bluuebunny/crossref_metadata_2025 huggingface dataset:
I Like to Move It, Move It: The Role of Kif3B of Kinesin II in Primary Cilia Cilia
Authors: Kohlwey Kyle , Kuhn Meredith , Lang Nicole , Tiffany Kathryn , Tiffany Laura , Albrecht Jacqueline , Clapp Sarah , Fenton Kayla , Gallogly Austin , Jankowski Rebecca , Tiffany Karen , Willardsen Minde
Published: June 2021

Primary cilia are structures found on the surface of most cells and are important for cell signaling. For cilia to develop and function properly, materials must be transported in and out of these structures by motor proteins that travel along microtubules in the cilia. The Kinesin II motor protein transports cargo toward the tip of the cilia. Our interest lies in a motor subunit of Kinesin II, Kif3B. Kif3B binds to both ATP and microtubules; hydrolysis of ATP causes Kif3B to change its shape and move up the microtubules. Cilia development depends on the movement of materials into the cilia, and research indicates that if Kif3B is not functioning, cilia formation will not occur properly. Diseases called ciliopathies result if primary cilia production is altered. These diseases include Bardet‐Biedl syndrome (BBS), Joubert syndrome, and MORM syndrome. Using 3D printing technology, the Cedarburg SMART (Students Modeling a Research Topic) Team has designed a model of Kif3B to investigate the interaction of amino acid residues 96 through 104 of Kif3B with ATP and to visualize the neck region in Kif3B important for dimerization with Kif3A and for microtubule motility. Supported by grants from NIH‐SEPA and NIH‐CTSA
Cholera toxin: Vibrio cholera's dehydration machine
Authors: Anderson C. , Assyia D. , Bannerman K. , Fehr A. , Hermanson J. , Hron N. , Hsu J. , Larson N. , Long G. , Long J. , Mann D. , McLean R. , Menon V. , Mohar G. , Moret F. , Nguyen V. , O'Leary K. , Pedersen K. , Pohl R. , Reynolds H. , Rice A. , Rodriguez T. , Rottinghaus A. , Schaefer T. , Scott T. , Strombom K. , Vollick J. , Wehemyer G. , Whitcomb B. , Taghizadeh A. , Whalen J. , Kessler E.
Published: June 2021

The Blue Valley Center for Advanced Professional Studies (BVCAPS) SMART Team members researched the actions of the microbe Vibrio cholera in causing the water born disease cholera, which infects millions of humans annually and causes death to thousands of individuals through extreme fluid loss. During this project, students used JMOL to model cholera toxin, 1XTC.pdb, the structure of which was determined in 1995. Cholera toxin, or choleragen, is an enterotoxin that forms a heterohexameric AB5 complex. The pentameric B subunit binds to the intestinal surface of epithelial cells and the A subunit, once cleaved, infiltrates the host cell and catalyzes reactions that ultimately activate sodium pumps resulting in movement of sodium and water into the intestinal lumen. Structurally, the pentameric B subunit contains a central pore lined by α‐helices, and each of the five domains contains a GM1 ganglioside binding site. The A subunit, tethered above the B pentamer, is wedge‐shaped and connected to the enzymatic A1 chain by the A2 chain. Understanding choleragen structure is important for the development of drugs and vaccines for combating this disease. BVCAPS SMART Team members generated a physical model of the protein using our in‐house Z‐Corp 450 3D printer. This project was supported by a grant awarded to the Center for BioMolecular Modeling at the Milwaukee School of Engineering by NIH‐SEPA.
Structured self‐directed learning using whole‐body dissection
Authors: Kuo Kuo-Hsing
Published: June 2021

The decreased use of whole‐body dissection has led to variations in students’ anatomical knowledge therefore hindering student learning during their clinical years, particularly those who continue on to surgical disciplines. To address the diverse needs of anatomical knowledge we piloted a project allowing students to self direct their learning according to their clinical interests.

In the design of structured self‐directed learning, anatomical knowledge is grouped into two levels. General knowledge is aimed at training in the medical disciplines, thus encompassing surface anatomy and associated structures for clinical physical examination. Advanced knowledge is aimed at training in the surgical disciplines, thus learning anatomical structures encountered during invasive procedures. General knowledge is obtained by adopting mini tutorials utilizing pre‐dissected human cadavers and advanced knowledge through whole‐body dissection.

Final assessment was conducted using spot examination. Results indicate that structured self‐directed learning is able to address the diverse needs of learning anatomical knowledge. A large scale investigation is needed to confirm the impact on students’ performances in the clinical setting.
Short chain fatty acid absorption in lean vs obese humans
Authors: Rozenbloom Sari Rahat , Fernandes Judlyn , Wolever Thomas MS
Published: June 2021

Short chain fatty acids (SCFA) produced during the colonic fermentation of dietary fiber may reduce risk for obesity and type 2 diabetes. Our objectives were to determine if fecal SCFA concentration and absorption differ and/or are correlated in lean and overweight/obese (OW) subjects. Lean (n=11) and OW (n=11) subjects were recruited. After completing a 3‐day diet record subjects provided a fresh fecal sample to measure SCFA concentration. Subjects then inserted a rectal dialysis bag filled with 10ml of SCFA solution for 30 min. The procedures were repeated after two weeks. Fecal SCFA was higher in OW than lean subjects (80±6 vs 56±6 mmol/kg, p=0.02). The rate of SCFA absorption was similar and proportional to the SCFA concentration in the dialysis bag in both groups. There was a significant positive correlation between the SCFA absorption and fecal SCFAs for the OW group (r 0.44, p=0.04), but not for the lean group (r −0.22, p= 0.33) (difference in slopes, p=0.03). Since SCFA absorption from the dialysis bag was concentration dependent in the groups, OW people may absorb more colonic SCFA from their own feces. Within the OW group, the positive correlation between SCFA absorption and fecal SCFA concentration may be a result of upregulation of transporter expression for ionic diffusion.

