected DNA amplicons, thus allowing the removal of unexpanded arrays from the sample pool and enabling 1,000-times more sensitive detection of new spacers than alternative PCR protocols.

Group Psychological Abuse and Psychopathological Symptoms: The Mediating Role of Psychological Stress.

In the context of the long-term effects of psychological abuse, this study examined psychological stress and psychopathological symptoms in survivors of groups with cultic dynamics where abusive behaviors take place on an ongoing basis. We specifically tested the mediating role of psychological stress suffered after the departure from the group between the group psychological abuse experienced and current psychopathology. An online questionnaire was administered to 608 Spanish-speaking former members of different groups.

Psychometric properties of the Spanish version of the Experience of Caregiving Inventory (ECI) among caregivers of individuals with an eating disorder.

Purpose: The aim of this study was to examine the psychometric properties of the Spanish version of the Experience of Caregiving Inventory (ECI-S), which is designed to assess the caregiver's appraisal of the impact of caring for a relative with a serious mental illness.

Methods: A cross-sectional study was conducted among 320 caregivers of a relative with an eating disorder to examine: (a) descriptive statistics; (b) internal consistency reliability; (c) the fit of the original ten-factor structure of the ECI through exploratory factor analysis, using a semi-confirmatory approach, for each subscale individually, and (d) concurrent validity. A total of 307 caregivers completed the scale.

Bereaved relatives' decision about deceased organ donation: An integrated psycho-social study conducted in Spain.

Rationale: Family refusal to organ donation of a deceased relative represents one of the most important barriers to organ transplantation. Although a large literature about family decisions has amassed, the existing evidence needs further integration and structuring.

Objective: This study seeks to analyse relationships between bereaved relatives' decisions and a wide range of factors that converge in the family decision process, including interactions and complex relationship patterns, and taking psychosocial theoretical frameworks as reference to conceptualize empirical findings.

Cas4 Facilitates PAM-Compatible Spacer Selection during CRISPR Adaptation.

CRISPR-Cas systems adapt their immunological memory against their invaders by integrating short DNA fragments into clustered regularly interspaced short palindromic repeat (CRISPR) loci. While Cas1 and Cas2 make up the core machinery of the CRISPR integration process, various class I and II CRISPR-Cas systems encode Cas4 proteins for which the role is unknown. Here, we introduced the CRISPR adaptation genes cas1, cas2, and cas4 from the type I-D CRISPR-Cas system of Synechocystis sp.

Persuasion strategies and sexual solicitations and interactions in online sexual grooming of adolescents: Modeling direct and indirect pathways.

Online sexual grooming and exploitation of adolescents is receiving increasing social attention. Drawing on a social influence framework, the aim of this study was to test a model of the direct and mediated relationships between an adult's use of persuasion strategies and online sexual grooming of early adolescents. The initial sample of the study consisted of 2731 early Spanish adolescents between 12 and 15 years old (50.

Risky online behaviors among adolescents: Longitudinal relations among problematic Internet use, cyberbullying perpetration, and meeting strangers online.

Background and aims This study aims to analyze the cross-sectional and longitudinal relationship between three major risky online behaviors during adolescence: problematic Internet use, cyberbullying perpetration, and meeting strangers online. An additional objective was to study the role of impulsivity-irresponsibility as a possible explanatory variable of the relationships between these risky online behaviors. Methods The study sample was 888 adolescents that completed self-report measures at time 1 and time 2 with an interval of 6 months.

Anti-cas spacers in orphan CRISPR4 arrays prevent uptake of active CRISPR-Cas I-F systems.

Archaea and bacteria harbour clustered regularly interspaced short palindromic repeats (CRISPR) loci. These arrays encode RNA molecules (crRNA), each containing a sequence of a single repeat-intervening spacer. The crRNAs guide CRISPR-associated (Cas) proteins to cleave nucleic acids complementary to the crRNA spacer, thus interfering with targeted foreign elements.

Development and validation of the scale of psychological abuse in intimate partner violence (EAPA-P).

Background: In the context of intimate partner violence, psychological abuse (PA) has progressively gained scientific relevance. Even so, a greater effort is needed to define and evaluate psychological intimate partner abuse. A new exhaustive and operative taxonomy of PA strategies leads to the contribution of a new evaluation instrument.

CRISPR Content Correlates with the Pathogenic Potential of Escherichia coli.

Guide RNA molecules (crRNA) produced from clustered regularly interspaced short palindromic repeat (CRISPR) arrays, altogether with effector proteins (Cas) encoded by cognate cas (CRISPR associated) genes, mount an interference mechanism (CRISPR-Cas) that limits acquisition of foreign DNA in Bacteria and Archaea. The specificity of this action is provided by the repeat intervening spacer carried in the crRNA, which upon hybridization with complementary sequences enables their degradation by a Cas endonuclease. Moreover, CRISPR arrays are dynamic landscapes that may gain new spacers from infecting elements or lose them for example during genome replication.

Exploring CRISPR Interference by Transformation with Plasmid Mixtures: Identification of Target Interference Motifs in Escherichia coli.

Plasmid transformation into a bacterial host harboring a functional CRISPR-Cas system targeting a sequence in the transforming molecule can be specifically hindered by CRISPR-mediated interference. In this case, measurements of transformation efficacy will provide an estimation of CRISPR activity. However, in order to standardize data of conventional assays (using a single plasmid in the input DNA sample), transformation efficiencies have to be compared to those obtained for a reference molecule in independent experiments.

Spanish validation of the Family Questionnaire (FQ) in families of patients with an eating disorder.

Background: The objective of this study is to evaluate the psychometric properties of the Spanish version of the Family Questionnaire (FQ) and to further examine the differences between mothers' and fathers' emotional response to an eating disorder (ED).

Method: A total of 382 carers of patients with an ED participated in the study, with ages ranging from forty to fifty-three years old.

Results: The use of confirmatory factor analysis according to gender supported both factor models of the FQ, with only minor differences in comparison to the original study performed in 2002 by Wiedemann and collegues.

CRISPR-Cas functional module exchange in Escherichia coli.

Unlabelled: Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (cas) genes constitute the CRISPR-Cas systems found in the Bacteria and Archaea domains. At least in some strains they provide an efficient barrier against transmissible genetic elements such as plasmids and viruses. Two CRISPR-Cas systems have been identified in Escherichia coli, pertaining to subtypes I-E (cas-E genes) and I-F (cas-F genes), respectively.