bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of hyperthermia on cognitive and auditory processing
Authors: Shibasaki Manabu , Nanba Mari , Morimoto Keiko , Nakata Hiroki
Published: June 2021

Hyperthermia is thought to cause a significant strain on either the brain function or the locomotive system. In fact, hyperthermia impairs psychological and working memory performances, but the effect of hyperthermia on either cognitive or auditory processing remains unknown. We hypothesized that a passive heat stress impairs the cognitive function when the internal temperature was excessively increased. Thirteen healthy males performed an auditory oddball paradigm before and after heat stress (Pre and Post) and when esophageal temperature was increased by 0.8 °C and 2.0 °C (Mild and Severe). The reaction time and event‐related potentials (ERPs) were recorded in these four sessions. As a time control, subjects performed the same sessions without heat stress. The reaction time was shortened while esophageal temperature was elevated relative to the Pre but did not change in the time control trial. However the peak latency and amplitude of N100, indicated auditory processing, did not change throughout the experiment. Although the latency of P300 was unaffected due to heat stress, the amplitude of P300, indicated cognitive processing, was significantly reduced at the Severe (10.5±5.9 μV) and Post (11.1±5.5 μV) relative to at the Pre (16.3±4.7 μV). These results suggest that excessive elevation of internal temperature impairs cognitive processing but not auditory processing.
Effects of Voluntary Exercise on Progression of Aortic Aneurysm in a Mouse Model of Marfan Syndrome
Authors: Gibson Christine , Alex Ramona , Broderick Thomas , Vallejo Johana , Esfandiarei Mitra
Published: June 2021

Aortic aneurysm is the leading cause of death in Marfan syndrome (MFS), a connective tissue disorder caused by mutations in the fibrillin‐1 gene. MFS aneurysm is characterized by weakening of the aortic wall due to elastin fibers fragmentation and aortic aneurysm. The above‐average height and distinct physical features make young adults with MFS, desirable candidates for competitive sports; but little is known about the exercise limit at which they will be at risk for aortic rupture. On the other hand, voluntary cardiovascular exercise has been shown to have a protective effect on the aortic wall structure and function. In this study, we investigated whether a voluntary aerobic exercise routine would have any effects on structure and function of the aorta in a mouse model of MFS. Four‐week old MFS and control mice were subjected to voluntary wheel exercise or sedentary life‐style for five months. Aortic tissue were isolated at the age of 24‐weeks, and subjected to structural and functional studies using histological staining and myography. During the study period, running activity (Km/day) in MFS mice was reduced by 37 percent, as compared to control mice. However, compared to non‐exercise MFS mice, their aortic wall structure (elastin fibers organization) and blood vessel function (contractility and vasodilation) were significantly improved. These findings provide new insights into the potential protective effects of a mild exercise routine in Marfan patients, especially in young adults affected by cardiovascular complications and aortic aneurysm. This work was funded by Midwestern University Graduate Research Fund.
Elevated Core and Skin Temperatures Independently Attenuate Simulated Hemorrhagic Tolerance
Authors: Pearson James , Lucas Rebekah , Schlader Zachary , Gagnon Daniel , Crandall Craig
Published: June 2021

The separate influences of increased core (Tcore) and skin (Tskin) temperatures upon tolerance to a simulated hemorrhagic challenge (Lower Body Negative Pressure; LBNP) are unknown. Nine participants underwent progressive LBNP to pre‐syncope on four occasions: 1) Control: Neutral Tskin and Tcore (34.3±0.5 and 36.8±0.2 ºC), 2) Combined: High Tskin and high Tcore (38.8±0.6 and 38.1±0.2 ºC), 3) Hot Skin: High Tskin and neutral Tcore (37.6±0.2 and 37.1±0.2 ºC) and 4) Hot Core: Neutral Tskin and High Tcore (35.0±0.5 and 38.3±0.2 ºC). LBNP tolerance was quantified via a cumulative stress index (CSI; LBNP level * time). Cutaneous vascular conductance (CVC) was measured throughout. LBNP tolerance (CSI) was not different between Hot Skin and Hot Core (569±151 vs. 563±194; P = 0.92), while both were greater than Combined (257±106; both P 蠄 0.05) and all were lower than Control (1010±246; all P 蠄 0.05). Prior to LBNP CVC was not different between Combined, Hot Skin and Hot Core (2.3±0.6, 2.2±0.5 and 2.1±0.7 AU/mmHg, respectively; all P > 0.05) but all were higher than control (0.6±0.4 AU/mmHg; all P蠄0.05). Relative to pre‐LBNP, the decrease in CVC at pre‐syncope was not different between Combined and Hot Skin (‐12±14 vs. ‐11±19 %; P = 0.90) but both were appreciably lower than Hot Core (‐65±9%; both P 蠄 0.05). These data indicate that tolerance to a simulated hemorrhagic challenge is similarly reduced by elevations in Tskin and Tcore alone despite large reductions in CVC with increased Tcore.
GPS2 at the crossroad of lipid metabolism and inflammation in mouse adipose tissue
Authors: Cederquist Carly , Lentucci Claudia , Cardamone Maria , Perissi Valentina
Published: June 2021

