bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood Pressure Regulation Using Electroacupuncture in Middle‐Aged Hypertensive Women Associated With Mitochondrial Beta‐Oxidation
Authors: Tjen‐A‐Looi Stephanie C , Fu Liang‐Wu , Xie Lifang , Li Peng , Malik Shaista
Published: June 2021

Acupuncture at specific acupoints reduces elevated blood pressure. We have shown that 8‐week electroacupuncture (EA) at P5‐6 and St36‐37 decreases blood pressure in men (38–75 year) with mild to moderate hypertension (HTN) more effectively (70% responsiveness, ≥ −6 mmHg). In contrast, the majority (64%) of middle‐aged hypertensive women were not responsive to this treatment. We decided to add acupoints that target the endocrine system as well (LI4, CV3‐4, SP 6, LV3, K3). However, the underlying mechanisms of how acupuncture lowers blood pressure remain unclear. Metabolomics provides a powerful method to assess the biochemical signature involved in cellular processes that are involved in human health and disease. By using untargeted metabolomics, therefore, we aimed to identify metabolic biomarkers and assess pathways involved in sympathoinhibitory EA and endocrine‐sympathoinhibitory EA treatment lowering blood pressure in middle‐aged perimenopausal females with mild to moderate HTN. Twenty four enrolled women ages 45 to 70 year with hypertension were randomly assigned to sympathoinhibitory EA treatment (P5‐6+St36‐37, n=10) or endocrine‐sympathoinhibitory EA treatment (P6‐LI4+St36‐Sp6+CV3‐4+Li3‐K3, n=14). Participants were not on antihypertensive medications and hormonal therapy. The blood pressure lowering responsiveness (≥ −6 mmHg) to endocrine‐sympathoinhibitory EA was 80% while only 33% of the subjects treated with sympathoinhibitory EA lowered their blood pressures. Peak systolic blood pressure (SBP) during 24 hours dropped from 169±4.1 to 157±3.3 mmHg (P=0.002) while averaged 24‐hour SBP dropped from 138±2.3 to 131±2.3 mmHg (P=0.001) in 14 subjects treated with endocrine‐sympathoinhibitory EA. The averaged 24‐hour mean blood pressure (MBP) and peak MBP decreased respectively with P values of 0.002 and 0.025 in the 14 subjects. On the other hand, 8‐week course of sympathoinhibitory EA treatment did not decrease SBP, MBP, peak SBP, and peak MAP in 10 other subjects, consistent with our prior study. Concurrently, metabolomics analysis indicated that the plasma acylcarnitines (short (P=0.03) and medium chain fatty acids (P=.008)) were reduced significantly in 14 subjects following 8‐week endocrine‐sympathoinhibitory EA treatment in contrast to sympathoinhibitory EA. Previous studies have shown that arterial stiffness and hypertension are associated with higher acylcarnitine levels which indicate higher beta‐oxidation associated with mitochondrial dysfunction. Endocrine‐sympathoinhibition EA enhances blood pressure lowering responsiveness in middle‐aged hypertensive women likely through decreasing acylcarnitine production, reflecting lower beta‐oxidation at the mitochondrial level.

Support or Funding Information

Adolph Coors Foundation

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic Cannabis Use and Circulating Bio‐Markers of Neural Health, Stress, and Inflammation in Physically Active Individuals
Authors: Lisano Jonathon , Kisiolek Jacob , Phillips Kristina , Smoak Peter , Stewart Laura
Published: June 2021

Purpose

The purpose of this cross‐sectional study is to compare measures of general health, circulating concentrations of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), c‐reactive protein (CRP), interleukin‐6 (IL‐6) and cortisol in physically active cannabis users (CU) and physically active non‐cannabis users (NU).

Methods

Participants (N=30; n=20 male) were defined as CU (n=15; n=10 male) if they were using cannabis products at least once a week for the past 6‐months or NU (n=15; n=10 male) if they had not used any cannabis products in the past 6‐months. Age, height, weight, and body composition were assessed. Resting heart rate (HR) was obtained using a Polar heart rate monitor and VO2max, as a measure of cardiovascular fitness, was assessed with a ParvoMedics metabolic cart using a graded treadmill protocol. Fasted and rested intravenous blood samples were collected and ELISAs were used to obtain NGF, BDNF, CRP, IL‐6, and cortisol concentrations. Data are presented as mean ± SD and analyzed with a student's t‐test (alpha=0.05).

Results

CU used cannabis products an average of 4.5±2.5 times per week. There were no significant differences between CU and NU age (23.4±4.4 yrs), height (177.6±8.2 cm), weight (75.7±15.5 kg), body composition, or VO2max (50.3±7.4 ml/kg/min). HR was significantly higher in CU (72.7±15.9 bpm) compared to NU (62.5±7.9 bpm) (p=0.04). Concentrations of BDNF were significantly lower in CU (5.6±0.8 ng/mL) compared to NU (6.3±0.8 ng/mL) (p=0.02). There were no differences in concentration of NGF (193.7±71.6 pg/mL), CRP (CU=1.0± 1.83; NU=0.5±0.4 mg/L), IL‐6 (1.28±0.6 pg/mL), or cortisol (19.8±7.0 ng/mL).

