bluuebunny/crossref_metadata_2025 huggingface dataset:
Hippocampal Dihomo‐γ‐Linolenic Acid Partially Mediates the Relationship Between Serum Lauric Acid and Hippocampal Choline Metabolites in the Young Pig
Authors: Mudd Austin T , Waworuntu Rosaline V , Berg Brian M , Donovan Sharon M , Dilger Ryan N
Published: November 2023

Fatty acids are pivotal for growth and development of the neonate, and the brain is especially susceptible to their presence or absence. There is interest in understanding the mechanisms by which specific fatty acids in the circulation influence development, but relatively little research has explored medium chain fatty acids (MCFA) in this context. Therefore, the objective of this exploratory study was to identify novel relationships whereby circulating MCFA levels modulate hippocampal metabolites. Beginning at 2 d and continuing until 30 d of age, 24 vaginally‐derived male piglets were provided custom milk replacer formulated to meet piglet nutrient requirements. At 30 d of age, piglets underwent magnetic resonance spectroscopy (MRS) procedures to quantify metabolite concentrations in the hippocampus. Following MRS, samples from the right hippocampus and serum were harvested from pigs at 31 d of age for lipidomic profiling. Linear regressions were used to assess the relationship between serum MCFA and hippocampal MRS metabolites, as well as between serum MCFA and hippocampal FA. Subsequently, a mediation model was used to investigate whether the relationships between serum MCFA and hippocampal MRS metabolites were mediated by hippocampal FA. The mediation analysis revealed that hippocampal free dihomo‐γ‐linolenic acid (FA‐20:3n6) partially mediated the relationship between serum free lauric acid (FA‐12:0) and brain glycerophosphocholine plus phosphocholine (GPC‐PCh). First, increased levels of serum FA‐12:0 related to decreased levels of hippocampal FA‐20:3n6 (P = 0.011). Second, increased levels of serum FA‐12:0 related to decreased brain GPC‐PCh (P = 0.002). We observed a significant (P < 0.05; 95% CI: −0.245 to −0.019) indirect path of the mediation (i.e., effect of serum FA‐12:0 though hippocampal FA‐20:3n6 on brain GPC‐PCh). Additionally, the direct path (i.e., effect of serum FA‐12:0 on brain GPC‐PCh, accounting for hippocampal FA‐20:3n6) was also significant (P < 0.05; 95% CI: −0.435 to −0.012). Therefore, hippocampal FA‐20:3n6 partially mediated the relationship between serum FA‐12:0 and brain GPC‐PCh. Overall, these results suggest the levels of circulating FA‐12:0 may be related to hippocampal metabolism of previously unrelated complex lipid moieties. Future research should test for direct effects of altering circulating FA‐12:0 on these metabolites and associated roles in brain development and functions.

Support or Funding Information

This project was funded by Mead Johnson Nutrition.
Diminished Abundance of Translation Initiation Signaling Components in Skeletal Muscle of Low Birthweight Neonatal Pigs
Authors: Chen Ying , McCauley Sydney R. , El‐Kadi Samer W.
Published: November 2023

Low birth weight (LBWT) increases the risk of fetal mortality and neonatal morbidity and is consistently associated with impaired postnatal muscle growth. Our previous data suggested that local insulin‐like growth factor‐I (IGF‐I) expression is lower and mammalian target of rapamycin / ribosomal protein S6 kinase 1 (mTOR/S6K1) signaling is downregulated in skeletal muscle of LBWT neonatal pigs. We hypothesized that protein translation initiation is compromised in skeletal muscle of LBWT pigs as a consequence of dysfunction of IGF‐I signaling cascade, which lead to impaired muscle growth in LBWT pigs. Twelve pairs of newborn male LBWT (816 ± 55 g) and normal birth weight (NBWT; 1,642 ± 55 g) littermates were euthanized to collect longissimus dorsi muscle. Muscle protein abundance and phosphorylation were measured by western blot. There was no difference in abundance of extracellular signal‐regulated kinase 1/2 (ERK1/2) in muscle of LBWT and NBWT groups. In addition, phosphorylation of ERK1/2 did not differ in LBWT pigs compared with their NBWT littermates. However, abundance of both MAPK‐interacting kinase (MNK) 1 and MNK2 was lower (P ≤ 0.05) in muscle of LBWT pigs compared with their NBWT siblings, while phosphorylation of MNKs did not differ between two groups. With decreased abundance of MNKs, protein expression and phosphorylation of eukaryotic initiation factor (eIF) 4E were not different in muscle of LBWT and NBWT pigs. However, abundance and phosphorylation of eIF4G were lower (P ≤ 0.05) in muscle of LBWT pigs compared with their NBWT siblings. In addition, abundance of eIF3e was lower (P ≤ 0.05) in LBWT compared with NBWT group. Lower abundance of eIFs in skeletal muscle of LBWT pigs suggested impaired formation of translation initiation complex, which would contribute to decreased protein synthesis and restricted muscle growth in LBWT neonatal pigs.
Low Omega‐3 Fatty Acid Status Found in Wellness Screening Participants
Authors: Bird Julia K , Salem Norman , Yurko‐Mauro Karin , Marshall Keri , McBurney Michael I
Published: November 2023

OBJECTIVE

96% of Americans nationally have a sub‐optimal omega‐3 status. The objective of this study was to determine the omega‐3 index and its relationship to diet and dietary supplement use in by volunteers participating in the Healthyroads™ biometrics wellness screening administered at their work site by their employer.

