Cause for optimism in mild hypoxic ischaemic hypoxic encephalopathyWilliam M. Curtin, MD, corresponding authorDivision of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology & Pathology and Laboratory Medicine, Penn State College of Medicine, Penn State Health, Milton S. Hershey Medical Center, Hershey, PA, [email protected] code H103500 University Dr, PO Box 850Hershey, PA 17033-0850Maternal-Fetal Medicine, Rm C3620Phone 717-531-8142/Option #5Fax 717-531-0947Acknowledgements: noneDisclosure of Interests: The author has no conflicts nor competing interests to disclose, financial or otherwise, in connection with this manuscript.Contribution to Authorship: Dr. Curtin alone completed review of the Törn et al. final manuscript entitled “Outcomes in children after mild neonatal hypoxic ischaemic encephalopathy: A population-based cohort study” The writings and opinions expressed in his mini commentary are solely the work of Dr. Curtin.Details of Ethics Approval: not applicable at our institution’s IRB as a commentary is not considered research and does not involve human subjects.Funding: Dr. Curtin neither received nor utilized any funds in writing this mini commentary.The objective of the study by Törn et al. was to determine if mild hypoxic ischaemic encephalopathy (HIE) was associated with severe neurological outcomes utilizing a population-based approach facilitated by five linked Swedish national databases. The rationale given was that while moderate to severe HIE is known to be associated with significant neurological morbidity, long-term disability, and mortality in children, less is known about mild HIE. The authors note that half of the cases of HIE are mild and they cite a systematic review of 250 infants (Conway JM, et al. Early human development . 2018; 120:80-7) showing a 22% prevalence of abnormal neurological outcomes in this disorder. There is therefore potential for significant burden of disease in mild HIE. Törn et al. chose a primary composite outcome that included cerebral palsy, epilepsy, mental retardation and death in children with mild HIE and non HIE cohorts followed up to 6 years of age. With a median follow-up of 3.3 years of age, 17 of 414 (4.1%) and 4786 of 504,661 (0.95%), in the mild HIE and non HIE cohorts respectively, had the composite outcome with an adjusted hazard ratio of 3.85 (95% CI: 2.27-6.50)In 1976 Sarnat and Sarnat reported clinical and EEG features of 21 neonates at term who experienced ischaemic-anoxic encephalopathy (Sarnat HB et al. Arch Neurol. 1976; 33:696–705). This temporal classification divided the infants into three progressively deteriorating stages. Fast forward to the current millennium and Sarnat’s original classification is used to differentiate between infants with mild and moderate/severe HIE, the latter two categories benefiting from therapeutic hypothermia (Jacobs SE et al. Cochrane Database Syst Rev. 2013, Issue 1. Art. No.: CD003311). Seven infants with moderate or severe HIE is the number needed to treat (NNT) to prevent one adverse neurological outcome.Therapeutic hypothermia is not standard of care in mild HIE; however, in a survey of neonatal clinicians from 35 countries the vast majority would support a large randomized controlled trial to examine neurodevelopmental outcomes (Singla M, et al. Neonatology. 2022; 119:712-718). The results from this methodical Swedish cohort study provide data that can be used to direct further research. The composite outcomes in HIE are lower than expected, and one could infer similar outcomes in high resource settings. The data are reassuring and will be useful for clinicians in counseling and reassuring parents with infants affected by mild HIE. Regarding a randomized controlled trial of therapeutic hypothermia in mild HIE: it would appear, based on the data provided by Törn et al., if we hypothesize this therapy would result in a 50% reduction in the composite outcome, the NNT would be approximately 50. This compares unfavorably to the NNT of 7 in moderate/severe HIE. Perhaps, further insight could be gained by review of individual patient data, particularly with respect to antenatal and neonatal course, imaging, EEG, and biochemical data in order to identify a subset that might benefit from therapeutic hypothermia or other novel therapy.Word count: 490

A recurring question is whether rating scales should be considered metrically scaled or merely ordinally scaled. This has direct implications for the permissible statistical procedures for significance testing. Based on the results of a simulation study, it is shown that the use of parametric procedures for rating scales has distinct advantages over the non-parametric alternatives. It is also shown that the parametric procedures are robust to violations of the assumption of normality, which only result in a modest loss of power compared with continuous variables. This loss should be taken into account when calculating the optimal sample size. The results suggest that sample sizes about 25% larger should be chosen for discrete rating scales than for continuous variables.