This work was supported by a grant from the Canadian Institutes for Health Research.
Impact of weight loss through calorie restriction on iron status, inflammation and hepcidin expression in circulation and adipose tissue of obese individuals
Authors: Dao Maria Carlota , Ruth Megan R , Karastergiou Kalypso , Apovian Caroline M , Fried Susan K , Meydani Simin Nikbin
Published: June 2021

In obesity, hepcidin (Hep) is upregulated systemically and in adipose tissue (AT), which may lead to iron status impairment. It is not known how weight loss (WL) through calorie restriction (CR) affects Hep and iron status in obese adults. We assessed the effect of WL through CR in 24 obese adults, hypothesizing that systemic Hep and inflammation (Infl) would decrease with WL while iron status would be enhanced, and that WL would modulate Infl and Hep in AT. Serum Hep, CRP and iron status were assessed before and after WL. Subcutaneous AT biopsies were collected at both time points. Hep and macrophage marker expression was assessed by RT‐PCR. WL correlated with decreased serum Hep (r=−0.7, p=0.01) in subjects with 5–10% body WL (n=14). There was a trend for decreased serum CRP and increased iron status. Expression of CD68, a macrophage marker, was highly correlated with Hep expression before (r=0.7, p<0.001) and after (r=0.8, p<0.0001) WL. This data shows a significant effect of WL on Hep, and suggests a tight association between AT macrophages and Hep. Further research is needed to determine the role of inflammatory dysregulation of AT in obesity‐related iron status impairment. Support: Atkins Foundation, Boston University Clinical and Translational Science Institute M01 RR000533 and U54 RR025771, USDA HNRCA Pilot grant, NIH T32 and P30DK046200.
Acute airway access and maintenance during anatomy dissection: An interprofessional teaching exercise integrating basic and clinical sciences
Authors: Berglund Andy , Benninger Brion
Published: June 2021

INTRODUCTION

Acute airway access and management is a critical skill for healthcare professionals. Majority of healthcare professional trainees are classically taught on manikin and animal models, but rarely on human tissue. The objective of this study is to investigate whether the access and management of temporary airways could be taught during an anatomy course.

METHODS

Literature search was conducted on journals, texts, and websites regarding temporary access and maintenance of emergency airway management using cadavers during the first year anatomy course. Medical students conducted surgical airway access, maintenance and intubations on cadavers. Upon course completion, a questionnaire was used to assess the efficacy of the activity.

RESULTS

Literature search revealed no studies addressing airway acquisition and management to first year medical students (MS1) during anatomy cadaver lab. Anatomy faculty, flight surgeons, paramedics and flight nurses provided airway tutorials and demonstrations to 108 MS1's.

DISCUSSION

Airway management is a critical skill. In this study, students reported that airway access and management was one of the most intimidating procedures. MS1's successfully acquired airway access and management under supervision of the emergency experts.

CONCLUSION

This study suggests that airway management should be taught to first year medical students during cadaver lab.
Alternate day fasting combined with exercise: An effective treatment for weight loss and cardio‐protection in obese humans
Authors: Bhutani Surabhi , Varady Krista A , Klempel Monica C , Kroeger Cynthia M
Published: June 2021

Background

The effect of alternate day fasting (ADF) combined with endurance exercise (Ex) on body weight, body composition, and lipid levels, has never been tested.

Objective

Thus, this study examined whether the combination of ADF plus exercise produces greater changes in the above‐mentioned variables, when compared to each treatment alone.

Methods

Sixty‐four obese subjects were randomized to 1 of 4 groups: 1) ADF+Ex, 2) ADF, 3) Ex, or 4) control, for 12 weeks.

Results

Body weight was reduced (P < 0.05) by 7.1 ± 1.0%, 5.8 ± 1.0% and 1.4 ± 0.5% in the ADF+Ex, ADF and Ex group, respectively. Fat mass decreased (6.3 ± 1.5%, P = 0.001) and fat free mass increased (6.7 ± 2.3%, P = 0.015) in the ADF‐Ex group only. Waist circumference was reduced (P < 0.001) in all the intervention groups. LDL cholesterol decreased (7.0 ± 8.0%, P = 0.04) while HDL cholesterol increased (12.0 ± 10%, P = 0.04) in the ADF‐Ex group only.

Conclusion

These findings suggest that the combination of ADF plus exercise is the most effective treatment in reducing body weight, and improving body composition and plasma lipids, when compared to each intervention alone. This project was funded by The American Heart Association.
Inhibititory Effect of Gastrointestinal Lipolysis by Green Tea, Coffee, and Gomchui (Ligularia fischeri) Tea during Simulated Digestion
Authors: Kang Kyungsu , Cha Kwang Hyun , Song Dae-Geun , Kim Sang Min , Lee Eun Ha , Lee Hee Ju , Nho Chu Won , Pan Cheol-Ho
Published: June 2021

