bluuebunny/crossref_metadata_2025 huggingface dataset:
Cross‐Reactivity of Rabbit Anti‐Bovine Thrombin IgGs with a Human Plasma Derived Thrombin Preparation (Evithrom™)
Authors: Zhu He , Hoppensteadt Debra , Cunanan Josephine , Fareed Jawed
Published: June 2021

Studies have shown that rabbit anti‐bovine thrombin IgGs cross‐react with human alpha‐thrombin and a recombinant version of human thrombin (Recothrom™). This study characterized the cross‐reactivity of anti‐bovine thrombin antibodies with a human plasma‐derived topical thrombin preparation (Evithrom™). Gel electrophoresis revealed that in addition to a human α‐thrombin protein band, Evithrom™ also contained a large amount of human albumin and a few faint high‐molecular weight bands which were absent in bovine thrombin, human α‐thrombin, human γ‐thrombin and Recothrom™. Western blotting results demonstrated no cross‐reactivity with Evithrom™ when anti‐bovine crude thrombin IgGs and thrombin 4B IgGs were used. However, Evithrom™ was recognized by rabbit anti‐bovine thrombin 4A IgG. Furthermore, the results revealed the cross‐reactivity of anti‐bovine thrombin 4A IgG with Evithrom™ was time‐dependent. These results indicate that besides human α‐thrombin and Recothrom™, rabbit anti‐bovine thrombin IgGs can also cross‐react with Evithrom™ suggesting that human antibodies against bovine thrombin may also cross‐react with human thrombin as well as human recombinant thrombin. Thus, the use of Evithrom™ and RecothromTM may not be recommended in patients who have been previously treated with/exposed to bovine thrombin preparations.
Role of Periostin in Epithelial Injury
Authors: Reaves Titus Antonio , Lebel David P. , Suber Jarod
Published: June 2021

Inflammatory Bowel Disease (IBD) is an inflammatory condition affecting the gastrointestinal tract. IBD is commonly represented by Crohn's Disease and Ulcerative Colitis. A major characteristic of IBD is intestinal epithelial injury resulting from the release of proteases by circulating and attached polymorphonuclear leukocytes (PMN). Periostin (90 kDa protein) is expressed by fibroblasts, up‐regulated during cellular injury, and we show that periostin is also expressed by T84 intestinal epithelial cells. Thus, we investigated epithelial repair following the combined culture of epithelial cells (underside) and intestinal fibroblasts (upper side) on 5 micron filters to resemble an in vivo configuration. “Scratch” injuries were made, but only to the epithelial side. Prior to epithelial injury, the filters were exposed to the inflammatory cytokine (IL‐6) and the non‐inflammatory cytokine (IL‐10). Results show periostin is expressed at the leading edge of an epithelial injury and Il‐6 treated cells increased expression of periostin while IL‐10 reduced expression of periostin. Taken together, these results suggest that periostin can play a role in the inflammatory responses of epithelial cells and we hypothesize that periostin serves as a chemoattractant for PMN to areas of inflammation, which contributes to the dysregulated cycle of intestinal inflammation observed in IBD.
An Analysis of 1 year versus 3 month alpha‐Interferon Treatment in Patients with Melanoma Stages IIB, IIC, and IIIA
Authors: Frankenthaler Amy , Atkins Michael B
Published: June 2021

Adjuvant therapy like alpha‐interferon is a common treatment for pts with melanoma staged IIB, IIC, or III. Standard therapy involved a 1 year course of trtmnt, recent studies suggest that the benefit of IFN in preventing relapse occurred during the first months of trtmnt. Our objective was to study the trtmnt course and outcome of in pts with stages IIB, IIC and IIIA melanoma treated with adjv IFN for 1 year vs 3 mnths. A registry of pts of the Cutaneous Oncology Program (COP) at the Beth Israel Deaconess Medical Center was used to identify pts with melanoma staged IIB, IIC, and IIIA who received IFN from Jan 2002 to Jan 2009. Of 1924 melanoma pts from the COP Registry, 86 received IFN from Jan 2002 and Jan 2009. Baseline demographic, disease characteristics, amount of IFN administered and disease outcome were compared. Multivariate analysis controlling for disease related prognostic factors that influence outcome in this non‐randomized comparison is pending. These prelim results suggest that trtmnt of melanoma staged IIB, IIC, or IIIA with interferon for 3 mnths may result in disease control equivalent to 1 year of therapy.
Inhibition of TNF‐α‐induced ROS generation, VCAM‐1 expression and NF‐κB activation by chloroform extract of aged black garlic in human umbilical vein endothelial cells
Authors: Park Jin Kyeong , Kim Hye Kyung , Kim Myoung June , Kim Hyo Jin , Lee Eun Na , Oh Sae‐Ock , Baek Sun‐Yong , Kim Bong‐Seon , Choi Young‐Whan , Yoon Sik
Published: June 2021

The current study investigated if the chloroform extract of aged black garlic (CEABG) could attenuate the activities of adhesion molecules in tumor necrosis factor‐α (TNF‐α)‐stimulated human umbilical vein endothelial cells (HUVECs). The study was performed on HUVECs that were pretreated with 30 μg/ml of CEABG before TNF‐α treatment. When pretreatment of HUVEC with CEABG for 30 min, CEABG suppressed reactive oxygen species (ROS) production induced by TNF‐α. HUVECs treated with CEABG exhibited remarkably suppressed TNF‐α‐induced mRNA expression of VCAM‐1, but little alteration in ICAM‐1 and E‐selectin mRNA expression. CEABG treatment also significantly decreased the TNF‐α‐induced cell surface and total protein expression of VCAM‐1 without affecting ICAM‐1 and E‐selectin expression. Furthermore, CEABG significantly inhibited NF‐κB transcription factor activation in TNF‐α‐stimulated HUVECs. Collectively, our findings suggest that CEABG can be an efficient new anti‐inflammatory agent. It may have a potential therapeutic use for the prevention and treatment of vascular disease such as atherosclerosis.
Ginger oleoresin supplementation lowers blood pressure and triglyceride levels in hypercholesterolemic rats
Authors: Henry Tricishanna Racquel , Simon Oswald , Gossell‐Williams Maxine
Published: June 2021