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Characteristics and treatments of patients with significantly elevated creatine kinase levels induced by seizures: case report and literature reviewKai Wang, Jinwei Yang, Wenhao Xu, Lei Wang, Yu Wang*The First Affiliated Hospital of Anhui Medical UniversityCorrespondence: Yu WangDepartment of Neurology, The First Affiliated Hospital of Anhui Medical UniversityJixi Road 218, Shushan District, Hefei, Anhui Province 230032,People’s Republic of China. Email [email protected]

Clival Ectopic Pituitary Prolactinoma was Successfully Managed by Transsphenoidal Surgery: A Rare Case Report

Chlamydomonas reinhardtii has been successfully engineered to produce compounds of interest following transgene integration and heterologous protein expression. The advantages of this model include the availability of validated tools for bioengineering, its photosynthetic ability and its potential use as biofuel. Despite this, breakthroughs have been hindered by its ability to silence transgene expression through epigenetic changes. Histone deacetylases (HDAC) are main players in gene expression. We hypothesized that transgene silencing can be reverted with chemical treatments using HDAC inhibitors. To analyze this, we transformed C. reinhardtii, integrating into its genome the mVenus reporter gene under the HSP70-rbcs2 promoter. From 384 transformed clones, 88 (22.9 %) displayed mVenus positive (mVenus+) cells upon flow-cytometry analysis. Five clones with different fluorescence intensities were selected. The number of integrated copies was measured by qPCR. Transgene expression levels were followed over the growth cycle and upon SAHA treatment, using a microplate reader, flow cytometry, RT-qPCR, and western blot analysis. First, we observed that expression varies with the cell cycle, reaching a maximum level just before the stationary phase in all clones. Second, we uncovered that supplementation with HDAC inhibitors of the hydroxamate family, such as vorinostat (suberoylanilide-hydroxamic-acid, SAHA) at the initiation of culture increases the frequency (% of mVenus+ cells) and the level of transgene expression per cell over the whole growth cycle, through histone deacetylase inhibition. Thus, we propose a new tool to successfully trigger the expression of heterologous proteins in the green algae C. reinhardtii, overcoming its main handicap as an expression platform.

Pharmacoepidemiology and Drug Safety’s Core Concepts in Pharmacoepidemiology Section at One Year: Where Do We Go from Here? Jennifer L. Lund,1,2 Vincent Lo Re III3,41University of North Carolina at Chapel Hill, Department of Epidemiology, Gillings School of Global Public Health, Chapel Hill, NC, USA2Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA3Division of Infectious Diseases, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA4Division of Epidemiology, Center for Real-World Effectiveness and Safety of Therapeutics, Center for Clinical Epidemiology and Biostatistics, Department of Biostatistics, Epidemiology, and Informatics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USARunning title: Core Concepts in Pharmacoepidemiology – Year 2Conflicts of interest: The authors report no conflicts of interest.Acknowledgements: The authors appreciated the critical input on the Core Concepts in Pharmacoepidemiology section from Brian Strom, Olaf Klungel, BJ Park, and the members of the International Society for Pharmacoepidemiology’s Publications and Communications Committee, particularly Elena Rivero, Patricia Saddier, Joan Largent, and Blànaid Hicks.

Transcription therapy is an emerging approach that centers on identifying the factors associated with the malfunctioning gene transcription machinery that causes diseases and controlling them with designer agents. Until now, small molecule drugs targeting the epigenetic enzymes and critical signaling pathways have been the primary research focus in therapeutic gene modulation. However, nucleic acid-based small molecules have gained popularity in recent years as they could be pre-designed on demand to achieve operative control over the dynamic transcription machinery that governs how the immune system responds to diseases. Pyrrole-imidazole polyamides (PIPs) are well-established DNA-based small molecule gene regulators that overcome the limitations of their conventional counterparts owing to their sequence-targeted specificity, versatile regulatory efficiency and biocompatibility. Here, we emphasize the rational design of PIPs, their functional mechanism and their potential as targeted transcription therapeutics for diseases by regulating the immune response. Furthermore, we also discuss the challenges and foresight of this approach in personalized immunotherapy in precision medicine.

