bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of tumor suppressor gene p16, mutant p53 and c‐erbB‐2 protein in patients with gastric carcinoma
Authors: Qin Wen‐Bo , Zhang Shi‐Ling , Kang* Yu‐Ming , Wang* Shu‐Qiu , Ma Xiao‐Ru
Published: June 2021

The present study was undertaken to investigate the expression of tumor suppressor gene p16, mutant p53 and c‐erbB‐2 protein (corresponding protein product of oncogene) in gastric carcinoma. The expression of p16, mutant p53 and c‐erbB‐2 protein were examined in oncologic tissues from 92 patients underwent surgical treatment of gastric carcinoma and mucosa samples around gastric carcinoma from 28 patients using immunohistochemistry. These 92 patients consist of 34 ones with intestinal‐type gastric carcinoma and 58 ones with diffuse‐type gastric carcinoma; 31 ones of these patients at early stage of gastric carcinoma and the other 61 at advanced stage. The positive rate of p16, p53 and c‐erbB‐2 in these patients was 44% (40/92), 53%(49/92) and 32.6% (30/92), respectively while in mucosa samples around gastric carcinoma from 28 patients, the positive rate of p16 is 67.9% (19/28), and mutant p53 and c‐erbB‐2 are all negative. A positive correlation was observed between the expression of p16, p53 and Lauren type of gastric carcinoma while no correlation between c‐erbB‐2 and Lauren type of gastric carcinoma. A correlation was observed between the expression of p16 and clinical stages. The expression of p53 and cerbB‐2 might be reliable indicator to evaluate severity of gastric carcinoma and their prognosis.
The Essential Role of the L4F‐Adiponectin Regulatory Axis: Leading to Improvements in the Metabolic Profile of Diabetes Mellitus
Authors: Burgess Angela Patricia Haynes , Peterson Stephen J , Tsenovoy Peter , Li Ming , Kim Dong Hyun , Abraham Nader G
Published: June 2021

Apolipoprotein A1 mimetic peptide (L‐4F), a natural component of HDL‐ has been shown to have both anti‐atherosclerotic and vasoprotective effects. Therefore, we hypothesized treatment with L4‐F in an obese‐ diabetic mouse model, would lead to improvements in the metabolic consequences of diabetes: inflammation, weight gain, and insulin resistance. L‐4F (1mg/kg.d) was administered to obese‐diabetic mice for 6 weeks. L‐4F improved HO activity and decreased superoxide production (P <0.001) in ob/ob mice. L‐4F treatment decreased visceral fat content and prevented weight gain (P<0.02), and decreased inflammatory cytokine IL‐1β (p<0.05) compared to wild type, c‐ob/+. L‐4F treatments also led to increased insulin sensitivity, glucose tolerance and adipose tissue remodeling (P <0.001). L‐4F inhibited adipogenesis in bone marrow and increased total adiponectin levels (P<0.02) only in obese mice. Therefore, suggesting a mechanism by which L4‐F increases HO‐1 activity and adiponectin, leading to decreased inflammation, increased insulin sensitivity, maintenance of normal weight, and an overall improvement in the diabetic profile of obese‐diabetic mice. This work was supported by NIH grants DK068134, HL55601 and HL34300 (NGA).
Resuscitation with hydroxyethyl starch 130/0.4 versus lactated Ringer on hemorrhage following a liver trauma in swine
Authors: Zaar Morten , Lauritzen Brian , Secher Niels H. , Krantz Torben , Johansson Pär I.
Published: June 2021

Background

: The effect of resuscitation fluids on hemorrhage remains disputed. This study evaluated the effect of resuscitation with hydroxyethyl starch 130/0.4 versus lactated Ringer on hemorrhage in a pig liver trauma model.

Methods

: A grade III crush and avulsion liver injury was inflicted on the left lateral lobe and, to simulate a pre‐hospital setting, resuscitation began after 7 min: phase 1, 0.7 mL kg−1 min−1 for 10 min; phase 2, 1.65 mL kg−1 min−1 for 20 min; and phase 3, according to the blood loss. Initial resuscitation was with hydroxyethyl starch 130/0.4 or lactated Ringer while in phase 3 it was with hydroxyethyl starch 130/0.4, and lactated Ringer was also evaluated for a grade IV laceration liver injury.

Results

: With a grade III liver injury, initial resuscitation with hydroxyethyl starch 130/0.4 (n=5) caused progressive bleeding and exsanguination after ~75 min, while lactated Ringer (n=5) elicited a slow bleeding that ceased after ~45 min. With a grade IV injury (n=5), initial resuscitation with lactated Ringer caused exsanguination after ~75 min.

Conclusion

: For a mild liver injury, initial resuscitation with hydroxyethyl starch 130/0.4 appeared to provoke uncontrolled hemorrhage, while hemostasis was achieved in animals resuscitated with lactated Ringer. In contrast, hemostasis was not achieved in animals with a severe liver trauma and resuscitated with lactated Ringer.
Shock survival improved by combined protease and lipase inhibition in the intestinal lumen
Authors: Penn Alexander Hayes , Schmid‐Schönbein Geert W.
Published: June 2021

Shock and multiple‐organ failure remain one of the leading causes of death. Our evidence shows it is triggered by a failure of the mucosal barrier in the intestine that is followed by cell death in the intestine and elsewhere. We have recently shown that homogenates of ischemic, but not non‐ischemic, intestines or intestinal wall homogenates digested with proteases from the intestinal lumen contain cytotoxic mediators. We have identified these mediators as free fatty acids (FFAs), already present or created by the action of a lipolytic enzyme present in the intestinal wall. The FFAs are bound to FFA‐binding proteins, such as albumin, unless those proteins are digested, as may occur in shock when the mucosal barrier fails and proteases enter the walls. To test this hypothesis in vivo, we injected saline, aprotinin (protease inhibitor), orlistat (lipase inhibitor), or a combination of the two into the lumen of the small intestine of Wistar rats and subjected them to 120 min of splanchnic arterial occlusion followed by up to 180 min of observed reperfusion. Saline treated animals all died after 86±43 min of reperfusion. Only the combination aprotinin and orlistat treatment significantly improved survival time to 164±11, suggesting that in severe models of shock both luminal protease and luminal lipase inhibition should be explored.
Differences among inbred rat strains in hematological and coagulation measures
Authors: Calderon M. , Ryan K. L. , Kheirabadi B. , Klemcke H. G.
Published: June 2021

