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Abstract Background While families headed by same-sex couples have achieved greater public visibility in recent years, there are still many challenges for these families in dealing with legal and community contexts that are not supportive of same-sex relationships. The Work, Love, Play study is a large longitudinal study of same-sex parents. It aims to investigate many facets of family life among this sample and examine how they change over time. The study focuses specifically on two key areas missing from the current literature: factors supporting resilience in same-sex parented families; and health and wellbeing outcomes for same-sex couples who undergo separation, including the negotiation of shared parenting arrangements post-separation. The current paper aims to provide a comprehensive overview of the design and methods of this longitudinal study and discuss its significance. Methods/Design The Work, Love, Play study is a mixed design, three wave, longitudinal cohort study of same-sex attracted parents. The sample includes lesbian, gay, bisexual and transgender parents in Australia and New Zealand (including single parents within these categories) caring for any children under the age of 18 years. The study will be conducted over six years from 2008 to 2014. Quantitative data are to be collected via three on-line surveys in 2008, 2010 and 2012 from the cohort of parents recruited in Wave1. Qualitative data will be collected via interviews with purposively selected subsamples in 2012 and 2013. Data collection began in 2008 and 355 respondents to Wave One of the study have agreed to participate in future surveys. Work is currently underway to increase this sample size. The methods and survey instruments are described. Discussion This study will make an important contribution to the existing research on same-sex parented families. Strengths of the study design include the longitudinal method, which will allow understanding of changes over time within internal family relationships and social supports. Further, the mixed method design enables triangulation of qualitative and quantitative data. A broad recruitment strategy has already enabled a large sample size with the inclusion of both gay men and lesbians.

Abstract Background Glycolipid transfer protein is the prototypical and founding member of the new GLTP superfamily distinguished by a novel conformational fold and glycolipid binding motif. The present investigation provides the first insights into the organization, transcriptional status, phylogenetic/evolutionary relationships of GLTP genes. Results In human cells, single-copy GLTP genes were found in chromosomes 11 and 12. The gene at locus 11p15.1 exhibited several features of a potentially active retrogene, including a highly homologous (~94%), full-length coding sequence containing all key amino acid residues involved in glycolipid liganding. To establish the transcriptional activity of each human GLTP gene, in silico EST evaluations, RT-PCR amplifications of GLTP transcript(s), and methylation analyses of regulator CpG islands were performed using various human cells. Active transcription was found for 12q24.11 GLTP but 11p15.1 GLTP was transcriptionally silent. Heterologous expression and purification of the GLTP paralogs showed glycolipid intermembrane transfer activity only for 12q24.11 GLTP. Phylogenetic/evolutionary analyses indicated that the 5-exon/4-intron organizational pattern and encoded sequence of 12q24.11 GLTP were highly conserved in therian mammals and other vertebrates. Orthologs of the intronless GLTP gene were observed in primates but not in rodentiates, carnivorates, cetartiodactylates, or didelphimorphiates, consistent with recent evolutionary development. Conclusion The results identify and characterize the gene responsible for GLTP expression in humans and provide the first evidence for the existence of a GLTP pseudogene, while demonstrating the rigorous approach needed to unequivocally distinguish transcriptionally-active retrogenes from silent pseudogenes. The results also rectify errors in the Ensembl database regarding the organizational structure of the actively transcribed GLTP gene in Pan troglodytes and establish the intronless GLTP as a primate-specific, processed pseudogene marker. A solid foundation has been established for future identification of hereditary defects in human GLTP genes.

Accurate time series forecasting often requires higher temporal resolution than that provided by available data, such as when daily forecasts are needed from monthly data. Existing temporal disaggregation techniques, which typically handle only single, uniformly sampled time series, have limited applicability in real-world, multi-source scenarios. This paper introduces the Iterative Shifting Disaggregation (ISD) algorithm, designed to process and disaggregate time series derived from sensor-sourced low-frequency measurements, transforming multiple, nonuniformly sampled sensor data streams into a single, coherent high-frequency signal. ISD operates in an iterative, two-phase process: a prediction phase that uses multiple linear regression to generate high-frequency series from low-frequency data and correlated variables, followed by an update phase that redistributes low-frequency observations across high-frequency periods. This process repeats, refining estimates with each iteration cycle. The ISD algorithm's key contribution is its ability to disaggregate multiple, nonuniformly spaced time series with overlapping intervals into a single daily representation. In two case studies using natural gas data, ISD successfully disaggregates billing cycle and grouped residential customer data into daily time series, achieving a 1.4-4.3% WMAPE improvement for billing cycle data and a 4.6-10.4% improvement for residential data over existing methods.