Conclusions

Although aspects of general health appeared to be similar between NU and CU groups, this study revealed elevated resting HR and reduced BDNF concentrations in CU compared to NU. Although there were no differences between groups with respect to bio‐markers of stress (IL‐6, CRP and cortisol); CU were classified as moderate risk for cardiovascular disease (CVD) compared to low CVD risk for NU as defined by CRP concentrations. This study suggests that chronic cannabis use is associated with altered central and peripheral synaptic plasticity, as well as the potential for increased CVD risk classification in physically active individuals.

Support or Funding Information

This research was funded by the University of Northern Colorado New Projects Program Grant, University of Northern Colorado Graduate Student Association and Natural and Health Sciences Student Grants.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bioengineered VEGF Therapy Following Angioplasty in Renovascular Disease: More and Better Microvessels
Authors: Guise Erika , Engel Jason , Williams Maxx , Bidwell Gene Lee , Chade Alejandro
Published: June 2021

Asymmetric dimethylarginine (ADMA) is a pro‐atherogenic marker of endothelial dysfunction and inhibitor of endothelial nitric oxide synthase (eNOS) that may contribute to progression of renal injury. We designed a novel bioengineered fusion of an elastin‐like polypeptide with vascular endothelial growth factor (ELP‐VEGF). We showed that ELP‐VEGF therapy in renovascular disease (RVD) restores renal hemodynamics by improving microvascular (MV) rarefaction in the stenotic kidney. Our preliminary data suggest that co‐adjuvant ELP‐VEGF therapy with renal angioplasty (PTRAs) enhances renal recovery compared to PTRAs alone. We aimed to elucidate underlying mechanisms of renoprotection and test the hypothesis that co‐adjuvant ELP‐VEGF+PTRAs decreases ADMA and thus, its effects on eNOS inhibition, causing augmented renal bioavailability of NO and improvements of MV endothelial function, renal hemodynamics, and reversal of MV rarefaction.

RVD was induced by unilateral renal artery stenosis and high cholesterol feeding in 14 pigs. Six weeks later, single‐kidney blood flow (RBF) and filtration (GFR) were quantified in vivo using multi‐detector CT at baseline and during an endothelium‐dependent challenge (with acetylcholine, Ach). Then pigs were randomized to either PTRAs+placebo or PTRAs followed by intra‐renal ELP‐VEGF therapy (RVD+PTRAs+ELP‐VEGF). Pigs were observed for four additional weeks; in vivo CT studies repeated, then pigs were euthanized, and ex vivo renal MV micro‐CT quantification, plasma ADMA concentration, and stenotic kidney eNOS, VEGF, and Flk‐1 expression were measured.

Pre‐treatment renal hemodynamics were equally reduced and PTRAs similarly resolved renal artery stenosis in all pigs with RVD. However, PTRAs+ELP‐VEGF led to significantly greater recovery of RBF and GFR and responses to Ach than PTRAs alone. Improvements in renal function were accompanied by decreased plasma ADMA and enhanced stenotic kidney expression of eNOS, VEGF, and its Flk‐1 receptor, and improved MV density and remodeling (Figure).

This study extends our previous findings by identifying a potential mechanism of enhanced renal recovery after combined PTRAs+ELP‐VEGF therapy. Reductions in circulating ADMA may have contributed to the improved renal expression of eNOS in tandem with VEGF therapy, resulting in augmented renal bioavailability of NO and ultimately improving functional and structural renal MV rarefaction. Furthermore, decreased ADMA may suggest a slowing down of the atherosclerotic process in the RVD kidney, which may have also helped contribute to the greater improvements in MV density and overall renal function after PTRAs+ELP‐VEGF therapy.

Support or Funding Information

R41DK109737, R01HL095638, P01HL51971, P20GM104357, R01HL121527, EIA18490005, IPA34170267, and T32HL105324

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interleukin‐4 supplementation improves the proinflammatory cell ratios, autoantibodies and blood pressure in response to placental ischemia
Authors: Amaral Lorena , Cottrell Jesse , Harmon Ashlyn , Cornelius Denise , Ibrahim Tarek , Cunningham Mark , Vaka Venkata , Wallukat Gerd , Herse Florian , Dechend Ralf , LaMarca Babbette
Published: June 2021

