Prenatal Choline Supplementation Improves Glucose Tolerance and Reduces Liver Fat Accumulation in Mouse Offspring Exposed to Ethanol during the Prenatal and Postnatal Periods.

Prenatal alcohol exposure (AE) affects cognitive development. However, it is unclear whether prenatal AE influences the metabolic health of offspring and whether postnatal AE exacerbates metabolic deterioration resulting from prenatal AE. Choline is a semi-essential nutrient that has been demonstrated to mitigate the cognitive impairment of prenatal AE. This study investigated how maternal choline supplementation (CS) may modify the metabolic health of offspring with prenatal and postnatal AE (AE/AE). C57BL/6J female mice were fed either a Lieber-DeCarli diet with 1.4% ethanol between embryonic day (E) 9.5 and E17.5 or a control diet. Choline was supplemented with 4 × concentrations versus the control throughout pregnancy. At postnatal week 7, offspring mice were exposed to 1.4% ethanol for females and 3.9% ethanol for males for 4 weeks. AE/AE increased hepatic triglyceride accumulation in male offspring only, which was normalized by prenatal CS. Prenatal CS also improved glucose tolerance compared to AE/AE animals. AE/AE suppressed hepatic gene expression of peroxisome proliferator activated receptor alpha (Ppara) and low-density lipoprotein receptor (Ldlr), which regulate fatty acid catabolism and cholesterol reuptake, respectively, in male offspring. However, these changes were not rectified by prenatal CS. In conclusion, AE/AE led to an increased risk of steatosis and was partially prevented by prenatal CS in male mice.

Seed mucilage in temperate grassland species is unrelated to moisture requirements.

Myxospermy, the release of seed mucilage upon hydration, plays multiple roles in seed biology. Here, we explore whether seed mucilage occurs in a suite of temperate grassland species to test if the prevalence of species producing seed mucilage is associated with habitat type or seed characteristics. Seventy plant species found in wet or dry North American temperate grasslands were tested for the presence of seed mucilage through microscopic examination of seeds imbibed with histochemical stain for mucilage. Mucilage production was compared among species with different moisture requirements and seed mass. In this study, 43 of 70 of species tested produced seed mucilage. Seed mucilage did not differ based on habitat type, species moisture requirements, or seed mass. Most seed mucilage was non-adherent and did not remain stuck to the seed after extrusion. Seed mucilage was a common trait in the surveyed temperate grassland species and was observed in 61% of evaluated species. Surprisingly, seed mucilage was more common in temperate grasslands than in previous ecological surveys from arid/semiarid systems, which found 10%-31% myxospermous species. Given the high prevalence, seed mucilage may influence seedling ecology in temperate grasslands and requires further investigation.

Overlapping Localization of MAP65-2, -6, and -7 in Arabidopsis Hypocotyl Cells.

Microtubules are essential components of eukaryotic cells. Myriad proteins associate with microtubules to facilitate the organization and operation of microtubule arrays. Various M icrotubule A ssociated P roteins (MAPs) assist the assembly and function of mitotic spindles and interphase arrays. Nine MAP65 genes exist in the genome of the acentrosomal model plant, Arabidopsis thaliana, and the function of majority of these proteins is unclear. To address this knowledge gap, we demonstrate the localization of A. thaliana MAP65-6 and MAP65-7 fusion proteins expressed from native promoters in interphase cells of developing A. thaliana seedlings. Analyses of these fusion proteins co-expressed with alpha-tubulin 6 reporters indicate that MAP65-6 and MAP65-7 bind a subset of interphase microtubules. Co-expression of GFP: MAP65-6 with mCherry: MAP65-2 from native promoters in A. thaliana showed overlapping localization patterns on interphase microtubule bundles. Collectively, these data suggested that MAP65-2 , -6, and -7 bind cortical microtubule bundles in plant interphase microtubule arrays.

Stomatal pore width and area measurements in Zea mays.