As part of ongoing studies to characterize responses to controlled hemorrhage (Klemcke et al., Shock, in press, 2007), we conducted hematological and coagulation evaluations of 15 different inbred rat strains (n = 8–11 rats/strain). Rats were catheterized and, 24 hours later, 55% of their blood volume was removed. Complete blood counts (CBC), thromboelastographic (TEG) measures, plasma fibrinogen concentrations and prothrombin times (PT) were measured in samples taken during the initial phase of hemorrhage. Two rat strains tended to be the most divergent for many of these measures: Brown Norway/Medical College of Wisconsin (BN/Mcwi), and Munich Wistar Fromter (MWF). BN/Mcwi rats had greater PT (10.8 ± 0.2 vs 9.4 ± 0.1 sec), RBC (7.5 ± 0.1 vs 5.4 ± 0.2 106/mm3) and WBC (12.6 ± 0.4 vs 1.4 ± 0.1 103/mm3), but less fibrinogen (277 ± 21 vs 412 ± 13 mg/dl), platelets (408 ± 21 vs 776 ± 47 103/mm3), and lower clot strength based on TEG measures (64.7 ± 0.9 vs 75.9 ± 0.7 mm) than did Munich Wistar Fromter (MWF) rats (P <0.05). Hence, MWF rats appeared to exceed BN/Mcwi rats in many aspects of coagulation. Should the extremely low WBC counts of the MWF be shown to reflect an altered immune system function, these data suggest a potential utility for BN/Mcwi and MWF rats as models for investigation of the important interactions between coagulation and immunity (Am J Transplant 7:499, 2007).
In isolated cardiomyocytes NF‐κB activation is modulated by alterations in protein O‐GlcNAc levels
Authors: Zou Luyun , Champattanachai Voraratt , Marchase Richard B , Chatham John C
Published: June 2021

O‐Linked N‐acetylglucosamine (O‐GlcNAc) modification of proteins plays an important role in transcription, translation, nuclear transport and cytoskeletal assembly. We have shown that glucosamine (GlcN) increased O‐GlcNAc levels and attenuated activation of NF‐κB signaling following trauma hemorrhage and resuscitation in vivo and in response to lipopolysaccharide (LPS) treatment of in cardiomyocytes in vitro. Therefore the goal of this study was to determine whether the effect of GlcN on NF‐κB activation was mediated via O‐GlcNAc transferase (OGT), which catalyzes O‐GlcNAc synthesis. Neonatal rat ventricular myocytes(NRVMs) were untreated, treated with 5mM GlcN, transfected with OGT adenovirus or OGT siRNA and then stimulated with LPS (2μg/ml) for 6h. In untreated cells LPS significantly increased IκB‐α phosphorylation, TNF‐α and ICAM1 expression and increased nuclear NF‐κB levels. Both GlcN and OGT overexpression significantly increased O‐GlcNAc levels, attenuated LPS‐induced TNF‐α and ICAM1 expression and decreased IκB‐α phosphorylation and nuclear NF‐κB levels. In contrast, OGT siRNA treatment decreased OGT and O‐GlcNAc levels and enhanced LPS‐induced increase in IκB‐α phosphorylation. These results demonstrate that modulation of cellular O‐GlcNAc levels alters the response to activation of NF‐κB signaling pathway, and may contribute the protective effect of GlcN in vivo.
Blockade of α‐adrenergic receptors in the preoptic area/anterior hypothalamus prevents lipopolysaccharide evoked hypotension
Authors: Yilmaz Mustafa Sertac , Myer Brian S , Feleder Carlos , Millington William R
Published: June 2021

The mechanisms responsible for endotoxic hypotension are not fully understood although previous data from this laboratory indicate that lipopolysaccharide (LPS) initially lowers arterial pressure (AP) through a central mechanism. Specifically, we showed that LPS hypotension could be prevented by inhibiting neuronal activity in the preoptic area/anterior hypothalamus (POA) with the local anesthetic lidocaine. This study tested the hypothesis that LPS lowers AP by activating α‐adrenergic receptors in the POA. Bilateral injection of the α‐adrenergic receptor antagonist phentolamine (5 ug; 1 ul) into the POA completely blocked the fall in arterial pressure evoked by LPS (1 or 15 mg/kg i.v.) in isoflurane‐anesthetized rats. Phentolamine had no effect on AP in control animals and neither LPS nor phentolamine influenced heart rate. The β‐adrenergic receptor antagonist propranolol (5 ug) was ineffective. Bilateral lidocaine injection into the nucleus tractus solitarius (NTS) also effectively prevented LPS hypotension suggesting that medullary A1/A2 noradrenergic neurons mediate the response. Importantly, phentolamine prevented the late‐developing, decompensatory phase of the hypotension induced by both low (1 mg/kg) and high (15 mg/kg) doses of LPS. These data are consistent with the hypothesis that LPS initially lowers AP by stimulating norepinephrine release in the POA.
Increases in brain insulin normalize baroreflex gain in conscious, pregnant rats
Authors: Azar Afaf S. , Brooks Virginia L.
Published: June 2021

