bluuebunny/crossref_metadata_2025 huggingface dataset:
Minimal effect of lifetime low phylloquinone intake on bone status in aged rats
Authors: Ferland Guylaine , Blasko Melissa , Park Lara , Weiler Hope
Published: June 2021

The specific action of vitamin K in bone, whether in the form of phylloquinone (K1) or menaquinone‐4, remains elusive. While there has been reports pointing to beneficial effects of VK in bone health, others have not confirmed such contribution. Furthermore, most research has focussed on the higher ranges of intakes, the potentially detrimental effect of low VK intakes having been neglected. To gain insight on this issue, we conducted a study in which bone status was investigated in 22‐month female SD rats which had been fed diets containing either very low (VL: 80; n=10), low (L: 500; n=8) or moderately high (H: 2000; n=15) levels of K1 (μg/Kg diet) since weaning. A fourth group (n=9) were fed the VL diet until the age of 14 months and the H diet until sacrifice (VL‐H). Measurements, included: area, weight, bone mineral content (BMC), and density (BMD) of whole body, tibiae and femurs; anthropometrics, BMC, BMD and biomechanical testing (Young's modulus, yield and maximum load) of excised femurs. Total body area (p=0.034) and weight (p=0.05) were higher in the VL‐H than in the L group. With respect to bone status, only the BMC of the tibiae were affected by diet, values being higher in the VL‐H than in the VL group (0.374 vs 0.329 mg; p=0.004). All other bone composition or biomechanical measures were unaffected by diet. In this model of normal aging, lifetime low K1 intake had minimal impact on bone status in old age. Supported by CIHR.
Abundant expression of SPARC, birc2 and boca mRNAs in goldfish scale
Authors: Li Li Hua , Katsuyama Hironobu , Do Son Ngoc , Saito Masayuki , Tanii Hideji , Saijoh Kiyofumi
Published: June 2021

The existence of scales in teleost is related to the protective functions of the vertebrate integument and each scale is considered to be a counterpart of hair and/or nail. However, the scales can contain about 20% of the total body Ca so that they also appear to represent an internal Ca reservoir. Thus, in order to confirm whether scales are available for calcified tissue research or keratinized tissue research, characteristics of cells in the scale were examined by identifying differentially expressed genes. Subtraction cloning was subjected to scale ‐ gill (SG) and gill ‐ scale (GS). Six clones from SG and 4 from GS were isolated. Among them, SG1, SG2, and SG3 showed the high homology to SPARC, baculoviral IAP repeat containing 2 (birc2), and bcl2‐related ovarian killer a (boka), respectively. In situ hybridization confirmed birc2 and boca expression in flat cells along the ridge. Birc2 also expressed in large cells between the ridges. SPARC expressed in flat and small cells between the ridges. Although all the cells in the scale were designated as scleroblasts, there were several types of cells with different gene expression. Co‐expression of birc2 and boka along the ridge suggested that anti‐apoptotic mechanism worked to maintain the ridges. Since the cells did not always express SPARC, Ca homeostasis and/or osteocytic manifestation seemed to be maintained by the limited number of the cells.
Lipid profile and total bone mineral content in an experimental model of nutritional hypercholesterolemia
Authors: Macri Elisa Vanesa , Zago Valeria , Ramos Cecilia , Gamba Carlota Alicia , Rodriguez Patricia Noemi , Schreier Laura , Lifshitz Fima , Friedman Silvia Maria
Published: June 2021

Studies showed a strong relationship between fat diets and CVD but not, cholesterol levels involving bone health.

Objective: to investigate in an experimental model of nutritional hypercholesterolemia, the association between lipid profile and total bone mineral content (BMC) under high‐oleic‐acid sunflower oil (HOSO) and sunflower oil (SO) diets. Weaning male Wistar rats (n=56) were randomly assigned to one of 7 groups: a) control group (C) or b) six experimental (E 1–6) for 6 wk. C received rodent diet for 6 wk. E were fed a high cholesterol diet (HCD) (pellets+biliary salts+colesterin+butter) for 3 wk (stage 1), followed by stage 2: E1, E3, E6 replaced the HCD by HOSO, SO or pellets, respectively. E2, E4: replaced butter by HOSO or SO, respectively. E5 was kept on HCD. After 6 wk, blood was drawn for cholesterol (CHO, mg/dl), triglycerides (TG, mg/dl), high density lipoprotein‐cholesterol (HDL‐CHO), HDL‐non‐cholesterol (N‐HDL‐CHO) and N‐HDL/HDL ratios. BMC by DXA (Lunar DPX‐L) was measured. Pearson's correlation coefficients were calculated, p was set at 5%.