Preeclampsia (PE), new onset hypertension, is associated with chronic inflammation, autoantibodies to the AT1 receptor (AT1‐AA) and placental ischemia. Currently, there is no effective treatment for PE except for early delivery, making PE the leading cause for premature births worldwide. Evidence indicates a shift towards proinflammatory NK cell, CD4+TH1 vs CD4+TH2 profile exist in PE and in response to placental ischemia in pregnant rats (RUPP). Therefore, we hypothesized that decreasing cytolytic NK and CD4+ TH1 cells and increasing CD4+TH2 cells would improve blood pressure during pregnancy. To do so we infused IL‐4, an anti‐inflammatory cytokine strongly associated with the proliferation of TH2 cells, in to normal pregnant (NP) and RUPP rats. Interleukin‐4, 600 ng/day, was administered via a mini‐osmotic pump on day 14 of gestation, carotid catheters were inserted on gestation day 18, and blood pressure (MAP), AT1‐AA, NK cells, TH2 and B cells were measured on GD 19. MAP in NP rats (n=10) was 96 ± 2, 101 ± 2 in NP+IL‐4 rats (n=14), 130 ± 4 in RUPP rats (n=6), and 108 ± 2 mmHg in RUPP+IL‐4 rats (n=15), p< 0.05. Circulating TH2 cells were 17 ± 3 in NP rats (n=3), 9.0 ± 2.0 in RUPP rats (n=5), but improved to 13 ± 3 % gated in RUPP+IL‐4 rats (n=9). Total placental NK cells were 0.7±0.1 in NP rats (n=4), 0.7±0.3 in NP+IL‐4 rats (n=7), 5.0 ±2 in RUPP rats (n=3) which significantly decreased to 1.6±4 in RUPP±IL‐4 rats (n=7), p<0.05. Placental cytolytic NK cells were 6±2 % in RUPP rats but reduced significantly to 2±0.8 in RUPP+IL‐4 rats. Importantly, circulating B cells were 2.5±0.3 in NP rats (n=3), which increased to 7.5±2 in RUPP rats (n=4) and reduced to 1.4±0.5 in RUPP+IL‐4 rats (n=3). Similarly, AT1‐AA were 0.3±0.4 in NP rats (n=8), 18±0.3 in RUPP rats (n=9), and were blunted to 4±1 bpm in RUPP+IL‐4 rats (n=14), p<0.05. This study illustrates the importance of a balanced CD4+ TH1/TH2, B and NK cell profile during pregnancy and that this can be achieved by administration of IL‐4, which ultimately improved hypertension in response to placental ischemia during pregnancy.

Support or Funding Information

NIH grants R01HD067541‐06, T32HL105324

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
H1/H2 Histamine Receptor Blockade Alters the Glutathione Redox Status in Skeletal Muscle Following a Bout of Prolonged Exercise
Authors: Turnbull Patrick C , Wallace Helen Elizabeth , Davidson Laura R , Samson Erin M , Graham Amy PR , Bell Mackenzie CW , Brebner Karen , Perry Christopher GR , Kane Daniel A
Published: June 2021

The effects of an acute bout of prolonged exercise on muscle include increases in oxidative metabolism and altered cellular redox environment. Histamine mediates many acute exercise responses, including sustained vasodilation and associated delivery of substrates to muscle. Previously, we demonstrated that H1 and H2 histamine receptor blockade with oral antihistamines resulted in a diminished capacity for mitochondrial H2O2 emission in skeletal muscle following an acute bout of prolonged exercise. Here, we tested the hypothesis that along with diminishing mitochondrial H2O2 emission, H1 and H2 histamine receptor blockade would attenuate exercise‐induced changes in skeletal muscle glutathione redox status. Adult male Wistar rats (83 ± 5.5 days old; 440.04 ± 40.70 g) were assigned to one of four groups (n = 12/group): exercise, in the form of one hour of continuous treadmill running at 20 cm/sec (Group 1); mass‐specific oral gavage with H1 antagonist fexofenadine (7.11 mg/kg) and H2 antagonist ranitidine (3.95 mg/kg) one hour prior to the same exercise protocol (Group 2); a group receiving oral H1 and H2 antagonists, but not exercised (Group 3); and a control group, which received neither antihistamines nor exercise (Group 4). An hour post‐exercise, total, reduced (GSH) and oxidized glutathione (GSSG) were assessed in red gastrocnemius (mixed fiber types) and soleus (oxidative) skeletal muscles. Redox status was expressed as GSH/GSSG. While no changes were observed in total glutathione for any treatment in either muscle, in soleus, antihistamine treatment, but not exercise decreased GSH/GSSG (main effect; p < 0.05). In red gastrocnemius, exercise, but not antihistamine treatment decreased GSH/GSSH (main effect; p < 0.05). These observations trend with those of a blunted capacity for mitochondrial H2O2 emission in skeletal muscle with antihistamine treatment and suggest an altered redox response to exercise following H1/H2 histamine receptor blockade in soleus. Given the widespread and routine use of antihistamines, further research into the effects of these antagonists on redox status in muscle is warranted.

Support or Funding Information

Natural Sciences and Engineering Research Council of Canada, Nova Scotia Health Research Foundation, Canada Foundation for Innovation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
INTEGRATIVE QUANTUM ELECTROMAGNETIC ENERGY HOMEOSTASIS SYSTEM
Authors: Holets Hilary
Published: June 2021

Photonic energy measured and applied by individual healing frequencies to each individual's homeostasis system is a modern approach to determining food allergies and health imbalances. Throughout time and especially the last few decades there has been an uprise of food sensitivities, extending past the general peanut, lactose intolerance, and infant honey allergies. Gluten intolerance for instance is more than a health trend. While the standard blood test has been used by physicians to determine allergy sensitivities, it does not help with determining how to measure the energy of foods categorized by macronutrient, nor the energy frequency of that food prior to, nor its immediate and long‐term effects on the body's energy prior to and post‐consumption of that particular food. Modern companies' products such as Dexafit's Biomarker Analysis and Habit's personalized nutrition package promise they are more accurate than the archaic blood test by accounting for individuals' DNA and its effect on how foods are metabolized; however, there have been no significant testimonials for disease or allergy reversal. This thesis deals with measuring different foods' effects on the body's homeostatic system, thereby allowing quantitative evidence and overall awareness of what foods individuals are allergic or sensitive to, further causing infection or disease. With this quantitative approach it is possible to then take the poisonous or toxic energy levels of foods and determine proper healing frequency for safe consumption, allowing for homeostasis to either return, be maintained, or further improved to the most optimal level for that particular individual. All of this is to represent how food sources either maintain, improve, or disrupt our body's homeostasis. The methods are specifically analyzing vegan and raw food diets compared to other well‐known lifestyles such as the keto diet to assess proper amounts of macronutrients required for individuals as well as from which nutrient sources. These experiments disclose data such as disease causation, mental illness, weight gain, and others pressed upon our systems. Once data is derived and applied, the body can ultimately heal itself. Photonic energy is the future for determining how we can bring our bodies back to their ideal state by proper mind, body, and soul alignment, eliminating instability and sickness.