Our recent studies suggest that insulin resistance contributes to pregnancy‐induced decreases in baroreflex (BR) gain. The mechanism is unclear, but may involve the reduced brain insulin levels that also occur. To test this hypothesis, it was determined if intracerebroventricular (icv) infusion of insulin (100 μU/min) improves BR gain in late pregnant (P, n=7), but not nonpregnant (NP, n=10), conscious rats. BR function was quantified by fitting a 4 parameter sigmoidal curve to the heart rate (HR) responses to changes in blood pressure (BP) following 3–5 minute iv infusions of increasing doses of nitroprusside and phenylephrine. Pregnancy significantly decreased BP (80±4 mmHg, P; 107±3 mmHg, NP; P<0.01), BR gain (2.4±0.4 bpm/mmHg, P; 4.6±0.4 bpm/mmHg, NP; P<0.01), maximum BR HR (455±24 bpm, P; 514±15 bpm, NP; P<0.01), and BP50 (84±6 mmHg, P; 102±5 mmHg, NP; P<0.01). BR width was increased (21±3 mmHg, P; 15±1 mmHg, NP; P<0.05), but HR and minimum BR HR were unaltered. After at least 1 hr of icv insulin infusion in P rats, BR gain increased (to 3.9±0.5 bpm/mmHg, P<0.01) and BR width decreased (to 12±1 mmHg; P<0.01); BP, HR and all other BR parameters were unchanged. In NP rats, icv insulin failed to change BP, HR, BR gain (to 4.2±0.4 bpm/mmHg), and other BR parameters. In conclusion, icv infusion of insulin reverses the effect of pregnancy to impair the baroreflex, suggesting that reductions in brain insulin contribute to this impairment. Supported by HL70962, AHA and Murdock Fdn.
Vasopressin and neuronal nitric oxide synthase in hypothalamic paraventricular nucleus (PVN) of pregnant rats.
Authors: Heesch Cheryl M , Shoge Korryn , Zidon Terese M , Hasser Eileen M
Published: June 2021

Decreased baseline GABAergic sympathoinhibition within the PVN of pregnant (P) rats is associated with decreased neuronal nitric oxide synthase (nNOS) activity (NADPH Diaphorase), and presumably decreased GABA release. nNOS is colocalized with vasopressin (VP) in both parvo‐ and magno‐cellular PVN neurons. Our goal was to determine if pregnancy changed the number of spinally projecting PVN neurons (presympathetic) containing nNOS or VP. The retrograde tracer fluorogold (FG) was injected unilaterally into the spinal cord of female nonpregnant (NP, n=3) and P (n=3) rats. After 7 days, ipsilateral FG and immunohistochemical staining for VP and nNOS in PVN sections (30 ìm) was counted at 3 levels: rostral (lev‐1) and intermediate (lev‐2) PVN contained the posterior magnocellular cell group, while lev‐3 (caudal) contained the majority of spinally projecting (FG) neurons. In both groups, distribution of VP cells was: lev2 > lev 1 > lev 3. Approximately 7% of all VP cells were spinally projecting, with the majority at level 3 in both groups (~ 19%). Of spinally projecting cells the % also containing nNOS tended to be less in P (16 ±2.9 %) compared to NP rats (24 ±2.4 %). Although preliminary, these data are consistent with decreased nNOS in presympathetic neurons in the PVN of P rats which could contribute to the previously reported decrease in GABAergic sympathoinhibition in the PVN in pregnancy. (HL36245; HL55306)
Non‐phagocytic NADPH oxidase is present in dopaminergic neurons
Authors: Zawada W Michael , Banninger Gregg P , Jones Susan , Thornton Jennifer , Marriott Beth , Cantu David , Rachubinski Angela , Poczobutt Andy , Das Mita
Published: June 2021

Oxidative stress is a key contributor to the pathogenesis of Parkinson's disease. NADPH oxidase is a membrane enzyme responsible for the oxidative burst in activated phagocytes. NADPH oxidase also exists in many types of non‐phagocytic cells including vascular cells and neurons, with a wide distribution throughout the brain. Although the microglial NADPH oxidase is known to participate in dopaminergic neurotoxicity, whether NADPH oxidase also exists in the dopamine neurons in the substantia nigra pars compacta (SNpc) and whether it contributes to the ROS generation has not been explored. We have examined the role of NADPH oxidase in MPP+ induced oxyradical production in immortalized dopaminergic cell line (N27). MPP+ dose dependently led to upregulation of ROS in N27 cells, which was attenuated by 40% with treatment with NADPH oxidase inhibitors. N27 cells were observed to express mRNA for all the necessary components of the NADPH oxidase, Nox2, p22, p40, p47 and p67, as well as Nox1 and Nox4. SiRNA‐mediated knockdown of p22, a critical stabilizing subunit of the NADPH oxidase, reduced the amount of ROS generated. Immunocytochemical examination of mouse dopaminergic neurons in sections of the SNpc revealed that these neurons express Nox2, p47 and p67. While our results suggest that NADPH oxidase is a contributor to ROS generation in dopaminergic neurons, the functional role of this process remains to be defined. Supported by NIH AA016654 and The Neuroscience Program at the UCD.
Catecholamine neurons in the rostral ventrolateral medulla (RVLM) contribute to the cardiovascular effects of endotoxin challenge
Authors: Stedenfeld Kristen A. , Johnson Nichole M. , Maldovan Victoria L. , Rinaman Linda M. , Sved Alan F.
Published: June 2021

The rostral ventrolateral medulla (RVLM) is critical to neural control of cardiovascular function. Spinally‐projecting sympathoexcitatory RVLM neurons can be divided into two groups: C1 neurons that express catecholamine biosynthetic enzymes, and non‐C1 neurons that do not. Selective lesioning of RVLM‐C1 neurons in rats with saporin conjugated to an antibody against dopamine beta‐hydroxylase has provided evidence that C1 neurons contribute to cardiovascular reflexes, homeostatic responses to glucoprivation, pressor responses to restraint stress, and other sympathoexcitatory responses. We hypothesized that C1 neurons also play a role in cardiovascular responses to interoceptive stress induced by injection of endotoxin (lipopolysaccharide; LPS), a fragment of gram‐negative bacterial cell walls that presents an immune challenge by stimulating the release of pro‐inflammatory cytokines. Radiotelemetry transmitters were used to record blood pressure (BP) and heart rate (HR) in conscious rats after LPS challenge (200 ug/kg, i.p.). In rats with substantial loss of C1 neurons, BP was attenuated throughout the biphasic pressor response to LPS, and both HR and HR variability response patterns were altered. These data provide further support for an important role of RVLM‐C1 neurons in mediating sympathoexcitatory cardiovascular responses to homeostatic challenges. (Supported by HL55786 and MH59911).
Brain angiotensin or nuclear factor kappa B blockade attenuates cytokines, oxidative stress and decreases sympathoexcitation in hypertension
Authors: Kang Yu‐Ming , Ma Ying , Sriramula Srinivas , Zheng Jin‐Ping , Yang Zhi‐Ming , Francis* Joseph
Published: June 2021