The aim of this study was to investigate the effects of ginger oleoresin on blood pressure and serum lipids in hypercholesterolemic rats. Three groups of Sprague Dawley rats were fed a hypercholesterolemic diet (consisting 95% rat chow, 4% cholesterol and 1% cholic acid) for six weeks and a control group was fed rat chow for the same period. Two of the groups on the hypercholesterolemic diet were given (orally) ginger oleoresin (242μg/kg and 484μg/kg) daily for six weeks. The third group on the hypercholesterolemic diet and the control group received (orally) ginger oleoresin vehicle daily for six weeks. Weekly blood pressure measurements were taken, and at the end of the six weeks blood was taken from each animal for serum lipid analysis. The data was analyzed using unpaired student's t test. The results show that the hypercholesterolemic diet significantly increased blood pressure, total cholesterol, triglyceride and low density lipoprotein (LDL) concentrations in comparison to the results in the control group of rats. Treatment with ginger oleoresin significantly decreased blood pressure from week three. Triglyceride concentration was also significantly decreased by ginger oleoresin (484μg/kg), but total cholesterol, LDL and high density lipoprotein (HDL) concentrations were not significantly affected by the ginger oleoresin treatment. It is concluded that ginger oleoresin prevented blood pressure elevation produced by hypercholesterolemic diet. It also reduced serum triglyceride concentration without significantly affecting total cholesterol, LDL and HDL concentrations.
Acute nicotine attenuates reflex tachycardia in male rats via inhibition of baroreceptor‐mediated cardiac sympathetic control
Authors: El‐Mas Mahmoud M , El‐gowilly Sahar M , Fouda Mohamed A , Saad Evan I
Published: June 2021

The effect of acute nicotine on reflex chronotropic responses was tested in conscious male and female rats. Baroreflex curves relating changes in heart rate to increases or decreases in blood pressure (BP) evoked by increasing i.v. doses (1–16 μg/kg) of phenylephrine (PE) and sodium nitroprusside (SNP), respectively, were constructed and slopes of the curves were taken as a measure of the baroreflex sensitivity (BRSPE and BRSSNP). In male rats, nicotine (25 and 100 μg/kg) caused significant and dose‐dependent reductions in reflex tachycardic responses (BRSSNP, −2.05±0.41 and −1.70±0.16 beats/min/mmHg, respectively) compared with control values (−3.11±0.42 beats/min/mmHg). Reflex bradycardia tested with PE was not affected by nicotine. The blockade of β‐adrenergic (propranolol) and not muscarinic (atropine) receptors abolished the nicotine‐evoked attenuation of BRSSNP, suggesting a prominent role for impairment of reflex sympathoexcitatory responses in the nicotine‐BRSSNP interaction. On the other hand, neither BRSPE nor BRSSNP was affected by nicotine in female rats. The results indicate that the interaction of nicotine with baroreceptor function depends on rat gender and nature of the response. Nicotine preferentially impairs reflex tachycardia in male rats only, probably via interruption of cardiac sympathetic control.
Efficacy versus toxicity of docetaxel in Asian and Caucasian cancer patients from the pharmacogenomics perspectives: a review of the literature
Authors: Jabir Rafid Salim , Naidu Rakesh , Azrif Muhammad , Stanslas Johnson
Published: June 2021

Docetaxel is a common anticancer agent in the treatment of breast and lung cancers. Metabolism of docetaxel occurs through cytochrome P450 (CYP) and the function of the enzyme can be altered by genetic, environmental and physiological factors. Interindividual differences in pharmacokinetic profiles of anticancer agents due to changes in CYP function lead to variations in drug efficacy and/or toxicity. Unpredictable toxicities of taxanes have been reported, related to interindividual variability of pharmacokinetics of the drugs. It is evident from multiple studies that adverse events, which are mainly haematological amongst Asian patients receiving docetaxel are higher than as reported in the Caucasian population. Most clinicians who treat Asian patients thus prophylactically support patients with growth factors or reduce the dose of docetaxel. Polymorphisms in genes encoding transporters involved in the uptake of docetaxel will lead to accumulation of the drug in the circulation e.g. ABCB1. Polymorphisms in β‐tubulin (target of docetaxel) affect the taxane chemotherapy results. Presently we have embarked on a project to study the pharmacogenomics of docetaxel in Malaysian breast cancer patients for the very first time to address the above issues. This project is funded by UPM and Al‐Malak Universality, Iraq.
Histone Acetylation Up‐regulates Phosphatidylinositol‐3,4,5‐triphosphate‐dependent Rac Exchange Factor 1 (P‐Rex1) Expression to Promote Prostate Cancer Metastasis
Authors: Wong Chuu‐Yun Anny , Ha Da , Xie Yan , Wolff Dennis W. , Abel Peter W. , Tu Yaping
Published: June 2021

We have reported that P‐Rex1 up‐regulation promotes metastasis, the major cause of prostate cancer death (Oncogene, 28: 1853–1863). The objective of this study was to combine biochemical, molecular and cellular techniques to define mechanisms underlying P‐Rex1 up‐regulation. Quantitative real‐time PCR and Western blot analysis indicated that histonedeacetylase (HDAC) inhibitors but not DNA methylation inhibitors increased P‐Rex1 expression in non‐metastatic prostate cancer cells to levels similar to that in metastatic prostate cancer PC‐3 cells. HDAC inhibitors enhanced P‐Rex1 promoter activity and a transcription factor Sp1 binding site in this promoter is responsible for the effect of HDAC inhibitors. Chromatin immunoprecipitation assays demonstrated that Sp1 association with the P‐Rex1 promoter is essential for P‐Rex1 expression. HDAC inhibitors caused dissociation of HDAC from Sp1, leading to histone acetylation and P‐Rex1 up‐regulation. Interestingly, HDAC1 over‐expression in PC‐3 cells caused histone deacetylation and P‐Rex1 down‐regulation, which inhibited metastatic activity in Transwell chamber assays. Our results demonstrate that histone acetylation stimulates P‐Rex1 expression in a Sp1‐dependent manner and suggest that targeting HDAC and Sp1 may provide a novel strategy for suppressing prostate cancer metastasis. Supported by NIH 1R011CA125661 and Department of Defense.
Metabolism of prostaglandin glyceryl esters (PG‐Gs) by human carboxylesterases, CES1 and CES2, and its inhibition by bioactive metabolites of organophosphate (OP) insecticides
Authors: Ross Matt K. , Borazjani Abdolsamad , Potter Philip M , Xie Shuqi
Published: June 2021