Moult Terminology: Envisioning an Evolutionary ApproachThe study of avian moult has been inhibited not only by its complexity but by convoluted and often conflicting terminologies that have combined to cloud the subject. Over time, two nomenclatures have emerged with differing bases of definition. The ’life-cycle’ system is used widely in European literature (e.g., Cramp 1988, Jenni & Winkler 2020) and defines moult terms based on timing relative to current-day life-history events, primarily breeding. Conversely, the Humphrey-Parkes or ’H-P’ system (Humphrey & Parkes 1959, as modified by Howell et al. 2003), is used more widely in the New World and Australia (e.g., Higgins & Davies 1996; Pyle 2008, 2022a; Howell 2010), and defines terms based on the principle of how moults have evolved along avian lineages (Pyle 2022b).Recently, Kiat (2022) proposed that moult terminology be simplified, and advocated use of the life-cycle approach as a more understandable system to those not familiar with moult terminology. We appreciate and endorse Kiat’s (2022) plea for a simpler moult terminology, and we empathize with his frustration, but we disagree with his primary conclusion. We argue that if one intends moult to be a subject for study, then a standardized terminology based on the best scientific evidence is preferable to a system that, while perhaps more widely understood among ornithologists and the general public for passerines in boreal regions, is often imprecise or ambiguous and is difficult to apply to the majority of avian taxa on a global basis (Higgins & Davies 1996, Howell et al. 2004, Johnson and Wolfe 2018, Kiat 2022, Pyle 2022b).The debate over the best moult and plumage terminology to use has persisted since the H-P system was proposed by Humphrey and Parkes (1959) over 60 years ago (e.g., Howell et al. 2003, 2004; Jenni and Winkler 2004, 2020; Kiat 2022; Pyle 2022b; and references therein). At this point, certain opinions on which system should be favoured are calcified, and we do not wish to belabour the details yet again. In our experience, however, newer students grasp the H-P system quickly, whereas older ornithologists—including ourselves—who first learned life-cycle moult terminology, often have initial difficulty envisioning the H-P system’s evolutionary approach. Our goal with this perspective is to propose and illustrate how moults may have evolved in birds, with the hope that it will assist future ornithologists to envision and appreciate the H-P system.Adhering to the evolutionary H-P approach, we propose considering the prebasic moult, and perhaps the preformative moult, as ancestral to all modern bird lineages, having evolved from reptiles (Howell & Pyle 2015, Pyle 2022b; see also Kiat et al. 2020, 2021; Figs. 1, S1). The complete or near-complete prebasic moult occurs in all current-day avian taxa and, rather than simply a replacement of feathers, appears to be part of an endogenous restoration of body tissues (Voitkevich 1966, Murphy 1996, Kuenzel 2003) that may be ancestral to all vertebrates (King 1972). Considering the prebasic moult as homologous among modern birds is thus a parsimonious hypothesis, providing a robust nomenclatural framework that can be applied to all bird moults and plumages. The preformative moult also appears to be present in most if not all modern bird lineages, and may have evolved in reptiles as body size developed quickly in the first year of life; if not, it evolved early in avian evolution as it is found in most or all basal lineages (Figs. 1, S1). Assuming homology among all modern bird taxa, the prebasic and preformative moults can provide the framework for defining all subsequently evolved moults within the H-P system (Figs. 1, S1). Additional inserted moults, including prealternate and presupplemental moults, can evolve (both appearing and disappearing) along bird lineages such that, unlike prebasic and preformative moults, they should not be considered homologous across all birds (Howell et al. 2003, Johnson and Wolfe 2018, Pyle 2022b).Once an evolutionary basis is appreciated, moult strategies become substantially easier to compare across all species and latitudes. For example, the four underlying strategies identified by Howell et al. (2003), which are defined by the number of moults that occur within the first and later moult cycles, can be provisionally placed in an evolutionary context (Fig. 1; see Supporting information file and Fig. S1 for details). Although it is quite possible that inserted moults may have disappeared without trace along some bird lineages, envisioning how these four strategies may have evolved for current taxa has the potential to help inform a greater appreciation for the adaptive causes of inserted moults (Figure S1).We contend that the evolutionary (H-P) system is more applicable on a global basis for studying avian moult than is the life-cycle system, and we thus encourage those who use the life-cycle system to also attempt visualizing an evolutionary approach to moult terminology, rather than trying to simply synonymize H-P terms with life-cycle terms. We suggest first determining the prebasic moult cycle, then whether or not inserted moults occur in the first and/or later cycles, and using an evolutionary approach to infer the correct designation of each inserted moult. Once envisioned, learned, and appreciated, the evolutionary approach proposed by Humphrey & Parkes (1959), as modified by Howell et al. (2003), is scientifically more precise, allows the recognition of all inserted moults, and is easier to apply consistently, to all taxa and by all parties interested in the study of moult in birds. We provide more detail on taking an evolutionary approach in the Supporting information file.