Support or Funding Information

The University of Washington and other institutes are providing laboratory assistance in the following research conducted.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ultrasound Contrast Microbubble Targeted Cardiac‐specific Copper Delivery Improves Myocardial Recovery from Ischemic Injury
Authors: Feng Qipu , Fan Suzhen , Kang Y. James
Published: June 2021

Previous studies demonstrated that ischemic insult causes a decrease in myocardial copper concentrations but an increase in serum copper concentrations. In addition, copper supplementation to the ischemic heart improves the structural and functional recovery of this organ. However, the high level of serum copper is not available for the copper‐depleted ischemic heart. The present study was undertaken to develop an ultrasound contrast microbubble targeted cardiac‐specific copper delivery procedure. Copper‐albumin microbubbles (Cu‐MB) were produced. Copper sulfate solution was mixed with human albumin solution through a series of reactions to reach an optimal proportion to form a saturated copper‐albumin complex as the membrane of microbubble (MB). The copper‐albumin complex was then subjected to gas‐filling and sonication to generate copper‐albumin microbubbles (Cu‐MB). The quality and the stability of the Cu‐MB were tested and proved to be applicable in vivo. Mice were then used to test the efficacy of the ultrasound‐targeted and Cu‐MB‐mediated cardiac‐specific copper delivery. The mice were subjected to permanent ligation of left anterior descending (LAD) coronary artery to induce myocardial ischemic injury. Four weeks after the LAD ligation, Cu‐MB was infused to mice via tail vein injection, and ultrasound treatment was applied to target the ultrasound probe at the position where infarcted myocardium was identified. Copper concentration in the blood was significantly increased after Cu‐MB infusion. The hearts were harvested 3 min after the Cu‐MB and ultrasound treatment and were found to contain significantly higher concentrations of copper than that in control or those mice subjected to Cu‐MB infusion without ultrasound treatment. Importantly, this Cu‐MB and ultrasound treatment significantly improved the myocardial structural and functional recovery from ischemic injury. This study thus demonstrates that the ultrasound Cu‐MB targeted cardiac‐specific copper delivery system is effective and the treatment with this procedure leads to improved myocardial recovery from ischemic injury in the mouse model.

Support or Funding Information

Scientific research funds of West China Hospital of Sichuan University

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Melatonin on Vascular Health in Humans
Authors: Jeong Jinhee , Tucker Matthew A , Rodriguez‐Miguelez Paula , Looney Jacob , Thomas Jeffery , Blackburn Marsha , Derella Casandra , Harshfield Gregory A , Harris Ryan A
Published: June 2021

Melatonin, a circadian hormone released from the pineal gland, has recently been recognized to be involved in the pathophysiology of cardiovascular disease. Due to systemic localization of its receptors, melatonin is thought to play an important role in the regulation of vascular function. Data from animal models provide evidence that melatonin is protective against endothelial damage and the subsequent development of atherosclerosis. However, the effects of melatonin on vascular function in humans have yet to be determined.

Purpose

The present study tested the hypotheses that, in humans 1) endogenous melatonin production is associated with vascular health and 2) supplemental melatonin improves vascular health.

Methods

Upon arrival to the lab, 12 apparently healthy adult men (28±3 y) provided an overnight urine sample to measure 6‐sulfatoxymelatonin, the urine metabolite of melatonin. Subsequently, vascular endothelial function (flow‐mediated dilation; FMD) was assessed. In a follow‐up experiment, a comprehensive assessment of vascular health including brachial arterial blood pressure, FMD, and arterial stiffness (carotid‐femoral pulse wave velocity; PWV) was assessed in 4 individuals (31±6 y) at baseline and 1 hour following a single oral dose of melatonin (5 mg).

Results

FMD (%) was positively associated with total overnight melatonin production (ng; r=0.592, p=0.042) and melatonin excretion rate (ng/hour; r=0.539, p=0.071). Compared with baseline, PWV significantly decreased (5.9±1.3 to 5.7±1.3 m/s, p=0.003) and FMD increased (7.5±2.2 to 8.1±1.8 %, p=0.173) 1 hour following ingestion of supplemental melatonin. Supplemental melatonin did not change blood pressure (114±6 to 114±9 mm Hg, p=0.927).

Conclusion

These novel data in humans demonstrate that higher endogenous melatonin production is associated with greater vascular endothelial function. In addition, a single dose of supplemental melatonin is capable of reducing arterial stiffness and improving endothelial function. Together, these findings suggest that melatonin may serve a beneficial role in regulating vascular function in humans. Achieving adequate overnight melatonin production may represent an important therapeutic strategy for the maintenance of optimal vascular health.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bacterial modification of bile acids alters host physiology
Authors: Devlin Sloan
Published: June 2021

The human gut microbiome is known to play a crucial role in human energy harvest and homeostasis and has been implicated in the pathophysiology of obesity and metabolic syndromes. Defining the roles of specific microbial activities and metabolites on host phenotypes has proven challenging, however, due to the complexity of the microbiome‐host ecosystem. This lack of mechanistic understanding regarding the functions of microbial species has limited the effectiveness of both dietary and therapeutic approaches to improving host physiology. Using both genetic and chemical tools that target gut bacteria, we now report that we can selectively modulate the in vivo levels of a class of metabolites called bile acids, and that this controlled alteration of the bile acid pool exerts distinct effects on host physiology.