Stomatal pores are adjustable microscopic holes on the surface of photosynthetic tissues that help regulate multiple aspects of plant physiology. Stomatal pores facilitate gas exchange necessary for photosynthesis, water transport, and temperature regulation. Pore size is influenced by many intertwined environmental, molecular, cellular, and physiological cues. Accurate and precise measurements of pore size is important for understanding the mechanisms that adjust pores and plant physiology. Here we investigate whether conventional pore measurements of width are appropriate for the economically important crop plant Zea mays . Our studies demonstrate that pore area is a more sensitive measurement than width in this plant.

Chemical remodeling of a cellular chaperone to target the active state of mutant KRAS.

The discovery of small-molecule inhibitors requires suitable binding pockets on protein surfaces. Proteins that lack this feature are considered undruggable and require innovative strategies for therapeutic targeting. KRAS is the most frequently activated oncogene in cancer, and the active state of mutant KRAS is such a recalcitrant target. We designed a natural product-inspired small molecule that remodels the surface of cyclophilin A (CYPA) to create a neomorphic interface with high affinity and selectivity for the active state of KRASG12C (in which glycine-12 is mutated to cysteine). The resulting CYPA:drug:KRASG12C tricomplex inactivated oncogenic signaling and led to tumor regressions in multiple human cancer models. This inhibitory strategy can be used to target additional KRAS mutants and other undruggable cancer drivers. Tricomplex inhibitors that selectively target active KRASG12C or multiple RAS mutants are in clinical trials now (NCT05462717 and NCT05379985).

The Role of miRNAs in Childhood Acute Lymphoblastic Leukemia Relapse and the Associated Molecular Mechanisms.

Acute lymphoblastic leukemia (ALL) is the most common cancer in children worldwide. Although ALL patients' overall survival rates in wealthy countries currently surpass 80%, 15-20% of patients still experience relapse. The underlying mechanisms of relapse are still not fully understood, and little progress has been made in treating refractory or relapsed disease. Disease relapse and treatment failure are common causes of leukemia-related death. In ALL relapse, several gene signatures have been identified, but it is also important to study miRNAs involved in ALL relapse in an effort to avoid relapse and to achieve better survival rates since miRNAs regulate target genes that participate in signaling pathways involved in relapse, such as those related to drug resistance, survival signals, and antiapoptotic mechanisms. Several miRNAs, such as miR-24, miR-27a, miR-99/100, miR-124, miR-1225b, miR-128b, miR-142-3p, miR-155 and miR-335-3p, are valuable biomarkers for prognosis and treatment response in ALL patients. Thus, this review aimed to analyze the primary miRNAs involved in pediatric ALL relapse and explore the underlying molecular mechanisms in an effort to identify miRNAs that may be potential candidates for anti-ALL therapy soon.

Diverse alterations associated with resistance to KRAS(G12C) inhibition.

Inactive state-selective KRAS(G12C) inhibitors1-8 demonstrate a 30-40% response rate and result in approximately 6-month median progression-free survival in patients with lung cancer9. The genetic basis for resistance to these first-in-class mutant GTPase inhibitors remains under investigation. Here we evaluated matched pre-treatment and post-treatment specimens from 43 patients treated with the KRAS(G12C) inhibitor sotorasib. Multiple treatment-emergent alterations were observed across 27 patients, including alterations in KRAS, NRAS, BRAF, EGFR, FGFR2, MYC and other genes. In preclinical patient-derived xenograft and cell line models, resistance to KRAS(G12C) inhibition was associated with low allele frequency hotspot mutations in KRAS(G12V or G13D), NRAS(Q61K or G13R), MRAS(Q71R) and/or BRAF(G596R), mirroring observations in patients. Single-cell sequencing in an isogenic lineage identified secondary RAS and/or BRAF mutations in the same cells as KRAS(G12C), where they bypassed inhibition without affecting target inactivation. Genetic or pharmacological targeting of ERK signalling intermediates enhanced the antiproliferative effect of G12C inhibitor treatment in models with acquired RAS or BRAF mutations. Our study thus suggests a heterogenous pattern of resistance with multiple subclonal events emerging during G12C inhibitor treatment. A subset of patients in our cohort acquired oncogenic KRAS, NRAS or BRAF mutations, and resistance in this setting may be delayed by co-targeting of ERK signalling intermediates. These findings merit broader evaluation in prospective clinical trials.