The main differentials in cases of sudden elevation of hepatic enzyme levels during immunochemotherapy are reactivation of hepatitis B virus or drug-induced liver injury. Here, we report a case of acute liver injury caused by hepatitis E virus (HEV) during chemotherapy for malignant lymphoma, wherein the patient was successfully treated and completed chemotherapy. A 57-year-old woman visited her local doctor because she felt light and tired. The patient underwent lower gastrointestinal endoscopy and was diagnosed with a malignant lymphoma of the small intestine (diffuse large B-cell lymphoma). The patient had a history of oral consumption of undercooked pork liver to improve anemia and was diagnosed with acute hepatitis E. Since the patient responded to chemotherapy, she was treated with single-agent ribavirin while continuing chemotherapy, resulting in a sustained virological response. Even during treatment with immunosuppressive drugs, if appropriate treatment for hepatitis E can be administered, the patient can be fully treated without interruption. The patient was able to complete chemotherapy adequately without interruption of treatment, which was a clinically beneficial result.

Ticks pose a significant threat to horses, as they can cause harm and transmit diseases. This study aimed to assess the experiences, knowledge, and practices of equine veterinarians regarding ticks and tick-borne diseases (TBDs) in the United States. A survey was conducted, covering topics such as tick biology, identification, prevention, and veterinary education. The results revealed variations in veterinarians’ knowledge on ticks and TBDs, highlighting the need for improved tick control options, enhanced education, and comprehensive resources for tick and TBD prevention. The study emphasizes the importance of accurate tick identification for effective diagnosis and treatment of TBDs in horses. The limitations of the study include potential response bias and a limited sample size. To address these issues, collaborations between academia, industry, and the human medicine community are recommended to strengthen veterinarian training and education on ticks and TBDs. This research provides valuable insights for developing continuing education programs that focus on tick-borne illness prevention in horses.

Most proteins function by forming complexes within a dynamic interconnected network that underlies various biological mechanisms. To systematically investigate such interactomes, high-throughput techniques including CF-MS have been developed to capture, identify, and quantify protein-protein interactions (PPIs) in large-scale. Compared to other techniques, CF-MS allows the global identification and quantification of native protein complexes in one setting, without genetic manipulation and overexpression. Furthermore, quantitative CF-MS can potentially elucidate the distribution of a protein in multiple co-elution features, informing the stoichiometries and dynamics of a target protein complex. In this issue, Youssef et al. (Proteomics 2023, XX, XXXX-XXXX) combined multiplex CF-MS and an in-house algorithm to study the dynamics of the PPI network for Escherichia coli grown under ten different conditions. While the results demonstrated that while most proteins remained stable, the authors were able to detect disrupted interactions that were growth condition-specific. Further bioinformatics analyses also revealed biophysical properties and structural patterns that govern such a response.