Bile acids are steroidal metabolites that play crucial roles in host metabolism by acting both as detergents that aid in digestion and as ligands for host receptors, including nuclear hormone receptors and GPCRs. In the colon, bacteria chemically modify host‐produced primary bile acids, generating a group of approximately 50 metabolites called secondary bile acids. While imbalances in this bile acid pool have been correlated with metabolic disorders, it is unclear which bile acids exert beneficial or harmful effects on host metabolism. Here, we report that we can control the in vivo pool of bile acids in mice by inhibiting the keystone enzyme of bacterial bile acid metabolism, bile salt hydrolases (BSH). Germ‐free mice monocolonized with a BSH‐deleted strain displayed altered metabolism, including reduced weight gain and reduced respiratory exchange ratios, as well as transcriptional changes in metabolic, circadian rhythm, and immune pathways in the gut and liver. We also determined that the metabolic differences observed were not due to differential detergent abilities of the bile acid pools but rather to bile acid‐mediated signaling. These results indicate that the deletion of a single bacterial gene can profoundly alter host metabolism. In ongoing work, we have developed a small molecule pan‐inhibitor of BSH. We are currently using this inhibitor as a chemical tool to probe the effects of bacterial bile acid modifications on host metabolism in fully colonized animals.

Support or Funding Information

R35 GM128618, NIH/NIGMS; 5P30DK034854‐33, NIH/NIDDK (Harvard Digestive Diseases Center); Center for Microbiome Informatics and Therapeutics at MIT Innovation Award; Karin Grunebaum Cancer Research Foundation Faculty Research Fellowship; Quadrangle Fund for Advancing and Seeding Translational Research (Q‐FASTR).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Base Editing: Chemistry on a Target Nucleotide in the Genome of Living Cells
Authors: Liu David
Published: June 2021

Point mutations represent the majority of known human genetic variants associated with disease but are difficult to correct cleanly and efficiently using standard nuclease‐based genome editing methods. In this lecture I will describe the development, application, and evolution of base editing, a new approach to genome editing that directly converts a target base pair to another base pair in living cells without requiring DNA backbone cleavage or donor DNA templates. Through a combination of protein engineering and protein evolution, we recently developed two classes of base editors (BE4 and ABE) that together enable all four types of transition mutations (C to T, T to C, A to G, and G to A) to be efficiently and cleanly installed or corrected at target positions in genomic DNA. The four transition mutations collectively account for most known human pathogenic point mutations. Base editing has been widely used by many laboratories around the world in a wide range of organisms including bacteria, fungi, plants, fish, frogs, insects, mammals, and even human embryos. We have recently expanded the scope of base editing by enhancing its efficiency, product purity, targeting scope, and DNA specificity. By optimizing base editor expression, we developed “max” versions of cytosine and adenine base editors with greatly increase editing efficiency in mammalian cells. We also show that base editing can function in vivo in post‐mitotic somatic cells that do not support homology‐directed repair. To improve the targeting scope of base editing, we used our phage‐assisted continuous evolution (PACE) system to rapidly evolve Cas9 and base editor variants with both broadened PAM compatibility and higher DNA specificity. Finally, we integrated several of these developments to address cell and animal models of human genetic disease. Base editing can be used to correct pathogenic point mutations, introduce disease‐suppressing mutations, and create new models of genetic diseases.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anticancer effects of Qizhen Capsule in human colorectal cancer cells in vitro and in vivo
Authors: Guo Dandan , Guo Chenjie , Wang Yujie , Wu Kaikai , Fang Liu , Guo Cuiling , Wang Xingya
Published: June 2021

Qizhen capsule is a new type of anti‐cancer traditional Chinese medicine preparation. As an adjuvant therapy for cancer, it has been widely used in clinical practice with remarkable therapeutic effects in clinic. According to the current research reports, Qizhen capsule is mainly used for the treatment of lung, gastric and breast cancer. However, whether Qizhen capsule has an inhibitory effect on colorectal cancer has not been reported, and the specific molecular mechanism of its anti‐tumor activity has not been well studied. In this study, the effect of Qizhen capsule water extracts on the proliferation and apoptosis of human colon cancer HCT116 cells was studied in vitro and in vivo. MTT assay revealed that Qizhen capsule at 0.3 to 0.7 mg/ml significantly inhibited proliferation of HCT116 cells in a dose‐and time‐dependent manner. Furthermore, Qizhen capsule water extracts induced apoptosis in HCT116 cells that determined by flow cytometric, which is associated with deregulation of the expression of key genes and proteins (Bcl‐2, Bax, PARP, caspase‐3 and caspase‐9) that regulate apoptosis. Moreover, oral administration of 4.5 g/kg and combination treatment of 1.5 g/kg Qizhen capsule and 5‐Fu significantly inhibited HCT116 xenograft tumor growth in nude mice by reducing HIF‐1α, Bcl‐2, NF‐κB and c‐FOS at mRNA levels, and up‐regulating PARP, FoxO3a and NAG‐1 at protein levels, which was further confirmed by suppressed Ki‐67, caspase‐3, Bcl‐2 and PCNA staining in tumor samples as determined by immunochemistry. Taken together, our study suggests that Qizhen capsule could inhibit the development of colorectal cancer by inhibiting cell proliferation, inducing cell apoptosis, and suppressing tumor growth. These results also suggest that Qizhen capsule may be a promising anticancer drug for the prevention and treatment of colorectal cancer.