Shifting the Paradigm: A Population Health Approach to the Management of Direct Oral Anticoagulants.

Over the past decade, direct oral anticoagulants (DOACs) have contributed to a major paradigm shift in thrombosis management, replacing vitamin K antagonists as the most commonly prescribed anticoagulants in many countries. While DOACs provide distinct advantages over warfarin (eg, convenience, simplicity, and safety), they are frequently associated with inappropriate prescribing and adverse events. These events have prompted regulatory agencies to mandate oversight, which individual institutions may find difficult to comply with given limited resources. Veterans Health Administration (VHA) has leveraged technology to develop the DOAC Population Management Tool (PMT) to address these challenges. This tool has empowered VHA to update a 60-year standard of care from one-to-one provider-to-patient anticoagulation monitoring to a population-based management approach. The DOAC PMT allows for the oversight of all patients prescribed DOACs and leads to intervention only when clinically indicated. Using the DOAC PMT, facilities across VHA have maximized DOAC oversight while minimizing resource usage. Herein, we discuss how the DOAC PMT was conceived, developed, and implemented, along with the challenges encountered throughout the process. Additionally, we share the impact of the DOAC PMT across VHA, and the potential of this approach beyond anticoagulation and VHA.

Appearance of microtubules at the cytokinesis to interphase transition in Arabidopsis thaliana.

Microtubule arrays drastically reorganize during the cell cycle to facilitate specific events. Many cells contain a centrosome that dictates the assembly and organization of microtubule arrays. However, plant cells and many others do not contain centrosomes or discrete microtubule organizing centers. In plants, microtubules nucleate and polymerize from gamma-tubulin-containing complexes in the interphase cell cortex. During plant cell division, microtubules nucleate near nuclei to form the mitotic spindle and plant-specific phragmoplast required for cytokinesis. Therefore, during the plant cell cycle, microtubule nucleation shifts from cell cortex to the perinuclear region. While it is unclear how this shift occurs, previous studies observed microtubules that appeared to extend from nuclei into the cortex as cells transitioned into interphase in small cells. These data led to the hypothesis that microtubule nucleation complexes move from the nuclear surface to the cortex at the transition from cytokinesis into interphase. Here we document GFP labeled microtubules in living plant cells during the transition from cytokinesis to interphase. We observed apparent groups of microtubules spanning between the nucleus and cell cortex in large, vacuolated epidermal leaf cells. We also observed microtubules in the cell cortex that appeared separate from perinuclear-associated microtubules. While these cortical microtubules were not always seen, when present they were apparent before cytokinesis was complete and/or before nuclear-associated microtubules were obvious. These data add to and deepen the knowledge of microtubule reorganization at this cell cycle transition.

Peer Instruction no aprendizado da montagem de articulador semiajustável / Peer Instruction in learning to mount a semiadjustable articulator

As metodologias ativas de ensino-aprendizagem são um desafio tanto para o docente quanto para o discente e instituições de ensino. O presente estudo objetivou diagnosticar as condições mais favoráveis ao aprendizado, comparando diferentes métodos de ensino sobre montagem de articulador semiajustável (ASA) para estudantes de terceiro período da Faculdade de Odontologia São José, Rio de Janeiro, Brasil. Participaram deste estudo 140 estudantes divididos em dois grupos, Grupo I: Leitura de artigo e aula expositiva e Grupo II: Peer Instruction (PI). Foram avaliados o nível de participação nas atividades propostas, a forma como o conhecimento era construído, o número de erros e acertos obtidos nas atividades práticas, além do domínio do assunto abordado e a postura profissional do aluno. Observou-se que no Grupo I 6 (21,4%) estudantes realizaram correta montagem dos modelos e no Grupo II 26 (29,2%) apresentaram este resultado. Os erros mais prevalentes ocorreram nas fases compensação de cera (Grupo I: 18 - 35,3%; Grupo II: 26 - 29,2%) e registro intermaxilar (12 - 23,5% no Grupo I; 17 - 19,1% no Grupo II). O resultado obtido comprovou que a proposta de ensino PI, empregada no Grupo II, favoreceu o aprendizado, uma vez que houve aumento do número de acertos nas atividades e domínio dos conceitos que envolvem o manuseio e a operação dos equipamentos utilizados (AU).