Inflammatory and oxidative stress induced by angiotensin II (ANG II) contributes to the pathogenesis of hypertension. This study determined the effect of central blockade of angiotensin‐1 receptor (AT1‐R) and cytokine transcriptional factor NFκB on ANG II‐induced inflammatory, oxidative stress and hypertensive response. Rats were infused with ANG II (0.6μg/hr) for 4 weeks. Some rats also received ICV infusion of vehicle or AT1‐R antagonist losartan (LOS, 20μg/hr), or pyrrolidine dithiocarbamate (PDTC, 5μg/hr), an inhibitor of NFκB. ANG II infusion resulted in a significant increase in the mean arterial pressure (MAP), renal sympathetic nerve activity (RSNA), plasma norepinephrine (NE) and cytokines. These animals also showed increased expression of cytokines, AT1‐R and NADPH oxidase gene in the paraventricular nucleus (PVN) of hypothalamus. In contrast, ICV treatment with LOS or PDTC significantly decreased MAP, RSNA, circulating NE and cytokines in ANG II‐induced hypertensive rats. ICV Treatments also reduced the expression of cytokines, AT1‐R and oxidative stress gene in the PVN. These finding suggests that ANG II induces the expression of cytokines and oxidative stress in the PVN. The interaction between angiotensin and NFκB in the PVN contribute to the progression of ANG II‐induced hypertensive response.
Postprandial lipemia and endothelial function in active and sedentary menes
Authors: Winters Julie L. , Horvath Peter J. , Browne Richard W. , Burton Harold W. , Leddy John J , Dorn Joan M.
Published: June 2021

Postprandial lipemia's effect on endothelial function in active and sedentary men without the influence of a recent bout of exercise has not been studied. Nine active and 8 sedentary males (18–38 yr old with VO2 peaks 64.1 and 39.4 mL/kg/min) drank a high fat dairy shake after a 12 h fast and 36 h of inactivity. Blood was collected and brachial artery flow mediated dilation (FMD) was determined by b‐mode ultrasound at 0, 3 and 6 h. Serum glucose remained stable but, non‐esterified fatty acids increased in both groups over time. Total cholesterol (TC) and triglycerides (TG) were highest at 3 h, TG did not return to fasting levels after 6 h, both groups were not statistically different (TG: 67.7±6.5, 107.9±9.5, 83.5±8.9 mg/dL and TC: 158.2±7.2, 164.4±7.0, 162.7±7.0 mg/dL at 0, 3 and 6 h respectively). Height adjusted baseline brachial diameter at 0 h was larger in the active group (4.9±0.1 Active, 4.1±0.1 mm Sedentary). Age adjusted %FMD was not different except at 6 h with sedentary being greater. The %FMD peaked at 3 h for active and 6 h in the sedentary group (Active: 4.4±0.5, 4.9±1.0, 4.0±1.0, Sedentary: 7.1±1.2, 7.7±1.6, 8.8±2.0mm at 0, 3, 6 h). Studies on postprandial lipemia need to control for recent exercise as fitness level did not alter postprandial lipemia with 36 h of inactivity. Postprandial lipemia did not alter endothelial function differently but active males had a larger vessel at rest. MDRF Grant, Univ. at Buffalo
The Influence of an Antioxidant Cocktail on Vascular Function in COPD
Authors: Harris Ryan Alan , Wray D Walter , Nishiyama Steven K , Berkstresser Kimberly A , Richardson Russell S
Published: June 2021

Vascular function may be impacted by the known elevation of oxidative stress in COPD. Surprisingly, little is known about the importance of these phenomena on the pathology of this disease. This study aimed to characterize vascular function measured by flow‐mediated vasodilation (FMD) in patients with COPD and determine the effect of an antioxidant cocktail (AOC). Patients (ages 69–79 yrs) performed a pulmonary function test to determine disease severity. In a randomized, double‐blind, experimental design (2 days), FMD was evaluated following the ingestion of either a placebo or an AOC. Both the placebo (sugar pills) and the AOC consisted of two doses (AOC: totals = 1,000mg of vitamin C, 800 IU of vitamin E, and 600 mg of α‐lipoic acid). The FVC/FEV1 (%) was 58±7%. The FMD (%), absolute change in diameter, and FMD/shear was 1.1±0.1 vs. 2.3±1.1%, 0.003±0.000 vs. 0.007±0.003 cm, and 0.12±0.05 vs. 0.17±0.00 au, for the placebo and AOC respectively. Shear (AUC) to peak dilation was 27% greater following antioxidant supplementation when compared with placebo (13569± 6584 vs. 10683± 3697 s−1). In conclusion, there appears to be an improvement in vascular function, measured via FMD, with the ingestion of an antioxidant cocktail in patients with COPD. Supported by TRDRP 15RT‐0100 and Parker B Francis Fellowship.
Antioxidant response to postprandial lipemia in active and sedentary men
Authors: Ahn Heajoon , Winters Julie L , Horvath Peter J , Burton Harold W , Dorn Joan M , Leddy John J , Browne Richard W
Published: June 2021