Antisense oligonucleotides (ASOs) are emerging as a promising class of therapeutics for neurological diseases. When injected directly into cerebrospinal fluid, ASOs distribute broadly across brain regions and exert long-lasting therapeutic effects. However, many phosphorothioate (PS)-modified gapmer ASOs show transient motor phenotypes when injected into the cerebrospinal fluid, ranging from reduced motor activity to ataxia or acute seizure-like phenotypes. Using a behavioral scoring assay customized to reflect the timing and nature of these effects, we show that both sugar and phosphate modifications influence acute motor phenotypes. Among sugar analogs, DNA induces the strongest motor phenotypes while 2'-substituted RNA modifications improve the tolerability of PS ASOs. Reducing the PS content of gapmer ASOs, which contain a stretch of PS-DNA, improves their toxicity profile, but in some cases also reduces efficacy or duration of effect. We show that this acute toxicity is not mediated by major nucleic acid sensing immune pathways. Formulating ASOs with divalent ions before injection and avoiding phosphate-based buffers modestly improved tolerability through mechanisms at least partially distinct from reduced PS content. Overall, our work identifies and quantifies an understudied aspect of oligonucleotide toxicology in the CNS, explores its mechanism, and presents platform-level medicinal chemistry and formulation approaches that improve tolerability of this class of compounds., Competing Interests: Declaration of interests F.W., R.H.B., and J.K.W. are co-founders of Nucyrna Therapeutics. J.K.W. is advisory board member of EnPlusOne, PepGen, and Sixfold. R.H.B. is co-founder and Scientific Advisory Board member of ApicBio. The authors have filed patent applications on oligonucleotides and designs intended for use in the CNS., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Small interfering RNAs (siRNAs) represent a novel class of drugs capable of potent and sustained modulation of genes across various tissues. Preclinical development of siRNAs necessitates assessing efficacy and toxicity in animal models. While identifying therapeutic leads with cross-species activity can expedite development, it may compromise efficacy and be infeasible for certain gene targets. Here, we investigate whether deriving species-active siRNAs from potent human-targeting leads-an approach termed mismatch conversion-can yield potent compounds. We systematically altered potent siRNAs targeting human genes associated with diseases- SOD1 (ALS), JAK1 (inflammation), and HTT (HD)-to generate species-matching variants with full complementarity to their target in NHPs, mice, rats, sheep, and dogs. Variants potency and efficacy were measured in corresponding cell lines. We demonstrate that sequence, position, and number of mismatches significantly influence the ability to generate potent species-active compounds via mismatch conversion. Across tested sequences, mismatch conversion strategy ability to identify a species-active lead varied from 0% to 70%. For SOD1 , lead compounds identified from species-focus screening in mouse and dog cells were more potent than leads obtained from mismatch conversion. Thus, a focused screening of therapeutic lead and model compounds may represent a more reliable strategy for the clinical advancement of siRNAs.

Fetal alcohol exposure is a growing public health concern. However, ascertaining its true extent remains challenging as maternal self-reports may lack validity. Increasingly, interest has turned to more objective measures of prenatal alcohol use (PAU) of which one, meconium, is recognized as a valuable tool. This review assesses both the international prevalence of PAU obtained using meconium biomarkers in general maternity populations and, when feasible, the level of agreement between meconium biomarkers and self-reported PAU. A systematic literature search for studies reporting the prevalence of PAU, as determined by meconium biomarker testing, was conducted using multiple electronic databases from 1990 to 2023. Seventeen studies were identified for inclusion and evaluated for methodological quality. Using fatty acid ethyl esters (FAEEs) meconium biomarkers, PAU prevalence varied from 2.4% to 44%. Rates based on EtG (ethyl glucuronide) analysis ranged from 0% to 16.3%, and EtS (ethyl sulfate) analysis from 7.8% to 16.7%. Studies were of moderate quality with high heterogeneity. Prevalence rates based on self-report data ranged from 0% to 46.4%. When reported, none of the reviewed studies identified agreement between meconium-based and self-report-based PAU prevalence rates. Using both self-reports to detect early pregnancy alcohol use, and meconium biomarkers to detect the occurrence of alcohol use later in pregnancy, may provide a more complete picture of PAU prevalence. Furthermore, research is warranted to develop stringent guidance on the ascertainment, storage, analysis, and reporting required in this field., (© 2024 The Author(s). Alcohol, Clinical and Experimental Research published by Wiley Periodicals LLC on behalf of Research Society on Alcohol.)

