bluuebunny/crossref_metadata_2025 huggingface dataset:
Tolerance and Cognitive Function Effects of a Proprietary Spearmint Extract in Men and Women with Self‐Reported Memory Impairment ‐ A Pilot Study (LB402)
Authors: Nieman Kristin , Herrlinger Kelli , Sanoshy Kristen , Schild Arianne , Kelley Kathleen , Maki Kevin , Ceddia Michael
Published: June 2021

This pilot trial assessed the tolerance of a proprietary extract from Mentha spicata L. and its effects on cognition in subjects with self‐reported memory impairment. Subjects (N = 11) consumed 900 mg of the extract daily for 30 d. Participants were 73% female and 27% male with a mean age and body mass index of 58.7 y and 27.4 kg/m2. Tolerance was assessed by evaluation of gastrointestinal (GI) tolerability, adverse events, and clinical laboratory tests at baseline and the end of treatment (EOT). Acute cognitive effects were evaluated by comparing pre‐dose vs. 0.5 to 4 h post‐dose values at baseline and EOT. Subjective cognition was also assessed at the EOT. Consumption of the extract for 30 d did not significantly alter GI symptoms and subjective cognition improved (3.5 vs. 4.0; P = 0.063). No serious adverse events or clinically relevant findings in the safety parameters were observed. Cognitive function task scores suggest improved reasoning (24.7 vs. 33.4 points; P = 0.023), attention/concentration (18.0 vs. 40.5 points; P = 0.002), and planning (29.8 vs. 44.1 points; P = 0.088) from baseline after 30 d of treatment. Day one, subjects experienced acutely improved attention/concentration from pre‐ supplementation vs. 2 h (63.4 vs. 82.4 points, P = 0.042 and 18.0 vs. 34.8 points, P = 0.025) and 4 h (63.4 vs. 92.5 points, P = 0.001 and 18.0 vs. 39.8 points, P = 0.002). These results suggest the extract was well‐tolerated and may improve certain aspects of cognitive function. Further investigation in a randomized, controlled trial is underway.

Grant Funding Source: Kemin Foods, L.C.
Exercise training modifies gut bacterial composition in normal and diabetic mice (LB434)
Authors: Lambert Jennifer , Bomhof Marc , Myslicki Jason , Belke Darrell , Reimer Raylene , Shearer Jane
Published: June 2021

Background: Intestinal microbiota are increasingly recognized as potential modifiers of whole‐body energy metabolism. Previous research has described changes in bacterial composition in diabetes; this study examines whether these differences can be altered with lifestyle treatment. Methods: Six‐week‐old diabetic (db/db) and control littermates (db+) were randomized into sedentary or exercise training groups for 6 weeks (n=9‐10/treatment). All animals consumed an identical, chow‐based diet. Exercise consisted of low‐intensity treadmill running (5d/week), and was ceased 48h prior to sacrifice. Cecal matter was collected from individual mice at sacrifice. Total bacterial DNA was extracted and quantified, and bacterial species were detected and profiled using qPCR and group specific primers. Genomic data is reported as the log transformed 16S rRNA copy number normalized for the amount of cecal matter analyzed. Results: Compared to db+ mice, db/db had higher Clostridium Difficile and lower Clostridium Perfringens (P蠄0.05). However, exercise training increased Clos. Perfringens levels in both db+ and db/db (P<0.001); in this way, exercise mitigated the diabetes‐induced reduction in Clos. Perfringens such that exercised db/db mice had levels (5.30 ± 0.09) similar to sedentary db+ (5.19 ± 0.06; P=0.30) and approached that of exercised db+ mice (5.49 ± 0.05; P=0.094). Exercise training independently reduced Bacteroidetes (P=0.009) and increased Clostridium Leptum (P蠄0.05) levels in both db+ and db/db animals. Finally, exercise training increased Bifidobacterium in db+ animals but not db/db mice (P<0.001 for interaction). Conclusion: Diabetes is associated with alterations in the gut bacteria profile, particularly in the Firmicutes species. Exercise training exerts independent effects on bacterial composition in both diabetic and non‐diabetic animals, and may rescue some diabetes‐associated alterations.
Methylated Medium‐ and Long‐Chain Fatty Acids Provide Novel Sources of Anaplerotic Carbon for Fasting or Exercising Mice (LB438)
Authors: Drayton Josephine , Lin Xi , Jacobi Sheila , Shanely R. , McIntosh Michael , Odle Jack
Published: June 2021

We hypothesized that methylated fatty acids (e.g. 2‐methylpentanoic acid (2MeP), phytanic or pristanic acids) would provide anaplerotic carbon to the tricarboxylic acid (TCA) cycle and thereby enhance fatty acid oxidation, especially under conditions when TCA cycle intermediates are depleted. The optimal dose of 2MeP and pristanic acid for increasing in vitro [1‐14C]‐oleic acid oxidation in liver or skeletal muscle homogenates from fasted mice (n=4) was assessed using doses of 0, 0.25, 0.5, 0.75, or 1.0 mM. The minimum dose (0.25 mM) maximally stimulated liver tissue oxidation of [1‐14C]‐oleate to 14CO2 by 2‐fold (P < 0.05), but no stimulation was detected in muscle. Similar incubations with 0.25 mM 2MeP, hexanoate, palmitate, phytanic acid, or pristanic acid or 0.1 mM malate or propionyl‐CoA were conducted with liver and skeletal muscle homogenates from acutely exercised or sedentary mice. Regardless of exercise, incubation of liver homogenates with 2MeP increased [1‐14C]‐oleate oxidation to 14CO2 by 60% over control incubations containing hexanoate (P < 0.05). Phytanic acid treatment increased [14C]‐acid‐soluble product accumulation in liver tissue as compared to palmitate (P < 0.05). Anaplerotic stimulation was unaffected by exercised state (P > 0.05) and not evidenced in muscle homogenates. Results were consistent with our hypothesis that methyl‐branched fatty acids may provide anaplerotic carbon and thereby stimulate in vitro fatty acid oxidation.