Obesity‐induced defects in insulin signaling lead to the development of insulin resistance and Type II Diabetes (T2D). Currently, in the United States, the large majority of adults are overweight or obese, with roughly 60 millions of Americans being at a significantly higher risk for several diseases including diabetes and hypertension. G‐protein pathway suppressor 2 (GPS2) has recently emerged as a novel and important regulator of homeostasis and inflammatory responses in metabolic organs. Previous work from our lab and others indicates that GPS2 is a multi‐functional protein shuttling between the cytosol, where it inhibits activation of the stress kinase JNK as a modulator of ubiquitin signaling downstream of TNFα receptor, and the nucleus, where it regulates gene expression at the level of gene transcription by inhibiting pro‐inflammatory gene targets and activating key mediators of the lipolysis pathway. Characterization of adipose tissue‐specific knock‐out and overexpression mice will be presented to confirm the critical role played by GPS2 in the regulation of obesity‐induced inflammation and lipid metabolism in vivo. Interestingly, our results suggest a complete and unexpected uncoupling between systemic insulin sensitivity and presence of immune cells and pro‐inflammatory markers. Thus, indicating that a better understanding of the causal relationship between insulin resistance and chronic inflammation is required for the design of most effective therapeutic approaches against T2D.
CALCIUM SENSING RECEPTOR DELETION IN THE MOUSE ESOPHAGUS ALTERS THE EXPRESSION OF CELL‐CELL JUNCTIONAL PROTEINS
Authors: Abdulnour‐Nakhoul Solange , Tu Chia‐Ling , Que Jianwen , Islam M Toriqul , Brown Karen , Nakhoul Nazih
Published: June 2021

Calcium sensing receptor (CaSR) is the molecular sensor by which cells respond to small changes in extracellular Ca+2 concentration. It regulates Ca+2 homeostasis by modulating PTH secretion and Ca+2 reabsorption in the kidney. CaSR also plays a role in glandular and fluid secretion in the GI tract and regulates differentiation and proliferation of keratinocytes. CaSR is present in the esophageal epithelium, but its role in this tissue has not been defined. We deleted CaSR in the mouse esophagus by generating KRT5‐CreER;CaSRloxP/loxP compound mutants, where loxP sites flank exon 7 of CaSR gene. For lineage tracing of epithelial cells we bred‐in the reporter allele Rosa26R. Recombination was initiated with multiple tamoxifen injections. We demonstrated exon 7 deletion and Cre expression by PCR analysis of genomic DNA and LacZ staining. qRT‐PCR & Western blot (WB) analyses showed a significant reduction in CaSR mRNA & protein expression as compared to control CaSRloxP/loxP mice. Microscopic examination of esophageal tissues (H&E) did not show gross abnormalities. However, WB data revealed a significant reduction (~25%) in levels of adherens junction proteins E‐cadherin and β catenin and tight junction protein claudin‐1. Levels of Rac/Cdc42, small GTPase proteins involved in actin remodeling, were also reduced. Our data indicate that CaSR plays a role in regulating cell‐cell junctional complexes and is therefore important for the maintenance of the barrier properties in the esophagus.

Support: VA Merit & Tulane Enhancement Award
PTPN2 Gene Deficiency Leads to Increased Epithelial Permeability and Promotes STAT‐1‐dependent Claudin‐2 Expression
Authors: Krishnan Moorthy , Sayoc Anica , McCole Declan
Published: June 2021

Interferon gamma (IFN‐γ) is an inflammatory cytokine involved in inflammatory bowel disease (IBD), and is known to compromise the intestinal barrier. siRNA silencing of the IBD candidate gene protein tyrosine phosphatase non‐receptor type‐2 (PTPN2) in human intestinal epithelial cells (IEC) elevates expression of claudin‐2 causing increased cation permeability of tight junctions. The aim of this study was to identify the cellular and molecular mechanisms underlying claudin‐2 expression in the absence of PTPN2. We developed a HT‐29 IEC line stably expressing short hairpin RNA for PTPN2 using a lentiviral system to knockdown PTPN2 expression. RT‐PCR and densitometry analysis indicated that PTPN2 expression was significantly reduced (60 ± 4%, p<0.0001) in PTPN2‐depleted cells, paralleled with an increased expression in claudin‐2 protein (180 ± 4%, p<0.0005). Western blot and immunofluorescence studies revealed that basal phosphorylation of the PTPN2 substrate, STAT‐1, as well as claudin‐2 protein expression, were significantly increased in PTPN2 KD cells. This effect was potentiated following IFN‐γ treatment. siRNA silencing of STAT‐1 decreased claudin‐2 protein expression in PTPN2 KD cells. There was also a significant increase in in vivo intestinal permeability to FITC‐Dextran (4 kD) in 3‐week old Ptpn2 knockout (298 ± 5%, p<0.0001) and heterozygote mice (206 ± 12%, p<0.001) compared to wild‐type mice. STAT‐1 phosphorylation and claudin‐2 protein expression were increased in Ptpn2 KO mouse cecum vs. wild type. In summary, PTPN2 deficiency leads to increased intestinal permeability and elevated claudin‐2 expression through a STAT‐1‐dependent pathway.
Cinobufotalin Inhibits Ovarian Cancer Cells Proliferation, Migration and Invasion
Authors: Afroze Syeda , Co Mary Antonette , McDowell Anthony , McCormick Timothy , Beeram Madhava , Zawieja David , KUEHL Thomas , Uddin Mohammad
Published: June 2021

Objective

Cinobufotalin (CINO), a cardiotonic steroid, is extracted from the skin secretions of the traditional Chinese medicine giant toads (Chan Su). CINO has been used as a cardiotonic, diuretic and a hemostatic agent. Recently, it has been shown that CINO inhibits the lung cancer cells function. In the present study, We evaluated the effect of CINO on ovarian cancer, SK‐OV‐3 cell function in vitro.

Study Design

Cells were treated with 0.1, 1, 5, and 10 μM CINO. Cell proliferation, migration and invasion were measured using kits. The effect of CINO on p38 phosphorylation was evaluated by a cellular activation of signaling ELISA kit. Cell cycle arrest and apoptotic signaling were determined by fluorescence‐activated cell sorting (FACS) analysis.