METHODS

636 employees at 16 DSM worksites in the United States consented to have whole blood, finger stick fatty acid testing as part of their Healthyroads™ wellness biometrics screening, and a second omega‐3 test was offered 3 months later. The research protocol was designed in accordance with the Principles of the Declaration of Helsinki and was approved by the New England IRB (#20160533) before subjects underwent any study‐related procedures. Basic demographics, a validated food frequency questionnaire, and a whole blood sample were collected at baseline and follow‐up. Blood samples were shipped to OmegaQuant LLC for fatty acid analysis and questionnaires for data collation and anonymization. Data were analyzed with SAS version 9.3. At the time of abstract submission, 7 sites and 135 participants had completed a second testing.

RESULTS

40% (N=252) of participants were female, and 25% (N=161) reported use of dietary supplements containing omega‐3 fatty acids. Participants reported mean intakes of 244±206 (±SD) mg/d EPA+DHA from foods and 183±454 mg/d EPA+DHA from supplements. Mean omega‐3 index (% EPA+DPA+DHA of total fatty acid concentration) was 4.9±0.2%; 29% had an omega‐3 index below 4%, 67% had an insufficient status (4% to 8%) and 3.6% had an adequate status (8% and above).

Dietary supplement users' EPA+DHA intakes from food were higher (267±207 mg/d) than dietary supplement non‐users (236±205 mg/d), but this was not statistically significant. Total EPA+DHA intake were significantly higher in dietary supplement users (989±694 mg/d vs 236±205 mg/d, p<0.001).

Males had a significantly lower omega‐3 index (4.7±0.1%) than females (5.0±0.1%, p<0.01). Dietary supplement users had a significantly higher omega‐3 index (6.0±0.2%) than non‐users (4.5%±0.1, p<0.001). There was a significant, positive relationship between total EPA+DHA intake and omega‐3 index (β=0.0013, intercept=4.3, p<0.001). For 135 individuals with a follow‐up visit, omega‐3 index was 5.2±0.2%, significantly higher than at baseline (mean increase 0.2%, p=0.03).

CONCLUSIONS

Less than 5% of employees participating in the Healthyroads™ biometrics wellness screening had an adequate omega‐3 index (>8%). Both regular consumption of fatty fish and use of omega‐3 dietary supplements were associated with higher whole blood concentrations of EPA+DHA. Preliminary findings from a limited number of sites finds that participants who elected to undergo a follow‐up omega‐3 test, after receiving the results from their first test, had taken action to increase their omega‐3 index.
Diet, Nutritional Indicators, Infections and Inflammation Are Associated with Mean Arterial Pressure (MAP) in Indigenous Pregnant and Lactating Women in Rural Panama
Authors: González‐Fernández Doris , del Carmen Pons Emérita , Rueda Delfina , Sinisterra Odalis T , Murillo Enrique , Scott Marilyn E , Koski Kristine G
Published: November 2023

Background

Hypertensive disorders of pregnancy (HDPs) are the leading cause of maternal mortality in Latin‐America. Mean arterial pressure (MAP) has been used as an early indicator of HDPs. However, to understand its usefulness in early detection of HDPs, it is important to understand the range of factors associated with MAP in resource poor environments.

Objective

We explored associations of MAP or elevated MAP (eMAP) with wood smoke exposure, multiple nutrient deficiencies, infections and inflammation in indigenous Panamanian pregnant and lactating women living in extreme poverty.

Methods

A cross‐sectional study recruited 213 pregnant and 99 lactating indigenous women from the Ngäbe‐Buglé Comarca in rural Panama. Information on systolic and diastolic blood pressure (SBP, DBP), environmental and dietary factors, measures of inflammation (C‐reactive protein and cytokines), nutritional status (serum retinol binding protein ‐ RBP, folic acid, vitamins B12, A and D), and several infections (skin, oral, urogenital and intestinal nematodes) was collected. MAP was defined as DBP + 1/3 (SBP‐DBP). eMAP was defined as blood pressure >87 mmHg between 10–18 weeks of gestation, >84 mmHg in weeks 18–34, >86 mmHg after week 34; and ≥95 in lactating women. Bivariate comparisons were followed by adjusted linear and logistic regression models for MAP and eMAP, respectively.

Results

Prevalence of eMAP was 11.2% in pregnancy and 12.1% in lactation. Bivariate comparisons: In pregnancy, MAP was higher in women with folic acid<10 nmol/L, urinary specific gravity (USG)>1020 and hookworm infection, and lower in women with vaginal diplococcal infection and Ascaris. In lactation, MAP was higher in women with USG>1020, intake of animal source foods ≥7/wk and bacteriuria. Regression models: In pregnancy, wood smoke exposure, RBP<30 mg/L, presence of hookworm and TNF‐α were positively associated with MAP whereas basophil numbers, diplococcal infection score and presence of Ascaris were negatively associated with MAP. RBP<30 mg/L, folate deficiency, Trichuris and higher IL‐6 increased the odds of eMAP. During lactation, vitamins B12 and A and bacteriuria were positively associated with MAP whereas animal source foods and bacteriuria increased the odds of eMAP.