This study compared the postprandial lipemia antioxidant status in healthy males, active and sedentary. Nine active and 8 sedentary males (18–38 yrs old) were recruited based on health (ACSM Risk Criteria) and fitness (VO2 peak on a cycle ergometer, 64.1 and 39.4 mL/kg/min, respectively). Plasma antioxidant status was investigated after 36 h of inactivity and 12 h of fasting (0 h) and at 3 h and 6 h after consumption of a high fat dairy based shake. Plasma levels of thiobarbituric acid reactive substances (TBARS) and antioxidant enzyme activity of glutathione reductase (GSR), glutathioneperoxidase (GPx), glutathione transferase (GST), and superoxide dismutase (SOD) were determined. TBARS levels did not change over time in either group, but on average the active group was 21% higher than sedentary. At 0 h, GSR was 19% lower in the active group than sedentary. In the active group, GSR decreased by 17% from 3 h to 6 h. GPx was higher at 0 h and 3 h for the active group but not at 6 h. GST and SOD were similar between groups. Being physically active may be associated with elevated activity of TBARS and GPx, decreased activity of GSR, while other antioxidant activity remains unchanged. Postprandial lipemia caused by a high fat dairy meal did not increase TBARS, and had little effect on antioxidant enzymes. MDFR Grant, Univ. at Buffalo
Fat mass is increased in the acyl‐CoA synthetase‐isoform 1 (ACSL1) adipose‐specific knockout mouse
Authors: Ellis Jessica Mary , Li Lei , Wu Pei‐Chi , Coleman Rosalind
Published: June 2021

Long chain acyl‐CoA synthetase (ACSL) catalyses the conversion of fatty acids into their acyl‐CoA derivatives. Acyl‐CoAs have multiple fates including oxidation, and incorporation into phospholipids, cholesteryl esters, and triacylglycerol. ACSL1, one of five ACSL isoforms, is highly expressed in white adipose tissue and thought to be important for triacylglycerol formation. To investigate the role of ACSL1 in adipose tissue we have created an adipose‐specific knock out of ACSL1, the ACSL1A−/− mouse. Although we expected mice with reduced fat mass, we report the contrary: the ACSL1A−/− mice have increased fat mass on a low‐fat diet. Animal weights are similar to their littermate controls yet fat mass is increased ~30%. ACSL1A−/− mice gain weight and fat mass similar to controls when fed a high‐fat diet. ACSL1A−/− mice have a ~40% reduction in total ACS activity in white adipose depots on both low‐fat and high‐fat diet with no evidence of compensation. Interestingly, we observe a reduction in total ACSL activity in adipose of mice fed a high‐fat diet that inversely correlates with fat mass. Despite increased adiposity, the ACSL1A−/− mice respond normally to glucose tolerance tests. The data suggest that the role of ACSL1 in adipocytes is not limited to the formation of triacylglycerol.
Impact of natural and industrial trans fatty acids consumption on HDL metabolism
Authors: Grenier Geneviève , Motard‐Bélanger Annie , Charest Amélie , Clerc Élise , Paquin Paul , Chouinard Yvan , Lemieux Simone , Couture Patrick , Lamarche Benoit
Published: June 2021

Unlike dietary industrial trans fatty acids (iTFA), which are known to increase the risk of cardiovascular disease (CVD) through adverse modifications of blood lipids, the impact of dietary trans fatty acids from ruminants (rTFA) remains unknown. The purpose of the present study was to compare the effects of iTFA and rTFA on in vivo HDL metabolism. According to a double‐blind, randomized crossover controlled study design, 9 healthy men were fed each of 3 experimental 5‐week isoenergetic diets: 1‐ high in iTFA (10.2 g/2500 kcal), 2‐ high in rTFA (10.2 g/2500 kcal) and 3‐ a control diet low in TFA (2.2 g/2500 kcal). Apolipoprotein (apo) A‐I kinetic was investigated after each diet after administration of D3‐Leucine in the fasted state. The fractional catabolic rate (FCR) and production rate (PR) of apo A‐I were significantly higher after the diet rich in iTFA than after the control diet (+12.9% and +12.6% respectively, p=0.03). No significant change in apo A‐I FCR and PR were observed after the diet rich in rTFA. When analysing all 3 diets, the apo A‐I FCR was a strong correlate of on‐diet plasma HDL‐C concentrations (r=−0.50, p=0.008). This kinetic study of apo A‐I suggested that a high consumption of rTFA may not significantly modify HDL in vivo kinetics.

Supported by grants from the Dairy farmers of Canada, Novalait and the Canada Research Chair in nutrition and Cardiovascular Health.
Randomized trial of guava as source of ascorbic acid to reduce iron deficiency among indigenous schoolchildren in northern Mexico
Authors: Monárrez‐Espino Joel , Pizarro‐Chávez Salvador , Vázquez‐Mendoza Guillermo , Greiner Ted
Published: June 2021

This trial aimed at examining the effect of drinking 300 ml of guava juice containing 200 mg of ascorbic acid (AA) with the main meal for 10 weeks (5 d/wk) on the iron status of iron‐deficient children. A total of 295 children aged 6–9 years from 4 boarding schools were screened for anemia using altitude‐adjusted capillary hemoglobin (Hb) cut‐offs. The 95 anemic children were randomly allocated to a guava or placebo group, each with 16 and 14 iron‐deficient children (serum ferritin‐SF <12 ng/ml with negative C‐reactive protein), respectively. The placebo group was given guava‐flavored juice free of AA. Children with fecal parasites were treated. Dietary intake was collected for weeks 3, 5 and 7 at school and household level. Differences in mean Hb and SF between the groups at the end of the trial were the main outcomes. Dietary iron intake did not differ significantly between the guava and placebo groups at school (25 vs. 24 mg/d; p 0.3) and household level (19 vs. 20 mg/d; p 0.3). Baseline mean‐s.d. venous Hb (g/l) and SF (ng/ml) were 120‐6 and 8.2‐3.5 for the guava, and 115‐13 and 7.4‐4.6 for the placebo group, respectively (Hb: p 0.2; SF: p 0.6); at the end of the study these were 130‐7 and 17.9‐10.3, and 126‐15, and 15.4‐5.8 (Hb: p 0.4; SF: p 0.4). No differences on the iron status were found between the children who received guava and placebo juice after 10 weeks. Study supported by grant IMSS‐FOFOI/2005/1/I/029 and CDI‐Chihuahua
Colored Potato Bioactive Compounds Suppress Proliferation and Induce Apoptosis in Both Androgen‐Dependent and –Independent Prostate Cancer Cell Lines
Authors: Reddivari Lavanya , Vanamala Jairam , Safe Stephen H. , Miller J. Creighton
Published: June 2021