Weed species are ecological models that recently received considerable attention due to their particular strategies linked to their ruderal-competitive traits. They are known to have the potential to provide additional floral resources for insects in flower-poor agroecosystems. However, their floral traits are much more scarcely studied than those of plants found in other habitats, such as grasslands. The aim of this study was to describe the floral phenotype of weeds and to determine to what extent their floral traits match their ecological strategies as described on the basis of leaf traits. We therefore cultivated 19 forb weeds from perennial agroecosystems, previously identified in Mediterranean fields, in a greenhouse for seven months and collected data on 12 floral and 5 leaf traits. We tested whether these traits covaried and whether they exhibited an ecological strategy at the phenotype scale. We found that in matters of flower production, weed species face a trade-off: either numerous small, low-stature flowers with small quantities of pollen and nectar, or few, large, higher-held flowers with more pollen and nectar. The floral traits were found to reflect Grime’s CSR strategies: the weed species producing fewer but costlier flowers belonged to C-strategy species, whereas those producing more but less costly flowers belonged to species dominated by an R strategy These findings indicate that the potential of weeds as floral resources for insects is related to their ecological strategies, which are known to be affected by agricultural practices that filter species composition. This implies that, as for the provision of other ecosystem services, weed communities can be managed so as to select species with interesting floral traits for pollinators.

Insect herbivory can vary from an inconsequential biotic interaction to a factor that contributes substantially to the diversity of plants and animals and overall interaction diversity. As herbivory is the result of numerous ecological and evolutionary processes, including complex population dynamics and the evolution of plant defense, it has been difficult to predict variation in herbivory across meaningful spatial scales. In the present work, we characterize patterns of herbivory on plants in a species rich and abundant tropical understory genus (Piper) across forests spanning 44° of latitude in the Neotropics. We modeled the effects of geography, climate, resource availability, and Piper species richness on the median, dispersion, and skew of generalist and specialist herbivory. By examining these multiple components of the distribution of herbivory, we were able to determine factors that increase biologically meaningful herbivory at the upper ends of the distribution. Site level variables such as latitude, seasonality, and maximum Piper richness explained variation in herbivory at the local scale (plot level) better for assemblages of Piper congeners than for a single species. Predictors that varied between local communities, such as resource availability and diversity, best explained the distribution of herbivory within sites, dampening broad patterns across latitude and climate and demonstrating why generalizations about gradients in herbivory have been elusive. The estimated population means, skew, and dispersion of herbivory responded differently to abiotic and biotic factors, illustrating the need for careful studies to explore distributions of herbivory and their effects on forest diversity. Nevertheless, we observed a roughly two-fold increase in median herbivory in humid compared to seasonal forests, and this finding aligns with the hypothesis that precipitation seasonality plays a critical role in shaping interaction diversity within tropical ecosystems.

Adeno-associated viruses (AAVs) have acquired a central role in modern medicine as delivery agents for gene therapies targeting rare diseases. While new AAVs with improved tissue targeting, potency, and safety are being introduced, their biomanufacturing technology is lagging. The AAV purification pipeline, in particular, hinges on protein ligands for the affinity-based capture step: while featuring excellent AAV binding capacity and selectivity, these ligands require strong acid (pH <3) elution conditions, which can compromise the product’s activity and stability; additionally, their high cost and limited lifetime has a significant impact on the price tag of AAV-based therapies. Seeking to introduce a more robust and affordable – yet equally effective – affinity technology, this study introduces a cohort of peptide ligands that (i) mimic the biorecognition activity of the AAV receptor (AAVR) and anti-AAV antibody A20, while (ii) enabling product elution under near-physiological conditions (pH 6.0) and (iii) granting extended reusability by withstanding multiple regenerations. A20-mimetic CYIHFSGYTNYNPSLKSC and AAVR-mimetic CVIDGSQSTDDDKIC demonstrated excellent capture of serotypes belonging to distinct clones/clades – AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9 – corroborating the in silico models documenting their ability to target regions of the viral capsid that are conserved across all serotypes. CVIDGSQSTDDDKIC-Toyopearl resin features binding capacity (~1014 vp per mL) and product yields (~60-80%) on par with commercial adsorbents, and purified AAV2 from HEK293 and Sf9 cell lysates affording high recovery (up to 78%) and reduction of host cell proteins (up to 700-fold), and high transduction activity (up to 65%) of the purified vectors.

