Hydrogels for Local and Sustained Delivery of Bacteriophages to Treat Multidrug-Resistant Wound Infections.

Bacteriophages (phages), viruses that specifically target and kill bacteria, represent a promising strategy to combat multidrug-resistant (MDR) pathogens such as Pseudomonas aeruginosa ( Pa ). However, delivering sufficient concentrations of active phages directly to the infection site remains challenging, with current methods having variable success. Here we present "HydroPhage", an innovative hydrogel system for the sustained release of high-titer phages to effectively treat infections caused by MDR pathogens. Our injectable hydrogels, featuring dual-crosslinking of hyaluronic acid and PEG-based hydrogels through static covalent thioether bonds and dynamic covalent hemithioacetal crosslinks (DCC), encapsulate phages at concentration up to 10 11 PFU/mL, and achieves controlled release of 10 9 PFU daily over a week, surpassing levels of current clinical dosages, with more than 60% total phage recovery. In a preclinical mouse model of extended wound infection, compared to intravenous treatment, we demonstrate enhanced bacterial clearance by localized, high-dose, and repeated phage dosing despite the emergence of bacterial resistance to phages. This work advances the development of clinically practical wound dressings tailored for resistant infections.

Multi-year population-based analysis of Asian patients with acute decompensated heart failure and advanced chronic kidney disease.

BACKGROUND: Data on disparities in outcomes and risk factors in Asian patients with advanced chronic kidney disease admitted for heart failure are scare. METHODS: This was a retrospective cohort study that utilized data from the National Inpatient Sample between January 2016 and December 2019. Patients who had a primary diagnosis of acute decompensated heart failure and a concomitant diagnosis of advanced CKD were included. The primary outcome of interest was in-hospital mortality. Secondary outcomes include hospital cost, length of stay, and other clinical outcomes. Weighted multivariable logistic regression was used to adjust for comorbidities. RESULTS: There were 251,578 cases of ADHF with advanced CKD, out of which 2.6 % were from individuals of Asian ethnicity. Asian patients exhibited a higher burden of comorbidities in comparison to other UREM patients, but a lower burden than White patients. Regardless of differences in comorbidity burden, Asian patients exhibited a higher likelihood of experiencing severe consequences. After adjusting for comorbidies, White (OR:1.11; 95 % CI 1.03-1.20;0.009) patients had higher odds of mortality than Asian patients. However, Blacks (OR: 0.58; 95 % CI 0.53 to 0.63; p < 0.001) and Hispanics (OR: 0.69; 95 % CI 0.62 to 0.78; p < 0.001) had lower odds of mortality. CONCLUSION: This first population-based studies shows that Asian patients with advanced CKD admitted for ADHF have greater comorbidity burden and poorer outcomes Black and Hispanic patients. This data underscores the importance of comprehensive approaches in phenotyping, and ethnic specific interventions.

The impact of exclusion due to COVID-19 restrictions on partners' satisfaction with Swedish hospital postnatal ward care: A multi-methods approach.

BACKGROUND: To prevent the spread of SARS-CoV-2, hospitals around the world adopted protocols that, in varying ways, resulted in the exclusion of partners from hospital postnatal care wards. The objective of this study was to examine the effect this exclusion had on partners' satisfaction with postnatal care. METHODS: An online survey (the Swedish Pregnancy Panel) including free-text comments was conducted before and during the first wave of the COVID-19 pandemic; partners of pregnant women were recruited at an early ultrasound appointment and followed until 2 months after childbirth. Data were linked to the Swedish Pregnancy Register. RESULTS: The survey was completed by 524 partners of women who gave birth during the pandemic and 203 partners of women who gave birth before. Partners' satisfaction with hospital postnatal care dropped 29.8 percent (-0.94 OLS, 95% CI = -1.17 to -0.72). The drop was largest for partners of first-time mothers (-1.40 OLS, 95% CI = -1.69 to -1.11), but unrelated to clinical outcomes such as mode of birth and most social backgrounds, except higher income. The qualitative analysis showed that partners (1) felt excluded as partners and parents, (2) thought the strain on staff led to deficiencies in the care provided, and (3) perceived the decision about partner restrictions as illogical. CONCLUSIONS: The exclusion of partners from the hospital postnatal wards clearly impaired satisfaction with care, and partners of first-time mothers were particularly affected. Planning for future restrictions on partners from hospital wards should factor in these consequences.

The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems.