Background: Paramagnetic rim lesions (PRL) are an emerging biomarker for multiple sclerosis (MS). In addition to associating with greater disease severity, PRL may be diagnostically supportive., Objective: Our aim was to determine PRL specificity and sensitivity for discriminating MS from its diagnostic mimics using real-world clinical diagnostic and imaging data., Methods: This is a retrospective, cross-sectional analysis of a longitudinal cohort of patients with prospectively collected observational data. Patients were included if they underwent neuroimmunological evaluation in our academic MS center, and had an available MRI scan from the same clinical 3T magnet that included a T2*-weighted sequence with susceptibility postprocessing (SWAN protocol, GE). SWAN-derived filtered phase maps and corresponding T2-FLAIR images were manually reviewed to determine PRL. PRL were categorized as "definite," "probable," or "possible" based on modified, recent consensus criteria. We hypothesized that PRL would convey a high specificity to discriminate MS from its MRI mimics., Results: 580 patients were evaluated in total: 473 with MS, 57 with non-inflammatory neurological disease (NIND), and 50 with other inflammatory neurological disease (OIND). Identification of "definite" or "probable" PRL provided a specificity of 98% to discriminate MS from NIND and OIND; sensitivity was 36%. Interrater agreement was almost perfect for definite/probable identification at a subject level., Conclusions: PRL convey high specificity for MS and can aid in the diagnostic evaluation. Modest sensitivity limits their use as single diagnostic indicators. Including lesions with lower confidence ("possible" PRL) rapidly erodes specificity and should be interpreted with caution given the potential harms associated with misdiagnosis., Competing Interests: Potential Conflicts of Interest: The authors declare no conflicts of interest with the study. DSR has received research funding from Abata and Sanofi for separate projects related to therapeutic targeting of PRL.

A hexanucleotide (G 4 C 2 ) repeat expansion (HRE) within intron one of C9ORF72 is the leading genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). C9ORF72 haploinsufficiency, formation of RNA foci, and production of dipeptide repeat (DPR) proteins have been proposed as mechanisms of disease. Here, we report the first example of disease-modifying siRNAs for C9ORF72 driven ALS/FTD. Using a combination of reporter assay and primary cortical neurons derived from a C9-ALS/FTD mouse model, we screened a panel of more than 150 fully chemically stabilized siRNAs targeting different C9ORF72 transcriptional variants. We demonstrate the lack of correlation between siRNA efficacy in reporter assay versus native environment; repeat-containing C9ORF72 mRNA variants are found to preferentially localize to the nucleus, and thus C9ORF72 mRNA accessibility and intracellular localization have a dominant impact on functional RNAi. Using a C9-ALS/FTD mouse model, we demonstrate that divalent siRNAs targeting C9ORF72 mRNA variants specifically or non-selectively reduce the expression of C9ORF72 mRNA and significantly reduce DPR proteins. Interestingly, siRNA silencing all C9ORF72 mRNA transcripts was more effective in removing intranuclear mRNA aggregates than targeting only HRE-containing C9ORF72 mRNA transcripts. Combined, these data support RNAi-based degradation of C9ORF72 as a potential therapeutic paradigm., Competing Interests: J.W.G., B.MDC.G., and A.K. are named as inventors on a patent application filed (OLIGONUCLEOTIDE-BASED MODULATION OF C9orf72). A.K. is a founder of Atalanta Therapeutics., (© 2024 The Author(s).)