Green tea (GrT) and coffee are both rich in bioactive polyphenols (PP), such as EGCG or caffeoylquinic acids (CQA). PPs are beneficial to human body, especially, known to inhibit pancreatic lipase resulting in anti‐obesitic effect. Gomchui (Ligularia fischeri) has been used as herbal medicine or consumed as tea in Eastern Countries. It is also known to be rich in PPs, dicaffeoylquinic acid (DCQA). Despite of these beneficial effects of PPs, some are poor in bioavailability due to either degradation or difficulty of absorption in the GI track. Among them, GrT EGCG is easily degraded during digestion, thus making the beneficial effect doubtful in our body. We first investigated the inhibitory effect of three teas on lipid catabolism and found out Gomchui tea (GoT) was the most potent inhibitor. We assumed that the potency was due to both lipase inhibitory effect and the stability of PPs. Thus we monitored their stability during simulated digestion (SD) and assayed for the inhibitory effect on lipase. Among tested, EGCG was the most potent inhibitor (IC50 1.8 ± 0.57 μM) followed by DCQA (33.2 ± 4.2 μM). However, the stability was inverted. EGCG was not detected after the SD whereas DCQA was stable. Taken these together, it was concluded that despite of high potency on lipase inhibition, the overall effect of a PP on lipid catabolism should be considered as the sum of inhibitory effect as well as the stability during digestion.
Negative association between habitual dietary calcium intake, BMI, and body fat mass in postmenopausal women
Authors: Hooshmand Shirin , Metti Dina M. , Garcia Sofia F. , Vereda Yenina S. , Chai Sheau C. , Saadat Raz L. , Arjmandi Bahram H.
Published: June 2021

Obesity has become a national public health epidemic in the past several decades. The high prevalence of obesity in the world has provoked great interest in identifying strategies that can help prevent weight gain. Increasing calcium intake is one potential approach to weight management that has received much attention. To investigate the relationship between BMI, body fat, and habitual calcium intake, a diet history questionnaire designed to measure dietary calcium intake was administered to 120 postmenopausal women. Participants were divided into three groups based on their BMI values: 1) normal (18.5–24.9); 2) overweight (25–29.9); and 3) obese (≥30). Body fat and BMD were measured using DXA. Calcium intake was negatively correlated with BMI and body fat in this population. Overweight and obese individuals had lower calcium intake compared to individuals with normal BMI. Interestingly, BMD was not correlated with habitual calcium intake in this population except for the ulna BMD. However, when the ulna BMD was compared between individuals with lower (<1200 mg) and higher (>1200 mg) calcium intake, the differences were not statistically significant. These results are in agreement with previous hypotheses regarding calcium intake and improved weight management. Further investigation is necessary to better define the relationship between BMD and habitual dietary calcium intake.
Dental microwear texture analysis of mandibular molars from Medieval Ireland and England
Authors: Schmalz Naomi A. , Finan Thomas J. , Organ Jason M.
Published: June 2021

Excavation of the Medieval ecclesiastical site at Kilteasheen (Co. Roscommon, Ireland) recovered the skeletal remains of 131 individuals, which demonstrate poor dental health and significant physiological stress at the site. Historical records suggest the site was a stronghold for the O'Conor Kings of Connacht from the mid‐7th to 14th centuries, but little is known about the subsistence patterns practiced by its residents. This study uses white light scanning confocal microscopy and scale‐sensitive fractal analysis to quantify the occlusal microwear fabric of 32 adult mandibular molars from Kilteasheen and 3 other Medieval sites in England and Ireland. Results indicate a smaller enamel surface scale of maximum complexity (smc) at Kilteasheen than at All Saint's church (York, England), which suggests that wear‐causing particles may have been smaller at Kilteasheen than at York. Historical records suggest similar diets among sites, but potentially different food preparation techniques: Mill usage at York may have left behind larger non‐food abrasives than at Kilteasheen, where no evidence of similar practices is available. Funded by Saint Louis University (President's Research Fund).
A novel genetic risk factor linking choline to skeletal muscle fragility
Authors: Kohlmeier Martin , Contreras-Sesvold Carmen , Deuster Patricia A
Published: June 2021

Earlier controlled feeding studies have demonstrated that men with the common MTHFD1 variant 1958A have a much higher choline requirement than men without it (Kohlmeier et al., PNAS 2005). With choline intakes below their needs, creatine kinase (CK) activity increased greatly in some of these men as a sign of muscle dysfunction. We wanted to know whether physically active and fit men with elevated CK activity were more likely to carry the MTHFD1 1958A allele associated with increased choline requirement.

CK activities were measured in 145 Caucasian men prior to their entry in a physically challenging officer candidates’ training course and related to their MTHFD1 1958 genotypes. Genotype distribution (25 AA, 82 AG, 38 GG) indicated Hardy‐Weinberg equilibrium. Following our previous practice, we used the 90th percentile to distinguish between normal and high CK values. Men with high CK levels were much more likely to have one or two A alleles than men normal levels (odds ratio 6.03, p=0.04). The attributable risk for exceeding the 90th percentile related to the presence of an MTHFD1 1958 A allele was 81%.

Whether the men with signs of muscle damage were choline‐deficient is not known. Nonetheless, the more common presence of the A allele suggests that men with a high CK activity are more likely to be choline deficient. Higher choline intake may help them maintain muscle integrity during times with high‐intensity exercise.
Mathematical modeling of one‐carbon metabolism in trisomy 21
Authors: Chango Abalo , Gnimpieba Etienne Z. , Eveillard Damien
Published: June 2021

Cystathionine b‐synthase gene is overexpressed in Down syndrome. We have successfully used a mathematical model to study the impact of CBS overexpression on the one‐carbon metabolism. (150% of its normal Vmax) changed the profile of metabolites when compared with 100% of Vmax. The model allowed running the overexpression of RFC, FTCD and GART. The model may be a helpful tool in the future to study in silico metabolic and disease processes.
Evaluation of immunomodulatory properties of Bio‐Rice EL‐3000 in OVA‐induced mice
Authors: Yu-Yu Kao , Lin Ting-An , Yu Chi-Cheng , Yang Jo-Hsuan , Huang Hui-Yu
Published: June 2021