We recently reported that colored potato extracts induce apoptosis in LNCaP (androgen‐dependent) and PC‐3 (androgen‐independent) prostate cancer cell lines via caspase‐dependent and ‐independent pathways. In order to identify the most active growth inhibiting and pro‐apoptotic compounds in the potato extract, we have further investigated the effects of different phenolic compounds (chlorogenic acid, caffeic acid, gallic acid, catechin and malvidin) and glycoalkaloids (α‐ chaconine and solanine) found in these extracts on PC‐3 and LNCaP cells. α‐ chaconine (5 μg/ml) and gallic acid (10 μg/ml) showed potent antiproliferative properties and increased levels of the cyclin‐dependent kinase inhibitor p27 in both cell lines. Gallic acid decreased cyclin D1 levels in LNCaP cells. The growth inhibitory effects of α‐ chaconine were potentiated by SP600125, a c‐jun N‐terminal kinase (JNK) inhibitor, and atractyloside, an ADP phosphorylation inhibitor in both cell lines. Both α‐ chaconine and gallic acid induced caspase‐dependent apoptosis through PARP cleavage and activation of caspase‐3 in LNCaP cells. However, induction of apoptosis in PC‐3 cells was caspase‐independent. These results suggest that apoptosis induced by whole potato extracts in prostate cancer cell lines may be in part due to α‐ chaconine and gallic acid. CSREES special research grant.
The Effects of betulinic acid on microRNA‐27a regulated target genes in MDA‐MB‐231 breast cancer cells
Authors: Talcott Susanne U , Li Xiangrong , Chintharlapalli Sudhakar , Safe Stephen
Published: June 2021

Betulinic acid (BA), a pentacyclic triterpene isolated from tree bark is cytotoxic to cancer cells and tumors by inhibiting their growth and inducing apoptosis. There is evidence that specificity proteins (Sps), such as Sp1, Sp3 and Sp4, are overexpressed in tumors and contribute to the proliferative and angiogenic phenotype associated with cancer cells. This objective of this study was to determine the efficacy of BA in decreasing Sps expression and underlying mechanisms. Our results show that BA decreased proliferation of estrogen‐receptor negative breast cancer cell line MDA‐MB‐231 within a concentration‐range of 2.5–10 μmMol/L. BA also decreased Sp1, Sp3, and Sp4 expression and this was accompanied by increased zinc finger ZBTB10 gene expression, a putative Sp‐repressor (ZBTB10) and decreased microRNA‐27a levels, a microRNA involved in the regulation of ZBTB10. Similar results were observed in MDA‐MB‐231 cells transfected with antisense miR‐27a and ZBTB10 expression plasmid. Both as‐miR‐27a and ZBTB10 overexpression decreased the percentage of cells in S phase of the cell cycle; however, ZBTB10 increased the percentage of cells in G0/G1, whereas as‐miR‐27a increased the percentage in G2/M. This latter response was associated with induction of Myt‐1 (a miR‐27a target gene) which causes inhibition in G2/M by phosphorylation of cdc2. The effects to BA can be reversed by transient transfection with a mimic of miR‐27a. These results show that part of the underlying mechanism of the anti‐cancer effect of BA is based on a decreased miR‐27a, accompanied by increased expression of Myt‐1 and ZBTB10 resulting in ZBTB10‐dependent suppression of Sp proteins and dependent genes.
Use of duplicate plate collection to evaluate self‐reported calcium intake among adult women
Authors: Yang Jimin , Boushey Carol
Published: June 2021

Calcium has been voluntarily added to a variety of food products. However, dietary assessment methods may be limited in capturing calcium intake from fortified foods. The objective of this study was to compare self‐reported calcium intakes using food records (FR) with duplicate plate collection (DPC), an objective measure of nutrient intake. During a 5‐week study period, 50 Asian, Hispanic, or non‐Hispanic White women completed 2 consecutive days of FR twice at least 7 days apart. The DPC was randomly assigned to either occurrence of the FR. The Nutrient Data System for Research?was used to calculate calcium intake from FR. Homogenized samples of collected food were analyzed with a spectrometer for calcium. Out of 1072 foods recorded, eight were identified as calcium‐fortified based on FR description. Calcium‐fortified orange juice was specified by 7 participants. Twenty participants recorded at least one type of calcium‐fortified food at least once over the study period. The calcium content in DPC (985±445mg) was statistically equivalent to the average intake estimated from FR (983±404mg, p=0.972). This strong match between the DPC and FR was consistent with the collected foods being correctly described in the FR. Given adequate motivation and attention from the participants and the use of a comprehensive nutrient database, self‐reported intakes may match with objective measure of calcium intake.
Use of Dietary Supplements at Baseline is Similar across Ethnic Groups: Results from the SURE Study
Authors: Monroe Kristine Robinson , Wilkens Lynne , Yonemori Kim , Morimoto Yukiko , Steffen Alana , Shen Lucy , Murphy Suzanne
Published: June 2021

Errors in reporting supplement intake in nutritional epidemiology studies have not been evaluated. The Supplement Reporting (SURE) Study is evaluating the most common methods of collecting dietary supplement use data. We present here cross‐sectional data reported on a supplement frequency questionnaire in the five racial/ethnic populations participating in the study: Caucasians, Japanese Americans, Native Hawaiians, African Americans, and Latinos divided into two comparison groups (US‐born and non‐US‐born) to study the effect of acculturation on supplement use practices. The questionnaire asked about the different types of multiple vitamin preparations and the common single vitamins including selenium, iron, zinc, fish oil and garlic.