Grant Funding Source: North Carolina Agricultural Research Service and by USDA‐AFRI training grant # 2010‐65200‐20354
Proliferating fibroblasts suppress cancer cell epithelial‐to‐mesenchymal transition (EMT) and migration via control inhibition of p300 HAT activation by 5‐methoxytryptophan (LB492)
Authors: Cheng Huei‐Hsuan , Chang Tzu‐Ching , Chiang Li‐Yi , Kuo Cheng‐Chin , Wu Kenneth K
Published: June 2021

We recently identified 5‐methoxytryptophan (5‐MTP), a novel tryptophan metabolite, as an endogenous inhibitor of cyclooxygenase‐2 (COX‐2) expression. Our reported data show that cancer cells are defective in 5‐MTP production and exogenous 5‐MTP suppresses cancer cell COX‐2 overexpression. Furthermore, 5‐MTP inhibits cancer cell migration and invasion. However, it is unclear whether endogenous 5‐MTP controls cancer cell COX‐2 and COX‐2 mediated cancer growth and metastasis. Since human proliferative fibroblasts (pFb) produce robust 5‐MTP, we determined whether pFb controls cancer cell COX‐2 expression and behavior. pFb co‐cultured with A549 lung cancer cells in a two‐chamber system inhibited PMA‐induced COX‐2 expression and migration in a time‐ and cell number‐dependent manner. pFb blocked A549 epithelial mesenchymal transition (EMT) induced by transforming growth factor‐β (TGFβ). Hydroxyindole O‐methyltransferase (HIOMT) catalyzes the final step of 5‐MTP synthesis. pFb expressed abundant HIOMT proteins. Knockdown of HIOMT in pFb resulted in abrogation of control of A549 COX‐2 expression, migration and EMT in the two‐chamber co‐culture system. These results are consistent with control of A549 COX‐2 expression and EMT/migration by 5‐MTP. We next determined whether 5‐MTP targets p300 histone acetyltransferase (HAT) which is essential for COX‐2 transcriptional activation. Basal p300 HAT activity in A549 was stimulated by proinflammatory cytokines and phorbol esters, which was inhibited by 5‐MTP. 5‐MTP inhibited TGFβ. Co‐cultured with pFb suppressed A549 p300 HAT, TGFβ and COX‐2 which were abrogated by treating pFb with HIOMT siRNA. These findings suggest that fibroblasts control cancer cell COX‐2 expression via inhibition of p300 HAT and TGFβ activation which results in suppression of cancer cell EMT and migration. Fibroblast produced 5‐MTP may confer endogenous resistance to cancer growth and metastasis.
Chronic Liver Diseases Which Mallory‐Denk Body (MDB) Form Progress to Hepatocellular Carcinoma is Involved in Ufmylation and FATylation Modification (LB493)
Authors: Liu Hui , Li Jun , French Barbara A , French Samuel W
Published: June 2021

We previously reported the Mallory‐Denk body (MDB) formation mechanism in DDC induced mice. To further provide clinical evidence whether chronic liver diseases which MDB form progress to hepatocellular carcinoma (HCC) is involved in Ubiquitin‐like proteins (Ubls) modification, gene transcript expression of Ufmylation and FATylation were investigated in human archived formalin‐fixed, paraffin‐embedded (FFPE) liver biopsy and mice frozen liver section from DDC refeding. It showed that all Ufmylation molecules (Ufm1, Uba5, Ufc1, Ufl1 and UfSPs) were at least 50% down regulated both in DDC refed mice and patient liver except for HCC without MDB biopsy, indicating that gene transcript changes was limited in MDB‐forming cells for a signal of proteasomal degredation. Interestedly, FAT10 and subunits of the immunoproteasome (LMP2, LMP7 and MECL‐1) were all up regulated. Among which, a 45‐ and 7‐fold upregulation of FAT10 were observed in DDC refed mice liver and human alcoholic hepatitis with MDB biopsy respectively, implying an important role of this gene. The FAT10‐specific E1 enzyme UBA6, however, was found to down regulation both in patient and DDC refed mice liver. Our results constitute the first systematic demonstration of transcript regulation of Ufmylation and FATylation in liver patient which MDB form progress to HCC and mice MDB‐forming liver by DDC refeding, providing key clue for the investigation of human liver disease development and strategies for drug design.

Grant Funding Source: Supported by NIH(RO1 AA021898‐02)
Bordetella pertussis Fimbriae proteins Fim2 and 3 are Immunodominant in T cell responses to Acellular Pertussis Vaccine (LB509)
Authors: Dillon Myles , Kolla Ravi , Sidney John , Sette Alessandro
Published: June 2021

Objective: To map the response of T cells to the Bordetella pertussis acellular vaccine (aP).

Methods: PBMCs from boosted (within three years) and control volunteers were stimulated in vitro with B.pertussis proteins present in the aP. IFNγ and IL‐5 responses were finely mapped using ELISpot assays with a panel of 16‐mer peptides spanning the B.pertussis proteins.

Results: Boosted individuals had increased levels of IFNγ and IL‐5 secretion to fimbriae (Fim) 2 and 3, pertactin (PRN), and pertussis toxin (PTx) peptides, while responses to the filamentous hemagluttinin were lower and similar in both groups. The boosting effect induced by aP was most apparent at the level of IL‐5 responses. Within proteins present in the aP, responses to Fim2 and 3 in the boosted group were disproportionately large in relation to the number of peptides tested in comparison to PRN or PTx.

Conclusions: The increase in the IL‐5 secretion to Fim2 and 3 reveals that this response is dominant upon aP. Future studies will map the actual epitopes recognized, and compare T cell responses in natural infection to vaccines with and without Fimbriae, in terms of targets of the responses and phenotypes of responding T cells.