Results

CINO at 蠅 0.5 μM inhibited CTB cell proliferation, migration, and invasion (p < 0.05) but had no effect on cell viability. There was a higher (p < 0.05) percentage of G0/G1 phase cells in groups treated with CINO at 蠅 0.5 μM. CINO caused an increase in stress signaling p38 MAPK and a positive annexin‐V staining in SK‐OV‐3 cells, indicating the activation of apoptotic signaling.

Conclusion

This data demonstrates that CINO inhibits SK‐OV‐3 cell function via cell cycle arrest and apoptotic signaling. The results of this study suggest that CINO might be further investigated as a novel anti‐ovarian cancer agent.
Interleukin‐33/ST2 Axis Promotes Epithelial Cell Transformation and Breast Tumorigenesis via Upregulation of COT Activity
Authors: Choi Hong Seok
Published: June 2021

Cytokines of the interleukin‐1 (IL‐1) family, such as IL‐1α/β and IL‐18, have pleiotropic activities in innate and adaptive immune responses in host defense and diseases. Insight into their biological functions helped develop novel therapeutic approaches to treat human inflammatory diseases. IL‐33 is an important member of the IL‐1 family of cytokines and is a ligand of the ST2 receptor, a member of the IL‐1 receptor family. However, the role of the IL‐33/ST2 axis in tumor growth and metastasis of breast cancer remains unclear. Here, we demonstrate that IL‐33 is a critical tumor promoter during epithelial cell proliferation and tumorigenesis in the breast. IL‐33 dose‐ and time‐dependently increased Cancer Osaka Thyroid (COT) phosphorylation via ST2‐COT interaction in normal epithelial and breast cancer cells. The IL‐33/ST2/COT cascade induced the activation of the MEK‐ERK, JNK‐cJun and STAT3 signaling pathways, followed by increased AP‐1 and stat3 transcriptional activity. When small interfering RNAs of ST2 and COT were introduced into cells, IL‐33‐induced AP‐1 and stat3 activity were significantly decreased, unlike that in the control cells. The inhibition of COT activity resulted in decreased IL‐33‐induced epithelial cell transformation, and knockdown of IL‐33, ST2 and COT in breast cancer cells attenuated tumorigenicity of breast cancer cells. Consistent with these observations, ST2 levels were positively correlated with COT expression in human breast cancer. These findings provide a novel perspective on the role of the IL‐33/ST2/COT signaling pathway in supporting cancer‐associated inflammation in the tumor microenvironment. Therapeutic approaches that target this pathway may, therefore, effectively inhibit carcinogenesis in the breast.
The Different Substrate Specificities Of Human Influenza Virus PA And PA‐X Endonucleases Support Distinct Roles In The Viral Life Cycle
Authors: Maga Giovanni , Cucuzza Stefano , Bavagnoli Laura
Published: June 2021

The PA subunit of the RNA dependent RNA polymerase of influenza virus is an endonuclease required for the cap‐snatching mechanism leading to the synthesis of viral mRNA. Recently, the PA‐X protein has been discovered in human influenza, which is translated from an alternative open reading frame within the PA gene coding sequence, resulting from a ribosomal frameshifting. The PA‐X retains the N‐terminal endonucleolytic domain of PA, but has a different and shorter C‐terminal part. It has been proposed that PA‐X degrades cellular mRNA, contributing to the attenuation of viral pathogenesis in the host. In human isolates from the 2009 pandemic H1N1 virus, a naturally truncated form of PA‐X is present. Here we express the recombinant PA and PA‐X proteins, the latter both in its full length and deleted form, from human seasonal influenza virus and show that their endonucleolytic activities have distinct substrate specificities, which depend on the structure of the RNA to be cleaved. Collectively, our results support distinct roles of PA and PA‐X in the viral life cycle.
Efficient Isolation, Expression and Improvement Technique System for Feed Enzymes in China
Authors: Yang Peilong , Yao Bin
Published: June 2021

Supplementation of hydrolytic enzymes in feed can decrease the effect of anti‐nutritional factors and increase feed conversion rate by 5‐10%. Consequently, this practice can improve the efficiency of livestock production and reduce environmental contamination. The authors have developed a highly efficient platform to enrich the genetic resources of several enzymes, characterize and assess their application potentials, and achieve high‐yield production. These works make the production of novel enzymes cost‐effective and commercially valuable.

Firstly, a new technology system was set up, which can directly clone full‐length genes and screen objective genes from environmental metagenomic or transcriptomic nuclear acid. Using these methods, 184 genes coding for enzymes have been identified in short time. The enzymes showed different properties including high catalytic efficiency, thermophilic, acidophilic, or protease‐resistance. Secondly, great contributions were made in the structure and function relationship research of enzyme proteins. An efficient screening vector system was constructed based on extracellular fusion mutant proteins, optimized the protein engineering system for enzyme improvement, and obtained several improved enzymes with excellent properties and application potentials. Third, another great finding was identification of several key factors in Pichia expression system and verification of their function. Based on these knowledge, a high‐yield expression system of 5‐50 mg/mL was developed for large‐scale, cost‐effective production of enzymes, including phytase, xylanase, glucanase, mannannase, and α‐galactosidase.
Microbial Community Responses to Toxic Hydrocarbon Exposure
Authors: Morais Daniel , Pylro Victor , Tótola Marcos
Published: June 2021

