bluuebunny/crossref_metadata_2025 huggingface dataset:
Sympathetic Nerves Promote Hypertension‐Specific CD8+ Effector Memory T Cell Homing in the Bone Marrow of Mice with Angiotensin II Infusion
Authors: Xiao Liang , Itani Hana A , Foss Jason D , Harrison David G
Published: November 2023

We have recently identified a critical role of hypertension‐specific effector memory T lymphocytes (TEM cells) in the blood pressure elevation and renal dysfunction caused by repeated hypertensive stimuli. Formed during an initial immune challenge reside, a substantial portion of TEM cells reside in the bone marrow (BM) in a quiescent state for prolonged periods, and they can be reactivated upon re‐exposure to the hypertensive stimulus. Hypertension is associated with increased sympathetic outflow. We confirmed that expression of tyrosine hydroxylase, a marker for sympathetic innervation, was increased by 70% in the BM in mice received angiotensin (Ang) II infusion (490ng/kg/min) for two weeks. We therefore hypothesized that sympathetic nerves regulate accumulation and reactivation of hypertension‐specific TEM cells in BM. We performed unilateral superior cervical ganglionectomy (SCGx) in wild‐type C57BL/6 mice, causing selectively sympathectomy of the forelimb on the surgical side. After recovery from surgery, mice received Ang II infusion for two weeks. The number of TEM cells in the denervated BM following Ang II was slightly but significantly less than observed in the innervated limb as measured by flow cytometry. To determine T cells in the BM that were specific to hypertension, 5×106 BM cells were isolated from either the SCGx or control limbs, loaded with proliferation marker CFSE, and co‐cultured with 0.5×106 splenic dendritic cells isolated from another Ang II‐infused mouse. We found 30% less CD8+ T cell proliferation in the SCGx BM compared to control side (1.8±0.1 vs. 2.6±0.3×104), but no difference in CD4+ T cells. To further study the effect of sympathetic nerves on T cell homing in BM, 1×107 pan T cells were isolated from wild‐type mice (w/CD45.2 allele) after Ang II infusion, and adoptively transferred into CD45.1 mice that had previously received SCGx. Flow cytometry indicated that 7 days after transfer, 25% less CD8+ TEM cells from hypertensive donors homed to the SCGx BM than the innervated BM of recipients (20.8±2.5 vs. 27.8±2.6 per 103 total BM cells). Similar adoptive transfer experiments were performed between CD45.1 mice and β2 adrenoreceptor deficient mice. CD8+ TEM cell homing was dramatically attenuated by 70% if the recipient was deficient of β2 receptors. We conclude that sympathetic nerves promote TEM cell homing to the BM after an initial blood pressure elevation via β2 adrenergic signaling. This may contribute to the predisposition to hypertension and end‐organ damage for prolonged periods following an initial episode of hypertension.
Network Models: Connecting Anatomy to Systems Biology
Authors: Esteve‐Altava Borja
Published: November 2023

Systems biology studies systems composed of biological components, ranging from molecules to species, focusing on the complexity that emerges in living systems from the non‐linear interaction among parts. Quantitative tools to study the structure and dynamics of biological systems of interacting components include new experimental techniques, as well as theoretical approaches, such as the development and analysis of mathematical and computational models. Among these tools, network models stand out for their conceptual elegance, and mathematical power to highlight the organization of complex living systems as different as a brain and an ecosystem. Network models are currently used to study complex anatomical systems, offering a novel, transversal approach to the analysis of the evolution and development (and pathology) of form. An anatomical network model formalizes body parts, such as bones and muscles, and their interactions as the nodes and links of a network, a structural organization that is readily accessible through mathematical analysis using methods from the field of Network Theory. Here we will learn how to create and use anatomical network models to study the morphological organization of anatomical systems, with examples from the study of the morphological complexity and modularity of the skull, the key morphological re‐organizations that took place during the evolutionary transition from fins to limbs, and the development of human pathologies such as craniosynostosis.

Support or Funding Information

This project received funding from the European Union's Horizon 2020 research and innovation programme under Marie Sklodowska‐Curie grant agreement No 654155, and the American Association of Anatomists Innovations Program.
Specialized Ribosomes: A New Frontier in Gene Regulation, Organismal Biology, & Evolution
Authors: Barna Maria
Published: November 2023

The central dogma of molecular biology has for decades served as an explanation for the flow of genetic information within a biological system. In so far as the normal flow of biological information from mRNA to protein, the ribosome has been perceived to decode the genome with essentially machine‐like precision; serving as an integral but largely passive participant in the synthesis of all effector proteins across all kingdoms of life. Our research has changed this view, by demonstrating that not all of the millions of ribosomes within each cell are the same and that ribosome heterogeneity provides a novel means to diversify the proteins that can be produced in specific cells, tissues, and organisms. I will present our work centered on providing a roadmap for the absolute quantification of ribosome composition at a single cell level and during cellular differentiation that has identified subsets of ribosomes that are heterogeneous for core ribosomal proteins and interacting proteins. To further address the functional role of ribosome heterogeneity in translational control of the mammalian genome, we employed CRISPR/Cas9 to endogenously tag and purify heterogeneous ribosome populations to characterize specific preferences in transcripts translated by distinct ribosome subtypes. This led to the identification of subpools of transcripts, critical for key cellular processes including cell signaling, metabolism, growth, proliferation and survival, which are selectively translated by specific types of ribosomes. I will further present recent findings on the mechanisms by which ribosome‐mediated control of gene expression is encoded by structured RNA regulons within 5′UTRs. Together, these studies reveal a critical link between ribosome heterogeneity and specialized translational control of the mammalian genome, which adds an important layer of control to the post‐transcriptional circuitry of gene regulation and mammalian development.
QTL Analysis of a Trade‐off in Bone Length within the Mouse Zygomatic Arch
Authors: Percival Christopher J. , Green Rebecca , Gatti Daniel M. , Pomp Daniel , Roseman Charles C. , Marcucio Ralph , Hallgrímsson Benedikt
Published: November 2023

