Initial validation of the Health Environment Rating Scale-Early Childhood Consultation-Classroom (HERS-ECC-C).

The present study validated a newly developed easy-to-use observational instrument, the Health Environment Rating Scale-Early Childhood Consultation-Classroom version (HERS-ECC-C), to measure the quality of the classroom environment within early care and education centers participating in a mental health consultation program in a diverse area of the southeastern United States. Using a confirmatory factor analysis, three factors emerged capturing critical aspects of a high-quality classroom environment and demonstrated good reliability: (1) Supportive Practices, Positive Socioemotional Practices, and Classroom Management (α = .88), (2) Health and Family Communication (α = .79), and (3) Individualizing to Children's Needs (α = .80). Criterion-related validity was established through concurrent associations between the three HERS-ECC-C subscales and the domains of the Classroom Assessment Scoring System (CLASS) and predictive associations with the Childcare Worker Job Stress Inventory. The HERS-ECC-C Supportive Practices and Health and Family Communication subscales were associated with all three CLASS domains, and the Individualizing to Children's Needs subscale was associated with the CLASS Instructional support domain. Higher HERS-ECC-C subscale scores were associated with lower teacher-reported job stress. Findings provide initial evidence to support the use and continued development of the HERS-ECC-C as a tool to evaluate programs and classrooms engaged in mental health consultation professional development interventions.

Fixation-resistant photoactivatable fluorescent proteins for CLEM.

Fluorescent proteins facilitate a variety of imaging paradigms in live and fixed samples. However, they lose their fluorescence after heavy fixation, hindering applications such as correlative light and electron microscopy (CLEM). Here we report engineered variants of the photoconvertible Eos fluorescent protein that fluoresce and photoconvert normally in heavily fixed (0.5-1% OsO4), plastic resin-embedded samples, enabling correlative super-resolution fluorescence imaging and high-quality electron microscopy.

A naturally monomeric infrared fluorescent protein for protein labeling in vivo.

Infrared fluorescent proteins (IFPs) provide an additional color to GFP and its homologs in protein labeling. Drawing on structural analysis of the dimer interface, we identified a bacteriophytochrome in the sequence database that is monomeric in truncated form and engineered it into a naturally monomeric IFP (mIFP). We demonstrate that mIFP correctly labels proteins in live cells, Drosophila and zebrafish. It should be useful in molecular, cell and developmental biology.

Categorical spectral analysis of periodicity in human and viral genomes.

Periodicity in nucleotide sequences arises from regular repeating patterns which may reflect important structure and function. Although a three-base periodicity in coding regions has been known for some time and has provided the basis for powerful gene prediction algorithms, its origins are still not fully understood. Here, we show that, contrary to common belief, amino acid (AA) bias and codon usage bias are insufficient to create base-3 periodicity. This article applies the rigorous method of spectral envelope to systematically characterize the contributions of codon bias, AA bias and protein structural motifs to the three-base periodicity of coding sequences. The method is also used to classify CpG islands in the human genome. In addition, we show how spectral envelope can be used to trace the evolution of viral genomes and monitor global sequence changes without having to align to previously known genomes. This approach also detects reassortment events, such as those that led to the 2009 pandemic H1N1 virus.

Diverse Parents Decision-Making to Vaccinate Their Child under Five Attending Childcare Programs.

Background: On 18 June 2022, Moderna and Pfizer-BioNTech COVID-19 vaccines were authorized under an Emergency Use Authorization by the United States Food and Drug Administration to prevent severe coronavirus disease in children six months to four years of age. Despite approval of the COVID-19 vaccinations for young children, there remain ongoing challenges reaching widespread coverage due to parental decision-making. Parental decision-making plays a pivotal, yet understudied, role governing vaccine adoption among this priority demographic. Methods: This cross-sectional analysis examined COVID-19 vaccine intentions for 320 predominately Hispanic parents of two to five-year-olds attending Miami-Dade County childcare programs in Florida USA, several months following the June 2022 emergency authorization. Parent's self-reported survey data encompassed vaccine choices and rationales, social determinants of health, and parent immigrant status. Data analyses illustrate the associations between parent decision-making and these variables. Regression modeling and tests of independence identified predicting factors for parental vaccine decision-making. Results: Only 25% of parents intended to vaccinate their young child, while 34% resisted and 41% felt unsure, despite 70% personal vaccination rates. Household income under $25,000, identifying as a migrant, or testing COVID-19-positive significantly predicted unsure decision-making. The majority of hesitant groups expressed concerns around side effects (20%), safety (2.9%), and sufficiency of vaccine knowledge (3.3%). Conclusions: In this sample, the predominance of parents were unsure and resistant rather than accepting of having their child vaccinated despite emergency approval of the pediatric vaccine. Associations and predictive factors are examined.