PG‐Gs are derived from the oxidation of 2‐arachidonoylglycerol by COX‐2 and have roles in inflammation and pain. Currently, the enzyme(s) responsible for degrading PG‐Gs are not well defined. Carboxylesterases (CES) are nonspecific hydrolases and candidate enzymes. Two CES isoforms are found in human tissues, CES1 and CES2. Here, we determined if PG‐Gs are hydrolyzed by human CES and then investigated the hydrolytic metabolism of these compounds in human THP1 monocytes/macrophages. Further, we determined if metabolites of OP insecticides could inhibit PG‐G metabolism in these cells. We show that recombinant human CES1 and CES2 effectively hydrolyze PGF2α‐G and PGE2‐G [CES1: (kcat/Km)PGF2α‐G = 0.60 min−1 μM−1and (kcat/Km)PGE2‐G = 0.37 min−1μM−1; CES2: (kcat/Km)PGF2α‐G = 1.7 min−1μM−1and (kcat/Km)PGE2‐G = 0.23 min−1μM−1]. In THP1 monocytes/macrophages, we demonstrated using selective CES1 inhibitors, immunoprecipitation, and immunoblotting that CES1 is likely the main enzyme responsible for hydrolyzing PGE2‐Gs, and that MAGL and FAAH do not have significant roles. In addition, pre‐treatment of intact cells with OP metabolite (0.1–10 μM paraoxon) significantly reduced the hydrolysis rate of exogenously added PGE2‐G (~20–70%, p<0.05). These findings suggest that exposure of CES1‐ and/or CES2‐expressing cells to bioactive OP metabolites may perturb the PG‐G tone in tissues composed of these cells, thus potentially inducing a pro‐inflammatory state.

Supported by NIH R15 ES015348.
PHARMACOLOGICAL EVALUATION OF FL‐6, A NEW IMMUNOMODULATORY DRUG, ON COCCIDIOSIS IN CHICKENS
Authors: Corbalá‐Nava Martha , Tellez Guillermo , Hargis Billy , Rodríguez‐Fragoso Lourdes , Reyes‐Esparza Jorge
Published: June 2021

The aim of this work was to evaluate the effect of pretreatment with FL‐6, a new imunomodulatory drug, on coccidiosis in chickens. Broiler chickens were used with a day of born. Animals were maintained under controlled conditions and without any treatment. We included five groups (30 animals per group): group 1 was control (vehicle); group 2, was infected with Eimeria maxima (EM) on day 8 and maintained separately; group 3 was treated with FL‐6 control, group 4 was pretreated with vehicle, and group 5 was pretreated with FL‐6 and infected with EM. The groups 1, 3 to 5 were pretreated with vehicle or FL‐6 at 10th and 17th days. On day 24th the group 4 and 5 were mixed with group 2 (EM) in order to acquire natural infection, for a week. Chickens were weighed every week and underwent necropsy. Chickens with FL‐6 pretreatment, showed a gains on corporal weight, this effect was higher than the control group, without infection. The guts of the animals from group pretreated with FL‐6 showed less severe injuries as compared with other groups. Our results might suggest that the pre‐treatment with FL‐6 probably increased the nonspecific immune response, thereby minimizing the effects caused by EM.
Structural importance of residue 334 in the stability of cytochromes P450 2B6 and 2B11
Authors: Talakad Jyothi C , Wilderman P. Ross , Davydov Dmitri R , Kumar Santosh , Halpert James R
Published: June 2021

Human P450 2B6 and canine P450 2B11 play an important role in xenobiotic metabolism, bioremediation research, and activation of anti‐cancer prodrugs. Thorough biochemical and biophysical characterization of the enzymes has been impeded by relatively low stability compared to the extensively studied rat 2B1 and rabbit 2B4. Comparison of the amino acid sequences of N‐terminal truncated and C‐terminal His‐tagged constructs revealed seven sites that are conserved between the more stable 2B1 and 2B4 but different from those found in the less stable 2B6 and 2B11. Site‐directed mutagenesis at these positions showed that P334→ Ser substitution increased heterologous expression levels and thermal stability in both 2B6 and 2B11. The mechanism of this effect was explored with pressure‐perturbation spectroscopy. Compressibility of the heme pocket in the four P450 2B enzymes and their P334S (2B6 and 2B11) or S334P (2B1 and 2B4) mutants was assessed from the pressure induced displacement of the Soret band. The variants of all four enzymes containing proline at position 334 are characterized by lower compressibility than their more stable serine 334 counterparts. Therefore, the stabilizing effect of P334S is associated with increased conformational flexibility in the region of the heme pocket. Mutation of the non‐active site residue P334→ Ser could find applications in drug metabolism and gene therapy for cancer treatment.
Electrophysiological characterization of postnatal enteric neuron cell line
Authors: Hawkins Edward G , Dewey William L , Srinivasan Shanthi , Akbarali Hamid I
Published: June 2021