The microbial community within each living organism is essential for its healthy state. Even highly complex matrices like soil, depend on microorganisms to sustain and even improve plant development. Although it is a very robust and well‐equilibrated system, soil microbial populations are prone to changes of abundance caused by environmental changes. These community shifts may result in loss of groups related to biogeochemical cycles or soil structure. This work aimed to compare changes in soil microbial community driven by the amendment with crude oil. We used microcosms of two soils from Rothamsted Research UK agricultural station, being one a bare fallow soil, one a grassland soil and a pristine beach soil (tide line under Cyperus atlanticus growth) from Trindade Island ‐ Brazil. Soils were contaminated with crude oil at a concentration of 5% (w/w) and incubated at 26 °C for 20 days. We monitored CO2 emissions and extracted total DNA and total RNA. The DNA obtained was used for microbial profiling using the protocol suggested by Earth Microbiome Project (http://www.earthmicrobiome.org). The bioinformatics pipeline was executed as recommended by the Brazilian Microbiome Project (http://www.brmicrobiome.org).
The preliminary results showed that the three environments are highly divergent about its microbial community even when they presented very similar CO2 emissions. The pristine soil presented more archeal individuals and a bigger community shift between the treated and non‐treated state, a sign of fragility. As future research we intent to evaluate the expression of genes related to soil services and related to degradation of crude oil hydrocarbon.

We are thankful to Rothamsted Research for the support and to our funding agencies CAPES and CNPq.
Reconstitution of Adiposome and Artificial Lipid Droplets
Authors: Liu Pingsheng , Wang Yang , Zhou Xiao‐ming
Published: June 2021

The Lipid droplet (LD) is a cellular organelle that consists of a neutral lipid core with a phospholipid monolayer membrane associated with proteins. The presence of this organelle in organisms ranging from bacteria to humans demonstrates that it is highly conserved and may be the first membrane‐bound organelle in living organisms. Recent progress in LD research demonstrates its important roles in energy homeostasis and metabolic syndromes. However, the mechanisms governing its formation, dynamics, interaction with other cellular organelles, and association with proteins have remained elusive. Therefore, we developed an in vitro system to facilitate the elucidation of these mechanisms, based on the reconstitution of artificial LDs. First, we generated sphere‐shaped structures with a neutral lipid core and phospholipid monolayer membrane by mechanically mixing neutral lipids and phospholipids followed by two‐step purification. Since the artificial lipid vesicle, a double layer phospholipid membrane structure, is termed as “liposome”, we named this monolayer phospholipid membrane‐bound structure “adiposome”. We then recruited LD structure‐like/resident proteins to the adiposome, including the bacterial LD protein MLDS, C. elegans LD protein MDT‐28, or mammalian LD protein ADRP/PLIN2. We term the functional protein‐coated adiposomes, “artificial lipid droplets”. Using this experimental system, different proteins can be recruited to build artificial LDs for myriad biological and medical goals.
Suppression of ACSL6 in Skeletal Muscle Induces Alteration of Glucose Metabolism
Authors: Bu So Young
Published: June 2021

Understanding the role of various sources of fatty acids and genes involved in fatty acid metabolism provide core information how nutrition therapy can be applied to the people who has different metabolic trait or genetic variance in susceptibility of metabolic diseases. Numerous isoforms of long chain acyl‐CoA synthetase (ACSL) mediate intracellular fatty acids channeling and regulate energy metabolism. However the role of these genes in skeletal muscle need to be identified in order to understand how they contribute to whole body energy metabolism. The purpose of this study was to identify a role of ACSL6 in skeletal muscle energy metabolism. C2C12 mouse myoblasts were cultured and expression of genes that involving glucose and fatty acids metabolism were analyzed. Also siRNA mediated knockdown of ACSL6 was performed to characterize the role of ACSL6 in skeletal muscle. In mouse skeletal muscle cells mRNA and protein expression of ACSL6 were increased over the time course of cellular differentiation. ACSL6 was up‐regulated by high‐glucose but its expression was not influenced by insulin treatment. Knockdown of ACSL6 decreased glucose without affecting insulin signaling or mitochondrial beta oxidation suggesting that ACSL6 is involved in glucose uptake or utilization independent of early stage of insulin signaling. On‐going study to investigate the effect of tissue specific knockdown of ACSL6 on glucose uptake and insulin signaling pathway in animal model will provide further insight into the role of ACSL6 in skeletal muscle.

This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education, Science and Technology (2012R1A1A1019253
miR‐142‐3p is a regulator of the TGFβ‐mediated vascular smooth muscle cell phenotype
Authors: Kang Hara , Kim Sunghwan
Published: June 2021

The transforming growth factor β (TGFβ) signaling pathway is critical for the promotion and maintenance of the contractile phenotype of vascular smooth muscle cells (VSMCs). Though multiple microRNAs (miRNAs) implicated in the regulation of the VSMC phenotype have been identified, the modulation of miRNAs in the VSMCs by TGFβ signaling has not been fully described. In this study, we identified microRNA‐142‐3p (miR‐142‐3p) as a modulator of the VSMC phenotype in response to TGFβ signaling. We showed that miR‐142‐3p is induced upon TGFβ signaling, leading to the repression of a novel target, dedicator of cytokinesis 6 (DOCK6). The downregulation of DOCK6 by miR‐142‐3p is critical for cell migration. Thus, this study demonstrates that miR‐142‐3p is a key regulator of the TGFβ‐mediated contractile phenotype of VSMCs that acts through inhibiting cell migration through targeting DOCK6.
Anti‐Inflammatory Dietary Intervention Selectively Improves Insulin Sensitivity in Metabolically Unhealthy Overweight Adolescents
Authors: McMorrow AM , Connaughton RM , McGillicuddy FC , Healy ML , Roche EF , Lithander FE , Roche HM
Published: June 2021

Anti‐inflammatory nutritional approaches may attenuate obesity‐induced insulin resistance; however results from clinical studies are inconsistent. Baseline metabotype may partially discriminate responders from non‐responders.