This study was purposed to examine the immunomodulatory properties of Bio‐Rice (fermented rice) in OVA‐induced mice model. Total forty BALB/c mice were categorized into four groups, ten in each, and considered as control, and different doses of Bio‐Rice treated groups (1X, 2X and 4X). After 8‐week treatment, the concentrations of interferon‐£^ (IFN‐£^) was increased with 1X, 2X and 4X doses, interleukin‐2 (IL‐2) was increased with 2X and 4X, and IL‐10 was elevated only with 4X dose of Bio‐Rice in the cultured splenocytes of ConA stimulate type. Nevertheless, all the inflammatory cytokines (IL‐2, IL‐10, IL‐5 and IFN‐£^) were elevated withal 4X dose in OVA‐induced mice, IL‐2 and IL‐10 increased with 2X, and no change with 1X. Estimated phagocytosis capacity was increased with 4X dose, and elevated natural killer cell (NK‐cell) activity was found with both 2X and 4X doses. Splenocytes subtype population, such as total T cells (CD4+ and CD8+), N K1.1 and F4/80 cells have been found to significantly higher in 2X and 4X doses of Bio‐Rice. Evidences from this study clearly demonstrating that 8‐week treatment of Bio‐Rice can modulate the immune system in OVA‐induced mice model. Our findings conclude that a fermented food product, such as Bio‐Rice is beneficial to boost the immune system.
Assessment of iatrogenic fat embolism with Nile red
Authors: Rubal Bernard J. , Muderhwa Jean M. , Rushing S. Renee
Published: June 2021

Fat embolism frequently occurs as a consequence of trauma, long bone injury and orthopedic procedures and is known to cause lung, brain and cutaneous lesions. There are currently no standardized laboratory tests for quantifying fat intravasation. This study investigates the use of a lipophilic fluorescent stain (9‐ diethylamino‐5‐benzo[α] phenoxazinone, Nile red) for assessing the fat embolic burden associated with intraosseous infusion of saline into the proximal tibia (N = 14) of swine (Sus scrofa
). After intraosseous cannulation, blood samples were obtained from the common iliac vein at baseline and following saline (0.9% NaCl) infusions (10 ml bolus, infusion at 300 mmHg, and at 0.3–4.0 ml per second). The effluents were centrifuged at 2000 × g for 10 minutes and supernatant aspirated and stained with 200 μl Nile Red (500 μg/ml in acetone). The buffy coat was separated with 8.0μ cellulose filters and lipids quantified by fluorescent imaging (excitation: 480 nm, emission: 535 nm). A >12 fold increase in relative fluorescence was noted with initial saline flush compared with baseline (P<0.05). The relative fluorescence was >20% baseline in 93% of effluent samples during saline infusion at 300 mmHg and at infusion rates ≥1.0 ml per sec. Significant (P<0.05) increases in relative fluorescence were observed with infusion rates of 1.0, 2.0, 3.0 and 4.0 ml per sec. These data suggest that Nile red may prove useful for quantifying iatrogenic fat embolism.
Differential genome‐wide DNA methylation patterns in young and old mice on calorie restricted or ad libitum diets
Authors: Park Lara Kristin , Schnitzler Gavin , Saltzman Edward , Choi Sang-Woon
Published: June 2021

Calorie restriction (CR) increases longevity and healthspan. We investigated how DNA methylation patterns change with aging and CR to gain insight into epigenetic mechanisms underlying healthspan extension. 14 week old C57BL6 mice were randomized to NIH31 Ad libitum diet (AL) or NIH31/NIA fortified CR diet to age 6 months (young) or 20 months (old). CR was initiated at 10% and increased to 40% over 2 weeks. DNA methylation was investigated in liver by the methylated DNA immunoprecipitationchip method on arrays covering all RefSeq genes and CpG islands. Data was normalized with the charm package in Bioconductor. Modified t‐tests and mean group differences were calculated per probe. We defined a differentially methylated probe (DMP) as p≤0.01 and mean difference ≥0.5. The age*diet interaction was investigated by a 2‐way ANOVA with DMP defined as the interaction p≤0.01. We identified 2918 DMPs in old vs young CR, 2256 in AL vs CR old, 1449 in young vs old AL, 943 in AL vs CR young and 1423 in the interaction. Unsupervised hierarchical clustering of interaction DMPs demonstrated that old CR and young AL cluster together and distinctly from the old AL and young CR. 1kb regions will be investigated, with differential methylation defined as ≥1‐DMPs and Wilcox Rank Sum FDR<0.1. This data supports that aging and CR modify DNA methylation independently and through an interaction. Supported by HNRCA Pilot grant.
Deletion of NALP3 protects against hyperoxia‐induced acute lung injury
Authors: Fukumot Jutaro , Fukumoto Itsuko , Huynh Bao , Cox Ruan , Parthasarathy Prasanna Tamarapu , Ho Matthew , Ramanathan Gurukumar Kollongod , Venugopal Rajan Babu , Lockey Richard F. , Kolliputi Narasaiah
Published: June 2021

Objectives

To determine whether NALP3 deletion has the ability to ameliorate hyperoxia‐induced acute lung injury.

Methods

C57BL/6 mice and age‐matched NALP3 KO mice of the same genetic background were exposed to 100% oxygen. 24–72 hours after exposure to hyperoxia, mice were sacrificed to collect bronchoalveolar lavage fluid (BAL fluid) for cell counts, cytokine measurement with ELISA, and TUNEL staining on BAL fluid cells. Additionally, after 48–72 hour exposure, lung tissues were collected for H&E and TUNEL staining on lung tisue sections, and also for Western blot analysis of Ceaved Caspase‐3 expresssion in lung tissue.