Grant Funding Source: Supported by: NIH HHSN27220300044C
In silico Assessment of Antihypertensive Potential of Sweet Proteins (LB542)
Authors: Rotimi Solomon , Rotimi Oluwakemi , Omotosho Elizabeth , Iyanda‐Joel Wisdom , Chinedu Shalom
Published: June 2021

The consumption of low‐calorie artificial sweeteners by patients affected by diseases linked to sugar consumption may be associated with diverse side effects. Hence, sweet proteins derived majorly from under‐utilized plants have been proposed as good replacements. The ability of sweet proteins to release Angiostensin‐Converting Enzyme (ACE)‐inhibitory peptides was investigated. ACE mediates arterial vasoconstriction and elevation of its activity is an important pathogenic mechanism of hypertension. The protein sequences of six sweet proteins, Thaumatin from Thaumatococcus danielli (NCBI accession number, gi|209473), Brazzein from Pentadiplandra brazzeana (NCBI accession number, gi|218218145), Monellin from Dioscoreophyllum cumminsii (NCBI accession number, gi|381144434), Madinlin from Capparis masaikai (NCBI accession number, gi|1817546), Curculin from Curculigo latifolia (NCBI accession number, gi|11225520) and Miraculin from Richadella dulcifica (NCBI accession number, gi|253735645) were selected for sequence alignment using Basic Local Alignment Search Tool (BLAST) analysis and biological activity search using BIOPEP. Although BLAST analysis gave no homologous similarity among the proteins, BIOPEP analysis showed that they demonstrated either di‐ or tri‐peptide with a total of 51, 14, 40, 28, 30 and 59 potential ACE inhibitory peptides from Thaumatin, Brazzein, Monellin, Madinlin, Curculin and Miraculin respectively. The combined digestion with pepsin, trypsin and chymotrypsin A, a simulation of human gastrointestinal digestion released 8, 2, 9, 2, 5 and 11 ACE inhibitory peptides from Thaumatin, Brazzein, Monellin, Madinlin, Curculin and Miraculin respectively. These results add value to these proteins by demonstrating their innate nutraceutical potential in their ability to reduce hypertension.

Table of Contents
Immunocharacterization of the atypical basal lamina at cell‐tooth interfaces in laminin‐332 deficient mice (LB548)
Authors: Wazen Rima , Moffatt Pierre , Adair‐Kirk Tracy , Nanci Antonio
Published: June 2021

Unlike elsewhere in the body, the atypical basal lamina (BL) at epithelial‐tooth interfaces must bind to mineral rather than connective tissue. Therefore, this BL has adapted by integrating specialized components. It is enriched in laminin‐332 (Lm‐332), and recently amelotin (AMTN), odontogenic ameloblast‐associated (ODAM), and secretory calcium‐binding phosphoprotein‐proline‐glutamine‐rich 1 (SCPPPQ1), members of the SCPP gene family, have been identified as novel constituents. To explore how these three small molecular weight proteins participate in BL structure and mediate cell‐mineral adhesion, we used a mouse model of Lm‐332 deficiency, which expresses a doxycycline‐controllable human Lm γ2 transgene under the cytokeratin 14 promoter on the Lm γ2 knockout background. Mandibles and maxillae from wild‐type and Lm‐332 deficient mice were paraffin‐embedded and sections were immunostained for AMTN, ODAM, or SCPPPQ1. In the maturation stage of amelogenesis, all three proteins localized very discretely in the BL at the ameloblast surface of wild‐type mice. However in Lm‐332 deficient mice, all three proteins accumulated as focal patches within the cell layer rather than at the cell apex. Thus, it is concluded that AMTN, ODAM, and SCPPPQ1 require Lm‐332 for their proper positioning in the BL and for mediating adhesion to the tooth surface. As such, the defects in teeth of patients with junctional epidermolysis bullosa resulting from mutation in Lm‐332 likely reflect altered interactions among these proteins.

Grant Funding Source: Supported by CIHR, NIH/NHLBI P01HL029594, Shriners of North America
Synergistic effect of palmitate and nicotine treatments in diabetic heart (LB560)
Authors: Arunachalam Gnanapragasam , Samuel Samson , Ghosh Suparna , Ding Hong , Machaca Khaled , Triggle Chris
Published: June 2021

Background: Metabolic disorders (hyperglycemia, dyslipidemia, hypertension) contribute to the development and progression of cardiovascular disease in individuals with type 2 diabetes. Risk factors such as obesity and cigarette smoking are also strongly associated with exacerbation of diabetes‐associated heart failure. Obesity is associated with hypertriglyceridemia and elevated circulating free fatty acids (FFA), result in endothelial dysfunction and heart failure. Recent findings have demonstrated that alterations in cardiac energy metabolism are primarily connected with the development of cardiomyopathy in the diabetic heart.

Methods: In the current study, the synergistic effects of FFA flux and nicotine treatment were investigated in‐vitro, using endothelial (MMECs), myoblast (C2C12) and cardio myoblast (H9C2) cell culture models under hyperglycemic conditions.

Results: Our data indicate that cells treated with palmitate/nicotine showed increased E2F1 transcription with altered PPAR‐α expression‐a transcription factor involved in lipid metabolism. In addition, combined effects of palmitate/nicotine on Rb‐phosphorylation and activation of cyclin‐dependent kinases (CDKs) and their cyclin partners (A/E/D), the upstream regulators of E2F1‐transcription were also studied. Conclusion: Our data demonstrate that elevated FFA and nicotine change PPAR‐α/CDK/E2F1 gene expression and promote apoptosis and genomic instability thus revealing a potential molecular mechanism in obesity and cigarette smoking‐associated heart failure in diabetic milieu.

Grant Funding Source: This project was funded by grant NPRP 9‐413‐3‐104
Singular and combined effects of nebivolol and lifestyle modification on large artery stiffness in hypertensive adults (LB567)
Authors: Werner Tim
Published: June 2021

Background: We hypothesized that the combination of nebivolol and lifestyle modification would reduce large artery stiffness in middle‐aged and older hypertensive adults more than either intervention alone.