Virtual robotic telepresence early childhood mental health consultation to childcare centers in the aftermath of COVID-19: training approaches and perceived acceptability and usefulness.

Introduction: Childcare center closures during COVID-19 impacted education for approximately 40 million children nationwide. Unfortunately, COVID-19 restrictions significantly limited the extent that outside personnel could provide in-person support to educators, resulting in the need for innovative approaches to meet childcare centers' needs. A virtual robotic telepresence approach was applied to early childhood consultation models to promote child resilience while mitigating COVID-19 risks. The goal of this study was to examine how training influenced consultants' and childcare staff uptake of the virtual robotic telepresence consultation approach and their acceptance of this technology. Methods: Ten early childhood consultants received multimedia/simulation training and weekly communities of practice related to virtual telepresence robotic consultation. Telepresence robotic consultation equipment was deployed to 16 childcare centers in a diverse multilingual metropolitan area as a part of a larger randomized controlled trial. Consultants trained childcare staff (14 center directors and 58 teachers) on how to receive virtual telepresence robotic consultation. Demographic information and measures of technology acceptability and uptake were collected from childcare staff and consultants. A mixed methods approach was used including multilevel modeling and focus groups to examine consultation uptake, acceptability, barriers, and facilitators of virtual telepresence robotic consultation implementation. Results: Consultants and childcare staff generally perceived the virtual telepresence consultation approach to be useful and easy to use. Consultant perceptions of the acceptability of technology did not change over time. Childcare staff, center, and consultant factors impacted the uptake of the virtual robotic telepresence consultation approach and childcare staff acceptance of the technology. Focus groups revealed that consultants believed that additional hands-on training with childcare staff would have benefited implementation and expressed a desire for a hybrid approach for consultation. Discussion: Perceptions of telepresence robotic consultation acceptability are discussed, including future recommendations for training.

Effectiveness of education and attitudes toward different types of deceased donor kidneys: Survey analysis of single-center experience.

Background: We lack data on the effectiveness of education and the patient's attitude toward different deceased donor kidney types. A prospective study was performed to evaluate patient attitudes, baseline knowledge, and effectiveness of our kidney transplant education process. We also analyzed the knowledge retention of our waitlist patients. Design: We prospectively surveyed a patient cohort using a paired analysis pre and post education with initial evaluation visit. Knowledge retention among waitlist patients was assessed with annual waitlist visit. Results: One hundred four patients received paired surveys to assess the baseline knowledge and effectiveness of education. Forty-three patients received a single survey with their annual waitlist evaluation to assess knowledge retention. Paired survey showed mixed results, with no statistically significant improvement in the kidney donor profile index domain. Significant improvement was seen in the hepatitis C virus-positive donor domain and the Public Health Service (PHS) increased-risk donor domain. For the waitlist cohort, overall knowledge retention ranged from excellent to fair, with a decline in knowledge for the PHS increased-risk donor domain. Conclusion: Our study suggests that the education intervention regarding different deceased donor kidney types is effective overall and transplant candidates retain the knowledge while waiting for transplant.

Type-specific prevalence and persistence of human papillomavirus in women in the United States who are referred for typing as a component of cervical cancer screening.

OBJECTIVE: The purpose of this study was to report type-specific prevalence and persistence of human papillomavirus (HPV) in women who underwent cytologic screening. STUDY DESIGN: We examined HPV prevalence in 73,371 women who had type-specific HPV testing in 1 of 23 clinical laboratories in the United States. Persistence was evaluated in 963 women who were tested within 8-16 months of their index test. RESULTS: HPV was detected in 31% of the women, and high-risk HPV was detected in 23% of the women. HPV-16, -53, -52, and -31 were the most prevalent types. Of the 953 women with 2 tests, 39% of the women had persistent HPV infection. High-risk HPV persistence was detected in 34% of the women who were positive initially for high-risk HPV. CONCLUSION: Approximately one-third of our sample had HPV; of those women who were retested within 8-16 months, more than one-third had persistent infection. Among women with high-risk HPV infections, the likelihood of persistence was highest with HPV genotypes that were phylogenetically similar to HPV-16.

Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.