Conclusion

Given that we identified both positive and negative associations of infections and nutrient deficiencies with MAP and eMAP during pregnancy and lactation, a strategy that simultaneously reduces all infections and all nutrient deficiencies may not be advisable. Rather, a reduction in eMAP may be achieved by reducing wood smoke exposure, treating intestinal nematodes, addressing folate and protein deficiencies in pregnancy, and by treating bacteriuria during lactation.

Support or Funding Information

SENACYT (Panama), McGill Vitamin Fund
Diet Macronutrient Composition Impacts White Matter Microstructure in the Brain
Authors: Sawin Emily A , Gonzalez Jose G , Ong Irene M , McIlwain Sean J , Adluru Nagesh , Alexander Andrew L , Yu John‐Paul J
Published: November 2023

Background

Diffusion tensor imaging (DTI) is a noninvasive magnetic resonance imaging‐based method of examining neural structure, organization and white matter integrity.

Objective

To determine if (1) diet‐dependent macronutrient composition can impact white matter microstructure in the brain and (2) to determine if changes in diet‐dependent gut microbiome populations correlate with these observed alterations in white matter microstructure.

Methods

40 weanling male Sprague Dawley rats were randomized to one of four diets, a control diet, a high fat diet, a high fiber diet, or a high protein‐low carbohydrate diet and were fed for three weeks. For each animal, a fecal sample was collected at the beginning of the diet and at the culmination of the study to assess for changes in gut microbiome populations. Neural structure and white matter integrity were assessed by DTI, and transcriptomic changes were assessed with an Affymetrix GeneChip Rat Gene 2.0 ST array.

Results

Diffusion tensor imaging shows diet‐dependent alterations in neural structure, organization, and white matter integrity (Figure 1). The use of advanced modeling and predictive analytics further demonstrate that these observed changes in neural structure, organization, and integrity can be associated with specific microbial populations.

Conclusion

Diet‐dependent macronutrient composition and corresponding changes in the gut microbiome can influence neural structure and white matter integrity.
Protein Intake and Risk of Prediabetes and Type 2 Diabetes
Authors: Voortman Trudy , Chen Zhangling , Franco Oscar H.
Published: November 2023

Objective

Studies indicated that certain amino acids could affect glucose metabolism. However, few epidemiological studies examined the association between protein intake and risk of type 2 diabetes (T2D), and reported mixed findings. Furthermore, no study examined the association between protein intake and risk of prediabetes and for different types of protein are unclear. Therefore, we aimed to examine the associations of intake of different sources of protein on the development of T2D in large population‐based cohort study.

Methods

The Rotterdam Study is a prospective cohort study of subjects aged 45 years or older in the Netherlands. Prediabetes and T2D were diagnosed on the basis of WHO criteria using medical records and fasting glucose and insulin concentrations, measured in our research center. We included 6805 subjects who were free of T2D at baseline, of whom 5794 were without prediabetes. Protein intake was assessed at baseline with the use of validated food‐frequency questionnaires and expressed in energy percentages (E%). We used multivariable cox proportional hazard regression to analyze the associations between protein intake and risk of prediabetes and T2D. We used nutrient density substitution models to examine macronutrient substitution effects.

Results

During a mean follow‐up of 8.0 years, we documented 643 incident cases of T2D among 6805 subjects. For prediabetes, we documented 931 incident cases among the 5794 subjects who were free of prediabetes at baseline, during a median follow‐up of 7.6 years. In multivariable models including age, sex, physical activity, smoking status, education level, diet quality and family history of diabetes, a higher intake of total protein was associated with a higher risk of prediabetes (hazard ratio (HR)=1.19, 95% CI: 1.05–1.34, per 5E%) and a higher risk of T2D (HR=1.35, 95% CI: 1.18–1.54 per 5E%). Results were similar for a higher protein intake at the expense of of lower fat or at the expense of a lower carbohydrate intake. The association of protein was driven by a higher animal protein intake (prediabetes: HR=1.35, 95%CI: 1.20–1.51; and T2D: HR=1.35, 95%CI: 1.18–1.53); and not protein from plant sources. In line with this, a higher intake of animal protein at the expense of plant protein was positively associated with risk of prediabetes and T2D. After additional adjustment for body mass index or waist circumference associations attenuated, but still remained significant.

Conclusions

Our results suggest that a higher intake of animal protein, but not plant protein, is associated with a higher risk of developing prediabetes and type 2 diabetes.
Factors Associated with Food Safety Behaviors in Cancer Patients Seeking Treatment
Authors: Paden Holly , Hatsu Irene , Lustberg Maryam , Grenade Cassandra , Kane Kathleen , Mo Kevin , Ilic Sanja
Published: November 2023

Introduction

Over 14 million Americans suffer from cancer and are at a greater risk of foodborne disease due to weakened immune systems. However, food safety practices and factors that may lead to risky food safety behaviors are not well understood.

Objective

The objective of this study is to determine the risk perception and demographic factors associated with food safety behaviors in cancer patients seeking chemotherapy and radiation treatment.

Methods

This was a cross‐sectional study that recruited participants from two cancer‐specific hospitals in Ohio. A 173‐item questionnaire assessed the food acquisition and preparation practices and attitudes toward different aspects of food safety in the cancer population. It included sociodemographic factors, food insecurity status, and disease factors. The data were analyzed in SPSS.