Support or Funding Information

National Natural Science Foundation of China (Grant No. 81473397).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
THE TOY AS ANATOMY TEACHING AND LEARNING TOOL
Authors: Campos Letícia Cavalcante , Filho Cezar Nilton Rabelo Lemos , de Menezes Victor Matheus Lean , Carioca Levi Costa , Filho Luiz Alexandre Porto Castro , da Silva Ivan do Nascimento , Barreto Joao Erivan Facanha , Santos Cerqueira Gilberto S
Published: June 2021

Playing is a rich, complex human experience that in the process of teaching and learning anatomy promotes meaningful learning. Among these activities in anatomical teaching we use the dolls and dolls as a playful tool for students of medicine and pharmacy courses. Nowadays the task of teaching is a big challenge, especially when we compete with the problems of using the telephone in the classroom. In this way we elaborate an active and playful method as a tool for learning the muscular system in the teaching of anatomy. Based on these premises the objective of this work was to investigate if the use of toys (dolls) as a tool in the process of teaching and learning the anatomy of the muscular system promotes learning. An analytical and cross ‐ sectional study was carried out with 74 pupils who used dolls and pins to identify muscles. Then, based on atlas images of human anatomy, the students performed 100 anatomical structure markings with pins with the purpose of identifying the muscles and their action, developing the spatial notions of the muscles that make up the human body. To collect data, a semi‐structured questionnaire on the perception of learning was applied. For statistical analysis, the Mann Whitney test was considered significant when presented at p <0.05. The mean age of the interviewees was 19.34 ± 0.53 for males and 19.22 ± 0.2 for females. A statistically significant difference was observed in pupils' perception of learning before (5.60 ± 0.31) and after (8.08 ± 0.2) the use of toys as an anatomy learning tool. Regarding the preference for traditional teaching and play teaching no statistically significant difference was observed, most of the students reported that they prefer hybrid teaching with a mixture of active and traditional methodology. However, we observed a reasoning and correctness of the muscles in the students who performed the activity of markings with the muscular system. It was verified that the toy was demonstrated as a methodology with great acceptability, promoting significant and playful learning of the muscular system among the students for the study and learning of the human anatomy.

Support or Funding Information

CNPQ

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Highly efficient capture of circulating tumor cells by microarray in a microfluidic device
Authors: Liu Hui‐Yu , Hille Claudia , Haller Anna , Kumar Ravi , Pantel Klaus , Hirtz Michael
Published: June 2021

We established a versatile microarray‐based platform that is able to capture single target cell from large background populations. To show the efficacy of the device, we consider the challenging application of detecting circulating tumor cells (CTCs) – where the target cell appears in a concentration of about one cell in a billion. The cells were incubated with a biotinylated antibody cocktail, targeted cells are extracted on a streptavidin microarray in a herringbone design microfluidic chip. The accessibility to the captured target cell on our device facilitates subsequent recovery of the targets for further analysis. Our previous results (Brinkmann et al., 2015) have shown that analyzing blood samples from cancer patients with our platform reaches, and partly outreaches, gold standard performance, demonstrating feasibility for clinical application. Currently, we aim to optimize the design of our device to further increase the capturing rate. We tried 3 different herringbone designs of our microfluidic chips (which we will refer to as the HA, HB and HC chips), 3 different patterns of streptavidin depositions (dots with a pitch of 25 μm, 50 μm, and a homogenous coating). The combination of 50 μm pitch dots, the HC chip showed the highest capturing rate. In the viability test, 90% tumor cells were alive after capturing. Furthermore, we can easily open the PDMS cap to access the alive tumor cells, which allows us to do further downstream analysis. Our device allows clinical researchers to freely choose the desired antibody and detect different rare bio‐targets. This facilitates virtually arbitrary capturing of target species and therefore a wide spread application in the biomedical sciences.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic reprogramming of the stromal epigenome in ovarian cancer metastasis
Authors: Eckert Mark A , Coscia Fabian , Chryplewicz Agnieszka A , Chang Jae Wong , Hernandez Kyle M , Pan Shawn , Tienda Samantha M , Nahotko Dominik A , Li Gang , Blaženović Ivana , Lastra Ricardo R , Curtis Marion , Yamada S. Diane , Perets Ruth , McGregor Stephanie , Andrade Jorge , Fiehn Oliver , Moellering Raymond E , Mann Matthias , Lengyel Ernst
Published: June 2021