Type III CRISPR-Cas systems employ multiprotein effector complexes bound to small CRISPR RNAs (crRNAs) to detect foreign RNA transcripts and elicit a complex immune response that leads to the destruction of invading RNA and DNA. Type III systems are among the most widespread in nature, and emerging interest in harnessing these systems for biotechnology applications highlights the need for detailed structural analyses of representatives from diverse organisms. We performed cryo-EM reconstructions of the Type III-A Cas10-Csm effector complex from S. epidermidis bound to an intact, cognate target RNA and identified two oligomeric states, a 276 kDa complex and a 318 kDa complex. 3.1 Å density for the well-ordered 276 kDa complex allowed construction of atomic models for the Csm2, Csm3, Csm4 and Csm5 subunits within the complex along with the crRNA and target RNA. We also collected small-angle X-ray scattering data which was consistent with the 276 kDa Cas10-Csm architecture we identified. Detailed comparisons between the S. epidermidis Cas10-Csm structure and the well-resolved bacterial (S. thermophilus) and archaeal (T. onnurineus) Cas10-Csm structures reveal differences in how the complexes interact with target RNA and crRNA which are likely to have functional ramifications. These structural comparisons shed light on the unique features of Type III-A systems from diverse organisms and will assist in improving biotechnologies derived from Type III-A effector complexes.

A Longitudinal Dyadic Study of Six Leisure Activities in Swedish Couples During the Transition to Parenthood.

Parents are not only caregivers to their children; they also have leisure routines that can impact their own well-being. However, little is known about how leisure activities change within the context of a couple during the transition to parenthood. This study uses latent growth curve models and data from the Swedish Pregnancy Panel to examine how often 918 first-time, heterosexual couples participated in six leisure activities from around pregnancy week 19 to one year postpartum. Compared to fathers, mothers less frequently exercised and listened to news, and more frequently read newspapers, spent time for themselves, and spent time with friends. Over time, mothers increased their frequency of praying to God and decreased spending time for themselves and with friends. Fathers decreased frequency of exercise. Within couples, there was a positive correlation between mothers' and fathers' frequency of engaging in leisure activities, although most changes over time were not associated. Our finding that two individuals within a couple may change their leisure activities independently of each other during the transition to parenthood can help healthcare professionals and researchers prepare expectant couples for upcoming changes (or lack thereof) and promote parent well-being. Our findings also highlight the possibility that in contexts with more state support for families, parenthood may not exacerbate gender gaps in leisure.

Critical roles for 'housekeeping' nucleases in type III CRISPR-Cas immunity.

CRISPR-Cas systems are a family of adaptive immune systems that use small CRISPR RNAs (crRNAs) and CRISPR-associated (Cas) nucleases to protect prokaryotes from invading plasmids and viruses (i.e., phages). Type III systems launch a multilayered immune response that relies upon both Cas and non-Cas cellular nucleases, and although the functions of Cas components have been well described, the identities and roles of non-Cas participants remain poorly understood. Previously, we showed that the type III-A CRISPR-Cas system in Staphylococcus epidermidis employs two degradosome-associated nucleases, PNPase and RNase J2, to promote crRNA maturation and eliminate invading nucleic acids (Chou-Zheng and Hatoum-Aslan, 2019). Here, we identify RNase R as a third 'housekeeping' nuclease critical for immunity. We show that RNase R works in concert with PNPase to complete crRNA maturation and identify specific interactions with Csm5, a member of the type III effector complex, which facilitate nuclease recruitment/stimulation. Furthermore, we demonstrate that RNase R and PNPase are required to maintain robust anti-plasmid immunity, particularly when targeted transcripts are sparse. Altogether, our findings expand the known repertoire of accessory nucleases required for type III immunity and highlight the remarkable capacity of these systems to interface with diverse cellular pathways to ensure successful defense.

A Simple, Robust, and Cost-effective Method for Genotyping Small-scale Mutations.