Subjects are members of the Multiethnic Cohort who reported regular use of at least one dietary supplement (n =1,109). In general, baseline supplement use practices were similar across ethnic groups. A multivitamin with minerals was reported most often (55% of subjects) followed by calcium, vitamin C, and vitamin E. Significantly fewer Caucasians and Japanese Americans reported use of iron and significantly more African Americans and non‐US‐born Latinos report use of garlic.

Future analyses, using more detailed instruments, will compare supplement use across ethnic groups to determine if the homogeneity of use persists. Supported by NCI, grant # R01 CA106744.
Comparison of a one‐month recall and a one‐month diary of dietary supplement use: Results from the SURE Study
Authors: Wilkens Lynne Ross , Steffen Alana , Morimoto Yukiko , Monroe Kristine , Yonemori Kim , Albright Cheryl , Murphy Suzanne
Published: June 2021

Information on dietary supplement intake has rarely been compared across methods. The Supplement Reporting (SURE) Study evaluated the most common methods of collecting dietary supplement use data among participants of the Multiethnic Cohort Study in Hawaii and Los Angeles who reported use of at least one supplement. In the first year of the study, 205 participants completed a one‐month diary of supplement use and an open‐ended recall at the end of the period covering the month. The supplements were categorized into 22 broad supplement types (4 multivitamins, 12 single nutrients, and 6 herbals). The percentage reporting usage ranged from <1% for vitamin A, ginseng and wheat grass to 67% for regular multivitamins with minerals (MVM) on the one‐month recall. The percentages were similar, but slightly higher, for the diary (e.g., 71% reported use of MVM). Kappa coefficients comparing use on the two instruments were good to excellent, ranging from 0.49 for multivitamins with no minerals to 0.91 for folic acid as a single nutrient; there was complete agreement for ginseng and wheat grass. Most (86%) of the kappas exceeded 0.60. The recall and diary methods agreed well over a 1‐month period. Future analyses will compare these methods for a shorter time period (2 weeks) as well as compare them to an inventory of supplements. Supported by NCI, grant # R01 CA106744.
Clinicopathological correlations in a large Alzheimer's Disease Center autopsy cohort: utility of plaque and tangle counts and application to specific topics.
Authors: Nelson Peter T. , Markesbery William R.
Published: June 2021

Research subjects who underwent longitudinal clinical evaluation and postmortem quantitative neuropathological assessment at the University of Kentucky Alzheimer's Disease (AD) Center were studied to address topics related to AD pathogenesis and therapies. Patients without superimposed brain diseases showed strong correlations between AD‐type pathology counts in isocortex and premortem Mini‐Mental State Examination scores. A theoretical model is proposed in which our results are interpreted to support the “amyloid cascade hypothesis” of AD pathogenesis. With this dataset we also addressed two issues of current interest to the AD research field: 1. Imaging studies (PET for metabolism and amyloid ligands, as well as structural MRI studies) have shown that the precuneus in the parietal lobe is among the first areas affected by AD, however, there have not been prior rigorous studies of the evolution of precuneus neuropathology in AD; 2. The effect of acetylcholinesterase inhibitors on human brains has been a matter of debate, and there has not been extensive analyses of differences in the clinical and pathological progression of the disease among persons who have or have not received the medications.
An Audit Of Single CNS Masses In Patients With Known Systemic Malignancy
Authors: Keohane Catherine , Qassim Abdulla , Bermingham Niamh , O'sullivan Michael
Published: June 2021

The objective of this audit was to determine the nature of symptomatic single CNS space‐occupying masses in patients referred to neurosurgery who had a prior diagnosis of systemic malignancy outside the CNS. The records and histological slides including immunohistochemistry of 78 patients aged between 22 and 80 with a variety of systemic malignancies and a confirmed tissue diagnosis of an intracranial or intradural spinal mass were reviewed. The study period was 8 years (1999–2006). Results: In 20 of the 78 patients the CNS mass was a new primary tumour of the CNS, (16 gliomas, 4 meningiomas). In 57 patients, the CNS mass was a metastasis from the known systemic malignancy, and one patient had a metastasis from a new undiagnosed primary systemic cancer. In all 14 patients with lung carcinoma, the solitary brain mass was a metastasis from lung. In 20 patients with breast carcinoma, 5 were new primary CNS tumours, 15 were metastases from their primary breast tumour. In 12 patients with melanoma 8 were metastases from melanoma, 4 were new gliomas. We conclude that in this study group, almost 26% of patients referred to our neurosurgery department had a new primary CNS tumour, although the proportion is very different for specific cancer types.
Glioma invasion: Identification of determinants of invasion using time‐lapse imaging
Authors: Phillips Joanna J. , Dijkgraaf Gerrit P. , Huillard Emmanuelle , Rowitch David H. , Werb Zena
Published: June 2021

Glioblastoma multiforme (GBM) is a malignant glial tumor that is nearly uniformly fatal despite use of current surgical, radio‐ and chemotherapeutic modalities. GBM infiltrate into the surrounding brain, making them highly resistant to treatment. To design improved therapeutics for GBM, we need to understand the factors that drive tumor cell invasion. Using a robust model for glioma we are studying how alterations in the tumor microenvironment influence tumor cell invasion. During tumor development we have identified specific patterns of glioma cell invasion using both immunohistochemistry and ex vivo brain/glioma slice cultures. We monitored the dynamic interaction between fluorescently tagged tumor cells and components of the microenvironment (blood vessels and microglia/macrophages), at various stages of tumor development using four‐color spinning disk confocal microscopy. We determined that tumor cells can invade along blood vessels, and we are examining the role of microglia/macrophages in this mode of invasion. This mechanism of infiltration emphasizes the importance of the perivascular niche, as it may provide crucial factors for brain tumor progenitor cells. By understanding the factors that influence the interaction of tumor cells and blood vessels we hope to elucidate potential new targets for tumor therapy.
Innate immune modulation as a mechanism for therapeutic action of early EPCs in a nude rat model of pulmonary hypertension
Authors: Ormiston Mark , Deng Yupu , Stewart Duncan J. , Courtman David
Published: June 2021