High grade serous ovarian cancer (HGSOC) is characterized by early metastasis throughout the peritoneal cavity and a poor clinical outcome. During the metastatic process, ovarian cancer cells interact with diverse cell populations, including mesothelial cells, adipocytes, and cancer‐associated fibroblasts (CAFs). Ovarian cancer CAFs have well‐described roles in promoting the growth and progression of ovarian cancer by secreting pro‐proliferative cytokines and oncogenic extracellular matrix components (ECM). To better understand the proteomic changes that occur during metastasis in both tumor and stromal compartments, laser‐capture microdissection was coupled to label‐free, quantitative proteomics to independently characterize the proteomes of both tumor and stromal compartments in a cohort of HGSOC patients. We identified a conserved stromal proteomic signature associated with metastasis which included high expression of nicotinamide N‐methyltransferase (NNMT) in the metastatic stroma. NNMT is a metabolic enzyme that catalyzes the methylation of nicotinamide and consumes S‐adenosyl methionine (SAM), thus depleting cellular methylation potential and reducing methylation of DNA and proteins. NNMT regulated the expression of thousands of genes and was necessary and sufficient for the reprogramming of normal fibroblasts to CAFs. Stromal NNMT expression altered multiple metabolic pathways and led to a significant reduction in cellular SAM pools. Attenuated DNA methylation and histone H3 lysine 27 trimethylation (H3K27me3) were associated with global changes in gene expression. Using genetic and small‐molecule inhibitor strategies, we found that stromal NNMT activity promoted the progression and metastasis of ovarian cancer. This study provides support to the idea that stromal methyltransferase activities can be targeted to normalize the metastatic stroma and should be further explored as treatment targets for cancer.

Support or Funding Information

This work was supported by a Marsha Rivkin Foundation award (MAE), National Cancer Institute (NCI) grants CA111882 and CA211916 (EL), the Harris Family Foundation (MAE and SDY), the Ludwig Institute for Cancer Research (EL), NIH grant CA175399 and DP2GM128199 (REM), V Foundation for Cancer Research (V2016‐020 to REM), the Körber Foundation/Körber European Science Prize (MM), the Max‐Planck Society for the Advancement of Science (MM), the Novo Nordisk Foundation (Grant agreement NNF14CC0001 and NNF15CC0001; FC and MM), and University of Chicago Cancer Center Support Grant P30CA014599.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of Genes that Mediate Persistence in Halophilic Microbes Subjected to Osmotic Shock
Authors: Gregory Abby M , Miranda‐Katz Margot R. , Miranda‐Katz Margot R
Published: June 2021

All organisms are subject to stress from environmental changes, and understanding the specific molecular responses to allow cells to maintain viability is a fundamental biological problem. We explore this process in the halophilic (salt‐loving) microbe Haloferax volcanii. This species was originally isolated from the Dead Sea and grows optimally at 12–14% sodium chloride. However, these organisms are regularly subjected to rapid changes in their osmotic environment during rainfall or flooding events, and we hypothesize that they have evolved so that a subset of their population consists of cells prepared to survive dramatic stress events. As a first step to examine how these ‘persisting’ cells endure, we used RNA sequencing to examine transcriptome differences in cells that survived osmotic shock compared to unstressed cells. Our data revealed that approximately half of the protein‐coding genes exhibited statistically significant differences in transcriptome abundance with a nearly equal distribution of more abundant and less abundant transcripts. There were about twice the number of transcripts exhibiting a 10‐fold or greater decrease (5.3% of protein‐coding genes) in abundance than those with the same magnitude of increased abundance (2.7% of protein‐coding genes). This result may indicate that cells primed for survival reduce production of proteins required for active metabolism. We are currently conducting experiments to determine if deletion or overexpression of some of these genes affects susceptibility of H. volcanii to osmotic shock. Our work may provide insight into why some cells in a microbial population survive stress events despite being genetically identical to cells that are killed. Similar mechanisms may also allow disease‐causing bacteria that persist even after being exposed to antibiotic agents. Potentially, the factors that carry out these responses could be targeted to enable antibiotics to effectively eliminate persister cells.

Support or Funding Information

This project was supported by an Institutional Development Award from the National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health under grant P20GM103423. Additional funding was provided by the Colby College Natural Science Division.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Three Foodborne Bacteria using Hyperspectral Microscopy
Authors: Riggs Rebecca Dobson , Chen I‐Hsuan , Barbaree James , Zinner Bertram , Sorokulova Iryna , Vodyanoy Vitaly
Published: June 2021

The field of hyperspectral imaging (HSI) has progressed exponentially since its inception in 1972 and is applied to various research fields including agriculture, food quality and archeology. The purpose of the technology is to acquire images containing spatial and spectral data that can be used for detection and/or identification purposes. Our goal was to use HSI technology to differentiate between 3 different bacteria that are common foodborne organisms in 3 phases of growth: log, late log and stationary.

For our study we used the Cytoviva hyperspectral imaging system to extract the individual hyperspectral profiles produced by Salmonella enterica serotype Typhimurium, Escherichia coli O157: H7, and Listeria monocytogenes. All organisms were cultured in Brain Heart Infusion (BHI) broth, pelleted at 10,000× rpm and washed in 1× PBS buffer prior to imaging. 0.8 μL of each sample was transferred to a poly‐l‐lysine coated slide for immobilization and topped with a glass cover slip to extract the profiles of each organism. Our protocol was enhanced to extract spectral profiles of intact live microorganisms.

Data is collected by the pushbroom approach through which an image is made that has both hyperspectral and spatial data by moving the sample pixel line by pixel line to produce distinct profiles for each specimen. The use of a patented enhanced darkfield condenser in conjunction with high resolution optical and hyperspectral imaging creates sharp spectral profiles based upon the unique surface chemistries of each organism. The hyperspectral profiles of the cells were composed from each image through the ENVI software.