Background and objectives: Genotyping is an important tool for studying gene functions in animals or detecting genetic variants in humans. Various methods using low to high concentrations of agarose or polyacrylamide gel electrophoresis have been developed for genotyping. These methods rely on the detection of large-size differences (20-2,000 bp) of targeted PCR products between a wild-type gene and a mutant gene. Endonuclease digestion was introduced to identify heterozygous mutations, but it was not possible to differentiate the wild-type from the homozygous mutants with the same or similar size. This study thus developed a novel, simple, and reliable test for genotyping animals or cells following genetic modifications. Methods: We developed an improved and simple method that used 2% agarose gel electrophoresis following T7E1 or Surveyor endonuclease digestion to firstly separate the heterozygous mutations from the wild-type or homozygous mutations. By adding a wild-type PCR product to a potentially homozygous product, which would form heteroduplexes, we could then separate the wild-type from a homozygous mutation with a nearly identical size or only a single base pair substitution without Sanger sequencing. Results: We verified this method in genotyping zebrafish mutants with a 2-8-bp deletion or insertion and mouse mutants with a 1- or 8-bp substitution. The wild-type, heterozygous, and homozygous mutations ranged 1-8 bp were clearly differentiated on agarose gel. Sanger sequencing also confirmed our genotyping results. Conclusions: This novel and improved genotyping method may have a broad application in many clinical and research laboratories for rapid and economical genotyping of patients and animals with a small area deletion or single base pair substitution, particularly in the era of gene editing or in those with naturally occurring mutations.

Expectant parents' emotions evoked by pregnancy: A longitudinal dyadic analysis of couples in the Swedish Pregnancy Panel.

RATIONALE: Holistic antenatal care requires knowledge of individuals' emotional response to pregnancy. Little is known about how a pregnant woman and her partner influence each other emotionally during a pregnancy. OBJECTIVE: This study examines six discrete emotions that expectant couples experience during pregnancy, how these emotions change mid-to late-pregnancy, and whether the partners' emotional responses influence each other. METHODS: A longitudinal dyadic study where pregnant women and their partners (1432 couples) rated the extent to which the pregnancy evoked joy, strength, security, worry, shame, and anger at pregnancy week 12-19, 22-24, and 36. Latent curve models with structured residuals identify levels of and change in these emotions over time, while accounting for between- and within-couple variance. RESULTS: Pregnancy evoked mainly joy, strength, security, and worry, and lower levels of anger and shame. Pregnant women and partners felt similar levels of joy, strength, and security, but pregnant women felt more worry, shame, and anger. There was little to no mean-level change in all six measured emotions evoked by pregnancy (between-couple change), and no reciprocal effects between the partners (within-couple change). CONCLUSIONS: Emotions in mid-pregnancy were also felt in late pregnancy. Furthermore, the pregnant woman and her partner have individual emotional trajectories. The results can assist healthcare professionals and researchers target interventions to expectant mothers and partners, specifically by understanding emotional response to pregnancy as a stable confound and by not approaching the couple as one emotional unit.

Association Between Educational Attainment and EFNEP Participants' Food Practice Outcomes.

OBJECTIVE: Examine the association between educational attainment and improvement in food practice outcomes of the California Expanded Food and Nutrition Education Program (EFNEP) participants. DESIGN: Secondary data analysis. PARTICIPANTS: A total of 19,089 participants, 92.3% female, 77.2% Hispanic, 19.7% with ≤ sixth-grade education, and 68.9% with incomes ≤ 100% of the federal poverty level. MAIN OUTCOME MEASURES: Improvement in food resource management practices (FRMP), nutrition practices, and food safety practices (FSP). ANALYSIS: Wilcoxon signed rank tests examined pre-post outcomes. Mann-Whitney U tests compared whether participants in the lowest and highest educational attainment quartiles had similar levels of improvement. RESULTS: California EFNEP is associated with improved FRMP (z = -95.33), nutrition practices (z = -94.91), and FSP (z = -92.37); (P < 0.001). Lowest educational quartile was associated with more improvement in FRMP and FSP (P < 0.001). CONCLUSIONS AND IMPLICATIONS: California EFNEP contributed to improved food practice outcomes for low and high educational attainment participants. Program content and instruction are effective across the education continuum.

A unique mode of nucleic acid immunity performed by a multifunctional bacterial enzyme.

The perpetual arms race between bacteria and their viruses (phages) has given rise to diverse immune systems, including restriction-modification and CRISPR-Cas, which sense and degrade phage-derived nucleic acids. These complex systems rely upon production and maintenance of multiple components to achieve antiphage defense. However, the prevalence and effectiveness of minimal, single-component systems that cleave DNA remain unknown. Here, we describe a unique mode of nucleic acid immunity mediated by a single enzyme with nuclease and helicase activities, herein referred to as Nhi (nuclease-helicase immunity). This enzyme provides robust protection against diverse staphylococcal phages and prevents phage DNA accumulation in cells stripped of all other known defenses. Our observations support a model in which Nhi targets and degrades phage-specific replication intermediates. Importantly, Nhi homologs are distributed in diverse bacteria and exhibit functional conservation, highlighting the versatility of such compact weapons as major players in antiphage defense.