Methods: To address this, 45 men and women (age 40‐75 years) with stage I hypertension were randomized to receive either nebivolol (NB; forced titration to 10 mg OD; n = 15; age 57.2 ± 11.4 years; body mass index [BMI] 30.8 ± 5.8 kg/m2), lifestyle modification (LM; 5‐10% weight loss via calorie restriction and physical activity; n = 15; age 52.7 ± 8.5 years; BMI 33.9 ± 7.2 kg/m2) or nebivolol plus lifestyle modification (NBLM; n = 15; age 58.9 ± 9.4 years; BMI 32.5 ± 4.9 kg/m2) for 12 weeks. β‐stiffness index, a blood‐pressure‐independent measure of arterial stiffness, and arterial compliance were measured via high‐resolution ultrasound and tonometry at baseline and after the 12‐week intervention. There was no difference between groups in age, body weight or composition, blood pressure, or in β‐stiffness index or arterial compliance at baseline (all p > 0.05).

Results: Following the 12‐week intervention, body weight decreased ~5% (p < 0.05) in the LM and NBLM groups but did not change from baseline in the NB group (p > 0.05). Supine brachial and carotid systolic and diastolic blood pressure declined following treatment in each of the groups (p < 0.05). However, the magnitude of reduction was not different (p < 0.05) between groups. β‐stiffness index declined (‐2.03 ± 0.60, ‐1.87 ± 0.83 and −2.51 ± 0.90 U) and arterial compliance increased similarly (both p > 0.05) in the NB, LM and NBLM groups, respectively.

Conclusion: In summary, our findings indicate that the combination of nebivolol and lifestyle modification reduced large artery stiffness to a similar degree as either intervention alone in middle‐aged and older hypertensive adults.

Grant Funding Source: Supported by Forest Laboratories, Inc
Comparative effects of cinnamaldehyde and cinnamyl‐containing compounds on the viability of two human melanoma cell lines, SK‐MEL19 and SK‐MEL23 (LB611)
Authors: Muhammad Yosra , Kerr Stephen
Published: June 2021

Trans‐cinnamaldehyde (CA) is an α,β‐unsaturated carbonyl compound, derived from cinnamon, and contains a “Michael acceptor” functionality capable of alkylating nucleophilic residues. Such Michael acceptors are believed to interact with cellular nucleophiles through a Michael addition reaction, making them potential anticancer alkylating agents. Methyl trans‐cinnamyl ketone (MeK), methyl trans‐cinnamic ester (MeCE), and trans‐cinnamic acid (CAC) are cinnamyl‐containing compounds, and are structurally similar to CA, where they have the α,β‐unsaturated carbonyl moiety. This study investigated the effects of these cinnamyl‐containing compounds, alone and in the presence of 100µM glutathione (GSH) and/or 50µM ethacrynic acid (EA), compared to that of CA on human melanoma cell lines: SK‐MEL23 and SK‐MEL19. The viability of the cells was evaluated by MTS assay following 24h of incubation with the treatments. The results show that CA and MeCK exhibited a concentration‐dependent cytotoxicity, with MeCK being less cytotoxic. In the co‐treatment studies, the presence of GSH with the cinnamyl‐containing compounds did not provide protection to the cells, while co‐treatment of EA with the test compounds enhanced the cytotoxicity on the treated cells as compared to the control. In the combination treatment studies, the viability of the cells did not improve when GSH and EA were combined with the test compounds (triple combination) as compared to the co‐treatment of the test compounds with EA. These results indicate that CA was the most cytotoxic among the test compounds, and CAC caused the least cytotoxicity. The results suggest that both the reactivity of the “Michael acceptor” functionality and the lipophilicity of the cinnamyl‐containing compounds play a role in their cytotoxicity.

Grant Funding Source: MCPHS Faculty Development Fund
Physiological aspects of the development of mastication in early childhood (LB63)
Authors: Edelson Lisa , Le Révérend Benjamin , Alder Marc , Loret Chrystel
Published: June 2021

The development of feeding skills is a complex process influenced by four main factors: (1) Anatomy, (2) Neurophysiology, (3) Environment, (4) Society and culture. Feeding skills are therefore investigated by two fields of science: the behavioral science of feeding and the biomechanics of feeding (e.g. chewing and swallowing), with a particular emphasis on the first approach.

Food intended to be fed to infants and toddlers are currently recommended based on motor and eating skills described by Speech Language Pathologists (SLPs) and feeding specialists. The biomechanical characterization of mastication and its development has been less addressed, even though it could bring a new insight in the design of developmentally appropriate food.

In this work, we focus on the development of mastication in infants and toddlers. We aim to describe changes in the oral physiology (e.g. bone and muscle structure, teeth and soft tissues) of infants and toddlers, and its relationship to mastication abilities. Finally, we reviewed previous works on the effect of the food consistency on children’s mastication abilities and on their level of texture acceptance.

Recommending products that are adapted to children’s mastication during weaning could facilitate their acceptance of new textures and support the development of healthy eating habits.
Interleukin ‐ 17 ‐ mediated immune responses in triptolide ‐ induced liver injury in mice (LB637)
Authors: Wang Xinzhi , Jiang Zhenzhou , Zhang Luyong
Published: June 2021

The mechanisms of drug ‐ induced liver injury (DILI) have not been fully clariﬁed. Many factors, including oxidative stress and hepatic immune response, can contribute to the pathogenesis of DILI. Interleukin (IL) ‐ 17 secreted by helper T (Th) 17 cells are critical components of immune response in DILI. Triptolide (TP) has been reported to induce hepatocyte damage with inflammatory cells inﬁltration in the liver. To investigate the involvement of IL – 17 ‐ mediated immune response in the TP ‐ induced hepatotoxicity, we examined the plasma transaminase, histopathological changes, hepatic frequencies of Th17 cells, hepatic expression of transcriptional factors and cytokines genes and plasma IL ‐ 17 levels after oral administration of TP (600 μg/kg) to female C57BL/6 mice for 24 h. Mice treated with TP displayed acute liver injury with signiﬁcantly increased hepatic frequencies of Th17 cells, mRNA expression of retinoid ‐ related orphan receptor (ROR) ‐ γt and plasma IL ‐ 17 level as well as the plasma ALT and AST. Neutralization study using anti – IL ‐ 17 antibody ameliorated TP – induced liver injury. In contrast, when challenged by co ‐ administration of recombinant IL ‐ 17, hepatotoxicity was exacerbated in the triptolide ‐ administered mice. In summary, this report was demonstrated for the first time that IL ‐ 17 – mediated immune response is involved in the pathogenesis of TP ‐ induced liver injury in mice.