Results

Participants (n=120) were mostly breast cancer patients (48.1%), female (84.6%), older than 50 (73.1%), and white non‐Hispanic (83.5%). About 38% had a college degree or more and 26.2% had at least one year of college education. A total of 2.9% participants had low food security while 8.7% were marginally food insecure. Low income was associated with risky food acquisition practices, such as buying damaged packages (r=0.318, P<0.01) and expired foods (r=0.310, P<0.01). Other high risk food acquisition practices identified were eating other people's leftovers (33%), eating roadkill (3%), or eating spoiled foods (2%). Low income was associated with unsafe food storage and preparation behaviors, such as storing eggs at room temperature (r=0.434, P<0.01) and leaving cooked foods on the stovetop overnight (r=0.380, P<0.01). Half of the participants were aware that they are at increased risk of contracting foodborne diseases due to cancer. Food insecurity in cancer patients was associated with a decreased risk perception (r=0.262, P<0.01). In general, cancer patients with more children (<18) in the household were more concerned about maintaining their refrigerators at adequate temperatures (r=0.252, P<0.05).

Conclusion

The findings can be used to develop effective food safety education programs for cancer patients and highlight the need for food safety education of patients during cancer treatment.

Support or Funding Information

Food Innovation Center New Faculty Initiative Award, The Ohio State University
Regulation of Bi‐organellar CHCHD2 in Response to Hypoxia is Dependent upon EGFR Molecular Subtype in Glioblastoma
Authors: Lumibao Jan , Chen Emily , Harley Brendan A , Gaskins H. Rex
Published: November 2023

Glioblastoma (GBM) is the most common and malignant form of brain cancer in adults, with a median survival time of 12–15 mo. Tumors are heterogeneous and characterized by hypoxic foci. Amplification, overexpression, and mutation of the receptor tyrosine kinase (RTK) EGFR are prevalent in GBM, and the constitutively active EGFRvIII mutant shows the poorest clinical outcomes. However, RTK inhibitors have failed clinically against GBM, with EGFR subtype poorly predicting outcome. We are investigating compensatory signaling pathways active within GBM tumor margins as a possible source of resistance to molecular subtype therapies. Previous studies have reported coiled‐coil‐helix‐coiled‐coil‐helix domain‐containing protein 2 (CHCHD2) as a bi‐organellar protein localized predominantly in the mitochondrial inner membrane but also present in the nucleus where it acts as a transcription factor. We hypothesized that this co‐localization enables GBM cells to adapt to hypoxic stress in the tumor microenvironment, particularly via mitochondrial retrograde signaling. We examined CHCHD2 localization by immunocytochemistry in U87wt and U87vIII (constitutively active EGFRvIII mutant) GBM cells subjected to 24, 48, and 72 h hypoxic (1% O2) incubation. Time‐lapse imaging revealed significantly greater nuclear accumulation of CHCHD2 in U87vIII cells compared to U87wt cells at all hypoxic time points. We next used engineered gliomas formed by microfluidic templating as a model of the GBM microenvironment. RT‐PCR and Western blot analyses revealed increased CHCHD2 gene expression but decreased CHCHD2 protein levels in U87wt cells in 72 h hypoxic incubation compared to normoxic conditions. No changes in CHCHD2 gene expression or protein content were observed in U87vIII cells after 72 h hypoxic incubation. Using the aforementioned engineered gliomas, we observed hypoxia significantly increased GBM invasion and pERK1/2 activation in both EGFRwt and EGFRvIII variants. These data indicate more stable levels of CHCHD2 and greater translocation to the nucleus in U87vIII cells under hypoxic stress, thus impacting downstream genes and pathways and potentially contributing to compensatory oncogenic signaling capable of overcoming RTK inhibition. Additional work will focus on subcellular fractionation and mitochondrial isolation to measure CHCHD2 protein levels in nuclei and mitochondria. EGFRwt and EGFRvIII variant GBM cells with CRISPR‐silenced CHCHD2 are being created to determine the extent to which differential mitochondrial‐to‐nuclear translocation of this bi‐organellar protein in hypoxic U87wt and U87vIII cells contributes explicitly to their enhanced invasion potential and therapeutic resistance.

Support or Funding Information

Funding: RO1CA197488
Soft Tissue and Protein Preservation in Dinosaur Fossils: Evidence, Criteria and Implications
Authors: Schweitzer Mary Higby , Zheng Wenxia
Published: November 2023

Evolutionary change first occurs in the genes or the proteins they encode; thus molecular investigations of extant organisms have transformed our understanding of evolutionary biology. DNA and protein sequence data have been used to test hypotheses of evolutionary relationships, paleobiogeographic trends, rates of molecular change, population mixing, and timing of dispersals. Molecular data have provided insight into population bottlenecks that place species at risk for extinction, and have shed light on species origins, radiations and extinctions, as well as organismal response to climate change at the molecular level. However, the extension of these investigations into deep time is extremely limited; estimates of rate and direction of molecular evolution are currently based upon data derived from living organisms and extrapolated to extinct ones, not directly derived. Remnants of these molecules in fossils allow observation of molecular evolution and diversity across deep time and can reveal molecular traits contributing to adaptation and resilience in the face of these challenges.