The active teaching-learning methodologies are challenging to both the professor and student, and to teaching institutions. The aim of the present study was to diagnose the conditions most favorable to learning, by comparing different methods of teaching third semester students about mounting a Semi-Adjustable Articulator, at the São José Dental School, Rio de Janeiro, Brazil. The participants in this study were 140 students, divided into two groups, Group I: Reading the article, and a lecture demonstration, and Group II: Peer Instruction (PI). The following aspects were evaluated: the level of participation in the proposed activities, the way in which knowledge was constructed, the number of errors and correct outcomes obtained in the practical activities, in addition to command of the subject approached and the student's professional posture. In Group I, 6 (21.4%) students were observed to perform correct mounting of the models, and in Group II, 26 (29.2%) presented this result. The most prevalent errors occurred in the stages of compensation with wax (Group I: 18 - 35.3%; Group II: 26 - 29.2%) intermaxillary register (12 - 23.5% in Group I; 17 - 19.1% in Group II). The result obtained proved that the PI teaching proposal used in Group II favored learning, because there was an increase in the number of correct outcomes obtained in the activities, and command of the concepts that involved the handling and operation of the equipment used (AU).

Genome Architecture and Evolution of a Unichromosomal Asexual Nematode.

Asexual reproduction in animals, though rare, is the main or exclusive mode of reproduction in some long-lived lineages. The longevity of asexual clades may be correlated with the maintenance of heterozygosity by mechanisms that rearrange genomes and reduce recombination. Asexual species thus provide an opportunity to gain insight into the relationship between molecular changes, genome architecture, and cellular processes. Here we report the genome sequence of the parthenogenetic nematode Diploscapter pachys with only one chromosome pair. We show that this unichromosomal architecture is shared by a long-lived clade of asexual nematodes closely related to the genetic model organism Caenorhabditis elegans. Analysis of the genome assembly reveals that the unitary chromosome arose through fusion of six ancestral chromosomes, with extensive rearrangement among neighboring regions. Typical nematode telomeres and telomeric protection-encoding genes are lacking. Most regions show significant heterozygosity; homozygosity is largely concentrated to one region and attributed to gene conversion. Cell-biological and molecular evidence is consistent with the absence of key features of meiosis I, including synapsis and recombination. We propose that D. pachys preserves heterozygosity and produces diploid embryos without fertilization through a truncated meiosis. As a prelude to functional studies, we demonstrate that D. pachys is amenable to experimental manipulation by RNA interference.

Hornwort Stomata: Architecture and Fate Shared with 400-Million-Year-Old Fossil Plants without Leaves.

As one of the earliest plant groups to evolve stomata, hornworts are key to understanding the origin and function of stomata. Hornwort stomata are large and scattered on sporangia that grow from their bases and release spores at their tips. We present data from development and immunocytochemistry that identify a role for hornwort stomata that is correlated with sporangial and spore maturation. We measured guard cells across the genera with stomata to assess developmental changes in size and to analyze any correlation with genome size. Stomata form at the base of the sporophyte in the green region, where they develop differential wall thickenings, form a pore, and die. Guard cells collapse inwardly, increase in surface area, and remain perched over a substomatal cavity and network of intercellular spaces that is initially fluid filled. Following pore formation, the sporophyte dries from the outside inwardly and continues to do so after guard cells die and collapse. Spore tetrads develop in spore mother cell walls within a mucilaginous matrix, both of which progressively dry before sporophyte dehiscence. A lack of correlation between guard cell size and DNA content, lack of arabinans in cell walls, and perpetually open pores are consistent with the inactivity of hornwort stomata. Stomata are expendable in hornworts, as they have been lost twice in derived taxa. Guard cells and epidermal cells of hornworts show striking similarities with the earliest plant fossils. Our findings identify an architecture and fate of stomata in hornworts that is ancient and common to plants without sporophytic leaves.

Reproductive success through high pollinator visitation rates despite self incompatibility in an endangered wallflower.

