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Primary cilia are membrane-covered microtubule-based structures that protrude from the cell surface and are critical for cell signaling and homeostasis during human development and adulthood. Dysregulation of cilia formation, length, and function can lead to a spectrum of human diseases and syndromes known as ciliopathies. Although some genetic and chemical screens have been performed to define important factors that modulate cilia biogenesis and length control, there are currently no clinical treatments that restore cilia length in patients. We report that the microtubule-targeting agent MI-181(mitotic inhibitor-181) is a potent modulator of cilia length and biogenesis. Treatment of retinal pigment epithelial-1 cells with MI-181 induced an increase in the average size of cilia and in the percent ciliated cells under nonstarved conditions. Importantly, MI-181 was effective at rescuing cilia length and ciliation defects in cells that had been treated with the intraflagellar transport inhibitor Ciliobrevin D or the O-GlcNAc transferase inhibitor OSMI-1. Most importantly, MI-181 induced an increase in cilia length and restored ciliation in cells with compromised shortened cilia at low nanomolar concentrations and did not show an inhibitory response at high concentrations. Therefore, MI-181 represents a lead molecule for developing drugs targeting ciliopathies characterized by shortened cilia.

Understanding plant-microbe interactions requires examination of root exudation under nutrient stress using standardized and reproducible experimental systems. We grew Brachypodium distachyon hydroponically in fabricated ecosystem devices (EcoFAB 2.0) under three inorganic nitrogen forms (nitrate, ammonium, and ammonium nitrate), followed by nitrogen starvation. Analyses of exudates with liquid chromatography-tandem mass spectrometry, biomass, medium pH, and nitrogen uptake showed EcoFAB 2.0's low intratreatment data variability. Furthermore, the three inorganic nitrogen forms caused differential exudation, generalized by abundant amino acids-peptides and alkaloids. Comparatively, nitrogen deficiency decreased nitrogen-containing compounds but increased shikimates-phenylpropanoids. Subsequent bioassays with two shikimates-phenylpropanoids (shikimic and p-coumaric acids) on soil bacteria or Brachypodium seedlings revealed their distinct capacity to regulate both bacterial and plant growth. Our results suggest that (i) Brachypodium alters exudation in response to nitrogen status, which can affect rhizobacterial growth, and (ii) EcoFAB 2.0 is a valuable standardized plant research tool.

Abstract In this study, we analysed the mutation spectrum in subjects with suspected lipodystrophy using a targeted Next-generation sequencing (NGS) approach. Subjects with suspected lipodystrophy were for screened six genes (AGPAT2, BSCL2, LMNA, PPARG, ZMPSTE24, INSR) and the variants identified were confirmed through Sanger sequencing. The clinical and biochemical parameters were compared among the mutation positive and negative subjects. We identified eight individuals with pathogenic or likely pathogenic mutations, including both homozygous and heterozygous variants. Homozygous variants included AGPAT2(NM_006412.4):c.493-2A>G, AGPAT2(NM_006412.4):c.254_258dup, and BSCL2(NM_001122955.4):c.570del, while heterozygous variants encompassed LMNA(NM_170707.4):c.1444C>T, LMNA(NM_170707.4):c.1456A>G, LMNA(NM_170707.4):c.1445G>A, and PPARG(NM_015869.5):c.949T>C mutations. In this cohort, three subjects were diagnosed with congenital generalized lipodystrophy, while the remaining five had familial partial lipodystrophy. Majority (7/8) of the patients with lipodystrophy had hepatic involvement. Notably, more than half of the subjects (5/8) achieved optimal glycemic control through insulin sensitizers (PPARγ agonist and Metformin). Interestingly, even with a limited gene panel test, mutation-positive individuals exhibited a higher prevalence of typical clinical features and biochemical characteristics associated with lipodystrophy compared to their mutation-negative counterparts. In subjects with lipodystrophy, targeted NGS based screening may establish a genetic diagnosis and aid in family screening and genetic counselling. Knowing the clinical and biochemical features typical to lipodystrophy may help in diagnosis especially in resource limited setting.