Examining correlates of feeding practices among parents of preschoolers.

Background: Parent feeding practices play a critical role in children's eating behaviors. Limited research has explored child-level correlates of parent feeding practices. Aim: To identify correlates of feeding practices (responsive and controlling) among parents of preschoolers US. Methods: Participants included parents (n = 273) of preschoolers (3-5 years), recruited from Early Care and Education settings (n = 24) located in a metropolitan city in the US. Analysis included descriptives, correlations, and multiple regression. Results: For responsive feeding practices, positive associations included child's weight with unintentional modeling (ß = .17, 95% CI [0.12, 0.53]), child vegetable consumption with behavioral role modeling (ß = 0.22, 95% CI [0.17, 0.44]), and parent monitoring with verbal modeling (ß = 0.21, 95% CI [0.12, 0.34]). For controlling feeding practices, parent restriction was positively associated with child weight concern (ß = 0.22, 95% CI [0.13, 0.39]) and parent monitoring (ß = 0.13, 95% CI [0.01, 0.19]), whereas child vegetable consumption was negatively associated (ß = -0.16, 95% CI [-0.27, -0.05]). Pressure to eat was negatively associated with child weight concern (ß = -0.18, 95% CI [-0.45, -0.09]), child fruit consumption (ß = -0.12, 95% CI [-0.37, -0.01]), household income (ß = -0.13, 95% CI [-0.30, -0.02]), and parent weight (ß = -0.14, 95% CI [-0.60, -0.05]), Conclusions: Findings highlight the importance of child characteristics when examining correlates of parent feeding practices, demonstrating bidirectional interactions between parent feeding practices and children's eating behaviors. Considering child-level correlates may improve the implementation of responsive feeding practices and reduce controlling feeding practices.

Mothers' Perceptions of the Climate of Their Children's Schools: Covariations With Children's Academic Adjustment in Families of Mexican Origin.

Although Latinx parents' perceptions of the climate of their children's schools may play a role in their children's academic adjustment, research examining this idea is sparse. Every 2 years beginning when children were in fifth grade (Mage  = 10.86 years) until they were in 11th grade, Mexican-origin mothers (N = 674) reported on their perceptions of the climate of their children's schools; information on children's academic adjustment was collected from children and mothers. Multilevel modeling indicated that when mothers had more positive school climate perceptions, their children valued school more and performed better in school, but did not necessarily hold higher educational expectations. The findings suggest the importance of schools in creating welcoming environments for Mexican-origin parents.

Are self-esteem and academic achievement reciprocally related? Findings from a longitudinal study of Mexican-origin youth.

OBJECTIVE: Previous research has shown that self-esteem is associated with academic achievement. However, few studies have used longitudinal data to examine how self-esteem and achievement co-develop over a long time span, and even fewer have focused on ethnic minority youth. METHOD: We used data from a longitudinal study of Mexican-origin youth (N = 674) to examine the bidirectional associations between self-esteem and academic achievement from 5th to 11th grade. Global and domain-specific self-esteem (academic, honesty, peer relationships, appearance) were assessed at ages 10, 12, 14, and 16 using Marsh et al.'s (2005) Self-Description Questionnaire. Academic achievement was assessed at the same ages using self-reported grades and standardized test scores from school records. RESULTS: Youth with high global and academic self-esteem showed relative improvements in their grades (but not test scores), and youth who received higher grades and test scores showed relative increases in global and academic self-esteem. Youth with high honesty self-esteem showed relative increases in grades and test scores, and youth with higher grades showed relative increases in peer relationship self-esteem. CONCLUSION: Students who feel better about themselves tend to show improvements in their grades, and getting better grades and test scores promotes more positive self-views.

The Respiratory Phenotype of Pompe Disease Mouse Models.

Pompe disease is a glycogen storage disease caused by a deficiency in acid α-glucosidase (GAA), a hydrolase necessary for the degradation of lysosomal glycogen. This deficiency in GAA results in muscle and neuronal glycogen accumulation, which causes respiratory insufficiency. Pompe disease mouse models provide a means of assessing respiratory pathology and are important for pre-clinical studies of novel therapies that aim to treat respiratory dysfunction and improve quality of life. This review aims to compile and summarize existing manuscripts that characterize the respiratory phenotype of Pompe mouse models. Manuscripts included in this review were selected utilizing specific search terms and exclusion criteria. Analysis of these findings demonstrate that Pompe disease mouse models have respiratory physiological defects as well as pathologies in the diaphragm, tongue, higher-order respiratory control centers, phrenic and hypoglossal motor nuclei, phrenic and hypoglossal nerves, neuromuscular junctions, and airway smooth muscle. Overall, the culmination of these pathologies contributes to severe respiratory dysfunction, underscoring the importance of characterizing the respiratory phenotype while developing effective therapies for patients.