Conditionally immortalized fetal and postnatal enteric neuronal cell lines (IM‐FEN and IM‐PEN) have been developed from the H‐2kb‐tsA58 mouse (immortomouse), however, their electrophysiological properties are unknown. The goal of the present study was to characterize the electrophysiological properties of IM‐PEN. We confirmed the neuronal background of the cell line with positive staining of neuron specific β‐III tubulin. The cells exhibited depolarized resting membrane potentials of ‐24.7 ± 4.7mV (n=6) and input resistances of 672 ± 29MΩ. Depolarizing pulses in current clamp failed to yield action potentials from rest or from hyperpolarized holding potentials (n= 6). This was consistent with the absence of inward Na+ or Ca2+ currents to depolarizing pulses (n= 5). In voltage clamp, step depolarization from Vh ‐60 mV to +30 mV in 10 mV steps produced time‐independent currents which were blocked by replacing external chloride. These currents were not blocked by the Icl,swell blocker, DCPIB (10μM) (n= 3), however, niflumic acid (1μM and 500 uM), a chloride channel blocker resulted in increased outward currents and a significant shift in the reversal potential from −37.3 ± 2.8mV to −53.0 ± 3.6mV (n= 14). The inhibitory neurotransmitter γ‐aminobutyric acid (GABA) at 100μM increased the outward current in 3 cells. External application of acetylcholine (500μM) (n= 4), nicotine (1mM) (n= 3), and serotonin (10μM) (n= 3), were ineffective. These studies confirm the neuronal origin of IM‐PEN which have a depolarized resting membrane potential due to high internal chloride concentration. Supported by NIH DA007027.
Diabetic pregnancy impairs EDHF‐dependent uteroplacental vasodilation mediated by intermediate‐conductance Ca2+‐activated K+ channels
Authors: Gokina Natalia I , Kuzina Olga Y , Vance Amanda M , Phillips Julie K
Published: June 2021

Diabetes in pregnancy is associated with such adverse outcomes as hypertension, preeclampsia and stillbirth. Increased uteroplacental vascular resistance due to endothelial dysfunction may be an underlying mechanism. This study aimed to (1) Characterize the effect of diabetes on EDHF‐mediated uteroplacental vasodilation; (2) Explore the role of endothelial intermediate‐conductance (IKCa) K+ channels in impaired of EDHF responses. Diabetes was induced by injection of streptozotocin (STZ) on day 2 pregnant rats. Uteroplacental arteries from day 20 of pregnancy were cannulated, pressurized and loaded with fura 2. The vasodilator and smooth muscle cell (SMC) [Ca2+]i responses to ACh, EBIO or NS309 were studied in pre‐constricted arteries in the presence of L‐NNA and indomethacin. STZ injection produced sustained hyperglycemia in treated rats. ACh‐induced vasodilation and reduction in SMC [Ca2+]i were attenuated by diabetes. EDHF responses were not affected by iberiotoxin or apamin but abolished by charybdotoxin or TRAM‐34. EBIO‐ or NS309‐induced vasodilation was abolished by TRAM‐34 and was significantly impaired in hyperglycemic rats. Diabetes in pregnancy results in a marked impairment of EDHF‐mediated vasodilation most likely due to dysfunction of endothelial IKCa channels, which may serve as a target of future pharmacotherapy to improve uteroplacental blood flow.

Supported by NIH HL088245
Overexpression of HIF‐1α transgene in the renal medulla attenuated salt‐sensitive hypertension in Dahl S rats
Authors: Zhu Qing , Wang Zhengchao , Xia Min , Li Pin‐Lan , Zhang Fan , Li Ningjun
Published: June 2021

We have previously shown that HIF‐1α‐mediated gene activation in the renal medulla in response to high salt intake (HS) plays an important role in the control of salt sensitivity of blood pressure. The present study was to test whether impairment of the renal medullary HIF‐1α pathway is responsible for the salt sensitive hypertension in Dahl S (DS) rats. By Western blot analysis, HS significantly increased renal medullary HIF‐1α levels in control (SS13BN) rats but not in DS rats, indicating a defect in renal medullary HIF‐1α. Renal medullary HIF‐1α levels were overexpressed in DS rats by either transfection of HIF‐1α expression plasmid or chronic infusion of CoCl2 into the renal medulla, which was accompanied by increased expressions of local anti‐hypertensive genes, nitric oxide synthase‐2 and heme oxygenase‐1. Hypertension induced by 2‐week HS was significantly attenuated in rats treated with HIF‐1α plasmid or CoCl2. The mean arterial pressure in these DS rats was 114.7 ± 2.7 mmHg on low salt intake, 157 ± 6.5 on HS, 133 ± 2.1 on HS with HIF‐1α plasmid and 135 ± 2.8 on HS with CoCl2, respectively. These results suggest that an abnormal response of renal medullary HIF‐1α to HS in DS rats may represent a novel mechanism mediating salt‐sensitive hypertension and that induction of HIF‐1α expression in the renal medulla could be a therapeutic approach for the treatment of salt‐sensitive hypertension (support: NIH grant HL89563).
Inflammatory Markers in Isolated Endothelial Cells of the Spontaneously Hypertensive Rat
Authors: Xiu Ruijuan , DeLano Frank A , Hongwei Li , Ailine Li , Schmid‐Schönbein Geert W
Published: June 2021

Besides an elevated blood pressure, the spontaneously hypertensive rat (SHR) has inflammation, extensive cell injury, microvascular and parenchymal cell dysfunction and vascular lesions. Current evidence in‐vivo shows that both excessive oxidative stress and proteolytic activity are involved, but their source is uncertain, We hypothesize that isolated endothelial cells of the SHR without the involvement of other sources (e.g. adrenal hormones) have elevated levels of oxygen free radical and degrading enzyme. To separate endothelial mechanisms from others, we isolated and cultured rat aortic endothelial cells (RAOEC) of adult SHR and control Wistar rats and propagated them with minimal loss of morphologic and phenotypic characteristics. The expression of oxidative enzymes (xanthine oxidase, XO; nicotinamide adenine dinucleotide phosphate, NADPH), apoptotic enzymes (BAX; BCl‐2) and matrix metalloproteinase (MMP‐9) was determined in cultured second passage RAOEC by immunoperoxidase detection of RAOEC using a standardized avidin/biotin immunolabeling technique and digitally measured by optical intensity. Compared to Wistar endothelium measurements, protein levels of XO, NADPH, MMP‐9 as well as the apoptotic marker BAX, were significantly enhanced in the SHR endothelium with attenuated BCL‐2 protein levels. These results suggest that even isolated aortic endothelium of the SHR has upregulation of proteins as seen previously in‐vivo that may promote inflammation and lesion formation and even in the absence of an elevated blood pressure.