Abstract Background Children’s active travel to school is associated with physical activity and thus health. Safe Routes to School (SR2S) programs identify ‘safe routes’ to promote children’s active travel to school. No field audit tool exists specifically to assess the microscale built environment of these ‘safe routes’ within Australia. This study describes the reliability assessment of the Field Audit for Children’s Active Transport to School (FACTS) tool. Methods The FACTS tool was developed using a multi-step process, including a literature search, expert opinion, and pilot testing. For the reliability assessment, two trained auditors collected data at eight schools over three weeks in April 2021. For inter-rater reliability, auditors conducted audits on the ‘safe routes’ for the same six schools. For intra-rater reliability, auditors conducted repeat measures on the ‘safe routes’ for four schools each (eight schools total), including three different schools each from the six used for inter-rater reliability and one additional school. Item-by-item reliability was assessed using Cohen’s Kappa, Cohen’s Weighted Kappa, and percentage agreement. The reliability of calculated domain scores was assessed using intraclass correlation coefficients. Results For inter-rater reliability, 31 of the 45 (68.9%) items had moderate to almost perfect agreement, seven items (15.6%) had below moderate agreement, and a Kappa statistic could not be calculated for seven items (15.6%) due to constant values. For intra-rater reliability, 37 of the 45 (82.2%) items had moderate to almost perfect agreement, two items (4.4%) had below moderate agreement, and a Kappa statistic could not be calculated for six items (13.3%) due to constant values. For inter- and intra-rater reliability of the segment domain scores, three of the four domains had substantial to almost perfect agreement. For inter- and intra-rater reliability of the crossing domain scores, all four domains had moderate to almost perfect agreement. For inter- and intra-rater reliability of the segment, crossing, route, and school scores, all had substantial to perfect agreement. Conclusions The FACTS tool can reliably characterise the microscale built environment of promoted ‘safe routes’ for their use within SR2S programs, and should be considered for use in future SR2S programs within the suburban Australian context.

Abstract Cyclization provides a general strategy for improving the proteolytic stability, cell membrane permeability and target binding affinity of peptides. Insertion of a stable, non-reducible linker into a disulphide bond is a commonly used approach for cyclizing phage-displayed peptides. However, among the vast collection of cysteine reactive linkers available, few provide the selectivity required to target specific cysteine residues within the peptide in the phage display system, whilst sparing those on the phage capsid. Here, we report the development of a cyclopropenone-based proximity-driven chemical linker that can efficiently cyclize synthetic peptides and peptides fused to a phage-coat protein, and cyclize phage-displayed peptides in a site-specific manner, with no disruption to phage infectivity. Our cyclization strategy enables the construction of stable, highly diverse phage display libraries. These libraries can be used for the selection of high-affinity cyclic peptide binders, as exemplified through model selections on streptavidin and the therapeutic target αvβ3.

Abstract Background Key functions of Ca2+ signaling in rodent microglia include monitoring the brain state as well as the surrounding neuronal activity and sensing the danger or damage in their vicinity. Microglial Ca2+ dyshomeostasis is a disease hallmark in many mouse models of neurological disorders but the Ca2+ signal properties of human microglia remain unknown. Methods We developed a novel genetically-encoded ratiometric Ca2+ indicator, targeting microglial cells in the freshly resected human tissue, organotypically cultured tissue slices and analyzed in situ ongoing Ca2+ signaling of decades-old microglia dwelling in their native microenvironment. Results The data revealed marked compartmentalization of Ca2+ signals, with signal properties differing across the compartments and resident morphotypes. The basal Ca2+ levels were low in ramified and high in ameboid microglia. The fraction of cells with ongoing Ca2+ signaling, the fraction and the amplitude of process Ca2+ signals and the duration of somatic Ca2+ signals decreased when moving from ramified via hypertrophic to ameboid microglia. In contrast, the size of active compartments, the fraction and amplitude of somatic Ca2+ signals and the duration of process Ca2+ signals increased along this pathway.