Grant Funding Source: Supported by National Natural Science Foundation of China（No. No.81173651）
In vitro characterization of the immunotoxic potential of pH‐sensitive microspheres on macrophage Raw 264.7 (LB639)
Authors: Youan Bi Botti , Miezan Ezoulin, Miezan , Zhang Tao
Published: June 2021

The objective of this study is to design pH‐sensitive microspheres (MS) for vagina delivery of microbicides to prevent HIV transmission. The MS were prepared by spray‐drying technique using pH‐sensitive polymethacrylate polymer (S‐100). The MS had a particle size of 76 ̶ 172 nm (in phosphate buffer, pH 7.3) and 6 ̶ 10 µm (in acetate buffer, pH 3.7) along with respective zeta potential in these media of ‐20 and ‐2 mV. It was hypothesized that because of the biocompatibility of the native materials, the MS will be non‐immunotoxic and suitable for the intended application. Methods: cultured murine macrophages RAW 264.7 were exposed for 24h to MS at 1‐1000 μg/mL. Cell membrane integrity, mitochondrial respiration, mitochondrial mass and membrane potential (∆Ψ), reactive oxygen species (ROS), glutathione (GSH) content, and the pro‐inflammatory mediator nitric oxide (NO°) were assessed using standardized in vitro assays. Results: the MS were shown to elicit no cell death at all concentrations tested. The mitochondrial functions were maintained intact except an increase in ∆Ψ (40%) at 1000 μg/mL MS. The MS had no effect on the cellular redox status reflected by the maintenance of ROS and GSH basal levels. Moreover, the MS did not activate the macrophages as evidenced by the basal level of NO°, suggesting an absence of iNOS‐induced inflammation. Conclusions: these findings further support the above working hypothesis, showing the MS as potential nanocarriers for HIV microbicides.

Grant Funding Source: RO1A1087304
In vitro avian pro‐epicardial cell spreading challenges the notion of how epicardial cells spread during cardiac morphogenesis (LB65)
Authors: Vilches‐Moure Jose , Erickson Carol
Published: June 2021

In the developing heart, the epicardium is derived from the pro‐epicardial organ (PEO), which spreads over the cardiac tube (purportedly as an epithelium), and then undergoes what is believed to be an epithelial‐to‐mesenchymal transition (EMT) to generate the subepicardial mesenchyme. We are interested in the process of epicardial cell spreading and the mechanisms that regulate the epicardial EMT. To study these processes, we have developed fluorescent labeling techniques to visualize the PEO and the spreading epicardium using time‐lapse confocal imaging in intact hearts and on a planar substratum. When the PEO is labeled and then explanted before it begins to spread in vivo, we saw that each cell has numerous filopodia and lamellipodia around its borders. Fixed images of spreading PEO cells labeled in ovo confirm the presence of numerous filopodia and lamellipodia in the spreading epicardium in vivo. Although the PEO has been assumed to spread as an epithelium with the force being generated at the free edge, our live‐cell imaging suggests that each cell in the epicardium is generating tractional force. These results show that real‐time behavior studies are critical since morphology alone does not reveal behavior. Our observations challenge the notion of how the PEO‐derived cells spread over the cardiac tube during cardiac morphogenesis.

Grant Funding Source: Supported by NIHT32OD011147 (JV‐M), UCD GSSP Fellowship (JV‐M), UCD GGCP Fellowship (JV‐M), IOS1146480 (CAE)
Impaired cardiac parasympathetic control in healthy young people with type 1 diabetes (LB658)
Authors: Wilson Luke , Coffey Sean , Hoye Neil , Murray Angus , Lamberts Regis , Williams Michael , Wilkins Gerard , Baldi J,Chris
Published: June 2021

Diabetes reduces heart rate (HR) reserve during maximal exercise. We measured HR during a low‐ and high‐dose (0.1 and 0.4 mg·kg‐1) atropine bolus followed by incremental dobutamine infusions (5 ‐ 40 μg·kg‐1·min‐1) up to 85% maximal HR to determine the roles of the parasympathetic and sympathetic nervous system in the diabetic HR response. Five 18 – 30 year old individuals with type 1 diabetes (T1D), but without cardiovascular disease or hypertension and four age‐, sex‐ and training‐matched non‐diabetic subjects (CON) performed a VO2max test (with ECG) on a cycle ergometer. On a second day, parasympathetic control was determined by comparing the change in HR during atropine doses using ANOVA. Sympathetic responsiveness was compared using the slope of the change in HR (after high‐dose atropine) across dobutamine infusion rates. VO2max and maximal HR during the exercise test were the same between groups (P > 0.05). Resting HR was greater in the T1D vs. CON (75 ± 4 vs. 62 ± 6 beats·min‐1; P < 0.05). Heart rate after high‐dose atropine was the same in T1D and CON (111 ± 19 vs. 123 ± 19 beats·min‐1; P > 0.05), but the change in HR after low‐ (18 ± 12 vs. 43 ± 9 beats·min‐1) and high‐dose atropine (40 ± 8 vs. 60 ± 11 beats) was smaller in the T1D vs. CON respectively. The percentage of maximal HR achieved with dobutamine (85 ± 2% vs. 84 ± 2%), dobutamine dose at 85% maximal HR (24 ± 11 vs. 20 ± 12 μg·kg‐1·min‐1) and sympathetic responsiveness (2.6 ± 1.0 vs. 2.6 ± 1.4 beats·μg·kg‐1·min‐1) were not different between T1D and CON respectively (P > 0.05). These data suggest that parasympathetic nervous activity is already impaired in healthy young individuals with type 1 diabetes, but the chronotropic response to sympathetic stimulation is unaffected.