This 8‐wk randomized, placebo‐controlled, cross‐over trial assessed the determinants of inter‐subject variability in response to an anti‐inflammatory supplement containing LC n‐3 PUFA, vitamins C, E and polyphenols in overweight adolescents (n=58; age 15.9±1.6y; BMI 32.1±6.5kg/m2). Subjects who demonstrated >12.5% improvement in HOMA‐IR were considered responders.

Anti‐inflammatory supplementation selectively reduced HOMA‐IR in 38% of subjects (responders; supplement ‐33.01±17.84% v placebo 14.57±54.91%, p<0.001). In comparison with non‐responders, responding subjects demonstrated an adverse pre‐treatment metabotype characterized by increased HOMA‐IR, total and LDL cholesterol and soluble CD163 (p<0.05), despite similar BMI. Additionally, changes in adiponectin concentration significantly predicted HOMA‐IR response to intervention (p=0.04). PBMC mRNA expression of adiponectin receptor 2 increased in responders only (p=0.03).

Despite similar BMI to non‐responders, the insulin resistant and dyslipidemic phenotype of responders enhanced the impact of the anti‐inflammatory intervention. This illustrates potential efficacy optimization within the context of personalized nutrition.

Funding

National Children's Research Centre, Ireland and SFI 11/PI/1117
Femoral Condyle Curvature and the Chondral Modeling Theory
Authors: Harris Emma , Sylvester Adam
Published: June 2021

The chondral modeling theory argues that the cartilage of a developing joint surface responds to mechanical loading during ontogeny, resulting in regions of low curvature where the joint is heavily loaded. This theory has been used to explain the relatively lower curvature exhibited by the anterior portions of human femoral condyles. In contrast, it has been argued that the femoral condyles of non‐human primates have constant curvatures, potentially in conflict with the chondral modeling theory, although this has never been rigorously quantified. The goal of this study was to measure the curvature across the medial femoral condyles of chimpanzees and gorillas, comparing them with humans.

Three‐dimensional surface models of human (n=9), chimpanzee (n=9) and gorilla (n=9) femora were created using a white light surface scanner. Landmarks were collected on the attachment point for the collateral ligaments and across the femoral head. These points were used to estimate functional/mechanical axes of the femur which were then used to align all bones to a standardized orientation. The profile of each medial condyle, relative to the axis of knee flexion/extension, was extracted from digital images of the models and curvature was quantified across the profile.

Measurements of curvature across the human profile match previous work, demonstrating a region of low curvature at the anterior portion of the condyle. While gorilla femora did not appear to exhibit an obvious low curvature region, the chimpanzee medial femoral condyle was found to show a consistent region of relatively low curvature. Furthermore, in support of the chondral modeling hypothesis, the position of low curvature regions appears to be associated with knee positions used during the stance phase of chimpanzee walking.
A Novel Infant Nutrition System Precisely Delivers Hygienic Reconstituted Formula
Authors: Yan Jian , Pecquet Sophie , Wyss Heinz , Steenhout Philippe
Published: June 2021

Powdered infant formula is conventionally supplied in multi‐serve packaging and reconstituted following multiple manual preparation steps. An advanced infant nutrition system, combining a dedicated dispenser with single‐serve capsules of powdered formula, was designed to precisely deliver hygienic reconstituted formula in < 1 min. To test the performance (technical reliability and quality of reconstituted formula), a field study was conducted at three hospitals in Switzerland with a prototype system, where hospital staff used the system (n = 4) to reconstitute formula when needed by infants. Only 22 (0.9%) of the 2445 bottles prepared over the 4‐mo period encountered preparation difficulties. After reconstitution, 93% of the 2445 capsules showed no solid residue, and 5% had only a few small lumps, suggesting good solubility. Additional preparations were conducted in hospitals to assess the dilution precision and microbiological quality of reconstitutes. The target for reconstituted formula weight (±3g) was met by 89% of the 252 bottles tested, suggesting good technical precision of formula dilution. 100% of the reconstituted formula tested negative for Enterobacteriaceae (n=281) and Salmonella (n=262), and Aerobic Mesophilic Count was <100 cfu/ml in 83% and <1000 cfu/ml in 98% of 276 bottles, which met CODEX good quality specification after reconstitution. Interviews with hospital staff (n=74) revealed that the system was preferred over conventional formula preparation partially due to its ease of use. In sum, these data indicate that the new infant nutrition system ensures a hygienic, safe, precise and convenient preparation for each feeding.

Funded by Nestec S.A.
Modulation of adipogenesis using citric acids cycle metabolites and vitamin C via controlling TET protein activity
Authors: Yoo Yeongran , Park Joo Hyun , Park Yoon Jung
Published: June 2021

Adipogenesis refers to the cellular differentiation of preadipocyte into adipocyte in the presence of adipogenic stimuli. In this study, we showed the roles of vitamin C and two citric acids cycle (CAC) metabolites, succinate and fumarate, during 3T3‐L1 adipocyte differentiation. TET proteins are Fe(II) and α‐ketoglutarate (α‐KG) dependent dioxygenases, which can convert 5‐methylcytosine (5‐mC) to 5‐hydroxymethylcytosine (5‐hmC) in DNA. They have been described as key epigenetic modulators for cellular development by removing DNA methylation in locus specific and genome‐wide levels and generating 5‐hmC, an intermediate for oxidative demethylation. Vitamin C enhances TET protein activity presumably by maintaining the ferrous state of iron while succinate and fumarate inhibit its activity by competiting with α‐KG. We found that treatment of vitamin C in 3T3‐L1 cells enhanced TET protein activity while succinate and fumarate repressed it showing increased and reduced level of global 5‐hmC, respectively. Furthermore, we found that treatment of vitamin C directly promoted the adipogenesis by upregulating the Pparγ expression, a master regulator for adipogenesis via enhanced demethylation of Pparγ region. On the other hands, treatment of fumarate directly repressed the accumulation of lipid droplets by preserving the methylated status at the Pparγ region. Together, the results demonstrated that development of adipocyte may be epigenetically modulated by CAC metabolites and vitamin C via controlling the TET protein activity.
Effects of a Unique Vitamin C Ingredient on Chronic Knee Complaints
Authors: Mitmesser Susan , Zhang Zhiyling , Ye Qian
Published: June 2021