PREMISE OF THE STUDY: Self incompatibility (SI) in rare plants presents a unique challenge-SI protects plants from inbreeding depression, but requires a sufficient number of mates and xenogamous pollination. Does SI persist in an endangered polyploid? Is pollinator visitation sufficient to ensure reproductive success? Is there evidence of inbreeding/outbreeding depression? We characterized the mating system, primary pollinators, pollen limitation, and inbreeding/outbreeding depression in Erysimum teretifolium to guide conservation efforts. METHODS: We compared seed production following self pollination and within- and between-population crosses. Pollen tubes were visualized after self pollinations and between-population pollinations. Pollen limitation was tested in the field. Pollinator observations were quantified using digital video. Inbreeding/outbreeding depression was assessed in progeny from self and outcross pollinations at early and later developmental stages. KEY RESULTS: Self-pollination reduced seed set by 6.5× and quadrupled reproductive failure compared with outcross pollination. Pollen tubes of some self pollinations were arrested at the stigmatic surface. Seed-set data indicated strong SI, and fruit-set data suggested partial SI. Pollinator diversity and visitation rates were high, and there was no evidence of pollen limitation. Inbreeding depression (δ) was weak for early developmental stages and strong for later developmental stages, with no evidence of outbreeding depression. CONCLUSIONS: The rare hexaploid E. teretifolium is largely self incompatible and suffers from late-acting inbreeding depression. Reproductive success in natural populations was accomplished through high pollinator visitation rates consistent with a lack of pollen limitation. Future reproductive health for this species will require large population sizes with sufficient mates and a robust pollinator community.

A coherent transcriptional feed-forward motif model for mediating auxin-sensitive PIN3 expression during lateral root development.

Multiple plant developmental processes, such as lateral root development, depend on auxin distribution patterns that are in part generated by the PIN-formed family of auxin-efflux transporters. Here we propose that AUXIN RESPONSE FACTOR7 (ARF7) and the ARF7-regulated FOUR LIPS/MYB124 (FLP) transcription factors jointly form a coherent feed-forward motif that mediates the auxin-responsive PIN3 transcription in planta to steer the early steps of lateral root formation. This regulatory mechanism might endow the PIN3 circuitry with a temporal 'memory' of auxin stimuli, potentially maintaining and enhancing the robustness of the auxin flux directionality during lateral root development. The cooperative action between canonical auxin signalling and other transcription factors might constitute a general mechanism by which transcriptional auxin-sensitivity can be regulated at a tissue-specific level.

The FOUR LIPS and MYB88 transcription factor genes are widely expressed in Arabidopsis thaliana during development.

PREMISE OF THE STUDY: The FOUR LIPS (FLP) and MYB88 transcription factors, which are closely related in structure and function, control the development of stomata, as well as entry into megasporogenesis in Arabidopsis thaliana. However, other locations where these transcription factors are expressed are poorly described. Documenting additional locations where these genes are expressed might define new functions for these genes. METHODS: Expression patterns were examined throughout vegetative and reproductive development. The expression from two transcriptional-reporter fusions were visualized with either ß-glucuronidase (GUS) or green fluorescence protein (GFP). KEY RESULTS: Both flp and myb88 genes were expressed in many, previously unreported locations, consistent with the possibility of additional functions for FLP and MYB88. Moreover, expression domains especially of FLP display sharp cutoffs or boundaries. CONCLUSIONS: In addition to stomatal and reproductive development, FLP and MYB88, which are R2R3 MYB transcription factor genes, are expressed in many locations in cells, tissues, and organs.

The Arabidopsis leucine-rich repeat receptor-like kinase MUSTACHES enforces stomatal bilateral symmetry in Arabidopsis.