Management of metabolic syndrome and reduction in body weight in type II diabetic mice by inhibiting glycosphingolipid synthesis.

Metabolic syndrome is defined by hyperlipidemia and cardiovascular complications. We have examined whether inhibition of glycosphingolipid synthesis can interfere with metabolic syndrome in a male mouse model of type II diabetes (db/db). The db/db and control mice (C57/BL6) (n = 6) fed chow for 30 weeks received vehicle (5% Tween-80 in PBS; 100 µl), or a biopolymer-encapsulated D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (BPD) glycosphingolipid synthesis inhibitor daily via oral gavage for 6 weeks. Echocardiography revealed increased Ao-IMT in db/db mice compared to control. However, BPD decreased Ao-IMT, monohexosylceramide and dihexosylceramide, LDL, triglycerides, glucose, and raised HDL levels in db/db mice. This was due to increased gene expression of HMG-CoA reductase, LDLr, SREBP2, and bile acids: Cy7-a hydroxylase, LXR and FXR, lipoprotein lipase, VLDL receptor and PPAR. Treatment also increased the expression of superoxide dismutase-II to reduce the pro-oxidant status in these mice. We observed that decreased cholesterol levels correlated with decreased cholesterol sensing proteins e.g. NPC1 gene/protein expression and mammalian target of rapamycin (mTORC-1) and reduced body weight. Thus, glycosphingolipid synthesis inhibition is a novel approach to manage metabolic syndrome and reduce body weight in diabetic mice and with potential applications in humans.

A type III-A CRISPR-Cas system employs degradosome nucleases to ensure robust immunity.

CRISPR-Cas systems provide sequence-specific immunity against phages and mobile genetic elements using CRISPR-associated nucleases guided by short CRISPR RNAs (crRNAs). Type III systems exhibit a robust immune response that can lead to the extinction of a phage population, a feat coordinated by a multi-subunit effector complex that destroys invading DNA and RNA. Here, we demonstrate that a model type III system in Staphylococcus epidermidis relies upon the activities of two degradosome-associated nucleases, PNPase and RNase J2, to mount a successful defense. Genetic, molecular, and biochemical analyses reveal that PNPase promotes crRNA maturation, and both nucleases are required for efficient clearance of phage-derived nucleic acids. Furthermore, functional assays show that RNase J2 is essential for immunity against diverse mobile genetic elements originating from plasmid and phage. Altogether, our observations reveal the evolution of a critical collaboration between two nucleic acid degrading machines which ensures cell survival when faced with phage attack.

The codevelopment of effortful control and school behavioral problems.

Effortful control refers to the propensity to regulate one's impulses and behaviors, to focus and shift attention easily, and to motivate the self toward a goal when there are competing desires. Although it seems likely that these capacities are relevant to successful functioning in the school context, there has been surprisingly little longitudinal research examining whether youth with poor effortful control are more likely to act out in the classroom, get suspended, and skip school. Conversely, there is even less research on whether youth who exhibit these school behavioral problems are more likely to decline over time in effortful control. We used multimethod data from a longitudinal study of Mexican-origin youth (N = 674), assessed biennially from 5th to 11th grade, to examine the codevelopment of effortful control and school behavioral problems. Bivariate latent growth curve models revealed a negative association between the trajectories of effortful control and school behavioral problems, indicating that steeper decreases in effortful control were related to steeper increases in school behavioral problems. Furthermore, this codevelopmental pattern was bidirectional; cross-lagged regression analyses showed that low effortful control was associated with relative increases in school behavioral problems, and school behavioral problems were associated with relative decreases in effortful control. Gender, nativity status, Mexican cultural values, and school-level antisocial behavior had concurrent associations with effortful control and school behavioral problems, but they did not moderate the codevelopmental pathways. We discuss the theoretical implications for personality development, as well as the practical implications for reducing school behavioral problems during adolescence. (PsycINFO Database Record (c) 2019 APA, all rights reserved).