Abstract Research demonstrates the important role of genetic factors in attention-deficit/hyperactivity disorder (ADHD). DNA sequencing of families provides a powerful approach for identifying de novo (spontaneous) variants, leading to the discovery of hundreds of clinically informative risk genes for other childhood neurodevelopmental disorders. This approach has yet to be extensively leveraged in ADHD. We conduct whole-exome DNA sequencing in 152 families, comprising a child with ADHD and both biological parents, and demonstrate a significant enrichment of rare and ultra-rare de novo gene-damaging mutations in ADHD cases compared to unaffected controls. Combining these results with a large independent case-control DNA sequencing cohort (3206 ADHD cases and 5002 controls), we identify lysine demethylase 5B (KDM5B) as a high-confidence risk gene for ADHD and estimate that 1057 genes contribute to ADHD risk. Using our list of genes harboring ultra-rare de novo damaging variants, we show that these genes overlap with previously reported risk genes for other neuropsychiatric conditions and are enriched in several canonical biological pathways, suggesting early neurodevelopmental underpinnings of ADHD. This work provides insight into the biology of ADHD and demonstrates the discovery potential of DNA sequencing in larger parent-child trio cohorts.

Introduction: Thyroid storm is an uncommon but life-threatening presentation of thyrotoxicosis with a mortality rate of 10%. Our objective was to study the demographics, clinical and biochemical characteristics, and outcomes of inpatients diagnosed with thyroid storm in the Indian context. Methods: This retrospective study was conducted by analysing the institutional electronic medical records (EMR) of all patients admitted with thyroid storm from 2004 to 2020 with a Burch–Wartofsky score (BWS) of ≥45. Results: Thirty-five patients with a BWS ≥45 were included, of whom 71.4% were women, with a mean age of 44.9 ± 10.2 years. 43% did not have any prior history of thyrotoxicosis. Graves’ disease was the most common underlying aetiology (71.4%), followed by toxic multinodular goitre (14.3%). Cardiovascular (94.3%) and gastrointestinal-hepatic dysfunction (88.6%) were the most common clinical manifestations. Features of Central nervous system (CNS) dysfunction were seen in only 42.3% of patients diagnosed with a thyroid storm. The Japanese Thyroid Association (JTA) criteria diagnosed only 26 patients (74.3%) with “definite” thyroid storm. The mortality rate was 8.6%, and all three patients expired within 48 hours of admission. Conclusion: Nearly one in every two patients with thyroid storm had previously undiagnosed thyrotoxicosis. Toxic multinodular goitre is a notable aetiology in Indians. Features of CNS dysfunction, considered relatively specific for thyroid storm, were less prominent in our series. The JTA criteria might alter the classification of some patients diagnosed with a thyroid storm, when compared to the BWS score due to fewer CNS features among Indian patients.

Abstract Nest‐site fidelity is a common strategy in birds and is believed to be adaptive due to familiarity with local conditions. Returning to previously successful nest sites (i.e., the win‐stay lose‐switch strategy) may be beneficial when habitat quality is spatially variable and temporally predictable; however, changes in environmental conditions may constrain dispersal decisions despite previous reproductive success. We used long‐term (2000–2017) capture‐mark‐reencounter data and hierarchical models to examine fine‐scale nest‐site fidelity of emperor geese (Anser canagicus) on the Yukon–Kuskokwim Delta in Alaska. Our objectives were to quantify nest‐site dispersal distances, determine whether dispersal distance is affected by previous nest fate, spring timing, or major flooding events on the study area, and determine if nest‐site fidelity is adaptive in that it leads to higher nest survival. Consistent with the win‐stay lose‐switch strategy, expected dispersal distance for individuals that failed their nesting attempt in the previous year was greater (207.7 m, 95% HPDI: 151.1–272.7) than expected dispersal distance for individuals that nested successfully in the previous year (125.5 m, 95% HPDI: 107.1–144.9). Expected dispersal distance was slightly greater following years of major flooding events for individuals that nested successfully, although this pattern was not observed for individuals that failed their nesting attempt. We did not find evidence that expected dispersal distance was influenced by spring timing. Importantly, dispersal distance was positively related to daily survival probability of emperor goose nests for individuals that failed their previous nesting attempt, suggesting an adaptive benefit to the win‐stay lose‐switch strategy. Our results highlight the importance of previous experience and environmental variation for informing dispersal decisions of a long‐lived goose species. However, it is unclear if dispersal decisions based on previous experience will continue to be adaptive as variability in environmental conditions increases in northern breeding areas.