We use a powerful mouse resource to identify candidate genes associated with a negative correlation in the length of bones that contribute to the zygomatic arch. Because system‐wide growth factors contribute to all skeletal dimensions, most skull linear distances are positively correlated with each other. However, the lengths of two adjacent zygomatic arch bones are negatively correlated in our Diversity Outbred (DO) mice. This is evidence for an opposite pleiotropic effect or developmental constraint during zygomatic arch formation, which appears to limit variation in total zygomatic arch length. Determining the basis for this negative correlation will help us understand developmental mechanisms that limit phenotypic variation and underlie variation in how individual bones contribute to overall skull form. We completed high‐resolution genetic mapping with 563 DO mice to identify genomic regions and candidate genes associated with this negative correlation. DO mice are ideal for this, because each mouse carries a high degree of heterozygosity and a unique combination of alleles. We separately mapped 1) jugal bone length, 2) jugal process of maxilla length, and 3) total zygomatic arch length. Analysis was completed on raw linear distances without scaling or removing the allometric component of variation, because these procedures remove variation associated with overall size and artificially create negative associations between linear distance measurements. A genomic interval between 85 and 86 Mb on chromosome 17 was significantly associated with variation in jugal bone length and jugal process length, but not total zygomatic arch length. In addition, founder haplotypes under this peak displayed opposite phenotypic effects on individual bone lengths, suggesting it includes a genetic factor driving relative length of these zygomatic bones, but not total arch length. This interval includes Six2, Six3, and Six3os1, which are expressed in the pharyngeal arches or forebrain during facial development, as well as Camkmt, which is pivotal for calmodulin methylation. Ongoing real‐time PCR will show whether expression of these genes within the maxillary arch at E10.5 in three founder strains is correlated with measured founder haplotype effects. Understanding the genetic and developmental basis for this negative correlation is important because it results in a developmental constraint on total zygomatic arch length, which will act to limit evolvability of this structure.

Support or Funding Information

Funding graciously provided by the NIH (U01DE020054, 2R01DE019638, R01DE021708, R01GM070683), NSERC (238992‐12), and to CJP by the Alberta Innovates Health Solutions.
Movement Guided Learning as an Efficacious, Effective, and Evidence‐based Teaching Strategy Within the Undergraduate Anatomy Classroom
Authors: Bentley Danielle Christine
Published: November 2023

Within the anatomy classroom, physical movements and stretches can turn a student's own body into their personal educational tool. Such direct relevance can be useful for the undergraduate junior learner as they attempt to truly understand musculoskelatal anatomy in a meaningful way. This presentation describes the longitudinal, iterative scholarship that went in to developing, piloting, refining, and experimentally assessing Movement Guided Learning (MGL)©, a self‐directed learning workbook that guides students through applications of musculoskeletal anatomy including physical movements/stretches, surface palpations/visualizations, and extensions of knowledge with case‐based scenarios.

Most recently, components of the refined MGL workbook were fully integrated as adjunct teaching activities into a large undergraduate classroom within health sciences. Using a randomized control design, the entire curriculum of muscular anatomy was divided into three equal groups; students received MGL activities for 1/3rd of the taught muscles, students received multiple choice style review questions for 1/3rd of the taught muscles (as an active, time‐matched control), and students received no additional learning material for 1/3rd of taught muscles (as a non‐active control). To facilitate full integration of supplementary teaching materials, the course instructor was been made aware of group allocations, provided with all teaching materials, and embedded those materials during in‐class lecture time and during independent student activities. Final exam scores will be segmented and compared in agreement with the three experimental groups, allowing for an intra‐individual superiority assessment of MGL. Student survey responses will allow for MGL efficacy to be further assessed against student learning preferences as well as self‐reported strategy utilization. To control for inadvertent instructor‐bias, time spent on each muscle will be quantified (using video lecture capture software) and compared across the three groups

There are currently 187 undergraduate students enrolled in the course with the majority of them enrolled in a global health degree (~75%) in their first year of undergraduate studies (~90%). MGL efficacy will be determined using the aforementioned mixed‐methods assessment strategies following the fall semester final exam. Based on previously reported MGL success, it is hypothesized that the MGL activities will be a useful learning adjunct, especially for students who display kinesthetic learning preferences.