Grant Funding Source: Supported by Healthcare Otago Charitable trust
Transvenous vagal nerve stimulation (VNS) in acute myocardial infarction (AMI) markedly reduces the infarction size and improves chronic cardiac function (LB670)
Authors: Arimura Takahiro , Saku Keita , Kakino Takamori , Akashi Takuya , Takehara Takako , Nishizaki Akiko , Oga Yasuhiro , Ikeda Masataka , Fujii Kana , Ide Tomomi , Kishi Takuya , Sunagawa Kenji
Published: June 2021

Background: Although VNS is known to have a powerful anti‐infarct effect, the technical difficulty associated with VNS precludes its application under emergent settings of AMI. We developed a novel technique where we stimulate the vagal system transvenously and evaluated how the VNS affects the infarction size and cardiac function in the long term.

Method: We ligated a left anterior descending coronary artery for 3 hours, then reperfused. For transvenous VNS, we performed the field electrical stimulation by pacing catheter in the superior vena cava. VNS reduced mean heart rate about 20‐30% during I/R. One month after ischemia‐reperfusion, we compared the infarct size, hemodynamics and load insensitive cardiac function, i.e., end‐systolic elastance (Ees) (Millar pressure and sonomicrometric volumetry) with or without VNS treatment.

Results: In comparison with no VNS, transvenous VNS significantly decreased the infarction size more than 80% (1.1±1.2 vs. 7.8±1.2cm2, p<0.05), doubled left ventricular Ees (6.5±1.7 vs. 13.2±0.6 mmHg/ml, p<0.05), and decreased NT‐pro BNP (3667±1637 vs. 843±256 pmol/ml, p<0.05).

Conclusion: Transvenous VNS in AMI markedly reduces the infarct size and improves cardiac function in the chronic phase (4 weeks after AMI). Transvenous VNS would be a novel therapy of AMI to prevent heart failure in the long term.
An anatomical model in plaster of the trunk: BIAS of the middle of the 20th century (LB7)
Authors: LE FLOCH‐PRIGENT Patrice , GILLOT Jean‐Bernard
Published: June 2021

A polychromic plaster represented the main viscera of the trunk, the visceral axis of the neck and the arterial and venous structures of the proximal part of the limbs. The superficial vessels of the scalp mainly the veins were represented. The anatomical structures were well represented but in a quite simple manner; so probably this object was made for the secondary schools more than for an use in the schools of medicine. The dimensions of the basis were 94 x 48,5 x1,4 cm and the model was 88 cm high. The manufacturer was BIAS in France probably on the middle of the 19th century.
Molecular mechanism of lung oedema clearance by AP301: dependence of ENaC pore forming subunits (LB781)
Authors: Shabbir Waheed , Hasanovic Minela , Jenn Soja , Tzotzos Susan , Fischer Bernhard , Pietschmann Helmut , Fischer Hendrik , Lucas Rudolf , Lemmens‐Gruber Rosa
Published: June 2021

Pulmonary oedema is a life threatening condition, caused by acute lung injury. Although failure of gas exchange in the alveolar spaces causes most of the morbidity and mortality, it remained uncertain how epithelial sodium channel lose their liquid clearance ability. Furthermore, the subunit composition of native ENaC in diseased pulmonary epithelium is still unknown. AP301, a lectin‐like domain derived peptide of TNF‐α, increases amiloride‐sensitive current in A549 cells as well as in heterologously expressed hENaC, an effect which is lost after deglycosylation. Consequently, the present study was undertaken to determine the interaction of AP301 with hENaC glycosylation sites and subunit compositions of hENaC. We substituted asparagine to glutamine in α‐hENaC, and studied the AP301 induced increase in sodium current in different subunit compositions. The effect of AP301 in HEK and CHO cells heterologously expressing human ENaC WT and mutants in addition to different subunit compositions (α, ß, γ and δ) was measured in patch clamp experiments. Indeed, AP301 weakly activated single as well as multiple asparagine mutants of α‐hENaC. For maximal AP301 activity αβγ‐ or δβγ‐hENaC co‐expression was required. Conclusively, αβγδ‐hENaC elicited a robust AP301 response at very low EC50 values. These findings suggest that specific interaction of AP301 ENaC requires precedent binding to glycosylated extracellular loop and pore‐forming α or δ delta subunits.

Grant Funding Source: Supported by Austrian Research Promotion Agency (FFG).
The enhanced effect of nucleoprotein in the recovery process of disuse muscle atrophy (LB811)
Authors: Nakanishi Ryosuke , Hirayama Yusuke , Tanaka Minoru , Maeshige Noriaki , Fujino Hidemi
Published: June 2021

Mechanical stress such as reloading is the best way to recover muscle whereas reloading combined other approached such as nutritional strategies is needed to facilitate muscle recovery. In addition, the muscle protein synthesis rate and mTOR activity, which is a central regulator of protein synthesis, are enhanced when amino acids are provided. Therefore, the aim of this study was to investigate the effect of reloading combined nucleoprotein, which includes essential amino acids and arginine, on the enhancement of muscle recovery. The fiber cross‐sectional area (FCSA) in the hindlimb unloading (HU) group was significantly lower than that in the control group. Although the FCSA was significantly higher in the reloading after hindlimb unloading (HUR) group than in the HU group, that in the reloaded after hindlimb unloading with nucleoprotein supplementation (HUR+NP) group enhanced compare with that in the HUR. In addition, the phospho‐S6/ total S6, one of key regulatory proteins of mTOR pathway, was significantly higher in the HUR+NP group than in the HUR group. These results suggest that nucleoprotein can enhance muscle recovery and nucleoprotein combined reloading treatment is effective to recover atrophy. Supported by Grants‐in‐Aid for Science Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology, and the internal funds of Nissan Chemical Industries, Ltd.
Association Between Problem Gambling and Functional Disability: A Nationally Representative Study Conducted in the United Kingdom
Authors: Jacob Louis , López-Sánchez Guillermo F. , Oh Hans , Kostev Karel , Koyanagi Ai , Haro Josep Maria , Pardhan Shahina , Shin Jae Il , Smith Lee
Published: June 2021

Objectives:

Problem gambling is associated with multiple detrimental health outcomes. However, to date, no study has investigated the association between problem gambling and functional disability. Therefore, the aim of this study was to investigate the association between problem gambling and functional disability in a UK nationally representative sample.