Abstract Estimating correlations among demographic parameters is an important method in population ecology. A recent paper by Deane et al. (Ecology and Evolution 13:e9847, 2023) attempted to explore the effects of different priors for covariance matrices on inference when using mark‐recovery data. Unfortunately, Deane et al. (2023) made a mistake when parameterizing some of their models. Rather than exploring the effects of different priors, they examined the effects of the use of incorrect equations on inference. In this manuscript, we clearly describe the mistake in Deane et al. (2023). We then demonstrate the use of an alternative and appropriate method and reach different conclusions regarding the effects of priors on inference. Consistent with other recent literature, informative inverse Wishart priors can lead to flawed inference, while vague priors on covariance matrix components have little impact when sample sizes are adequate.

Investigating among-individual differences in reproductive success and survival is essential for understanding eco-evolutionary processes. We used 5 years of demographic data from 556 breeding barn owls (Tyto alba) to estimate associations between intrinsic and extrinsic covariates on survival and reproduction throughout the annual cycle. As males and females have distinct roles in reproduction, environmental conditions and individual quality may be differentially linked to their fitness at different time points. Males breeding early and inhabiting prey-rich areas experienced higher reproductive success but faced greater reproductive costs. Indeed, the number of offspring a male cared for was negatively associated with his body condition and survival. However, our results indicate that these influences can be mitigated in males experiencing favourable post-breeding environmental conditions. For female owls, early breeding and high food availability during the breeding period were linked with increased reproductive success. Prey availability during incubation and higher reproductive output were associated with higher survival into the next breeding period in females. Unlike males, females did not exhibit obvious trade-offs between reproductive success and survival. Our research demonstrates trade-offs between fecundity and survival, and that females paired with males able to provide sufficient food experience higher survival and reproduction.

* Context.--The pathologic diagnosis of usual interstitial pneumonia (UIP) remains a challenging area, and application of histologic UIP guidelines has proved difficult. Objective.--To understand current practice approaches by pulmonary pathologists for the histologic diagnosis of UIP and other fibrotic interstitial lung diseases (ILDs). Design.--The Pulmonary Pathology Society (PPS) ILD Working Group developed and sent a 5-part survey on fibrotic ILD electronically to the PPS membership. Results.--One hundred sixty-one completed surveys were analyzed. Of the respondents, 89% reported using published histologic features in clinical guidelines for idiopathic pulmonary fibrosis (IPF) in their pathologic diagnosis; however, there was variability in reporting terminology, quantity and quality of histologic features, and the use of guideline categorization. Respondents were very likely to have access to pulmonary pathology colleagues (79%), pulmonologists (98%), and radiologists (94%) to discuss cases. Half of respondents reported they may alter their pathologic diagnosis based on additional clinical and radiologic history if it is pertinent. Airway-centered fibrosis, granulomas, and types of inflammatory infiltrates were considered important, but there was poor agreement on how these features are defined. Conclusions.--There is significant consensus among the PPS membership on the importance of histologic guidelines/features of UIP. There are unmet needs for (1) consensus and standardization of diagnostic terminology and incorporation of recommended histopathologic categories from the clinical IPF guidelines into pathology reports, (2) agreement on how to incorporate into the report relevant clinical and radiographic information, and (3) defining the quantity and quality of features needed to suggest alternative diagnoses. (Arch Pathol Lab Med. 2024;148:168-177; doi: 10.5858/arpa.2022-0530-OA), The histologic features of usual interstitial pneumonia (UIP), the fibrotic interstitial lung disease (ILD) pattern seen in, but not limited to, clinical idiopathic pulmonary fibrosis (IPF), was first described by [...]