(Supported by: 1Grant of “Knowledge Innovation Project”, Academy of Science, China No. KJCX1‐SW‐07, 1Grant of IM0608, CAMS and 2NIH HL‐10081)
Quantification of dynamic mitochondrial morphologies in myoblasts
Authors: Iqbal Sobia , Hood David A
Published: June 2021

Mitochondria are vital organelles, critical for energy supply and cell survival. Such functional versatility is paralleled by the structural complexity of the organelle. Mitochondria can compensate for alterations in energy requirements by adjusting their size and distribution within skeletal muscle. To quantify this in living cells, we used C2C12 myoblasts transfected with fluorescently labeled pEYFP‐Mito in order to track mitochondrial dynamics. We captured mitochondrial movements at 2 second intervals for an observation time of 5 minutes using real‐time imaging. Approximately 60% of the mitochondria localized in the periphery of myoblasts were punctate spheres less than 0.5μm2 in area, while 19% of mitochondria were present as elongated reticular structures greater than 1.0μm2 in area. These dynamic organelles also varied in their displacement, moving an average of 3.4μm from the origin. However, the total path length travelled by these organelles was approximately 4–5 fold greater, averaging 15μm within the 5 minutes time frame. Thus, mitochondrial morphology in myoblasts is under dynamic control. Understanding the changes in mitochondrial morphology will allow us to elucidate the underlying basis for mitochondrial reticular formation in skeletal muscle, and the effects of contractile activity. Supported by NSERC.
Effects of ganoderma lucidum spores on changes of both caspase‐8 and nestin in epilepsy rats
Authors: Wang Shu‐Qiu , Kang Yu‐Ming , Li Xiao‐Jie , Liu Lei , Liu Jun‐Xing , Zhao Xiao‐Lian , Wang Fang‐Fang , Zhao Lu
Published: June 2021

To investigate the effects of ganoderma lucidum spores on caspase‐8 and nestin in brain tissue of rats by pentylenetetrazol (PTZ) induced epilepsy, and to further study the pathogenesis of epilepsy and the anti‐epilepsy mechanism of ganoderma lucidum spores. Thirty‐six male Sprague‐Dawley rats were divided into three groups randomly: control, PTZ epilepsy and ganoderma lucidum spores (GLS) groups. The results showed that caspase‐8‐positive cell's number in rats of PTZ model group were higher than that of normal control and GLS group, while the number of caspase‐8‐positive cells were decreased in the GLS group compared with that of the epileptic model group. The content of nestin were higher in PTZ model group than those of control group and GLS group in cortex and hippocampus, the level expression of nestin in GLS group were higher than control group.

Conclusion

Ganoderma lucidum spores can effectively reduce the expression of apoptosis cell and inhibit the caspase‐8 activations in the epileptic rat′s brain tissue, strengthen the function of anti‐oxidize in brain tissue, and GLS can supply recovering of epilepsy nerve cell with important basis by reducing the content of nestin neural internality protect mechanism can be strengthened to reduce the damage on nerve system induced by falling sickness. Ganoderma lucidum spores is an assistant medicine to cure epilepsy, it could be very useful in clinical application.
Cells Prioritize Responses When Faced With a Decision Between an Environmental Stress and a Developmental Cue
Authors: Nagiec Michal J , Dohlman Henrik G
Published: June 2021

When faced with competing signals cells must decide how to prioritize and coordinate their responses. Many signals are transduced via a mitogen activated protein kinase (MAPK) cascade. Yeast, Saccharomyces cerevisiae, respond to a developmental signal (mating pheromone) and an environmental stress (hyperosmotic stress) by two distinct MAPK pathways. It has been previously suggested that the two pathways are mutually exclusive. Here we find that cells respond to each stimulus and effectively prioritize the responses. The mating response was initially measured by the formation of a mating projection (shmoo). We found that shmoo formation is delayed by the presence of hyperosmotic stress and increasing stress concentration increases the delay period. We also measured mating transcription induction and mating MAPK activation and found both to be delayed by the presence of stress. The delay in shmoo formation correlates with induction and stabilization of a shmoo landmark protein Far1. Finally we establish that the stress MAPK is necessary for the delay in mating MAPK activation. We conclude that hyperosmotic stress delays the mating response by delaying MAPK activation. This delay allows cells to restore their osmotic balance before Far1 is induced, stabilized, and promotes shmoo formation. These findings illustrate how cells can prioritize multiple competing signals at the level of the MAPK activation.
CRH‐binding protein modulates CRH signaling in a time‐and signaling pathway‐dependent manner
Authors: Evans Ryan T , Seasholtz Audrey F
Published: June 2021

Corticotropin‐releasing hormone (CRH) and the related urocortins (Ucns) mediate the mammalian neuroendocrine stress response by signaling through two G‐protein‐coupled receptors, CRH‐R1 and CRH‐R2. CRH activity at these receptors is also modulated by a secreted CRH‐binding protein (CRH‐BP), which binds CRH/Ucns with equal or greater affinity than the receptors. Initial studies suggested that CRH‐BP predominantly functions to inhibit CRH‐R1 activation, especially at anterior pituitary corticotropes. However, the role of CRH‐BP in CRH signaling via multiple G‐protein pathways has not been fully examined, especially for CRH‐R2. Using purified CRH‐BP and cell culture systems stably‐expressing CRH‐receptors, we determine the modulatory effect of CRH‐BP on CRH‐R1/2 signaling though adenylyl cyclase and MAPK pathways. These studies demonstrate that the role of CRH‐BP is context sensitive, depending on both the specific signaling pathway and whether CRH encounters CRH‐BP prior or simultaneously to the CRH‐receptors. CRH‐receptor activation is inhibited only when CRH is pre‐incubated with CRH‐BP. Without pre‐incubation, CRH‐BP attenuates signal duration but does not prevent receptor activation.