Support or Funding Information

none to declare
Transforming A Large Anatomy Class Into A Student Centered Learning Experience
Authors: Mackenzie Leslie , Michels Alison , Anstey Lauren
Published: November 2023

Anatomy education is challenged to develop contemporary pedagogies that move beyond memorization and factual recall to integration of information eliciting from students meaningful and deeper understandings. Inquiry‐based learning is one such strategy that involves students' active and increasingly independent investigation of questions and problems that are of interest to them. This learning strategy aims to encourage learners to draw upon the broader contexts for learning and emphasizes the development of skills that extend beyond the confines of the classroom. This study explores undergraduate students' perspectives and experiences of an inquiry project and its learning utility over a five year period. At the conclusion of each iteration qualitative analysis of assessment surveys were indicative that the inquiry‐based learning activity was useful for developing teamwork skills, communication skills and achieving personal goals, while the responses from facilitators of the project reflected on deep and active learning as being the primary benefits to the students. Although the perceived benefits of the activity differed between the two groups, the overall reaction was positive from all participants. Furthermore, from the data collected each year modifications were made to the inquiry‐based activity and these changes were tracked to inform trends over time. Positive perspectives consistently highlighted the value of student‐selected topics of interest, being creative with research and presentations, and working collaboratively with peers. Negatives were present every year but less consistent and reflective of the modifications of the activity. While a common theme has been the desire for more guidance with this activity, changes in negative feedback demonstrate how modifications are working to address students' learning needs. In addition, students and facilitators were observed during group work sessions and presentations, activity documents and students' work were analyzed, and interviews were conducted. Data analysis indicated that outcomes of the learning experiences reflect that of authentic learning. These observations offer a favourable view of an inquiry‐based learning activity in a large‐scale undergraduate anatomy course designed to engage students through self‐directed education, peer interaction, and collaboration. Although inquiry‐based learning continues to remain a positive experience for students, further quantitative studies are required to determine the benefit of this active learning strategy in terms of learning anatomy and applying anatomy‐specific knowledge.
Strategies to Help Students Tackle Complicated Anatomical Terminology
Authors: Carnegie Jacqueline
Published: November 2023

The study of anatomy involves the acquisition of a new language as students familiarize themselves with the names of the components of organs and organ systems. Many of these names derive from Latin and Greek, languages with which students have limited experience and they struggle with the pronunciation and spelling of each new term. Furthermore, students gain little experience in actively recalling, speaking and writing this terminology (they are provided with PowerPoint (PPT) lecture slides) and high enrolments (250–300 students per course) make weekly laboratory sessions impossible. Interactive learning strategies are needed to enrich their educational experience. Videos narrated by peers allow students to hear the names of bones and muscles while simultaneously using visual pathways to situate their study of musculoskeletal (MSK) anatomy within applied contexts such as a fractured wrist or the assumption of a yoga pose. Interactive PPT‐based Hangman games (PPTAlchemyR) require students to both recall functional anatomy content and to focus on the letters composing a muscle's name when assigning letters to spaces. Indeed, gamification is a new educational trend that promotes interactive learning by giving students low‐stakes opportunities to practice and to learn from their errors. A final approach, the development of content‐specific crossword puzzles using EclipseCrosswordR, also uses clues to stimulate recall and forces students to focus on word structure. But these puzzles have the additional advantages of accommodating words of unlimited length (word limit for PPT‐based Hangman is 15 letters), alerting students to an answer with the wrong number of letters, allowing students to identify spelling errors following puzzle submission, and motivating students to produce a finished product. Despite the fact that marks were not assigned to these interactive activities, tracking data shows that they were well used by students when preparing for summative evaluation. Students in two undergraduate courses in MSK anatomy (n = 233 and 235) presented with a series of four yoga videos over the three weeks leading up to summative exams watched each video an average of 1.92–2.70 times with most students being sufficiently engaged to view the entire series at least once. Preliminary data tracking student use of crossword puzzles in a first‐year anatomy and physiology course (n = 296) revealed a high level of interest when each puzzle was released (immediately accessed by over 25% of students) and that many students (at least 25–30%) returned to try the puzzles more than once. These studies involving students of anatomy show that educational strategies that combine audio with visual and/or use interactive word‐based games can enrich learning and are welcomed by students tackling a new language.

Support or Funding Information

University of Ottawa Undergraduate Research Opportunity Program
Expansion of the Functional Border of the Audiovisual Cortex Following Partial Hearing Loss in Adulthood
Authors: Allman Brian
Published: November 2023

Following damage to a major sense (e.g., hearing loss), crossmodal plasticity occurs whereby there is an increased responsiveness in the deprived cortex to the remaining, intact senses. Using a rat model and single‐unit electrophysiological recordings, our recent studies on hearing‐impaired animals have revealed that crossmodal plasticity is not restricted to the core auditory cortex; higher‐order auditory regions as well as visual and audiovisual cortices show differential effects following noise‐induced hearing loss. To our surprise, the cortical area showing the greatest relative degree of multisensory convergence transitioned away from the normal audiovisual area toward a neighboring, predominantly auditory area. Thus, our collective results suggest that crossmodal plasticity induced by adult‐onset hearing impairment manifests in higher‐order cortical areas as an expansion in the functional border of the audiovisual cortex. Our ongoing studies seek to determine the implications of this crossmodal plasticity on the rats' ability to perceive the precise timing of audiovisual stimuli using novel behavioral tasks that are consistent with studies of perceptual judgement in humans.