Results

Cell count in BAL fluid showed that infiltration of macrophages and neutrophils into lung parenchyma was significantly suppressed in KO mice. The pro‐inflammatory cytokine IL‐1β, TNF‐α, MCP‐1 and MIP‐2 levels in BAL fluid are all significantly suppresed in KO mice. Semi‐quantitative histological evaluation of H&E‐stained lung tissue sections revealed that lung pathological severity score was notably less in KO mice. TUNEL staining on lung tissue sections showed that lung epithelial cell apoptosis in NALP3 KO mice was markedly suppressed when compared to WT mice.

Conclusions

The deletion of NALP3 gene protects against hyperoxia‐induced acute lung injury. This result suggest that inflammasome platform can be a novel target for the treatment of acute lung injury.
IFN‐alpha plays a moderate role in the host response to influenza in aged treadmill‐exercised mice
Authors: Warren Kristi Jo , Kohut Marian L
Published: June 2021

IFNα is an anti‐viral protein known for its immune‐modulating involvement in influenza infection. Previous work by our lab has shown that in BALB/c mice exercise training prior to influenza infection results in improvements in illness severity in comparison to non‐exercised mice. It is possible that the exercise‐associated improvements to influenza are mediated by IFNα. To test this hypothesis, mice were treadmill‐trained for 10 weeks, and infected with Influenza (A/PR/8/34) ~24 hours after the last exercise session. Anti‐IFNα antibody or irrelevant antibody was administered 24 hours prior to infection, and continued daily through the first 4 days of infection. On day 8 post‐infection, blood, bronchoalveolar lavage (BAL) fluid, and lung draining lymph nodes were collected to assess the immune response. The results showed that exercise treatment attenuated body weight loss, increased IFNγ, IL‐10, CD8+ cells and inflammatory monocytes in the BAL of the exercised mice whether they were treated with anti‐IFNα or not. The anti‐IFNα treatment had no effect on lung viral titer, but reduced BAL IL‐5 and IL‐15. Alternatively, anti‐IFNα treatment increased BAL IFNγ and IL‐12p70, along with lymph node CD19+ and CD11c+ cells. Serum anti‐influenza IgG and IgG1 were significantly elevated with anti‐IFNα in both exercised and non‐exercised mice, and exercise treatment reduced IgG and IgG1. The results generally suggest that the majority of exercise‐associated alterations to the host defense to influenza are not mediated by IFNα however it does appear that increased TNFα and monocytes in the BAL of exercised mice maybe regulated by IFNα. Overall, this study shows that IFNα modulates the immune response in an alternative way than initially anticipated, perhaps via TNFα/iNOS producing DCs infiltrating the lungs, however this would require further investigation before a conclusion could be drawn.
Mast cell role in pulmonary hypertension in rat models
Authors: Lawrenz Josh M , Baybutt Richard , Baidoo Cletus , Dim Daniel , Chang Quin , Herndon Betty , Poisner Alan , Molteni Agostino
Published: June 2021

Elevated numbers of mast cells were found in a rat model of fat embolism and pulmonary vasculitis and fibrosis where the vasculitis was related to increased secretion of angiotensin II. Our early work suggests that this increase could be consequent to an increase in renin secretion produced by an increased number of mast cells. This study reports mast cell response in another lung damage model also associated with severe vasculitis and fibrosis: Bleomycin administration.

Twenty four rats were divided between diets receiving 15% flax seed oil or corn oil controls. The groups were further divided to receive intratracheally 8 units/kg of bleomycin or vehicle. After 7 days, rats were euthanized and lungs harvested for histological studies. Arteries were measured for lumen patency and the ratio of media/adventitia diameter. Mast cells were counted in Evans Blue stained sections.

Bleomycin induced marked reduction of vascular lumen patency and media/adventitia ratio (p< 0.01), similar to that obtained with fat embolism. Mast cell number, however, was not increased. Flax seed oil treatment markedly reduced vasculitis and fibrosis without influencing mast cell number and location (mostly around peribronchial arteries and subpleural septa). Contrarily to what was observed in the fat embolism model mast cells did not correlate with vasculitis and fibrosis, suggesting a different pathogenic mechanism.

Wheaton College
Variability in the Anterior Extralaryngeal Branch of the Recurrent Laryngeal Nerve: Clinical Implications
Authors: Hessel Kara Marie , Wright Barth , Olinger Anthony
Published: June 2021

The location and variation in the branching pattern of Recurrent Laryngeal Nerve (RLN) is of particular relevance to surgeons performing thyroidectomy. The variable occurrence of extralaryngeal branches (ELB) is perhaps the most surgically relevant. It is believed that, when present, the Anterior Extralaryngeal Branch (AELB) carries all of the motor fibers to the laryngeal constrictors, while the Posterior Extralaryngeal Branch (PELB) carries only sensory fibers to the laryngeal mucosa. This study aimed to identify the AELB and PELB, measure these branches when present, and compare between gender, sidedness and neck length. ELB was found in 77.78% of our population, in 77.14% left RLNs and 54.29% of right RLNs. When present, AELB length was found to be significantly longer on the left. Those individuals that exhibited ELBs were found to have significantly longer necks, on average, than those without. We suggest that surgeons measure the necks of their patients to screen for those at high risk for ELB and begin searching for ELB earlier during dissection of the left RLN, but be aware that ELB can, and does, occur in all populations at variable lengths.
Rhythmic expression of vasoactive compounds in the rat brain and cerebral vasculature
Authors: Carver Koryn A , Lourim David , Besharse Joseph C , Harder David R
Published: June 2021