We present evidence that proteins are preserved in at least two dinosaur specimens that also evidenced still‐soft tissues, discuss the need for rigorous criteria for accepting the identity of biomolecules in deep time, and briefly discuss what these data may mean to our understanding of molecular evolution, species relationships and molecular adaptability over time.

Support or Funding Information

NSF INSPIRE Program EAR‐1344198

David and Lucile Packard Foundation
microRNAs in Cardiac Conduction System
Authors: Wang Jun
Published: November 2023

The cardiac conduction system (CCS) is required for initiating and maintaining regular rhythmic heartbeats and the CCS defects can give rise to cardiac arrhythmia, a leading cause for morbidity worldwide. MicroRNAs (miRs) are small non‐coding RNAs that repress gene expression post‐transcriptionally. We found that miR‐17‐92 is actively expressed in the CCS. Specific disruption of miR‐17‐92 in the CCS gave rise to cardiac arrhythmias in mice. We combined different screenings and identified miR‐17‐92 and its homologous cluster miR‐106b‐25 as candidate miR function in atrial fibrillation (AF), the most common adult arrhythmia. Intracardiac electrical stimulation revealed that mice with miR‐17‐92 and miR‐106b‐25 deficiency were predisposed to pacing‐induced AF. Mice with miR‐17‐92 cardiac‐specific conditional knockout (CKO) had spontaneous arrhythmias. miR‐17‐92 and miR‐106b‐25 directly repress expression of the sinoatrial node (SAN) program gene such as Shox2 and Tbx3. Notably, miR‐17‐92 also represses Hippo signaling, an ancient organ size control pathway represses cardiomyocyte proliferation and regeneration. miR‐17‐92 directly represses expression of the core Hippo component Lats2. Hippo disruption in the CCS also gave rise to cardiac arrhythmias in mice. Chromatin immunoprecipitation deep sequencing (ChIP‐Seq) data using antibody of Yap, the Hippo signaling effector, suggested that Hippo signaling regulates genes involved in the CCS homeostasis. Together, our data indicate a critical function of miR in maintaining the CCS homeostasis and avoiding arrhythmogenesis.
Hippo pathway in Heart Development and Regeneration
Authors: Martin James
Published: November 2023

Heart failure due to cardiomyocyte loss after ischemic heart disease is the leading cause of death in the United States in large part because heart muscle regenerates poorly. The endogenous mechanisms preventing mammalian cardiomyocyte regeneration are poorly understood. Hippo signaling, an ancient organ size control pathway, is a kinase cascade that inhibits developing cardiomyocyte proliferation but has not been studied postnatally or in fully mature adult cardiomyocytes. We investigated Hippo signaling in adult cardiomyocyte renewal and regeneration by inactivating Hippo pathway kinases and adaptor molecules in the post natal and adult heart and performed different types of heart injury. We found that unstressed Hippo‐deficient adult mouse cardiomyocytes re‐enter the cell cycle and undergo cytokinesis. Moreover, Hippo deficiency enhances cardiomyocyte regeneration with functional recovery after postnatal myocardial infarction and cardiac apex resection. In damaged hearts, Hippo mutant cardiomyocytes also have elevated proliferation. We conclude that Hippo signaling is an endogenous repressor of postnatal cardiomyocyte renewal and regeneration and that targeting the Hippo pathway in human disease may be beneficial to treat heart disease.

Support or Funding Information

NIH/NHLBI‐R01, Grant # 1R01HL127717‐01A1
Hematopoietic stem cell induction and development
Authors: Zovein Ann Capela
Published: November 2023

Hematopoietic stem cells (HSC) are generated during embryogenesis from specialized endothelium, termed hemogenic endothelium, in a transient process. We present our recent work dissecting the signaling pathways that regulate hematopoietic versus endothelial fate during hematopoietic stem cell emergence. We demonstrate cell intrinsic and extrinsic signaling are part of the hemogenic program. Arterial fate specifiers are required to be down regulated in hemogenic endothelium, with acquisition of hematopoietic fate determinants in emerging hematopoietic cells, which also exhibit changes in cell cycle dynamics and acquisition of inflammatory mediators. In addition, we explore a possible role of the supporting mesenchyme.

Support or Funding Information

The work was supported by the NIH 1DP2HL117743‐01 Award, and the California Institute for Regenerative Medicine New Faculty award RN3‐06479.
Tyrannosaurus rex Shows Histological Evidence For Avian‐Style Cranial Kinesis
Authors: Bailleul Alida M , Horner John R , Holliday Casey M
Published: November 2023

Modern avian cranial kinesis played a significant role in the adaptive radiation of birds as they evolved flexible skulls from their presumably less kinetic, non‐avian theropod ancestors. Avian cranial kinesis is partly mediated by novel synovial joints, which differ from those of other extant sauropsids in that they form secondary articular cartilage on dermatocranial elements. This tissue is different from the primary articular cartilage that is ubiquitous among the endochondral elements of all vertebrates. The evolutionary origin of secondary cartilage among the dinosaurian line still remains unclear, consequently limiting inferences of cranial kinesis in dinosaurs and other extinct taxa.

For the first time, we investigate the presence of secondary articular cartilage in the cranial joints of a non‐avian theropod dinosaur, Tyrannosaurus rex (MOR 1125), by comparing microanatomical features of a reference synovial joint, the jaw joint, and the lesser understood otic joint, which is responsible for streptostylic movements in the avian feeding apparatus. Five extracted fragments of these joint articular surfaces from the surangular, quadrate and squamosal were microCT‐scanned, and then serially sectioned via histological techniques to identify secondary cartilage. Complementary histological data from two extant archosaurs (Anas platyrhynchos and Alligator mississippiensis) were also examined.