Stomata display a mirror-like symmetry that is adaptive for shoot/atmosphere gas exchange. This symmetry includes the facing guard cells around a lens-shaped and bilaterally symmetric pore, as well as radially arranged microtubule arrays that primarily originate at the pore and then grow outwards. Mutations in MUSTACHES (MUS), which encodes a leucine-rich repeat receptor-like kinase, disrupt this symmetry, resulting in defects ranging from skewed pores and abnormally focused and depolarized radial microtubule arrays, to paired guard cells that face away from each other, or a severe loss of stomatal shape. Translational MUSproMUS:tripleGFP fusions are expressed in cell plates in most cells types in roots and shoots, and cytokinesis and cell plates are mostly normal in mus mutants. However, in guard mother cells, which divide and then form stomata, MUS expression is notably absent from new cell plates, and instead is peripherally located. These results are consistent with a role for MUS in enforcing wall building and cytoskeletal polarity at the centre of the developing stoma via signalling from the vicinity of the guard cell membrane.

Forecasting the student-professor matches that result in unusually effective teaching.

BACKGROUND: Two important influences on students' evaluations of teaching are relationship and professor effects. Relationship effects reflect unique matches between students and professors such that some professors are unusually effective for some students, but not for others. Professor effects reflect inter-rater agreement that some professors are more effective than others, on average across students. AIMS: We attempted to forecast students' evaluations of live lectures from brief, video-recorded teaching trailers. SAMPLE: Participants were 145 college students (74% female) enrolled in introductory psychology courses at a public university in the Great Lakes region of the United States. METHODS: Students viewed trailers early in the semester and attended live lectures months later. Because subgroups of students viewed the same professors, statistical analyses could isolate professor and relationship effects. RESULTS: Evaluations were influenced strongly by relationship and professor effects, and students' evaluations of live lectures could be forecasted from students' evaluations of teaching trailers. That is, we could forecast the individual students who would respond unusually well to a specific professor (relationship effects). We could also forecast which professors elicited better evaluations in live lectures, on average across students (professor effects). Professors who elicited unusually good evaluations in some students also elicited better memory for lectures in those students. CONCLUSIONS: It appears possible to forecast relationship and professor effects on teaching evaluations by presenting brief teaching trailers to students. Thus, it might be possible to develop online recommender systems to help match students and professors so that unusually effective teaching emerges.

Arabidopsis guard cell integrity involves the epigenetic stabilization of the FLP and FAMA transcription factor genes.

Arabidopsis guard cell (GC) fate is conferred via a transient pulse of expression of FAMA that encodes a bHLH transcription factor. Stomata often function for years, suggesting that the FAMA expression window stabilizes long-term GC identity or that additional factors operate. Transgenic lines harboring a copy of a FAMA transgene were found to induce the fate resetting of mature GCs to that of lineage-specific stem cells causing new stomata to arise within shells of the old, a Stoma-in-Stoma (SIS) phenotype. These lines disrupt the normal trimethylation on lysine 27 of histone3 (H3K27me3) on stomatal stem cell genes, a phenotype rescued by constitutive expression of the Polycomb Group (PcG) gene CURLY LEAF. Thus the stability of stomatal fate is enforced by a PcG-mediated reduction in the transcriptional accessibility of stem cell genes and by the endogenous FAMA gene itself. Moreover, a transgenic FOUR LIPS gene, which encodes a MYB protein that is not required for GC fate, also induces a SIS phenotype and disrupts H3K27 trimethylation. Thus FLP might indirectly enforce GC fate as well.

Deep functional redundancy between FAMA and FOUR LIPS in stomatal development.

Functional redundancy arises between gene paralogs as well as non-homologous genes that play a common role at a shared node. The bHLH transcription factor FAMA, along with the paralogous MYB genes, FOUR LIPS (FLP) and MYB88 all ensure that Arabidopsis stomata contain just two guard cells (GCs) by enforcing a single symmetric precursor cell division before stomatal maturity. Consistent with this function, FLP and FAMA exhibit the same expression pattern in which both translational GFP fusions emit fluorescence just before and after symmetric division; however, FAMA but not FLP is required to confer GC fate. Strikingly, swapping the genes and promoters of the FLP and FAMA genes results in the reciprocal complementation of respective loss-of-function mutants. Thus, an FLP transgene can restore GC fate to a fama mutant background. FAMA, FLP and the FLP paralog MYB88 were previously shown to influence higher order functions in stomatal development, including maintaining and stabilizing stomatal fate. Here we show that these overlapping functions are likely to also involve interactions between FLP and FAMA with the RETINOBLASTOMA-RELATED (RBR) protein.