Fibroblast growth factor 10 (FGF10) is a signaling molecule with a well-established role for lung branching morphogenesis. Rare heterozygous, deleterious variants in the FGF10 gene are known causes of the lacrimo-auriculo-dento-digital (LADD) syndrome as well as aplasia of lacrimal and salivary glands (ALSG). Previous studies indicate that pathogenic variants in FGF10 can cause lethal human developmental disorders of the lung, but reports on diffuse lung disease caused by pathogenic variants in the FGF10 gene are lacking. We describe four children with postnatal onset of severe diffuse lung disease and heterozygous variants in FGF10, each detected by whole exome or whole genome sequencing. All children presented with postnatal respiratory failure. Two children died within the first 2 days of life, one patient died at age of 12 years and one patient is alive at age of six years, but still symptomatic. One patient presented signs of severe dental caries suggestive for ALSG or LADD-syndrome. Histopathological analysis of lung biopies from the two children with early postpartum demise revealed diffuse developmental disorder representing acinar dysplasia. Sequential biopsies of the child with survival until the age of 12 years revealed alveolar simplification and progressive interstitial fibrosis. Our report extends the phenotype of FGF10-related disorders to diffuse developmental disorders of the lung and early onset lung fibrosis. Therefore, FGF10-related disorder should be considered even without previously described syndromic stigmata in children with postnatal respiratory distress, not only when leading to death in the neonatal period but also in case of persistent respiratory complaints.

13C-Breath testing is increasingly used in physiology and ecology research because of what it reveals about the different fuels that animals oxidize to meet their energetic demands. Here we review the practice of 13C-breath testing in humans and other animals and describe the impact that contamination that ambient/background CO 2 in the air can have on the accuracy of 13C-breath measurements. We briefly discuss physical methods to avoid sample contamination as well as the Keeling plot approach that researchers have been using for the past two decades to estimate δ 13C from breath samples mixed with ambient CO 2. Unfortunately, Keeling plots are not suited for 13C breath testing in common situations where 1) a subject’s VCO 2 is dynamic 2) ambient [CO 2] may change, 3) a subject is sensitive to hypercapnia, or 4) in any flow-through indirect calorimetry system. As such, we present a mathematical solution that addresses these issues by using information about the instantaneous [CO 2] and the δ 13CO 2 of ambient air as well as the diluted breath sample to back-calculate the δ 13CO 2 in the CO 2 exhaled by the animal. We validate this approach by titrating a sample of 13C-enriched gas into an air stream and demonstrate its ability to provide accurate values across a wide range of breath and air mixtures. Researchers can now instantaneously calculate the δ 13C of alveolar gas of humans or animals in real time without having to scrub ambient CO 2 or rely on estimated values.

Combining magnetic resonance imaging (MRI) data from multi-site studies is a popular approach for constructing larger datasets to greatly enhance the reliability and reproducibility of neuroscience research. However, the scanner/site variability is a significant confound that complicates the interpretation of the results, so effective and complete removal of the scanner/site variability is necessary to realize the full advantages of pooling multi-site datasets. Independent component analysis (ICA) and general linear model (GLM) based harmonization methods are the two primary methods used to eliminate scanner/site-related effects. Unfortunately, there are challenges with both ICA-based and GLM-based harmonization methods to remove site effects completely when the signals of interest and scanner/site-related variables are correlated, which may occur in neuroscience studies. In this study, we propose an effective and powerful harmonization strategy that implements dual-projection (DP) theory based on ICA to remove the scanner/site-related effects more completely. This method can separate the signal effects correlated with site variables from the identified site-related effects for removal without losing signals of interest. Both simulations and vivo structural MRI datasets, including a dataset from Autism Brain Imaging Data Exchange II and a traveling subject dataset from the Strategic Research Program for Brain Sciences, were used to test the performance of DP-based ICA harmonization method. Results show that DP-based ICA harmonization has superior performance for removing site effects and enhancing the sensitivity to detect signal of interest as compared with GLM-based and conventional ICA harmonization methods.