Support or Funding Information

Support provided by an NSERC Discovery Grant
Changes in Oligodendroglial and Microglial Cell Populations in the Embryonic Rat Spinal Cord Following Maternal Immune Activation
Authors: McDermott Kieran W , Anderson Rebecca C , Foley Tara , Radford Jay , O'Halloran Sophie , O'Keeffe Gerard W
Published: November 2023

Epidemiological studies suggest that maternal immune activation (MIA) may contribute to the development of neurological disorders such as cerebral palsy, schizophrenia, autism, epilepsy and potentially, neurodegenerative diseases. These studies, and those in animal models, have suggested that the effects of MIA on the developing nervous system, and thus the longer term consequences for the offspring, may be spatially and temporally regulated. However, much work remains in order to elucidate fully the CNS regions, cell populations and timing of gestational vulnerability.

The in vivo effects of MIA on cellular development in the spinal cord (SC) at embryonic day (E)12, 14 and 16 were investigated in this study. Dams received a single intraperitoneal injection of 50μg/kg lipopolysaccharide (LPS), 100 μg/kg LPS, or saline. Embryos were sacrificed either 5 hours post‐injection or, following their delivery, two weeks postnatally(P). Embryos and pups were fixed and frozen prior to cryosectioning at 15μm. Rostral, intermediate and caudal SC sections were immunofluorescently stained with Olig‐2 (an oligodendrocyte marker), Iba‐1 (a microglial marker), nestin (a radial glial marker) and reelin antisera.

Olig2‐positive cells were counted in white matter (WM) and grey matter (GM) regions of the SC. Olig2‐positive cell number in E16 embryos decreased 5 hours following treatment with 100μg/kg LPS but not with treatment with 50μg/kg LPS. At E14 Olig2‐positive cells showed no change at 5 hours in response to 50μg/kg LPS treatment.

Iba‐1+ cells decreased in GM and WM of the E16 SC 5Hr post treatment with 100μg/kg LPS.

Reelin fluorescence intensity is decreased in the GM of the E16 SC 5 hours after 100μg/kg LPS treatment. Reelin fluorescence intensity is unaltered in E16 spinal cord in response to 50μg/kg LPS. Reelin appears to co‐localise with a specific population of neurons at E16 and P14.

In conclusion, MIA appears to have an acute effect on the development of key cell populations at specific time points. Ongoing work is extending these analyses and will seek to identify how such perturbations affect ongoing SC development into the early postnatal period.

Support or Funding Information

Anatomical Society

Graduate Entry Medical School University of Limerick
Orphan Nuclear Receptor, GCNF, Is Required for Early Neural Crest Cell Induction and Survival
Authors: Munoz William A , Bhatt Shachi , Crane‐Dennis Jennifer , Achilleos Annita , Sakai Daisuke , Cooney Austin , Trainor Paul
Published: November 2023

Neural crest cells (NCC) are considered to be a vertebrate innovation that significantly contributed to the evolution, predation, radiation and adaptation of vertebrates to most niches of the planet. NCC comprise a unique vertebrate cell population that is frequently termed the “fourth germ layer” because they form in conjunction with the other germ layers and give rise to a diverse array of cell types and tissues including most of the craniofacial skeleton, peripheral nervous system, and pigment cells amongst many others. NCC development is dependent on gene regulatory network (GRN) control of several cellular mechanisms including induction, migration and differentiation. Defects in these processes result in clinical manifestations termed neurocristopathies.

In contrast to aquatic and avian species, we currently have a very poor understanding of the factors that regulate mammalian neural crest cell induction and specification. Here we describe molecular analyses in mouse models that reveal a critical role for the orphan nuclear receptor, Germ cell nuclear factor (Gcnf/Nr6a1) in mammalian neural crest cell formation and survival. Comparison of null and conditional Gcnf mutant embryos indicates that Gcnf is required prior to E8.0 for proper neural crest cell specification and induction. Furthermore, Gcnf functions as a bimodal switch to repress neural stem cell fate and promote the acquisition of neural crest cell identity. Thus our findings have identified a novel regulator of mammalian neural crest cell development and defined a temporal window for mammalian neural crest cell formation which is earlier than previously thought and raises important questions regarding the appropriateness of Wnt1Cre in studies of mammalian neural crest cell specification and induction.

Support or Funding Information

This work in the Trainor lab is supported by the Stowers Institute for Medical Research, the March of Dimes (#6‐FY08‐265) and the National Institute of Health ‐ National Institute for Dental and Craniofacial Research (DE016082). WIM is further supported by the American Association of Anatomist Postdoctoral Fellowship.
LHX2 is a necessary intermediate in FGF‐induced SHH expression during limb development
Authors: Watson Billy A. , Pira Charmaine U. , Oberg Kerby C.
Published: November 2023

During limb development, Fibroblast growth factors (FGFs) from the apical ectodermal ridge (AER) promote limb outgrowth and proximal‐distal patterning. The zone of polarizing activity, a cluster of mesodermal cells in the distal posterior limb bud, secretes Sonic hedgehog (SHH) which promotes anterior‐posterior outgrowth and patterning. Coordination of patterned development between these axes is facilitated by a reciprocal feedback loop and is necessary for normal limb formation as well as redevelopment/regeneration. The molecular mechanisms governing this feedback loop, however, are poorly understood, particularly the intermediates downstream of FGF that induce/maintain SHH expression. In order to elucidate potential target genes, we performed a DNA microarray analysis in SHH‐competent chicken limb bud cells following FGF treatment. The analysis identified a candidate intermediate molecule, the Lim homeobox 2 (LHX2) transcription factor, that displayed a robust response to FGF (>5 fold upregulation). Via comparative genomics, we found LHX2 to be highly conserved across vertebrate species. Additionally, the endogenous expression of LHX2 is in an FGF‐responsive domain which overlaps that of SHH. Thus, we hypothesize that LHX2 is an intermediate molecule downstream of FGF in the regulation of SHH expression.