Methods:

Cross-sectional data from the 2007 Adult Psychiatric Morbidity Survey were analyzed. Problem gambling was assessed using a questionnaire including 10 DSM-IV criteria, whereas functional disability referred to at least 1 difficulty in 1 of 7 activities of daily living and instrumental activities of daily living. Control variables included sociodemographic factors, smoking status, alcohol dependence, drug use, the number of chronic physical conditions, depression, and anxiety disorder. The problem gambling-functional disability relationship was studied using a logistic regression model.

Results:

This study included 6941 adults aged ≥16 years (51.2% women; mean [SD] age 46.3 [18.6] years). The prevalence of functional disability was significantly higher in the at-risk problem gambling/problem gambling group than in the no problem gambling group (46.2% vs 32.1%, P value < 0.001). After adjusting for control variables, both at-risk problem gambling (OR = 1.55, 95% CI = 1.03–2.35) and problem gambling (OR = 3.05, 95% CI = 1.09–8.52) were positively and significantly associated with functional disability.

Conclusions:

In this large representative sample of UK adults, problem gambling was associated with higher odds for functional disability. If confirmed with longitudinal studies, these results suggest that those suffering from problem gambling should receive targeted intervention to aid in the prevention of functional disability.
Participant Experiences of a Quit Smoking Attempt Through Either Nicotine Replacement Therapy (NRT) Methods or the Use of an E-cigarette
Authors: Jones Gareth , McIntosh Emma , Brose Leonie S. , Klonizakis Markos
Published: June 2021

Objectives

There is a lack of evidence exploring experiences of using e-cigarettes for smoking cessation. The study’s main aim was to explore participant experiences of e-cigarettes compared to nicotine replacement therapy (NRT) delivered through stop smoking services.

Methods

Semi-structured, face-to-face, and telephone interviews at 3-month post-quit follow-up in a randomized controlled trial comparing nicotine-containing e-cigarettes, nicotine-free e-cigarettes, and NRT for smoking cessation. N = 17 participants, 9 were male, mean age 44 years, 5 using nicotine-containing e-cigarettes, 7 nicotine-free e-cigarettes, and 5 NRT. Interviews were transcribed and analyzed using thematic analysis.

Results

Two global themes and 5 organizing themes were identified. Global themes included: (1) experiences of e-cigarette and NRT quit aids (e-cigarette positive impact and dilemmas, NRT perceptions and experiences), and (2) key mechanisms to support quit attempt (physical aids, advice and support, feedback and structure). E-cigarettes were viewed with caution, however, generally evaluated positively alongside NRT methods, finding e-cigarettes useful during a quit attempt due to their versatility in application. Nicotinecontaining e-cigarettes were favored due to their support with nicotine cravings. Participants were, however, wary of replacing smoking addiction with vaping habit.

Conclusions

Participant e-cigarette experience were generally positive; however, concerns over long-term application were noted. There was a noticeable preference for nicotine-containing e-cigarettes, but further research is required to better understand how nicotine is used in conjunction with e-cigarettes long-term as a quit aid alongside other NRT.
Pelvic Ring Injury Mortality: Are We Getting Better?
Authors: Marchand Lucas S. , Sepehri Aresh , Hannan Zachary D. , Zaidi Syed M.R. , Bangura Abdulai T. , Morrison Jonathan J. , Manson Theodore T. , Slobogean Gerard P. , O'Hara Nathan N. , O'Toole Robert V.
Published: June 2021

Objectives:

To determine if changes in pelvic trauma care and treatment protocols have affected overall mortality rates after pelvic ring injury.

Design:

Retrospective cohort study.

Setting:

Level I trauma center.

Patients/Participants:

A total of 3314 patients with pelvic ring injuries who presented to a single referral center from 1999 to 2018 were included in the study.

Intervention:

Pelvic ring management, years 1999-2006 versus years 2007-2018.

Main Outcome Measurements:

In hospital mortality. Other examined variables included change in patient demographics, fracture characteristics, date of injury, associated injuries, length of hospital stay, Abbreviated Injury Severity Score.

Results:

The composite mortality rate was 6.5% (214/3314). The earliest cohort presented a mortality rate of 9.1% [111/1224; 95% confidence interval (CI), 7.6%–10.8%] compared with the more recent cohort mortality rate of 4.9% (103/2090; 95% CI, 4.1%–5.9%). Overall mortality was significantly lower in the more recent period, a risk difference of 4.1% (95% CI, 2.3%–6.1%; P < 0.01). After adjusting for age and Abbreviated Injury Severity Score of the brain, chest, and abdomen, the mortality reduction was more pronounced with an adjusted risk difference of 6.4% (95% CI, 4.7%–8.1%; P < 0.01).

Conclusion:

Significant improvement in the mortality rate of pelvic ring injuries has been demonstrated in recent years (4.9% vs. 9.1%) and the difference is even large when accounting for known confounders. Improvement appears to coincide chronologically with changes in trauma resuscitation and implementation of adjuvant treatments for managing patients with severe hemorrhagic shock. Although the exact benefit of each treatment awaits further research, these data might indicate improved care over time for these difficult patients.