In mammals, circadian variation is evident in hormone levels, arterial pressure, heart rate and cerebral blood flow (CBF). In humans there is an increased stroke incidence in the morning. CYP2C11 and CYP4X1 are cytochrome P450 epoxygenases expressed in rat brain which convert arachidonic acid (AA) to vasodilatory epoxyeicosatrienoic acids (EETs). We have shown that these genes are expressed in a circadian or biphasic circadian manner in rat brain and vasculature. Here we investigate rhythms in cultured astrocytes and cerebral microvascular endothelial cells over 48 hours using RT‐PCR. Using Western blotting, we measured CYP2C11 and CYP4X1 protein levels over 24 hours and CYP450 enzyme activity by measuring EETs and 20‐HETE formation following addition of the substrate AA. We show Cyp2c11, Cyp4x1 and Cyp4a2 mRNA expression is rhythmic with a 12 hour period although the phase differs between genes. This pattern was also apparent at the protein level. Preliminary P450 activity data suggest rhythmic formation of EETs and 20‐HETE with 12 and 24 hour periods. Our data indicate CYP450 enzymes have rhythmic mRNA and protein expression and activity levels, suggesting their involvement in regulation of rhythmic physiological phenomena, such as CBF. Our findings will lead to a better understanding of the mechanisms underlying increased morning stroke incidence in humans.
Morphometric analysis of the intestinal villi in experimental administration os semicarbazide
Authors: Cabeças José Ricardo , Marques Pedro , Farinha Rodrigo , Silva Fernando Capela , Cabrita António Silvério
Published: June 2021

The objective of this study was to evaluate the morphologic changes of the small intestine related to lathyrism.

Thirty animals were randomly distributed by three groups of ten animals each.

The control group (CP), had no manipulation. In the groups 2 and 3 we incorporated semicarbazide in the diet, in the doses of 3 and 6mg/kg, respectively, during 4 weeks. By the 30th day, all animals were killed with an overdose of anesthetics and the complete necropsy was performed. Histopathological processing and analysis of the small intestine was made.

Pictures of routine stained slides were taken and the length of the small intestine villi was measured using the software ImageJ.

The intestinal villi were distributed over 5 classes (very small‐VS, small‐S, medium‐M, big‐B and very big‐VB) using the average and standard deviation, for the CP.

In the CP the percentage of each class was: 2 VS; 16 S; 64 M; 16 B; 2 VB. In the group 2: 87 VS; 13 S; 6 M; 1 B; 0 VB. In the group 3: 73 VS; 24 S; 3 M; 0 B; 0 VB.

Significant changes were found, with the increase of the number of small villis in groups 2 and 3. These results suggest that an adequate collagen synthesis is necessary to maintain the intestinal villi length.
Monitoring of the small animal cage environment at the bioterium
Authors: Cabeças José Ricardo , Farinha Rodrigo , Cabrita António Silvério
Published: June 2021

Monitorization of small animal's cages environmental variables is a major concern in animal experimentation. The access to variables such as the temperature and humidity of the cage allows greater control over animal welfare during an experimentation and can help improve the environmental conditions.

The purpose of this work is to create a means for controlling the environment in the animal's cage, using temperature, humidity and gas sensors connected to a microprocessor. The microprocessor will connect to a server where the data will be stored.

Using an array of sensors in each cage one can monitor the environment inside the cage individually, managing the information from each sensor, processing that information and triggering events that will help the researcher throughout the experiments.

With this project we want to create a system that can help the researcher monitor the animals without having to be physically present. This system will gather all the information in a server which will have limited access.

With the implementation of this system we expect to optimize the time and the resources spent in the maintenance of the small animal's cage, while getting real‐time information about the environment of each cage.
Chronic methamphetamine (METH) induces memory deficits that are associated with changes in hippocampal functions and plasticity factors
Authors: Aduonum Adwoa , Louis Vanessa , Blanc Adnouse , Aiyede Onize , Chirwa Sanika
Published: June 2021

METH users perform subpar than controls on tasks of verbal and nonverbal memory, recognition, attention, and decision‐making but the mechanisms that underlie these disturbances are unclear. Here we conducted memory tests in 20 male guinea pigs (200–250 g) using customized home cages and video‐tracking software. METH (10 mg/kg/day) or saline (24 μl/day) were infused using osmotic pumps for 7 days at the rate of 1.0 μl/hr. Following familiarization to 2 similar objects, in the novel object recognition test, subsequent discrimination for novel objects when tested after 3‐hours was impaired in METH‐treated guinea pigs (n=8). This impairment was still evident even after 7 days post‐drug infusion (n=8). By contrast saline controls showed discrimination for novel objects indicating good recognition memory (n=4). Molecular assays showed up‐regulation of the plasticity factors GAP‐43, BDNF and α‐synuclein in METH‐treated animals. Furthermore METH‐treated guinea pigs exhibited increased theta rhythms and decreased slow oscillations in hippocampus in vivo. Long‐term potentiation (a cellular correlate of memory) was reduced in METH guinea pigs relative to saline controls. These results suggest that METH affects some of the same processes undergirding memory functions and their drug‐induced activation partly leads to occlusion of mnemonic functions. Supported by NIH grant DA021471 & PCOM Seed Funds.
Particularities of blood vessels correlate with VEGF expression in human renal medulla
Authors: Alexa Aurora , Zahoi Delia Elena , Daescu Ecaterina , Baderca Flavia
Published: June 2021