To confirm FGF induction of LHX2, we performed in situ hybridization on chicken embryos implanted with FGF‐soaked beads. LHX2 is upregulated within 3 hours of bead implantation. Using cyclohexamide to globally block protein translation, we observed that LHX2's upregulation via ectopic FGF persisted. We therefore identified LHX2 as a primary response target of FGF signaling. Studies are underway to determine which of the canonical signaling pathways (MAPK/ERK1/2 or AKT) are involved in LHX2 induction.

In an effort to assess LHX2's ability to regulate SHH expression in the context of FGF signaling, we knocked‐down LHX2 protein around an FGF bead which lead to a decrease in SHH expression around the bead. We also overexpressed LHX2 in the presence of ectopic FGF and observed an increased in SHH expression. LHX2 alone, however, was insufficient to upregulate SHH expression.

Taken together, these data suggest that LHX2 is an intermediate factor downstream of FGF that is necessary but not sufficient for SHH expression in the FGF‐SHH arm of the reciprocal feedback loop.

Support or Funding Information

Department of Pathology and Human Anatomy
Propagating the Ovarian Cancer Stem Cells by Reprogramming the Tumor Cells
Authors: Chen Yu‐An , Lu Chen‐Yu , Cheng Wen‐Fang , Ho Hong‐Nerng , Cheng Hsin‐Fu
Published: November 2023

Drug resistance and metastasis are considered to be the major factors leading to tragic prognosis and high rate of mortality in cancer patients. Recent evidences suggest that cancer stem cells (CSCs) possess the essential property of self‐protection from chemotherapy, exhibit the ability to self‐renew and generate cancer cells by asymmetric cell division. Nevertheless, the currently known mechanisms likely explain only the tip of an iceberg about CSC property, acquisition and maintenance. The information from data mining is still limited due to the rarity of CSCs in clinical samples. Ovarian cancer is the most lethal malignancy of the female reproductive system. CSCs in ovarian cancers are considered to acquire the molecular armaments of stem cells, which carry stem cell property and can determine the level of malignant potential of the cancer cells, similar to the findings in other cancer cell types.

To overcome the limitation and assess the underlying mechanism, we introduced a set of defined factors (OCT3/4, SOX2, KLF4 and c‐Myc) into human serous ovarian cancer cells, which is one of the most frequently encountered ovarian cancer type in female, through Sendai virus reprogramming, aiming to induce CSC properties in the cancer cells. We then evaluated the CSC properties in the cells. In preliminary data, we observed that the ovarian cancer cells transduced with the four Yamanaka factors showed significantly enhanced pluripotent stem cell properties and drug resistance. We thus named the cells with CSC properties as induced ovarian CSCs (iOCSCs). These iOCSCs could be maintained and expanded up to at least 40 passages.

Notably, consistent with the original ovarian cancer cells, iOCSCs could be differentiated into ovarian lineage cells, displaying markers as CA125 and CK‐7, shows that we generate generated iOCSCs with lineage specificity directly from cancer cells. In addition, we also confirmed that the phenotypes of our iOCSCs were reproducible in organoid culture and serial transplantation. The next, we are going to analysis the gene expression, transcriptome and metabolomics in iOCSCs to fully understand CSCs and find out new therapies target. Intriguingly, iOCSCs also acquired the ability to differentiate into cells belonging to the three embryonic germ layers, similar to human iPS cells or ES cells, this finding is probably the dawn of uncovering the stem cell tumorigenecity.

It is worth to emphasize again, this great value present approach thus makes it feasible to obtain abundant CSCs to support our understanding and in‐depth research into CSCs and hopefully can help in developing new therapeutic strategies to treat ovarian cancers.

Support or Funding Information

National Science Council of the Republic of China (NSC) and National Taiwan University Hospital (NTUH), Taiwan
Deformation‐Induced Ingression of Transitional Pleural Myofibroblasts during Compensatory Lung Growth
Authors: Ysasi Alexandra , Bennett Robert , Valenzuela Cristian , Servais Andrew , Kienzle Arne , Wagner Willi , Ackermann Maximilian , Tsuda Akira , Mentzer Steven
Published: November 2023