Background

Chronic knee complaints are very common in athletes. Overuse injury can cause pain in the patellar tendon. Vitamin C is required for collagen synthesis, which is an important component of tendons. It has been reported that vitamin C can suppress cytokine production, which is indicative of an inflammatory response. Ester‐C®, calcium ascorbate plus vitamin C metabolites, has been clinically studied and may have a higher biological availability in body tissues than ascorbic acid.

Objective

To examine the effect of Ester‐C® on the improvement of injury complaints in athletes with patella tendonitis (also known as “jumper's knee”).

Methods

Basketball players with patella tendonitis were recruited to participate in this study. Each athlete received Ester‐C® in increasing daily doses for 6 weeks (Week 1: 1000 mg, Week 2: 2000 mg, Weeks 3‐6: 3000 mg). Improvement scores of six complaints ‐ pain, power, mobility, speed, jumping ability, and general condition ‐ were assessed on a scale of 0‐3 (0=no improvement; 1=noticeable improvement; 2=full improvement at rest; 3=full athletic improvement). The percentage of subjects that showed improvement in each category and the average improvement scores (AIS) were calculated.

Results

A total of 16 basketball players completed the study. Most complaints started to show improvement after Week 2. Dramatic improvements were observed in all injury criteria during Week 5 and were maintained at the end of Week 6. The greatest improvements were in pain reduction (AIS 1.75) and jumping ability (AIS 1.5), with the least improvement observed in mobility (AIS 0.38).

Conclusion

Ester‐C® has a positive effect on chronic knee complaints caused by overuse in high‐impact sports, such as basketball.
Dynamic imaging of the growth plate cartilage reveals multiple contributors to skeletal morphogenesis
Authors: Li Yuwei , Trivedi Vikas , Truong Thai , Fraser Scott
Published: June 2021

The diverse morphology of vertebrate skeletal system is genetically controlled, yet the means by which cells shape the skeleton remains to be fully illuminated. Here, we perform a quantitative analysis of cell behaviors in the growth plate cartilage, the template for long bone (limb skeleton) formation, to gain insights into this process. The proliferative zone (PZ) is comprised of elliptical chondrocytes aligned perpendicular to the proximal‐distal axis (PDA) of the tissue, suggesting some coordination between oriented cell behaviors and oriented tissue growth. Using a robust avian embryonic organ culture, we employ time‐lapse two‐photon laser scanning microscopy to observe PZ cell behaviors during cartilage elongation in 4D (xyz and t). These time‐lapse movies reveal distributed growth in the PZ, resulting in cell trajectories with a spreading displacement mainly along the PDA. We build a novel software toolkit of quantitative methods to segregate the contributions of various cellular morphogenetic processes that contribute to the cell trajectories. Surprisingly, convergent extension (CE) and cellular column formation are not the major contributors of the stages examined; instead, the analyses reveal important roles of extracellular matrix (ECM) deposition and cell volume enlargement.
Nutritional Composition of Complementary Infant Foods: Evidence from 109 Commercially Marketed Products in 22 Low‐Income Countries
Authors: Bell Winnie , Nene Marc , Masters William
Published: June 2021

To sustain healthy growth, infants from about 6 to 23 months of age need to complement breastmilk with solid foods that are more nutrient‐dense and prepared more frequently during the day than the family diet. Many products can be used as complementary foods, but the most widely consumed and nutritionally cost‐effective products are composite flours used for porridge, typically made from a starchy staple (e.g. maize, rice or wheat) and mixed in fixed ratios with a legume flour (e.g. roasted soya beans) and other ingredients. The global market for composite‐flour infant foods is dominated by one multinational brand, despite the fact that many competitors actively produce alternatives at the same or much lower cost. This study provides the first global study of such products, including laboratory tests of the actual nutrient content for 109 samples purchased in 22 countries across Africa, Asia and Latin America. We find the nutrient content of locally‐made products are highly variable with many products offering nutritional profiles comparable to international standards, while others have much lower density of both macronutrients and micronutrients. This finding helps explain why caregivers rarely buy products other than heavily advertised multinational brands, whatever their cost and despite the high quality of some local alternatives. Third party certification could help sellers signal quality and expand access to safe, nutritious and affordable complementary foods across the developing world.
The Role of Male Closeness on Black Adolescent Male Weight Status in Father‐Present Versus Male‐Kinship Households
Authors: Coleman Ailton , O'Neil James , Asencio Marysol , Caldwell Cleopatra , Ferris Ann
Published: June 2021