The membranous squamosal and surangular of T. rex possess a thin layer of mineralized, secondary articular cartilage above their subchondral bone. The quadrate fragments possess primary articular cartilage, but no clear histological differences between primary and secondary cartilage were identified. These results suggest that the jaw and otic joints of T. rex were synovial and identical in structure to those of modern birds, but substantively different from those of A. mississippiensis. This study provides for the first time clear paleohistological evidence for a synovial otic joint, which bolsters hypotheses of avian‐style streptostyly in a theropod dinosaur, and that birds inherited this tissue from their dinosaurian ancestors. This study has significant implications for our understanding of the origins of avian cranial kinesis, for inferences of kinesis in theropod dinosaurs, and increases our knowledge of the evolution of cranial skeletal tissues in vertebrates.

Support or Funding Information

This study was funded by: NSF‐IOS 1457319; Department of Pathology and Anatomical Sciences.
Antibiotics from the microbial dark matter
Authors: Lewis Kim
Published: November 2023

Our ability to discover novel compounds has diminished, and pathogens acquire and spread resistance largely unchecked, leading to a human health crisis. The main source of antibiotics – soil actinomycetes – has been overmined. The problem is compounded by the presence of dormant persister cells tolerant to killing by all antibiotics. As a result, chronic osteomyelitis or infections of patients with cystic fibrosis can be untreatable. There are then two main problems facing the field of antibiotic discovery – resistance; and tolerance. Ideally, we need compounds free of resistance; and antimicrobials capable of killing dormant cells with inactive targets.

We reasoned that antibiotic producers face similar problems that we do, and evolved anti‐persister compounds. We considered an old abandoned antimicrobial acyldepsipeptide (ADEP) as a possible anti‐persister antibiotic. The compound targets the ClpP/A,C,X protease which normally degrades misfolded peptides with the aid of its ATP‐dependent chaperones. In the presence of ADEP the chaperones are no longer needed. We found that ADEP causes massive degradation of mature proteins in S. aureus by the disregulated ClpP protease, killing persisters both in vitro and in a biofilm model of infection in a mouse.

In search of new antibiotics, we decided to target uncultured microorganisms, which make up 99% of the total diversity. We developed approaches to grow uncultured bacteria and find that this “microbial dark matter” harbors novel antimicrobials. Lassomycin produced by soil actinomycetes kills persisters of Mycobacteria by forcing the ClpC1 chaperone to hydrolyze ATP. Teixobactin, made by a β‐proteobacterium, is essentially free of resistance. It binds two immutable targets, lipid II and lipid III. These examples suggest that natural products that can address the dual problem of resistance and tolerance were evolved by producing microorganisms, and mining uncultured bacteria holds the promise of reviving the field of antibiotic discovery.

Support or Funding Information

T‐RO1 AI085585, Bill & Melinda Gates Foundation
Radical strategies for biological methylation
Authors: Booker Squire J.
Published: November 2023

A new and menacing mechanism for antibiotic resistance is on the rise, involving the C8 methylation of adenosine 2503 of 23S rRNA of the bacterial ribosome. Indeed, this seemingly minor addition, installed by the radical SAM protein, Cfr, impedes the binding of over five classes of antibiotics to their targets. Considering that one‐half of all antibiotics currently in use target the bacterial ribosome, this emerging mechanism of resistance has the potential to reduce the effectiveness of the world's current arsenal of antibiotics severely. Interestingly, C8 of adenosine is chemically resistant to biological methylation by all currently accepted mechanisms, which usually involves the attack of a nucleophile onto an electrophilic methylating agent. This lecture will detail a new and novel strategy for the biological methylation of sp2‐hybridized carbon centers, which involves radical intermediates generated via the reductive cleavage of S‐adenosylmethionine and the participation of five essential cysteinyl residues.

Support or Funding Information

NIH GM 101957
Nature's Imitation Game: Decipher the Combinatorial CTD Code for Eukaryotic Transcription
Authors: Zhang Y. Jessie
Published: November 2023

In eukaryotes, the C‐terminal domain of RNA polymerase II (CTD) orchestrates the temporal and spatial control of transcription and is involved in the epigenetic regulation of gene expression. Errors in CTD regulation can result in cell death, cancer and severe developmental defects. The CTD executes its function as a transcription template to recruite transcription regulators through various post‐translational modifications on its heptad repeats. Recently, novel modifications on CTD have been identified, setting the stage for the possibility of combinatorial mechanisms for transcription regulation. We focused on the eukaryotic phosphatases in the CTD regulation and how they affect the function of CTD binding proteins. Determination of the structures of these CTD phosphatases guide our structure‐function investigation. Using a collection of chemical biology techniques including analytical chemistry, spectroscopy, enzymology and structural biology, we dissect the multiple layers of regulation in CTD‐mediated transcription. In particular, we developed an innovative method of mass spectromety, Ultraviolet photodissociation (UVPD), to map the site of modification in S. cerevisiae and Drosophila CTDs. Overall, we probe how the identities and phosphorylation states of individual residues in the consensus heptad repeats of CTD result in differentiated post‐translational modification states within CTD that lead to various transcription outcomes.