In many mammals, including rodents and humans, removal of one lung results in compensatory growth of the remaining lung. Following left pneumonectomy in mice, growth in the remaining right lung involves an increase in lung volume (46.03% in 14 days, p<0.01), weight (45.56% in 14 days, p<0.01) and cell number (46.77% in 14 days, p<0.05). Despite the increase in lung volumes, there is no detectable change in the internal linear dimensions (mean linear intercept 59.70 +/− 2.00 μm) indicating an increase in the number of alveoli. However, the mechanism of compensatory neoalveolarization is largely unknown. We have recently shown that expression of α‐smooth muscle actin (SMA), a cytoplasmic myofibroblast marker, is induced in the visceral pleura following pneumonectomy. Here, we used a cell tracer (cell surface biotin) to investigate the migration of these transitional pleural cells into the subpleural alveolar ducts. After pneumonectomy was performed, NHS‐ester activated biotin was instilled into the pleural cavity to label primary amines on the surface of the remaining lung. The biotin tracer was used to detect cell migration (ingression) into heterogeneous subpleural regions within 3 days of pneumonectomy (11.83%, 97.89 cells/mm of pleura, p<0.05). Cells colocalizing the biotin tracer and SMA increased 4‐fold in the pleural and subpleural regions within 3 days of pneumonectomy (6.25%, 51.71 cells/mm of pleura, p<0.05). SMA+/biotin+ cells co‐expressing vimentin, a cytoskeletal marker of epithelial‐mesenchymal transition, increased 4‐fold in the pleura within 24 hours of pneumonectomy and 18‐fold in the subpleural region within 3 days of pneumonectomy (p<0.05). This pleural transition and ingression of biotin+ cells occurred preferentially in areas of increased post‐pneumonectomy deformation, as previously modeled by finite element analysis. Dependence of pleural transition and cell ingression on lung deformation was confirmed by the limited migration of biotin+ cells after pneumonectomy and plombage or pneumonectomy and phrenic nerve transection (p<0.05). These data indicate that visceral pleural cells undergo a deformation‐induced phenotypic transition with subsequent migration into subpleural regions of the lung—likely participating in compensatory remodeling of the lung.
SPARC and Hevin in Mustard‐Induced Skin Wound Healing
Authors: Chang Yoke‐Chen , Chang Hui‐Ying , Hahn Rita , Svobada Kathy K. , Gordon Marion K , Gerecke Donald R.
Published: November 2023

Sulfur mustard (SM) and its analog nitrogen mustard (NM) are potent bifunctional alkylating agents that induce skin blisters. They crosslink proteins and DNA and alter cell adhesion of basement membrane molecules. Dermal toxicity of the mustard agents includes separation of the epidermis and dermis, a strong inflammatory response, and prolonged wound healing. Matricellular proteins (MPs) including SPARC and hevin (aka SPARC‐like 1) are secreted proteins that are incorporated into the provisional matrix upon injury. MPs act as temporally dynamic signaling molecules to modulate cell‐matrix interactions during healing. Hevin can be cleaved by proteinase ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4). This cleavage produces a SPARC‐like fragment (SLF) which is essential to tissue development and wound healing. The expression and removal of MPs is controlled in normal healing, but may persist in chronic skin wounds. The temporal and spatial expression of MPs may play a role in the delayed repair of mustard injury. SPARC, hevin, ADAMTS4, and the basement membrane zone (BMZ) marker, laminin 332 (LM332), were examined over the course of 10 days after exposure of mouse dorsal skin to 5 μmoles NM. In unexposed controls, immunofluorescence (IF) analysis of LM332 showed continuous staining across the BMZ, with minimal staining of SPARC, and hevin. At 1 day post NM‐exposure, H & E staining showed characteristic dermal‐epidermal separations, while IF showed a spotty discontinuous pattern for LM332. Strong staining of SPARC appeared throughout the BMZ. Hevin staining was less intense than SPARC at this time point. Hevin became more apparent in the hyperplastic epidermis at 3 days post‐NM. By days 7–10, hevin stained throughout the hyperplastic epidermis. WB analysis confirmed the sequential deposition of SPARC and hevin in NM injured skin. SPARC showed a 43 kDa band at 1 day, and hevin showed both full length (130 kDa) and fragments (75, 66, and 43 kDa) at 1–10 days post NM exposure. In addition, activated forms of ADAMTS4 were observed in NM exposed skin samples with bands at 75, 60, and 50 kDa by WB. Dual IF staining showed ADAMTS4 co‐localized with hevin in the BMZ and in the basal keratinocytes. This suggests hevin may be processed by ADAMTS4 to produce SLF for wound repair in our injury model. Overall, we observed sequential expression of SPARC and hevin in the BMZ of NM induced skin wounds. This may play a role in modulating the delayed wound healing of vesicant induced injury.

Support or Funding Information

ES005022, T32ES007148, and NIAMS U54AR055073
Left‐Right Patterning and Congenital Heart Disease
Authors: Yost H. Joseph
Published: November 2023

It has been known for centuries that most external anatomical features of the human body are bilaterally symmetric, and that many of the internal organs have highly conserved Left‐Right (LR) asymmetries. However, we are only beginning to understand the developmental mechanisms that coordinate and execute bilateral symmetry and LR asymmetry. This lecture will provide a brief overview of LR developmental pathways that impact cardiovascular and neural development, and a view toward the future, in which the LR development field will impact basic sciences, human genetics and clinically relevant biomedical research.