In the normal kidney, VEGF is constitutively expressed in podocytes and tubular epithelial cells of the renal cortex and medulla. This study determined the particularities of renal vascularization and VEGF expression in renal medulla using immunohistochemical methods on normal kidneys samples taken from 28 patients. Sections were stained with routine haematoxylin‐eosin method and immunohistochemically for VEGF, CD34, CD31 and SMA. All cases presented an intense immunoreaction for VEGF in the cells lining the nephron tubular system, the higher immunoreaction intensity in the collecting and distal tubules, but weak or moderate in the proximal tubules. The Henle loop cells showed a negative immunoreaction. The VEGF immunostaining was significantly higher in outer than inner medulla. In outer portion of outer medulla we identified using vascular markers, two types of vascular plexus: peritubular plexus around contort and collector tubules of outer zone of outer medulla and segmentary capillary plexal nodes near vasa recta. The strong positive immunoreaction for VEGF in the collecting ducts and distal tubules epithelial cells suggest a relationship between VEGF expression and adjacent straight vessels (vasa recta), with important roles in gradient concentration, being known that VEGF was initially named vascular permeability factor.
Inhibition of JMJD2 histone demethylase by curcumin analogs
Authors: Choi Jaehwa , Li Chenglong , Li Pui-Kai , Ihnat Michael A. , Janknecht Ralf
Published: June 2021

Epigenetic mechanisms include changes in chromosomes through post‐translational modification of histones. The jumonji‐domain (JMJD)‐containing proteins are a family of histone demethylases. The JMJD2 genes appear to be proto‐oncogenes as these genes are frequently found to be amplified in various types of cancer. Therefore, inhibition of JMJD2 function potentially is a promising target for anticancer therapeutics. The current study focuses on the identification of curcumin analogs that show inhibition of both histone demethylase activity and cancer cell proliferation. The ability of each curcumin analog to inhibit the activity of the JMJD proteins was assessed by measuring the methylation level of histone substrates through Western blotting using antibodies to trimethylated histone. The ability of the curcumin analogs to inhibit cell proliferation was measured using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT). Several curcumin analogs showed inhibitory action towards histone demethylation by JMJD2A. These curcumin analogs also inhibited proliferation of several cancer cells, including colon cancer cells (EC50 = 2–6 μM). These data support our previous finding that JMJD2A is a novel promoter of colon cancer cell proliferation. In conclusion, we have identified several curcumin analogs that both have anticancer activity and inhibit histone demethylation.
Isoflurane Preconditioning Protects Cardiomyocytes Against Hypoxia/Reoxygenation Injury Via A Vascular Endothelial Growth Factor‐Dependent Mechanism
Authors: Liu Yanan , Procknow Jesse , Ge Zhi-Dong , Bienengraeber Martin , Xia Zhengyuan , Warltier David , Pratt Phillip , Kersten Judy
Published: June 2021

Isoflurane preconditioning (IsoPC) has been demonstrated to exert a cardioprotective effect. We aimed to examine the role of vascular endothelial growth factor (VEGF) during cardiomyocyte protection produced by IsoPC. Human coronary endothelial cells(ECs) were co‐cultured with cardiomyocytes isolated from neonatal rats. The co‐culture underwent 2 hours of hypoxia followed by 2 hours of reoxygenation in the presence or absence of isoflurane administered 60 mins prior to hypoxia. IsoPC significantly increased the production of nitric oxide from ECs and decreased the levels of lactate dehydrogenase release. The beneficial effect of IsoPC on cardiomyocytes was blocked by VEGF neutralizing antibody applied prior to IsoPC. To examine whether VEGF is a downstream target of hypoxia‐induced factor 1α (HIF1α), ECs were infected with lentivirus containing shRNA against HIF1α for 72 hours prior to co‐culture. The expression of total VEGF and VEGF receptor 2 proteins was significantly decreased by HIF1α knockdown. IsoPC failed to protect cardiomyocyte against hypoxia/reoxygenation (H/R) injury in HIF1α knockdown ECs. The detrimental effect of HIF1α silencing on IsoPC was restored by administration of recombinant VEGF during the period of co‐culturing. These results demonstrate that VEGF plays a critical role in cardiomyocyte protection against H/R injury. During IsoPC, VEGF may be a downstream mediator of HIF1α.
Endothelial Regulation of Calmodulin Expression in Vascular Smooth Muscle Cells
Authors: Stencel Michael , FRANCIS SARAH , GILES JENNIFER , BURGARD MICHELLE , TRAN Quang-Kim
Published: June 2021

Interactions between vascular endothelial cells and the underlying smooth muscle cells (VSMCs) are of paramount importance in maintaining vascular functions. Calmodulin (CaM) is involved in a wide variety of cellular functions and is a limiting factor in both cell types, with free cytoplasmic CaM constituting only a small fraction of the total cellular CaM. Currently nothing is known about potential interactions between ECs and VSMCs as it involves CaM. We have begun to investigate the possibility that vascular endothelial cells impact VSMC functions via CaM‐dependent activities. Using a co‐culture model of primary culture vascular endothelial cells and smooth muscle cells isolated from the same vessels, we have found that VSMCs in co‐culture with proliferating ECs express on average 90% more CaM than monocultured VSMCs. Media fractionation and eNOS inhibition experiments indicate that the CaM‐elevating effect was exerted by a soluble factor that is not nitric oxide. The CaM‐elevating effect exerted by ECs is strongly dependent on endothelial density, such that at a starting 50% confluency of VSMCs, a starting 20% endothelial confluency triggers the greatest CaM increase in VSMCs, whereas a starting 70% endothelial confluency yields no visible effect on VSMC CaM expression after 48 hrs. Pharmacological inhibition of cyclooxygenase‐1, vascular endothelial cell growth factor (VEGF), and endothelin‐1 receptors (ETA and ETB) does not affect the observed increase in CaM. The data suggest that proliferating endothelial cells produce a soluble factor that can regulate CaM‐dependent signaling in VSMCs via alterations in total cellular CaM expression.
Canine coronary microvascular resistance at vasodilation depends on coronary perfusion pressure
Authors: van den Wijngaard Jeroen , van Horssen Pepijn , Hoefer Imo , Spaan Jos , Siebes Maria
Published: June 2021

Objective

We tested the premise of a constant microvascular resistance at full vasodilation (Rm) that underlies the concept of fractional flow reserve.