Support or Funding Information

The work is funded by National Institute for Health, Welch Foundation and Alzheimer's Drug Discovery Foundation.
Desire to Eat and Intake of Sweet and Savory, High‐ and Low‐Calorie Foods in Male and Female Adolescents
Authors: Singh Jaishree , Papantoni Afroditi , Carnell Susan
Published: November 2023

Highly palatable, energy‐dense (ED), sweet and savory foods may contribute to the obesity epidemic by triggering desire to eat, experienced by some as an addiction‐like response. However, individual differences in appetitive responses, and their relationships with hunger level or food intake, are not well understood. Fifty‐nine 14–18 year‐olds (10 overweight M, 13 overweight F, 23 lean M, 13 lean F) from the Baltimore, MD area, attended two study visits, both of which involved consuming a meal‐replacement drink (474 ml, 480 kcal) or water (474 ml, 0 kcal) and then undergoing an fMRI scan 38±12 min later, during which they viewed pictures of different foods and reported their desire to eat each one. Following the scan, participants were allowed to eat ad libitum from a multi‐item dinner meal. Examples of high‐ED food pictures and buffet items were: chips (savory) and ice cream (sweet); examples of low‐ED food pictures and buffet items were: vegetable pieces (savory) and fruit (sweet). Analyses revealed that males reported desiring high‐ED, both sweet and savory, foods more than did females, in the fed condition only, with no sex differences emerging for high‐ED foods in the fasted condition, or for low‐ED foods in either condition. Males and females consumed equal amounts of high‐ED, sweet and savory, foods in both conditions. In both conditions, overweight adolescents reported desiring high‐ED, both sweet and savory, foods less than did lean adolescents, with no weight group differences for low‐ED foods. Lean and overweight adolescents consumed equal amounts of high‐ED, sweet and savory, foods across both conditions. Participants reported desiring high‐ED, both sweet and savory, and low‐ED, savory only, foods more when fasted than when fed. Participants consumed more high‐ED, both sweet and savory, foods when fasted than when fed, and consumed more low‐ED, both sweet and savory, foods when fed than when fasted. There were no sex differences in modulation of intake by fasted/fed condition. Males' desire to eat savory high‐ED foods was negatively correlated with savory high‐ED food intake (r=−.504, p=.009), but only in the fasted condition (r=−.537, p=.005); there were no correlations between desire and intake emerging for the other food categories in either condition. Our results suggest that adolescent reports of desire to eat certain foods and/or subsequent food intake depend on hunger state consistently across sex and weight groups. However, overweight and female participants may under‐report desire to eat certain foods due to social desirability bias. Further research should explore potential sex differences in appetitive responses and how they translate into actual intake.

Support or Funding Information

Research was funded by an R00 National Institute of Diabetes & Digestion & Kidney Disease grant.
Does the Recent Chilean Regulation on Food Labelling Improve Chilean Population Knowledge on Nutritional Facts? A longitudinal comparison between 2012 and 2016
Authors: Gregori Dario , Gafare Claudia Elena , Lorenzoni Giulia
Published: November 2023

Obesity epidemic is concerning worldwide, both in developed and newly industrialized countries. It represents a severe public health issue, since obese subjects are prone to develop metabolic, cardiovascular, and orthopaedic diseases. In recent years, several public health interventions have been developed in order to fight such epidemic. Several governments worldwide have implemented interventions for obesity prevention by regulating food labelling. Particularly, the Chilean government has recently approved a regulation (law 20606/2012), aimed at improving consumers' knowledge on the content of food products' nutrients, by including front‐of‐packages labelling in products high in sodium, total sugar, satured fat, and energy.

The aim of the present study is to assess knowledge about nutritional facts of Chilean population before and after the approval of the Chilean law on food labelling.

In 2012, a “Computer Assisted Telephone Interview” (CATI) was conducted on a Chilean population‐based sample. The survey's questionnaire consisted of four main sections (participants socio‐demographic characteristics; attitudes towards the concepts of nutrition and obesity; understanding of nutritional facts; attitude towards the potential development of a new front‐of‐pack labelling). The survey was undertaken on a sample of 1280 Chilean subjects, selected using random digital dialling. The same survey is now taking place (on the same sample of subjects interviewed in 2012) in order to asses if the awareness of Chilean population on nutritional facts has improved, after the recent introduction of the regulation.

Results of the 2012 survey revealed a concerning lack of knowledge about nutritional facts (most of subjects interviewed did not read the labels and reported a low interest in paying an additional fee for additional information about nutritional facts). Preliminary results of the survey, currently ongoing and planned to be over by early December, are based on 24% of the sample, and showed no improvements in labeling knowledge after 4 years with an increase about 0.97% (95% C.I 0.3%–1.7%).

Evaluation of effectiveness of policies need longitudinal information and definition of sensitive targets. Improving knowledge of consumers is showing, on the basis of this preliminary results, to be a target not being caught by current Chilean regulation.