The unique spectral graphs produced by each organism appear to align with increased growth, especially at the 24 hour time point. However, these organisms do exhibit different hyperspectral patterns at each time point and they are not the same for any organism. Hyperspectral imaging has the potential to be used as a rapid diagnostic tool that does not damage the sample while reporting data that can be shared and analyzed by multiple people at the same time.

Support or Funding Information

This work was supported in part by a grant from the Lake Erie College of Osteopathic Medicine, USA, Department of Biological Sciences and Department of Anatomy, Physiology and Pharmacology at Auburn University.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ganoderma lucidum polysaccharides induce cytotoxicity in colorectal cancer cells through inducing autophagosome accumulation and inhibiting autophagic flux
Authors: Pan Haitao , wang Yujie , Wang Ying , Li Mingyan , Li Zhenhao , Xu Jing , Wang Xingya
Published: June 2021

Ganoderma lucidum (G. lucidum), a traditional Chinese medicine, has been used in cancer prevention for many years. As one of the main components of G. lucidum, G. lucidum polysaccharides (GLPs) showed significant anti‐cancer activity. Previous studies in our laboratory show that GLPs extracted from sporoderm‐broken spores of G. lucidum, significantly inhibited the proliferation and induced apoptosis in colorectal cancer (CRC) cells in vivo and in vitro. However, whether GLPs regulate autophagy in CRC cells is unknown. In the present study, we found that GLPs enhance the level of LC3B and activating autophagy initiation both in HCT116 and HT29 CRC cells as determined by Western blot, transmission electron microscope, and confocal microscope. However, autophagy initiation induced by GLPs was accompanied by impaired degradation of SQSTM1/p62, which suggesting autophagic flux inhibition in CRC cells by GLPs. mRFP‐GFP‐LC3B adenovirus translocation and colocalization analyses confirmed that autophagosome‐lysosome fusion is blocked by GLPs treatment. Furthermore, GLPs inhibited autophagic degradation by inhibiting lysosomal acidification and downregulating the expression of cathepsin B and cathepsin D. Next, we found GLPs induced expression of MAPK/Erk in both CRC cells. Erk inhibitor PD98059 inhibited GLPs‐induced apoptosis and LC3B expression in CRC cells suggesting that Erk activation is partially, at least, responsible for GLPs‐induced autophagy initiation. Importantly, further analysis using flow cytometry revealed that GLPs induced autophagosome accumulation is required for GLP‐induced cytotoxicity. Similar results were obtained using a xenograft mouse model, in which GLPs reduced tumor growth and increased the levels of LC3B and p62 in tumors. Moreover, the levels of cathepsin B and cathepsin D were decreased in tumors from GLP treated mice in vivo. Collectively, these data demonstrate that GLPs inhibit CRC cell proliferation and induce apoptosis through regulation of autophagosome accumulation and autophagic flux inhibition, which may be dependent, at least in part, on MAPK/Erk activation.

Support or Funding Information

Supported by Science and Technology Planning Project of Zhejiang Province (Grant No. 2019C02100) and National Natural Science Foundation of China (Grant No. 81473397).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Re‐evaluation of Artemether‐Lumefantrine Combination Effectiveness in Malaria Treatment in Elele
Authors: Chukwu Leo Clinton , Agbasi Patrick Ugochukwu , Unekwe Prince Chiazor
Published: June 2021

Background

Nigeria adopted Artenisinin based combination therapy (ACT) for uncomplicated malaria treatment owing to ACT's efficacy in combating malaria. The need arises to protect ACT efficacy by scaling up: continued assessment, monitoring and evaluation in malaria endemic areas.

Purpose

Re‐evaluation of effectiveness of artemether‐lumefantrine (ALC) combination for malaria treatment in Elele.

Method

This is a cross sectional study carried out at Madonna University Teaching Hospital (MUTH) Elele involving qualitative and quantitative data. 100 patients were recruited after consent using the inclusion criteria: temperature of ≥ 37.5 °C, malaria symptoms positive parasitaemiaetc. Ethical certificate was obtained from MUTH ethical committee. Patient's symptoms at Day 0 was assessed while their disappearance was determined on days 2;4;6;8;10;14. The data gotten was analyzed using SPSS 16.0 version and Microsoft excel packages. The student t‐test was used to calculate the P‐value, <0.05 was considered significant.

Results

Patients majority identified mosquito bites as the means of malaria transmission, (70%). ACT types known and used by patients: artesunate/amodiaquine: 41%); artesunate/lumefantrine: (40%) etc. Fever was the predominant symptoms 92% followed by weakness (90%) etc. Day 10 of therapy, fever, vomiting and abdominal pains disappeared in all patients. By day 14, no patient reported any symptom giving an: “Effective clinical response (ECR) by day 14”.

Conclusion

Malaria is an most important infectious disease. Artemether‐lumefantrine combination therapies has been proven to be efficacious in malaria treatment. Our hereby reaffirm the effectiveness of ALC for uncomplicated malaria treatment in Elele. The knowledge gap of continuous monitoring and evaluation of this regimen in rural endemic areas like Elele has been satisfactorily done in this study. This will help to protect the component drugs from treatment failures and the menace of resistant strains.

Support or Funding Information

No funding

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.