Many genetically encoded biosensors use Förster resonance energy transfer (FRET) to dynamically report biomolecular activities. While pairs of cyan and yellow fluorescent proteins (FPs) are most commonly used as FRET partner fluorophores, respectively, green and red FPs offer distinct advantages for FRET, such as greater spectral separation, less phototoxicity, and lower autofluorescence. We previously developed the green-red FRET pair Clover and mRuby2, which improves responsiveness in intramolecular FRET reporters with different designs. Here we report the engineering of brighter and more photostable variants, mClover3 and mRuby3. mClover3 improves photostability by 60% and mRuby3 by 200% over the previous generation of fluorophores. Notably, mRuby3 is also 35% brighter than mRuby2, making it both the brightest and most photostable monomeric red FP yet characterized. Furthermore, we developed a standardized methodology for assessing FP performance in mammalian cells as stand-alone markers and as FRET partners. We found that mClover3 or mRuby3 expression in mammalian cells provides the highest fluorescence signals of all jellyfish GFP or coral RFP derivatives, respectively. Finally, using mClover3 and mRuby3, we engineered an improved version of the CaMKIIα reporter Camuiα with a larger response amplitude.

Three-dimensional acrylamide fluorescence in situ hybridization for plant cells.

Plant meiosis involves complex and dynamic processes that occur within the space inside the nucleus. Direct inspection of meiotic chromosomes by fluorescence microscopy has been used to investigate many of these processes. In particular, optical sectioning microscopy of fluorescence in situ hybridization (FISH)-stained nuclei provides three-dimensional spatial information about the organization and distribution of specific sequences and chromosomal loci within the nucleus. Here we provide a fully detailed three-dimensional (3D) acrylamide FISH method for the analysis of plant meiotic nuclei. Several examples illustrate the versatility of this technique for the investigation of meiotic telomere dynamics in maize, Arabidopsis, and oat. Additional examples of 3D FISH include chromosome painting in a maize chromosome-addition line of oat and telomere FISH with maize nuclei from plants expressing a fluorescently tagged fusion protein, histone H2B-mCherry.

An engineered monomeric Zoanthus sp. yellow fluorescent protein.

Protein engineering has created a palette of monomeric fluorescent proteins (FPs), but there remains an ~30 nm spectral gap between the most red-shifted useful Aequorea victoria green FP (GFP) variants and the most blue-shifted useful Discosoma sp. red FP (RFP) variants. To fill this gap, we have engineered a monomeric version of the yellow FP (YFP) from Zoanthus sp. coral. Our preferred variant, designated as mPapaya1, displays excellent fluorescent brightness, good photostability, and retains its monomeric character both in vitro and in living cells in the context of protein chimeras. We demonstrate that mPapaya1 can serve as a good Förster resonance energy transfer (FRET) acceptor when paired with an mTFP1 donor. mPapaya1 is a valuable addition to the palette of FP variants that are useful for multicolor imaging and FRET-based biosensing.

Maize histone H2B-mCherry: a new fluorescent chromatin marker for somatic and meiotic chromosome research.

Cytological studies of fluorescent proteins are rapidly yielding insights into chromatin structure and dynamics. Here we describe the production and cytological characterization of new transgenic maize lines expressing a fluorescent histone fusion protein, H2B-mCherry. The transgene is expressed under the control of the maize ubiquitin1 promoter, including its first exon and intron. Polymerase chain reaction-based genotyping and root-tip microscopy showed that most of the lines carrying the transgene also expressed it, producing bright uniform staining of nuclei. Further, plants showing expression in root tips at the seedling stage also showed expression during meiosis, late in the life cycle. Detailed high-resolution three-dimensional imaging of cells and nuclei from various somatic and meiotic cell types showed that H2B-mCherry produced remarkably clear images of chromatin and chromosome fiber morphology, as seen in somatic, male meiotic prophase, and early microgametophyte cells. H2B-mCherry also yielded distinct nucleolus staining and was shown to be compatible with fluorescence in situ hybridization. We found several instances where H2B-mCherry was superior to DAPI as a generalized chromatin stain. Our study establishes these histone H2B-mCherry lines as new biological reagents for visualizing chromatin structure, chromosome morphology, and nuclear dynamics in fixed and living cells in a model plant genetic system.

NSeq: a multithreaded Java application for finding positioned nucleosomes from sequencing data.

We introduce NSeq, a fast and efficient Java application for finding positioned nucleosomes from the high-throughput sequencing of MNase-digested mononucleosomal DNA. NSeq includes a user-friendly graphical interface, computes false discovery rates (FDRs) for candidate nucleosomes from Monte Carlo simulations, plots nucleosome coverage and centers, and exploits the availability of multiple processor cores by parallelizing its computations. Java binaries and source code are freely available at https://github.com/songlab/NSeq. The software is supported on all major platforms equipped with Java Runtime Environment 6 or later.