Level of Evidence:

Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.
Operative Versus Nonoperative Treatment of Acute Displaced Distal Clavicle Fractures: A Multicenter Randomized Controlled Trial
Authors: Hall Jeremy A. , Schemitsch Christine E. , Vicente Milena R. , Dehghan Niloofar , Nauth Aaron , Nowak Lauren L. , Schemitsch Emil H. , McKee Michael D. ,   on behalf of the Canadian Orthopaedic Trauma Society (COTS)
Published: June 2021

Objectives:

To evaluate the differences in patient outcomes after operative or nonoperative treatment of displaced, type II distal clavicle fractures.

Design:

Multicenter, prospective, randomized controlled trial.

Setting:

Level I trauma centers.

Patients/Participants:

Patients with completely displaced type II distal clavicle fractures were included. Fifty-seven patients were randomized: 27 to the operative group and 30 to the nonoperative group.

Intervention:

Patients randomized to nonoperative care received a standard shoulder sling, followed by pendulum or gentle range of motion shoulder exercises at any time as directed by the attending surgeon. Patients randomized to the operative group received plate fixation with a precontoured distal clavicular plate or a “hook” plate within 28 days from injury.

Main Outcome Measure:

Disabilities of the Arm, Shoulder and Hand scores at 1 year.

Results:

There were no between-group differences in Disabilities of the Arm, Shoulder and Hand or Constant scores at 1 year. More patients in the operative group went on to union (95% vs. 64%, P = 0.02) within 1 year. Twelve patients in the operative group underwent a second operation for implant removal (12/27, 44%). In the nonoperative group, 6 patients (6/30, 20%) subsequently underwent 8 operative procedures.

Conclusion:

Although this study failed to demonstrate a difference in functional outcomes between operative and nonoperative treatment of Neer type II distal clavicle fractures, nonoperative management led to more complications including a moderate rate of nonunion, which often required secondary surgery to correct, a higher rate of early dissatisfaction with shoulder appearance, and a delayed return to activities in the first 6 months. Operative management provided a safe and reliable treatment option with few complications, but often required secondary implant removal, especially with hook plate fixation.

Level of Evidence:

Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.
Myocardial strain and left ventricular geometry: a meta-analysis of echocardiographic studies in systemic hypertension
Authors: Tadic Marijana , Sala Carla , Carugo Stefano , Mancia Giuseppe , Grassi Guido , Cuspidi Cesare
Published: June 2021

Aim:

Evidence on systolic dysfunction, as assessed by left ventricular (LV) mechanics, in hypertensive heart disease phenotyped by abnormal LV geometric patterns is still limited. Thus, we performed a systematic meta-analysis of 2D/3D speckle-tracking studies in order to provide an updated comprehensive information on this topic.

Methods:

The PubMed, OVID-MEDLINE, and Cochrane library databases were analyzed to search for articles published from the inception up to 31 January 2021. Studies were identified by using MeSH terms and crossing the following search items: ‘myocardial strain’, ‘left ventricular mechanics’, ‘speckle tracking echocardiography’, ‘systolic dysfunction’, ‘hypertensive heart disease’, ‘left ventricular hypertrophy’, left ventricular remodeling”, ‘concentric hypertrophy’, ‘eccentric hypertrophy’.

Results:

Overall, data of 267 normotensive controls and 712 hypertensive patients (234 normal geometry, 97 LV concentric remodeling, 176 eccentric LVH, and 205 concentric LVH) from eight studies were considered. Pooled ejection fraction was similar in normotensives and in the whole group of hypertensive patients: (SMD 0.037 ± 0.05, confidence interval (CI): 0.14/0.07, P = 0.51) as well as in hypertensive subgroups. LVMI, and E/e’ gradually increased from hypertensive patients with normal geometry and concentric remodeling, to patients with eccentric and concentric LVH. LV global longitudinal strain (GLS) was lower in all hypertensive groups compared with normotensive controls, in particular, in the eccentric (SMD 1.14 ± 0.22, CI: 0.69/1.59, P < 0.0001) and concentric LVH group (SMD 1.37 ± 0.21, CI: 0.95/1.79, P < 0.0001). The meta-analysis restricted to hypertensive patients showed that GLS gradually deteriorated from hypertensive patients with normal LV geometry, across those with concentric remodeling, to participants with eccentric and concentric LVH (−19.5 ± 0.6 versus −18.8 ± 0.73 versus −17.6 ± 0.66 versus −16.5 ± 0.6%, P < 0.001 for trend). Statistical significance was reached when GLS of the concentric and eccentric LVH group but not the concentric remodeling was compared with the normal geometry one.

Conclusion:

The present meta-analysis confirms that GLS performs better than ejection fraction in identifying systolic dysfunction in the hypertension setting. More importantly, it suggests that this occurs in patients with altered LV structure and geometry, such as LVH.
Postpulmonary embolism syndrome
Authors: Fabyan Kimberly D. , Holley Aaron B.
Published: June 2021

Purpose of review

It is now recognized that more than half of patients with acute pulmonary embolism (APE) will have persistent symptoms beyond 3 months after their initial event. Persistent symptoms are referred to as post-PE syndrome, an umbrella term that covers a spectrum of patient complaints and underlying pathologies. Data published over the last 5 years have added significantly to our understanding of this syndrome and its management.

Recent findings

Underlying pathologies linked to post-PE syndrome include chronic thromboembolic pulmonary hypertension (CTEPH), chronic thromboembolic disease (CTED), cardiac dysfunction, and deconditioning. Treatment for post-PE syndrome will depend on the underlying causative pathologies found. Evaluation and treatment for CTEPH is well defined, but less than 10% of patients with post-PE syndrome will qualify as having this diagnosis.

A large percentage of patients will experience post-PE syndrome following APE. Strategies for identification and treatment for some pathologies are well studied, but the majority of patients will have subtle abnormalities on imaging and functional testing for which diagnostic criteria and management are not well defined. A number of active studies are designed to help optimize the management of post-PE syndrome and should help us improve intermediate and long-term outcomes for patients following APE.