Black males from the ages of 12‐19 years olds are the only adolescent group in the United States to experience a significant increase in obesity rates from 1999‐2010. This rise translates into an increase in the odds of obesity prevalence of 1.05 annually. This study used the National Survey of American Life‐Adolescent, a nationally representative sample of African American and Caribbean Black adolescents (n=434), to examine the relationship between male closeness and BMI in father‐present and male‐kinship households among Black adolescents. Obesity levels were similar between father‐present (19%) and kinship households (20%). However, a greater percentage of adolescent males in male‐kinship homes (68%) were normal weight than in father‐present homes (60%). Male‐kinship homes reported higher percentages of male closeness (49%) in comparison to father‐present homes (46%). Ethnic differences were not present between households. In the initial bivariate analysis, Paternal Closeness Scores were negatively correlated with BMI in adolescent males who reported excellent relationship with a male in their life (p=.05). Yet, after accounting for age, household income, ethnicity, and participation in sports, higher male closeness was only predictive of lower BMI in kinship homessage (p=<.01). Family structure has a macro level role on obesity in Black adolescent males. Future interventions among Black adolescents should focus on male bonding as a way to reduce obesity levels.
Semaphorin 6A Mediates Sensory Afferent Projection into the Developing Spinal Cord
Authors: Barry Denis , O'Malley Aisling , Kelly Albert , O'Sullivan Ruari , Mitchell Kevin
Published: June 2021

During vertebrate development, sensory axons project from the peripheral nervous system (PNS) into the spinal cord at the dorsolateral margin via the dorsal roots. They then ascend to the brain in separate tracts depending on sensory modality. This segregation of sensory axons is essential for appropriate perception and reflex activity. We show that radial glial cells support the formation of these tracks by forming scaffolds which channel various axon fasciculi through the spinal cord along the rostrocaudal axis. Semaphorins are the largest family of axon guidance cues expressed during central nervous system (CNS) formation. Class 6 semaphorins are found throughout the developing CNS and play vital roles in the guidance of growing axons in the brain, yet their roles in the spinal cord are relatively unknown. We investigated the role of Sema6A and its canonical receptor PlexinA2 during spinal cord development by comparing the organisation of radial glia and axon tracts in WT, Sema6A‐/‐and PlexinA2‐/‐ mice. This study shows that Sema6A is inhibitory to TAG‐1 expressing sensory axons entering the spinal cord from the dorsal roots and is a key regulator of sensory axon growth through the spinal cord during development. This effect is independent of its receptor PlexinA2. Sema6A is expressed on boundary cap cells in the dorsal root ganglion and the phenotype described here suggests that it acts as a barrier between the CNS and the PNS. The axonal growth guidance properties of Sema6A and the radial glial cell scaffold suggest they may be therapeutic targets for axonal regeneration following spinal cord injury.
MicroRNA‐122 (miR‐122) Regulates Postnatal Polyploidization in the Murine Liver
Authors: Hsu Shu‐hao , Otero Anthony , Meehan Kolin , Duncan Andrew
Published: June 2021

A defining feature of the mammalian liver is polyploidy, a numerical change in the complement of chromosomes. Hepatocytes are either mono‐ or binucleate, and ploidy is determined by the number of nuclei/cell, as well as the ploidy of each nucleus. The first step of polyploidization involves cell division with failed cytokinesis. Polyploidy is common, affecting ~90% of hepatocytes in mice and 50% in humans, but the specialized role played by polyploid cells in liver homeostasis and disease remains poorly understood. The goal of this study was to identify signals that regulate polyploidization, and we focused on microRNAs (miRNAs). First, to test whether miRNAs could regulate hepatic polyploidy we examined livers from Dicer1‐/‐ mice, which are devoid of mature miRNAs. Loss of miRNAs resulted in a 3‐fold reduction in binucleate hepatocytes, indicating that miRNAs could indeed regulate polyploidization. Secondly, we surveyed age‐dependent expression of >500 miRNAs (NanoString) in wild type mice and identified a subset of miRNAs, including miR‐122, differentially expressed at 2‐3 weeks, a period when extensive polyploidization occurs. Next, we examined Mir122‐/‐ mice and observed profound, life‐long depletion of polyploid hepatocytes, proving that miR‐122 is required for complete hepatic polyploidization. Finally, we showed that miR‐122 antagonizes pro‐cytokinesis genes, Iqgap1, Mapre1, Nedd4l and Slc25a34, which promoted cytokinesis failure and emergence of binucleate, polyploid hepatocytes. In summary, our data suggest a novel regulatory role for miR‐122 in liver polyploidization. These studies will serve as the foundation for future work investigating miR‐122 and polyploidy in liver diseases. Support provided to AWD from the NIH (R01DK103645) and Commonwealth of PA.
Neutrophilia Delays Equine Neutrophil Apoptosis
Authors: Anderson Stacy , Singh Baljit
Published: June 2021

Regulation of neutrophil apoptosis has assumed a critical role in inflammation. The objective of this study was to determine the effect of neutrophil concentration on their apoptosis in vitro and ex vivo in horses with experimental intestinal ischemia and reperfusion injury. Neutrophils were isolated from healthy horses randomized to the following treatments: sham celiotomy (CEL, n=4), intestinal ischemia and reperfusion (IR, n=6), intestinal ischemia and reperfusion with gadolinium chloride treatment to deplete pulmonary intravascular macrophages (IRGC, n=6). All horses experienced a significant neutrophilia following surgery without detectable plasma endotoxin concentrations (<0.06 EU/ml). Isolated neutrophils were cultured for 12 or 24 hours at concentrations of 2, 5, or 8 x 106 cells/ml and evaluated for the occurrence of apoptosis using Annexin V and propidium iodide staining. Caspase‐3, ‐8, and ‐9 activities were measured in cell lysates before and after treatment. Delayed neutrophil apoptosis was highly correlated (12 h incubation: r = ‐0.75, 24 h incubation: r = ‐0.57) with increasing neutrophil concentration in vitro. Neutrophil apoptosis was significantly delayed ex vivo after treatment, except in IRGC horses (12 h incubation: CEL: P =0.03, IR: P = 0.05;24 h incubation: CEL:P = 0.001, IR: P = 0.004). Caspase‐3, ‐8, and ‐9 activities were reduced in neutrophils isolated after surgery in all groups. The data show that equine neutrophil apoptosis is delayed in vitro and ex vivo via down‐regulation of caspase activity when neutrophils become more concentrated, which could have implications for the development and progression of systemic inflammatory responses.
Induction of Mindin is Associated with Pathological Changes in the Kidney of Cd151‐/‐ Mice
Authors: Naudin Crystal , Weidenhofer Judith , Roselli Severine , Keely Simon
Published: June 2021