This work was supported by NIH T32 GM07544, T32 HD07048, and U. Michigan Biological Chemistry (RTE), NIH DK42730 (AFS), and MDRTC Cell and Molecular Biology Core.
Nox1‐mediated activation of Slingshot phosphatase in VSMC
Authors: Martin Alejandra San , Maheswaranathan Mithunan , Griendling Kathy K
Published: June 2021

PDGF stimulates vascular smooth muscle cell (VSMC) migration by a NADPH oxidase 1 (Nox1)‐derived, H2O2 dependent mechanism. Migration is partially controlled by cofilin, an actin binding protein activated by Slingshot‐1L (SSH1L) phosphatase. We previously showed that SSH1L is activated and required for PDGF‐mediated cofilin activation and migration in VSMCs, but the mechanism of SSH1L activation is unclear. 14‐3‐3 proteins are one potential regulatory mechanism, and bind to partners that contain phosphorylated consensus motifs. We hypothesized that PDGF disrupts a SSH1L/14‐3‐3 inhibitory complex through Nox1/H2O2‐mediated SSH1L dephosphorylation in a 14‐3‐3 binding motif. We found that SSH1L is Ser‐phosphorylated at a 14‐3‐3 binding motif (probably S834) and co‐immunoprecipitates with 14‐3‐3‐β and ‐γ. In cells transfected with 14‐3‐3K49E DN, which fails to recognize phosphorylated partners, SSH1L is activated (94±6 vs. 45±1 A.U.p=0.01), and PDGF has no further effect (44.6±1.4 vs. 40.2±10.8 A.U.). Interestingly, PDGF reduces phospho‐SSH1L and 14‐3‐3 binding in wild type (29±11 vs. 100±8 A.U.p=0.01) but not in Nox1 knockout cells (101±18 vs. 86±14 A.U.), and the SSH1L‐S834A mutant is constitutively active. Our work shows that PDGF induces SSH1L/14‐3‐3 complex disruption and SSH1L activation via its Nox1‐dependent dephosphorylation in a 14‐3‐3 binding motif.
Reactive Oxygen Species Mediate Bi‐axial Strain‐Induced Fibroblast Proliferation
Authors: Xing Liyu , Wen Yuan , Culbertson Eric J , Franz Michael G
Published: June 2021

Mechanical force modulates fundamental short‐ and long‐term responses in various cell types. We sought to define a mechanism for rat abdominal wall musculo‐tendon fibroblast (abMTF) proliferative response during biaxial cyclic strain (BCS). abMTF were subjected to BCS (10%; 0.2 Hz) in the presence of 2.5–5% FBS up to 6 days. Proliferation of abMTF in response to BCS was measured by direct cell counts. BCS stimulated abMTF proliferation in a time dependent manner compared with control. Increased proliferation depended on the concentration of FBS, with no BCS‐induced proliferative response when cultured in 1% FBS. BCS‐induced proliferative response was blocked by selective MAPK inhibitor PD98059 (50 μM) with inhibiting BCS induced rapid ERK1/2 phosphorylation. Further, BCS‐induced MAPK activation and proliferation was inhibited in the presence of antioxidants, 1% DMSO, 5 μM ebselen and 25μM resveratrol, respectively. H2O2 (3–300 μM), exogenous reactive oxygen species (ROS), stimulated abMTF proliferation and ERK1/2 phosphorylation in a dose‐dependent manner. Antioxidant treatment inhibited BCS‐induced intracellular ROS production detected by DCF fluorescence, while 300 μM H2O2 stimulated abMTF proliferation and ERK1/2 phosphorylation. These results suggest that BCS‐induced abMTF proliferation depends on a MAPK/ERK pathway mediated by intracellular ROS in response to mechanical force.
Identification of curcuminoid analogs that down‐regulate expression of the early growth response protein 1 in prostate cancer cells
Authors: Aguiniga Lizath Monserrath , Moreno America Yajaira , Bisoffi Marco
Published: June 2021

The incidence of prostate cancer for US men is 1/6. It is important to identify therapeutic agents for chemoprevention. Our studies on histologically normal tumor adjacent human prostate tissues indicate an up‐regulation of transcription factor early growth response protein 1 (EGR‐1). We hypothesize that EGR‐1 mediates tumorigenesis and is a therapeutic target for chemoprevention. We aim to identify small molecule inhibitors with safe pharmacological profiles that can down‐regulate EGR‐1 expression in prostate cancer cells. We developed an EGR‐1 specific reporter system amenable to high‐throughput flow cytometric screening of small molecule libraries with combichemical design, including analogs of the natural product curcumin. We cloned a 522 bp EGR‐1 promoter fragment into a plasmid to regulate expression of green fluorescent protein (GFP) in transient transfections. Sequence analysis revealed recognition sites for several transcription factors. We report: (i) EGR‐1 specific GFP activation by mitogenic stimuli by fluorescence microscopy and flow cytometry; (ii) the inhibition of EGR‐1 promoter activation by curcuminoid compounds. We conclude that our reporter system is an ideal tool to identify and test therapeutic agents with chemopreventive potential for the risk of developing prostate cancer. This work is supported by NIH Grant RR0164880 and DOD PCRP Grant W81XWH‐07‐1‐0081.
The effects of Human placenta extract on a Murine Model of Chronic Obstructive Pulmonary Disease.
Authors: Park Soojin , Kim Young Eun , Kwon Young Joo , Kim Yong , Lee Hyo eun , Shin Minkyu , Hong Moochang , Kim Jinju , Bae Hyunsu
Published: June 2021