Support or Funding Information

Research in the Yost Lab is currently funded by a NHLBI Bench‐to‐Bassinet Consortium (http://www.benchtobassinet.com) grant to HJY (UM1HL098160), and has previously been funded by other grants from NHLBI and from the American Heart Association.
The Path to Anatomy via Molecular Biology
Authors: Gordon Marion K
Published: November 2023

Prior to the advent of Maxam and Gilbert DNA sequencing and Sanger, Nicklen and Coulson DNA sequencing, I was a student in an RNA sequencing laboratory. The experience taught me the proper paranoia for isolating and handling RNA. This was useful when, in the 1980s, I joined the lab of Bjorn Olsen at Rutgers Medical School, now called Robert Wood Johnson Medical School. There I learned the most current procedures in molecular biology, DNA sequencing and cloning. Bjorn was a great mentor, and he was an interesting guy. One day, another student and I found his histology notebook on his bookshelf‐‐he had hand drawn and colored every tissue section he had viewed so that he could study them for his histology exams. They were beautiful, accurate works of art, so intricate that I wished I could draw like that. Bjorn's passion about his work was contagious, and as a consequence, I fell in love with the family of collagens, which consisted of only 9 or 10 members then. A postdoc in the Olsen lab, Yoshifumi Ninomiya, had a role in determining the structure of type IX collagen by making a cDNA library from cartilage mRNAs and isolating the cDNAs corresponding to the collagen IX polypeptide chains. I was in awe of the idea that there were molecular biology strategies that could be used to identify and characterize new collagens, all because these triple helical molecules had repeating Gly‐Xxx‐Yyy sequences. The thrill of potentially being able to discover a new collagen made me love this type of science even more, and I put my whole heart into it. Around that time, a postdoc from Elizabeth Hay's lab visited Bjorn's lab, and this led to my meeting one of the best friends I'll ever have. That person, Kathy Svoboda, is not only partially responsible for Bjorn being hired by Betty Hay, chair of the Department of Anatomy and Cell Biology at Harvard Medical School, but her suggestion meant my life would change too! If Bjorn went to Boston, my husband Don and I would also get to go. And we did! It was an exciting experience to be in that department. Kathy, Betty, Bjorn and other faculty would talk about the American Association of Anatomists, and Kathy convinced me that AAA had investigators and educators that covered just about any topic in the biological sciences that could possibly interest me. She talked me into attending one of the annual meetings. I joined AAA sometime in the late 1980s or early 1990s, and, finding that what Kathy had said was true, I wanted to play a role in the society. I began serving on a committee as a guest member in 1999, and by 2001 I was committed to giving back to the Association since I had gotten so much from it. As I met more and more of the AAA members, I realized I now had greatly extended my “family” of scientists from this society, and that they played a big role in my being happy in my career.

Support or Funding Information

Supported by National Eye Institute EY009056, National Institute of Arthritis and Musculoskeletal and Skin Diseases U54AR055073, and National Institute of Environmental Health Sciences P30ES005022
From dividing cells to helping students overcome socio‐economic barriers
Authors: Robinson Douglas N.
Published: November 2023

Our lab investigates how cells form the shapes required for the specialized functions necessary for human health. Our effort is designed to decipher cellular morphogenesis from molecular and biophysical perspectives and to identify broad principles relevant for healthy and disease states. By applying forward genetics, quantitative cell biology, biochemistry and cell biophysics with computational approaches, we are developing understanding of cell morphogenesis that has predictive power. A major goal then is to be able to predict how cell and tissue systems will operate under diverse contexts and in response to a variety of insults. Because mechanics are central to cell behavior, we also seek to identify novel small molecule modulators of cell mechanics. Towards this goal, we have already utilized our amoeboid system to identify one such compound, 4‐HAP, which increases cortical myosin‐II and cortical elasticity. We are now testing 4‐HAP in animal models of three different cancers types. Finally, we are advancing our comparative effort, focusing on a range of mammalian systems and disease models, including pancreatic ductal adenocarcinoma, chronic obstructive pulmonary disease, and degenerative motor neuron disease, where cell mechanics and architecture play a prominent role.

Using our research environment, we have also built a pipeline initiative to develop scholars from economically and educationally disadvantaged backgrounds into future professionals in medicine and biomedical research. Our pipeline includes high school, undergraduate and postbaccalaureate programs and has now served ~110 scholars from across all levels with our first cohort starting to apply to medical and/or graduate schools. Our program provides scholars with intensive academic development, mentored research experience, and professionalism training.

Support or Funding Information

Baurenschmidt, DARPA, Hay Family, HRSA, JHU INBT, NHLBI, NIGMS, Sol Goldman Foundation, Thomas Wilson Foundation
Dynamics of autophagy and mitophagy in neurons
Authors: Holzbaur Erika L.F.
Published: November 2023

Neurons are large, polarized, and very long‐lived cells. In order to maintain cellular function over decades, neurons are highly dependent on homeostatic mechanisms such as autophagy. Autophagy is a cellular pathway for the degradation of aggregated proteins and dysfunctional organelles such as mitochondria, which are degraded by mitophagy. Mutations in proteins required for autophagy or mitophagy including PINK1, Parkin, Optineurin and TBK1 are sufficient to cause neurodegeneration, indicating the importance of this pathway to maintain neuronal function. We are using live cell imaging in cell models and in primary neurons to study the dynamics of autophagy and mitophagy, in order to define the underlying molecular mechanisms and to better understand how defects in this pathway lead to neurodegenerative diseases such as Amyotrophic lateral sclerosis (ALS) and Parkinson's disease.