Cardiac complications are frequently seen in patients with anorexia nervosa, with bradycardia being the most common. The degree of bradycardia can be a marker of disease severity as heart rate (HR) nadir is correlated with body mass index, total weight loss, and recent weight loss. Despite the prevalence and potential severity of bradycardia, there is limited literature evaluating the optimal management of severe bradycardia. This case report discusses a patient with a HR lower than previously reported in the literature and provides a valuable opportunity to examine the management of cardiac output in cases of severe bradycardia.

Abstract Background Rosa species are rich sources of polyphenols with physiological functions. In this study a polyphenol-rich Rosa multiflora (var. platyphylala) petal extract (RoseFit™) was investigated for weight loss in humans. Methods In a randomized, placebo-controlled, parallel-group, double-blind clinical trial seventy overweight male and female subjects (20–50 years) with body mass index (BMI) 25–30 kg/m2 were randomly allocated to the active treatment group (RoseFit) and placebo group in a 1:1 ratio. The subjects received 300 mg capsules twice daily for 12 weeks. The primary efficacy outcome measures included body weight, BMI, and body composition, as determined using Dual-energy X-ray absorptiometry (DEXA). Secondary measures consisted of serum lipid profile and appetite marker (leptin and ghrelin) analyses. Safety analyses included biochemical and hematological assessments. Results At the end of the study, a marked reduction in body weight (-1.20 ± 2.62 kg, p

Abstract Background While the impacts of atopic dermatitis (AD) on maternal and child sleep outcomes have been previously explored, less is known about the associations between infantile AD and sleep quality and quantity. Objective To describe the perceived causes of AD-associated maternal sleep disturbances and the association between AD severity and infant sleep outcomes. Methods Mothers with infants aged

ABSTRACTThere has been growing interest in the role of B cells in antitumour immunity and potential use in adoptive cellular therapies. To date, the success of such therapies is limited. The intrinsic capacity of B cells to specifically activate tumour-specific CD4+ T cells in vivo via TCR-dependent interactions remains poorly defined. We have developed an in vivo tumour model that utilizes MHCII I-E restriction which limits antigen presentation to tumour-specific CD4 T cells to either tumour-specific B cells or host myeloid antigen presenting cells (APCs) in lymphopenic RAG-/-mice. We have previously shown that these naive tumour-specific CD4+ T cells can successfully eradicate established tumours in this model when activated by host APCs. When naïve tumour-specific B cells are the only source of I-E+ APC, very limited proliferation of naïve CD4+ T cells is observed, whereas host I-E+ APCs are potent T cell activators. B cells pre-activated with an anti-CD40 agonistic antibody in vivo support increased T cell proliferation, although far less than host APCs. CD4+ T cells that have already differentiated to an effector/central memory phenotype proliferate more readily in response to naïve B cells, although still 100-fold less than in response to host APCs. This study demonstrates that even in a significantly lymphopenic environment, myeloid APCs are the dominant primary activators of tumour-specific T cells, in contrast to the very limited capacity of tumour-specific B cells. This suggests that future anti-tumour therapies that incorporate activated B cells should also include mechanisms that activate host APCs.

Metabolomics has a long history of using cosine similarity to match experimental tandem mass spectra to databases for compound identification. Here we introduce the Blur-and-Link (BLINK) approach for scoring cosine similarity. By bypassing fragment alignment and simultaneously scoring all pairs of spectra using sparse matrix operations, BLINK is over 3000 times faster than MatchMS, a widely used loop-based alignment and scoring implementation. Using a similarity cutoff of 0.7, BLINK and MatchMS had practically equivalent identification agreement, and greater than 99% of their scores and matching ion counts were identical. This performance improvement can enable calculations to be performed that would typically be limited by time and available computational resources.