Chronic obstructive pulmonary disease (COPD) refers to chronic bronchitis and emphysema, a pair of two commonly co‐existing diseases of the lungs in which the airways become narrowed. Human placenta extract (HP) is frequently used as folk medicine in Asian countries for treating various diseases. This study was conducted to evaluate the protective effects of HP on lipopolysaccaride (LPS)‐induced COPD animal model. In present study, Balb/c mice were sensitized with intranasal administration of LPS. six hours later, Balb/c mice were injected with HP daily for 2 days. The BALF cell numbers were counted using a hemacytometer and differential cell counts were performed on slides prepared by cytocentrifugation and Diff‐Quick staining. The levels of the pro‐inflammatory cytokines TNF‐a and IL‐6 were measured. For histologic analysis, hematoxyline and eosin (H&E) stained was evalutated. The levels of neutrophil elastease immunohistochemical (NE IHC) stain was also assessed. As a result, the number of neutrophil, lymphocyte, macrophage and total cells was significantly lower in the HP group than the COPD group. In addition, the IL‐6 and TNF‐a level were decreased in the HP group. These data suggest that HP exerts protective effects against LPS‐induced lung inflammation. Therefore, HP may represent a novel therapeutic agent for treating COPD.
Protective effects of Lindera obtusiloba blume treament on tert‐butyl hydroperoxide‐induced oxidative hepatotoxicity and structural analysis of its active compounds
Authors: HONG CHUNGOUI , KOO YUNCHANG , NAM MIHYUN , KIM JIHOON , WANG ZENG , YANG SUNGYONG , LEE KWANGWON
Published: June 2021

The Lindera obtusiloba blume (LO), a native plant in East Asia, trdionally used as a popular folk. The aim of this study was to evaluate the protective effects of LO extract (LOE) on the tert‐butyl hydroperoxide (t‐BHP)‐induced oxidative hepatotoxicity and confirmed the structure of its active compound with NMR. The study of liver protection showed that pretreatment with LOE (500 or 2000 mg/kg) by gabage to SD rats for 7 days before a single dose of t‐BHP (0.2 mmole/kg i.p.) significantly lowered the serum levels of the hepatic enzyme markers, aspartate aminotransferase and alanine aminotransferase and reduced the indicators of oxidative stress in the liver, such as lipid peroxidation. The active compounds were identified as quercetin 3‐O‐¥á‐rhamnoside (quercitrin) and kaempferol 3‐O‐¥á‐rhamnoside (afzelin). In antioxidant effects of quercitrin and afzelin, ferric‐reducing antioxidant power (FRAP) value were 3.05 ¡¾ 0.42 and 0.77 ¡¾ 0.04 mmol FeSO4․7H2O/§· DM, 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) EC50 value were 107.45 ¡¾ 4.12 and 438.71 ¡¾ 14.18 ¥ìM. FRAP value of quercitrin was 1.5‐fold higher than ascorbic acid (1.90 ¡¾ 0.26) which is a powerful antioxidant and also DPPH EC50 value of quercitrin was 2‐fold higher than ascorbic acid (192.90 ¡¾ 10.78). Thus, it has been suggested that these compounds might contribute to the health positive effects of LO against t‐BHP induced hepatotoxicity.
Comparison of antioxidant activity from different root part of 6‐year‐old Panax ginseng C.A. meyer(Yun‐poong).
Authors: Jo Ji eun , Yook Hong sun , Choi Jae‐eul
Published: June 2021

The objective of this study is to evaluation the total polyphenol and the antioxidant activity of freeze‐dried ginseng(Panax genseng C.A. meyer cv Yun‐poong) for root area and diameter. Ginseng root consists of the main root(MR), lateral root(LR) and fine root(FR). These roots were sorted 3~4 group(MR 1~4 group, LR 1~4 group, FR 1~3 group) depending on the diameter of each center region. Total polyphenol content of ginseng showed the highest in FR‐3 among the all samples. The antioxidant activity measured using DPPH radical‐scavenging activity and ABTS radical scavenging activity. DPPH radical‐scavenging activity of FR‐3(IC50: 7.03 mg/mL) showed the highest value in all samples but MR‐2(IC50: 37.02 mg/mL) showed the lowest value. On the whole, hydrogen donating activity of FR was significantly higher than for other groups. ABTS radical scavenging activity of FR‐3 was significantly higher than any of the other samples. Polyphenol content and antioxidant activity of ginseng showed the highest in FR‐3 and the result exhibited that polyphenol content and antioxidant activity of ginseng were related to root area and diameter.
Establishing heme iron values for the Canadian Nutrient File (CNF)
Authors: Cooper Marcia , Rondeau Isabelle
Published: June 2021

Objectives

Heme iron, primarily from the hemoglobin and myoglobin of meat, poultry and fish (MFP) is better absorbed than non‐heme iron. Monsen et al. (1978) assumed that the proportion of heme iron contained in various meats averages 40%, while the remaining 60% is found in the less bioavailable non‐heme form. More recent evidence suggests that this ratio may vary depending on the type and cut of MFP. Currently, the CNF lists only total iron values. The objective of our research was to match heme iron values from the literature to foods within the CNF.

Methods

An extensive literature search was conducted to identify the heme content of different souces of MFP. Values from the literature were then matched to CNF foods with consideration of cuts and similarities to North American cooking methods. In order to match heme values from the literature to MFP foods, average values were often generated across cuts and cooking methods eg. ground chicken was an average of white and dark meat.

Results and Discussion

A broad range of heme values were observed across MFP types. The following heme values were attributed: 33–70% for beef cuts, 21–40% for fish, 27–66% for pork cuts, 26–37% for poultry, and 40–63% for veal. When no literature value for heme iron was found eg., game meats, and many types of fish, a value of 40% was used as a default. The heme content of beef, veal and some pork cuts are higher than 40%, while poultry and fish might have lower heme iron contents.