Support or Funding Information

Supported by a Javits Neuroscience Investigator Award to E. Holzbaur (NIH R37 NS060698)
A novel enzymatic DNA modification on methylcytosine
Authors: Xu Guoliang
Published: November 2023

DNA methylation at cytosines plays various roles in both prokaryotes and eukaryotes. In the course of the search for novel DNA modifications, we found a bulky side chain of 90 Daltons conjugated to methylcytosine in the genomic DNA of an unicellular green alga, Chlamydomonas reinhardtii. The enzyme responsible for this modification, called Cmd1 (cytosine modifying enzyme 1) shows little homology to the proteins in databases. The modification reaction appears to require oxygen and Fe2+, as the dioxygenases of the Ten‐Eleven Translocation (TET) family do and depends on the presence of 5‐methylcytosine in the DNA substrate. 2‐oxoglutatate, an essential co‐substrate for TET enzymes, is however not required. Strikingly, vitamin C is used a donor for the modification. Data on biochemical and functional characterization of this new type of DNA modification will be presented. This modification and the Cmd1 enzyme might find applications in the detection of 5mC in DNA samples.

Support or Funding Information

National Science Foundation of China
A Shotgun Fingerprinting Approach to Document Compliance With an Almond Consumption Intervention
Authors: Dhillon Jaapna , Ferreira Christina R , Sobreira Tiago Jose Paschoal , Mattes Richard D
Published: November 2023

Introduction

Almonds are an extremely rich source of unsaturated lipids and flavonoids and increased consumption may be recommended to achieve various health benefits. However, there are no known analytical methods to document compliance to chronic almond consumption.

Objective

To develop an analytical approach that identifies metabolic profiles associated with long‐term almond intake to ascertain compliance with prescribed consumption.

Methods

A shotgun fingerprinting strategy was designed to isolate metabolic changes in erythrocytes after 12 weeks of almond consumption in a weight‐loss study. Representative samples (pools) of erythrocytes from individuals consuming almonds or no almonds were screened by flow injection mass spectrometry. Data based on scan modes for shotgun lipidomics, and m/z values compatible with almond flavonoid content were collected. Specific ion pairs and m/z values detected in pooled samples were combined into single ion monitoring and multiple reaction monitoring methods intended for individual sample screening. Values of ion intensities were normalized by total ion current to obtain relative ion intensities.

Results

Out of the 243 values of m/z monitored by both methods, 5 ratios and combinations of specific ions with receiver operating characteristic (ROC) curves AUC>0.89 had sensitivity of 74.2% and specificity of 90%. Eight out of the 31 participants (25.8%) in the almond group and 3 out of the 30 (10%) participants in the control group were misclassified by all 5 ratios.

Conclusion

Ratios and combinations of specific ions mainly related to membrane lipids were discriminatory of almond consumption from the nut‐free diet. The relationship between these metabolites indicates almond‐induced changes in the metabolomics profile and the misclassifications observed as a result of ratio performance evaluation could be indications of possible non‐compliance as supported by analysis of dietary intake data.

Support or Funding Information

Almond Board of California
Maternal Vitamin D supplementation significantly improved maternal plasma 25(OH)D but not infant 25(OH)D or bone turnover markers in Ethiopian lactating women and their infants
Authors: Girma Meron , Tilahun Birkneh , Mulugeta Biruk , Reta Fikadu , Stoecker Barbara J
Published: November 2023

We have reported Vitamin D insufficiency in Ethiopian women despite proximity to the equator, but responses to oral vitamin D supplementation in this population are not known. The aim of the study was to determine the effect of vitamin D3 supplmentation on maternal and infant plasma 25(OH)D concentrations and markers of bone turnover. We conducted a randomized, placebo controlled, double blind trial (NCT02210884) in Sidama Zone (7°N, 38°E), Southern Ethiopia. Lactating women (n=126) were enrolled within 2 weeks postpartum and were randomized to receive vitamin D3 (15,000 IU/week) or placebo for 12 months. Plasma 25(OH)D, bone specific alkaline phosphatase (BAP), osteocalcin (OC) and C‐telopeptide for type 1 collagen (CTX) were determined. Skin reflectance, dietary intake and individual UVB exposure also were measured. Median (IQR) 25(OH)D concentrations were not significantly different between vitamin D and placebo groups at baseline [47 (37, 57) nmol/L vs 45 (38, 55) nmol/L, p=0.78]. 25(OH)D concentrations were significantly higher in the vitamin D group at one year [109 (93, 121) nmol/L vs 62 (50, 81) nmol/L, p < 0.0001]. All women in the vitamin D group were vitamin D sufficient (> 50 nmol/L) by the end of the study. Furthermore, 95% had attained 25(OH)D concentrations > 75 nmol/L compared to 39% in the placebo group. Infant 25(OH)D concentrations at one year were not significantly different between groups. Maternal BAP and OC concentrations increased between 2 weeks post‐partum and 12‐month follow up in both groups (P< 0.001). No significant differences by group were seen for all bone turnover markers. Weekly Vitamin D3 (15,000 IU/week) supplementation safely eliminated vitamin D insufficiency in the study population.

Support or Funding Information

Supported by the Nutricia Research Foundation, OAES Regional Project W‐3002 and Pure Encapsulations Inc, Boston, USA.