Early and late‐outgrowth endothelial progenitor cells (EPCs) were cultured from human leukapheresis samples and tested for therapeutic efficacy in a nude rat, monocrotaline (MCT)‐induced model of pulmonary hypertension (PH). Three days after injection of MCT, 106 cells were administered to nude rats via central venous injection. Three weeks after cell injection, right ventricular systolic pressure was elevated in MCT treated animals (61 ± 3.8 vs. 27 ± 0.5 mmHg no MCT control). The administration of early‐EPCs prevented the onset of PH (33 ± 3.2 mmHg), while delivery of late‐EPCs produced no improvement (60 ± 4.0 mmHg). Analysis of cell retention by histology and PCR revealed that, despite the therapeutic benefit of early‐EPCs, neither cell type was retained within the lung at 24 hours post injection. A subsequent in vitro characterization of early‐EPCs revealed a capacity to modulate the innate immune response in a manner similar to that of dendritic cells. Early‐EPCs, which are positive for CD14, MHC II, CD86 and CD11c, and negative for CD1a and CD83, increased surface expression of CD86 and CD83 and decreased surface CD14 in response to stimulation with LPS. Exposure to LPS also induced the secretion of IL‐10 but not IL‐12. As with dendritic cells, co‐culture of early‐EPCs with autologous natural killer (NK) cells in the presence of LPS induced NK cytotoxicity, suggesting a potential mechanism of therapeutic action.
A faculty learning community model for preparing future faculty in anatomy and medicine
Authors: Terrell Mark A.
Published: June 2021

Doctoral programs are designed to prepare students to be competent researchers. However, the majority of faculty members (94%) work in non‐doctoral institutions in which the majority of the time is spent teaching. Concerns of both future and junior faculty include a lack of a comprehensible tenure system, lack of community, and issues not covered in most graduate programs (gaining teaching experience, mentoring, and learning about the academic triad of teaching, research, and service). A faculty learning community (FLC) on preparing future faculty for the medical and health sciences is an effective model to address these concerns and prepare alumni for success as assistant professors. This model consists of an interdisciplinary group of faculty and doctoral students who collaboratively engage in bi‐monthly seminars on all aspects of academia, including the scholarship of teaching and learning. In addition, each doctoral student gains part‐time faculty experience as an adjunct in another institutional setting different from their own. During the FLC meetings, faculty and students discuss, process, and analyze their faculty experiences. The FLC results in creating a strong, nurturing network in which both doctoral students and faculty contribute to a shared understanding about teaching and learning, academic life, and faculty roles and responsibilities.
Getting the message: Mechanisms of protein synthesis initiation
Authors: Doudna Jennifer A
Published: June 2021

The initiation of protein synthesis is a highly regulated process requiring recruitment and correct positioning of ribosomes on mRNA templates. We are investigating how initiation occurs on eukaryotic ribosomes, either through the actions of initiation factor proteins or via internal ribosome entry site RNAs (IRESs) that direct ribosomes to the translational start codon without requiring most initiation factors. Cellular IRESs are thought to play crucial roles in adaptation to environmental stresses that inhibit most protein synthesis. Recent results in yeast and human systems will be discussed.
mRNA control: PUF repressors and GLD2 activators
Authors: Wickens Marvin
Published: June 2021

Eukaryotic mRNAs often are controlled by regulatory elements in their 3’UTRs. Repressors bind those elements and turn down expression, either by affecting mRNA stability, translation, or both. Other proteins bind to the 3’UTR and enhance expression. The two families of proteins we will focus on – PUF protein repressors and GLD2 activators – have key biological roles, both in stem cells and in the nervous system.

Among repressors, we will focus on the PUF proteins. We will discuss how these proteins interact with RNA and exclusively bind to their cognate sites. The modularity of the interaction and the key role of spacer nucleotides in recognition are key features of these RNA‐protein interaction. PUF proteins achieve repression via their interaction with POP2, indirectly recruiting factors involved in decay, repression and deadenylation. Multiple PUF proteins regulate a single mRNA, providing versatility in control.

Among activators, we will focus the GLD2 family of poly(A) polymerase. Initially discovered in C. elegans, we have now identified members in many eukaryotic species, including mammals. Members of this family activate expression by extending the poly(A) tail. We will emphasize new biological roles of these enzymes, and their interactions with members of the PUF protein system.
Transition state analogues, purine salvage, cancer and malaria
Authors: Schramm Vern L.
Published: June 2021

Transition state theory proposes that chemically stable mimics of the transition state will bind more tightly than substrates by the rate acceleration factor imposed by the enzyme, typically 1010 to 1015 fold. Transition state structures of enzymatic targets for cancer and malaria have been explored by the systematic application of kinetic isotope effects and computational chemistry. Analogues of the transition states deduced by these methods bind up to 107 times tighter than substrate. Using these methods, three generations of transition state analogues have been produced for human purine nucleoside phosphorylase (PNP) and two of these are in clinical trials. Transition state analogues of human 5′‐methylthioadenosine phosphorylase show promise as anticancer agents in mouse xenografts. Inhibition of S‐adenosylmethionine recycling and DNA methylation are implicated in the anti‐cancer effect. Malaria parasites are purine auxotrophs and require parasite‐specific PNP and adenosine deaminase in the purine salvage pathway. Powerful and parasite‐specific transition state analogues were designed against both malarial PNP and adenosine deaminases. These inhibitors cause purine starvation (purine‐less death) in parasites cultured in human erythrocytes. Application of kinetic isotope effects and transition state have permitted synthesis of some of the most powerful enzymatic inhibitors.

Supported by GM41916 and AI49512.