Methods

In 7 anesthetized dogs, we measured aortic (Pa) and venous (Pv) pressure, flow in the left anterior descending (LAD) artery, distal pressure (Pd) in the left circumflex (LCX) artery. During LCX occlusion, wedge pressure (Pw) was measured and μspheres infused into the LAD at full vasodilation. In one dog, the RCA contribution was determined by left atrial injection of differently colored μsph. Hearts were analyzed using a high‐resolution 3D imaging cryomicrotome. The mass of LCX and LAD perfusion territories was obtained. Assuming equal resistance per gram tissue for LAD and LCX, the weight‐normalized Rm of LCX was determined at Pd and Pw. Collateral flow was calculated as the ratio of μsph in LCX and LAD territories multiplied by LAD flow.

Results

Accounting for the added 30%RCA contribution to the collateral flow obtained from LAD injection, Rm was 2.3 ± 1.9 times higher at low (Pw= 15.7+4.8 mmHg) compared to normal pressure (Pd=69.3+12.4 mmHg) (p<0.05). Collateral flow from LAD to LCX was 8.6% + 5.3% of LAD hyperemic flow.

Conclusions

Our data confirm the pressure‐dependence of coronary Rm at vasodilation, which suggests a reason for pressure‐based overestimation of collateral flow. Supported by the Netherlands Heart Foundation (2006B226)
A proteomic approach to identify proteins involved in Redox regulation of inflammation and immunity
Authors: salzano sonia , Coppo Lucia , Bowler Lucas , Cervellini Ilaria , Mengozzi Manuela , Ghezzi Pietro
Published: June 2021

To better understand the role of redox regulation in immunity, we studied the glutathionylation of proteins secreted in response to inflammatory stimuli such as lipopolysaccharide (LPS). These proteins were detected by cell labeling with biotinylated glutathione ethyl ester (BioGEE) and identified by one‐dimensional gel electrophoresis, subjected to tryptic digestion followed by mass spectrometry analysis. Several proteins were identified in supernatants from Raw 264.7 cells (murine macrophages) stimulated with 100 ng/ml LPS for 24 hours.

We focused our study on Peroxiredoxin 2 (Prx2), an antioxidant enzyme involved in cells protection against oxidative stress which breaks down H2O2. Following its identification, we studied secretion of Prx2 by Raw 264.7 cells using Western blot. In vivo studies were also performed to measure Prx2 levels in serum from mice injected intraperitoneally with saline (vehicle) or with 400 μg/mouse LPS (LPS‐treated mice). On the basis of our Western blot data, we confirmed higher levels of secreted Prx2 in Raw 264.7 cells after LPS stimulation. Increase in Prx2 levels were also observed in serum from LPS‐treated mice.

Supported by BSMS and interreg TC2N
Type 1 diabetes mellitus differentially regulates mitochondrially‐encoded proteins in cardiac mitochondrial subpopulations
Authors: Lewis Sara , Baseler Walter , Croston Tara , Thapa Dharendra , Jagannathan Rajaganapathi , Hollander John
Published: June 2021

Mitochondrial dysfunction is a contributing factor in diabetic cardiomyopathy. The mitochondrial genome encodes for thirteen proteins comprising essential constituents of the respiratory chain. Dysregulation of the mitochondrial genome has been shown to correlate with mitochondrial dysfunction. The purpose of this study was to determine whether the protein expression of mitochondrial genome‐encoded proteins were influenced during a type 1 diabetic insult in cardiac subsarcolemmal (SSM) and interfibrillar mitochondria (IFM). Male FVB mice were made diabetic through multiple low‐dose injections of streptozotocin and mitochondrial subpopulations were isolated from cardiac tissue five weeks post‐hyperglycemia. Western blot analyses revealed differential impact on mitochondrial genome encoded proteins from electron transport chain complexes I (ND2, ND3, ND4L, ND5, ND6), III, IV (CO1, CO2, CO3), and V (ATP6) (P<0.05 for all). Of particular interest, was observed significant decreases to mitochondrially‐encoded electron transport chain complex IV proteins CO1 and CO3 within diabetic IFM (P<0.05 for both). The results indicate that type 1 diabetes mellitus differentially regulates mitochondrially‐encoded electron transport chain complex constituents. Further, these effects appear specific for a given mitochondrial subpopulations.
Adaptation Leads to Loss of a Regulatory Site Affecting mRNA Levels and Fitness
Authors: ZHAO LUYI
Published: June 2021

Adaptive substitutions in regulatory regions of viruses often lead to problems. Our goal is to investigate the contribution of substitutions in regulatory regions to the adaptive evolution of a phage, ϕX174, to its host, Escherichia coli at different temperatures. During experiments where phages were grown in chemostats, the J terminator was deleted, and this deletion spread throughout the phage population. Phenotypic performance and RNA production at 37°C and 42°C was compared between a mutant strain whose J terminator was deleted and an isogenic wild type strain. The results showed that the deletion strain has a faster doubling rate but similar time to lysis as the wild type strain at 42°C, while they have similar doubling rate and time to lysis at 37°C. Capsid assembly assays showed that the deletion strain lags behind the wild type strain in producing virions at 37°C. At both temperatures, the amount of RNA for gene F relative to the amount of RNA for gene D is greater for the deletion strain than the wild type strain. Northern blot analysis showed that the transcript from gene D to gene J decreased after deletion of the J terminator. Based upon the phenotypic assays, the greater amount of RNA for gene F appears to be beneficial at 42°C. Our results suggest that the J terminator plays an important role in adaptive evolution.