Conclusion

Incorporating heme iron values into the CNF that are reflective of various MFP provides useful data for assessing iron bioavailability from foods/diets and for developing future nutritional policies targeted at reducing iron deficiency.
Impact of SprinklesTM on reducing childhood vitamin A deficiency in western Kenya
Authors: Ruth Laird J , Grant Frederick , Mandava Usha , Quick Robert , Patel Minal , Mbkaya Charles , Jefferds Maria Elena , Woodruff Bradley , Suchdev Parminder
Published: June 2021

In 2007–2008, we evaluated community‐based marketing of Sprinkles, a multiple micronutrient powder containing 375μg vitamin A, added to complementary foods.

Objective

To measure Sprinkles impact on the prevalence of vitamin A deficiency (VAD) among children 6–35 months.

Methods

In 30 intervention villages, vendors sold Sprinkles to neighbors. In 30 comparison villages, Sprinkles were not promoted. Biweekly household visits measured use. VAD (retinol binding protein <0.70 μmol/L) was measured among 500 children aged 6–35 months before distribution and 12 months later when the children were 18–47 months old.

Results

VAD Prevalence (excluding C‐reactive protein >10 mg/dL)

Intervention n=254 % (95% CI)
Comparison n=24 % (95% CI)
p‐value between groups

Baseline
17.7 (13.0, 22.4)
15.5 (10.9, 20.0)
0.14

12 month follow up
10.2 (6.5, 14.0)
13.0 (8.8, 17.2)
0.08

p‐value within group1
<0.01
<0.01

Change in VAD prevalence (%)
7.5
2.5
<0.012

Paired Chi‐square,

McNemar test

Children in intervention households consumed an average 0.9 sachets/week (~328 μg vitamin A). As Sprinkles use increased, follow up VAD prevalence decreased: no use=17.8%; infrequent use=10.6%; some use=12.0%; and frequent use=3.1% (p = 0.053; adjusted for age, sex, socio‐economic status, and maternal education).

Conclusion

Community‐based marketing of Sprinkles was associated with decreased prevalence of VAD in pre‐school age children.
Predicting epilepsy with peripheral blood transcriptome biomarkers during early latent period
Authors: Karsten Stanislav L. , Vi Nancy , Bragin Anatol , Mazarati Andrei , Kudo Lili C.
Published: June 2021

Status epilepticus (SE) is a common cause of hippocampal neurodegeneration and synaptic reorganization that may or may not lead to the development of epilepsy. Currently there is no method to predict whether chronic epilepsy would develop in post‐SE patients. We used microarrays to analyze expression changes in the blood of two rat models of chronic epilepsy to identify prognostic peripheral biomarkers of epilepsy.

Peripheral blood was collected from Adult Wistar male rats before SE and one, three, seven and thirty days after subjected to SE induced by either systemic pilocarpine or intrahippocampal kainic acid. Blood RNA was used for global transcriptome analysis using microarrays. Post‐SE animals underwent continuous monitoring to retrospectively correlate seizure syndrome with the changes in blood transcriptome profile.

We identified nearly 100 predictor/biomarker genes for later occurring chronic seizures regulated similarly in both models, which were confirmed in independent groups of post‐SE rats by semi quantitative RT‐PCR and by a custom designed prognostic microarray.

The molecular signature preceding the development of epilepsy present in the peripheral blood allows the design of a prognostic/diagnostic transcriptome biomarker chip for potential epilepsy patients and monitoring the effectiveness of antiepileptogenic therapy.

This work was supported by NIH Grant NS060476‐01 (SLK).
Impaired Homocysteine Transmethylation and Protein‐Methyltransferase Activity Reduce Expression of Selenoprotein P: Implications for Obesity and Metabolic Syndrome
Authors: Jackson Matthew I , Combs Gerald F
Published: June 2021

Obesity causes Metabolic Syndrome and Type‐II Diabetes, disrupting hepatic function, methionine (Met)/homocysteine (Hcy) transmethylation and methyltransferase (PRMT) activities. Selenoprotein P (SEPP1), exported from the liver, is the predominate form of plasma selenium (Se) and the physiological Se delivery mechanism to extrahepatic tissues. SEPP1 expression is effected by transfactors subject to post‐translational methylation and depends upon methylation of tRNA and methylation status of gene promoter regions. Se is capable of anti‐carcinogenic activity: we found plasma Se status to vary with body mass index. Therefore, we sought to know whether Se metabolism is affected by obesity and, specifically, whether methylation capacity dictates the expression of SEPP1. In order to test that hypothesis, we treated HepG2 cells with mechanistically distinct inhibitors of S‐adenosyl homocysteine hydrolase (AHCY), or PRMT. We found incubation with these agents to decrease the appearance of SEPP1 in HepG2 cell‐conditioned media, which served as a model for plasma, and that the decrease in SEPP1 was not due to sequestration within cells. Thus, deficiencies in the Met/Hcy cycle or in the activity of methyltransferases, can reduce the expression of SEPP1. This suggests that it may be useful to determine whether obesity‐impaired methylation result in decreased plasma levels of SEPP1 in animals and humans Se.
Risk Analysis‐Based Food Defense Certification Program for Professional and Academic Audiences
Authors: Yoe Charles , Parish Mickey , Eddy Doug , Lei K. Y. , Paleg Bradley , Buchanan Robert , Schwarz Jurgen G.
Published: June 2021

The post 9–11 awareness about food defense has evolved into a growing interest in food protection in the US, an interest that, until recently, remained nascent globally. Recent events including melamine in China and the unintentional contamination leading to the PCA recall have highlighted the vulnerability of the global supply chain to economic adulteration as well as intentional contamination. This ongoing research project provides an integrated multifunctional research, education and extension approach to better secure the safety and defense of the nation's food supply, utilizing the food safety risk analysis paradigm to empower the food protection community to better protect the food chain. The project has used the guidance of an expert panel from industry to define terminology in industry‐relevant ways and to identify emerging and the most pressing food defense issues. The grant team has conducted extensive applied research with the FDA's CARVER + Shock tool both domestically and internationally. Grant outputs include academic and professional development courses suitable for a practitioner's certificate in food defense and a series of short courses to fill a critical gap in the Nation's food defense by assisting small businesses in understanding and preparing effective food defense plans.