Support or Funding Information

Study has been done with a unrestricted grant from Prochild ONLUS, an Italian based ONG
The Efficacy of Silibinin to Reduce the Impact of Obesity on Invasive Prostate Cancer
Authors: Sherman Blaine , Hernandez Ana R , Alani Maraim F , Price Ramona S
Published: November 2023

Introduction

Obesity is a risk factor for therapeutic resistance, disease progression, and metastasis in prostate cancer (PCa) patients. Further, obesity results in a state of chronic inflammation, which has been linked, to several types of cancer including the prostate. Previously, we have shown that obesity‐related systemic factors promoted an invasive prostate cancer phenotype, in vitro. Specifically, interleukin‐6 (IL‐6) has been identified as a pro‐inflammatory cytokine, which may mediate obesity‐induced prostate cancer progression. Interestingly, silibinin, a polyphenol found in milk thistle seed, reduces inflammation and may have anti‐tumorigenic properties. The mechanisms involved in obesity‐induced PCa progression are not completely understood, therefore limiting the development of effective interventions to inhibit or reverse the tumor promoting effects of obesity. Therefore, we hypothesize that silibinin may be used to reduce or reverse the impact of obesity on invasive prostate cancer by using an in vitro approach of obesity and cancer.

Methods

To investigate the efficacy of silibinin to reduce obesity‐induced invasive prostate cancer, a novel in vitro model of obesity and human sera was used. Male human sera from each body mass index (BMI) category (n=5) of normal weight <25 (NW), overweight BMI 25–29.9 (OW), and obese BMI >30 (OB). The prostate cancer cell lines used for this study were PacMetUT1, 22RV1, and DU145. Matrigel chambers were used to measure the invasive capacity of prostate cancer cells. MTT was used to assess cell viability. Immunofluorescence was used to visualize the localization of e‐cadherin. Western blot analysis was used to detect IL‐6, phospho‐Androgen Receptor, total Androgen Receptor and COX‐2. qRT‐PCR was used to evaluate differences in gene expression of IL‐6, phospho‐Androgen, and COX‐2. Student's t‐test

Results

PacMetUT1, and 22RV1 cells exposed to serum from obese and overweight sera increased cell viability while silibinin inhibited cell viability. DU145 cell did not differ in cell viability upon exposure to sera from obese, overweight or normal weight with and without silibinin. Silibinin inhibited obesity‐induced invasion in 22RV1, DU145 and PacMetUT1 cells. At the time of the conference, qRT‐PCR and western data analyses will be presented on gene and protein expression of IL‐6, phospho‐Androgen, and COX‐2. Immunofluorescence was used to visualize localization of Ecadherin. In prostate cancer cells exposed to sera from obese sera resulted in dispersion of Ecadherin compared to normal and overweight sera which was modified by exposure to silibinin.

Conclusion

In summary, silibinin modulates several characteristics from the hallmarks of cancer. Silibinin was effective in selectively inhibiting obesity induced invasion, and suppressed proliferation in several prostate cancer cell lines. Future studies are needed to identify strategies to reverse the impact of obesity on prostate cancer progression.
Neonatal nutrition affects adolescent insulin sensitivity and the lineage allocation of bone marrow derived mesenchymal stem cells in Ossabaw pigs
Authors: Zhang Wei , Murray Robert , Stahl Chad
Published: November 2023

The Ossabaw pig is an excellent model for the study of nutritionally‐induced metabolic diseases due to similarities in their physiology and nutrient requirements to those of humans, and their voluntary consumption of excess dietary energy. In this study, we determined the impact of different neonatal feeding strategies on the later‐life development of obesity and insulin resistance. Additionally, we characterized alterations in the lineage allocation of mesenchymal stem cells (MSC) in response to these feeding strategies. Newborn ossabaw pigs were either sow‐reared or formula fed from 24h old until weaning (32.1±0.4 d of age). Sow‐reared pigs were then randomly assigned to either control diet or obesigenic diet. All pigs which received formula were then fed the obesigenic diet. After weaning, pigs were meal fed twice/day. The diets were formulated to provide equal protein, vitamins and minerals among the treatments and to provide 3X more calories in the obesigenic diet. Body weight was recorded weekly for 16 wks. At the end of the feeding study, IV glucose tolerance tests were conducted and MSC isolated. Pigs fed with obesigenic diet had 1.5‐fold heavier body weight than the control at the end of the study (P<0.05). This was coupled with significantly greater body length, neck, chest and waist size in the obesigenic groups. The obesigenic fed pigs had reduced glucose clearance capabilities compared to the control fed pigs. Among the obesigenic fed pigs, those that received formula had 1.7‐fold higher circulating glucose than sow‐reared pigs (P<0.05) and both were 2.8 and 1.6‐fold greater than the control, respectively (P<0.05). No significant difference in insulin area under the curve between the formula‐fed pigs and the sow‐reared pigs with obesigenic diet, while both had 1.9 and 2.1 folder higher insulin area under the curve than the control (P<0.05). The MSC isolated from the obese pigs had a greater ability to differentiate along an osteogenic lineage as observed by increased expression of osteocalcin and fibronectin (osteocalcin: 2.12±0.31 vs 1.04±0.31 and fibronectin: 3.42±0.30 vs 2.30±0.29, P<0.05). This alteration in MSC behavior was mirrored by increased bone size and mineral content (1.2 to 1.5 fold higher than the control (P<0.05). This work demonstrates that neonatal nutritional interventions can impact metabolic susceptibility to obesity, and that excessive dietary energy during pre‐pubertal life alters metabolism at both the systemic and cellular levels.