The presence of CD151, a transmembrane protein involved in cell adhesion complexes with α3β1 integrins, is essential for glomerular function. We have identified significant increases in mindin in the glomeruli of CD151 knock‐out mice (Cd151‐/‐)compared to wild‐types (Cd151+/+). Due to mindins known role in actin cytoskeleton and cell adhesion signalling, we hypothesised that aberrant expression of mindin in the glomerulus leads to the abnormal extracellular matrix deposition, and disruption to podocyte actin cytoskeleton structure, which occurs in this disease model.

Immunofluorescent labelling of mindin in 3‐week‐old Cd151+/+ and Cd151‐/‐ kidneys demonstrated specific accumulation in Cd151‐/‐ glomeruli, (absent in Cd151+/+). No expression of mindin was observed in 5 day old mice (when Cd151‐/‐ glomeruli first exhibit pathology) indicating it is unlikely to be involved in initial pathology.

As mindin is also an immune cell activator, we examined whether the induction of mindin leads to immune‐mediated glomerular injury as part of disease progression in the Cd151‐/‐ kidney. We also investigated mindin as a potential biomarker of glomerular disease in the broader sense and characterised immune cell infiltrates in Cd151‐/‐ kidneys.

Infiltration of lymphocytes, neutrophils, macrophages and eosinophils was observed as early as 3 weeks of age in Cd151‐/‐ but not Cd151+/+ kidneys by differential histological staining. This infiltration was progressive and more pronounced in 12 week old Cd151‐/‐ mice, associated with extensive inflammatory lesions. Together, these data suggest that in the absence of CD151, mindin is induced early in disease, associated with disruption to podocyte actin cytoskeleton homeostasis, and immune‐mediated glomerular damage.
The Effect of Croton lechleri on Prostate Cancer Progression
Authors: Glasscock Laura , Webb Stefanie
Published: June 2021

Prostate cancer (CaP) continues to be the second leading cause of cancer deaths in men in the US (Siegel 2014). Many of the cancer drugs currently on the market are from or are structurally similar to compounds found in natural plant products. We investigated the effect of a tree bark native to the Peruvian Amazon, Croton lechleri on the proliferation and invasion of two prostate cancer cell lines, DU‐145 and PC‐3. C. lechleri is commonly used by Peruvian healers to treat prostate conditions and is one of the most popular treatments purchased from herbal markets (Bussmann 2007). C. lechleri has antioxidant capabilities as a free radical scavenger (DeMarino 2008) and species in the same genus as C. lechleri decrease cell adhesion and induce apoptosis in stomach and colon cancer cell lines (Sandoval 2002). We used C. lechleri latex extracts, the form used by Peruvian Shaman for therapeutic use, to treat PC‐3 and DU‐145 CaP cell lines and evaluate their ability to proliferate using CellTiter 96® and invade using a modified Boyden Chamber. C. lechleri, at concentrations equal to and less than those used as Peruvian therapeutic treatments, significantly reduced proliferation of both PC‐3 and DU‐145 cells in a dose dependent manner. C. lechleri reduced invasion of DU‐145 cells but not the more invasive PC‐3 cells. Studies of the effect of C. lechleri on cancer progression have been contradictory, depending on the type of cancer under investigation. Here, we describe that in prostate cancer, C. lechleri decreases both the ability of prostate tumor cells to proliferate and invade. We are currently investigating the effect of C. lechleri on CaP cell apoptosis.

This project was funded NIH‐INBRE Grant from the National Center for Research Resources and Winthrop University Research Council
Using Protein Microarray Technology to Screen Anti‐E‐Cadherin (ECAD) Monoclonal Antibodies for Specificity and Applications in Pathology
Authors: Chen Caiwei , Wei Haitao , Yuan Kehu , Ma Donghui , He Wei‐wu
Published: June 2021

E‐cadherin (ECAD) is a calcium‐dependent cell‐cell adhesion molecule playing a crucial role in establishing epithelial architecture and maintaining cell polarity and differentiation. Reports showed that decreased ECAD expression has been associated with more advanced tumor stage and grade for lung tumors, gastric tumors, breast tumors and etc. Other research discovered that staining for ECAD may be useful for the differentiation between ductal and lobular breast carcinomas. Thus, a high quality monoclonal antibody validated for immunohistochemistry (IHC) is required to evaluate ECAD protein levels in formalin‐fixed paraffin‐embedded tissue samples.

To develop the a suitable monoclonal antibody for ECAD IHC analysis, 49 monoclonal antibodies were generated, of which 10 were selected for screening in IHC on breast and prostate cancer tissues. Further evaluation with a high density protein microarray chip containing 17,000 human proteins indicated that clone 3C11 is highly specific and only detected ECAD. Comparison to the monoclonal 4A2C7, showed that 3C11 is more sensitive on breast, prostate, and prostate cancer tissues. The new monoclonal antibody 3C11 will be a promising candidate to detect the expression and aberrant alterations of ECAD for the diagnosis, prognosis, and therapy of cancer.