Results: the correlation between lipid profile vs. BMC were: +0.18 (ns), ‐0.65 (p<0.0001), −0.66 (p<0.0001), +0.32 (p<0.01) and −0.65 (p<0.0001) for TG, CHO, N‐HDL, HDL‐CHO and N‐HDL/HDL, respectively.

Conclusion: there is a strong relationship between atherogenic lipid profile and bone formation. Awarded by UBACyT O 003–004
Development of a streptozotocin‐induced diabetic rat model for studies on the effects of cinnamon on glucose tolerance and insulin secretion
Authors: Striffler John Samuel , Anderson Richard A
Published: June 2021

A streptozotocin (STZ) dose response protocol using graded doses of STZ was utilized to develop a diabetic rat model to study the effects of cinnamon on glucose tolerance and insulin secretion. In addition to the presence of severe basal hyperglycemia, insulin responses to oral glucose showed no change from basal in rats given more than 45 mg of STZ/kg body wt. Oral glucose tolerance (OGT) in rats given 40 mg of STZ/kg body wt was abnormal but the pancreas was able to secrete significant amounts of insulin. In contrast, there was hyper‐secretion of insulin associated with near normalization of glucose tolerance at STZ doses of 20 mg/kg body wt or less. In light of these observations, effects of cinnamon dosing on OGT was characterized in rats made diabetic with 40 mg of STZ/kg body wt. The cinnamon dosing experiments compared OGT glucose and insulin responses in control diabetic rats (n=12) with diabetic rats administered cinnamon (n=12) added directly to the glucose test solution. Over the range of cinnamon levels tested (4 to 400 mg/kg body wt), a cinnamon dose of 65 mg/kg wt produced the most significant improvements of OGT, which was associated with enhanced insulin secretion. In conclusion, we have demonstrated that cinnamon can be used to control blood sugar under conditions of experimentally‐induced diabetes. (Funded by USDA/ARS).
Conjugated linoleic acids alter body composition differently according to physiological age in moulard ducks
Authors: Fesler Jeffrey Allen , Peterson Daniel
Published: June 2021

Recent developments in the field of lipid biology have shown conjugated linoleic acids (CLA) to have remarkable yet inconsistent metabolic effects in mice, rats, hamsters, chickens, cows, and humans. In particular, effects on lipogenesis vary with tissue, physiological state, and specie. In this study, sub‐adult (7 wk) and adult (11 wk) moulard ducks were grouped by age and fed a standard corn‐soy based diet supplemented with either 5% soybean oil (control) or 5% CLA isomer mixture (30% 9c,11t, 30% 10t,12c; CLA). Animals were harvested after 3 wk or 6 wk for assessment of body composition including liver, adipose, viscera, and empty carcass weight. After 3 wk, the sub‐adult CLA group exhibited a 24% decrease in dissectible adipose tissue (P<0.05) while adult animals showed no significant diet effect. After 6 wk, the sub‐adult CLA group exhibited a 20% increase in liver mass compared to the control (P<0.05), but no diet effect on adipose tissue, while the adult CLA group showed a 42% decrease in adipose tissue mass compared to the control (P<0.05). These data indicate that CLA have potent effects on lipid metabolism in ducks, but that these effects differ dependent on physiological age.
Estimating optimal true digestible Ca: P ratio for 20‐50Kg growing pigs fed a corn‐rough‐soybean based meals
Authors: Yin Yulong , Hu Qin , Huang Ruilin , Li Tiejun , Hou Rongqing , Wu Guoyao
Published: June 2021

This experiment was conducted to study the effects of a true digestible Ca: P ratio on growth and serum characteristics as well as the utilization of Ca,P,N and microelements on 20–50kg growing pigs. Total dietary contained 0.26% true digestible P, and the dietary true digestible Ca: P ratios were 0.89, 1.37, 1.82, 2.29, and 2.75, respectively. In experiment 1. 25 growing pigs were randomly assigned in five replicates to estimate the growth performance and serum metabolites in five dietary Ca: P ratios. Compared with the other groups, the group with the true digestible Ca: P ratio 1.82 had the best average daily gain(ADG), average daily feed intake(ADFI) and gain: feed(G:F), and was greater significant than the high Ca: P ratios(2.29 and 2.75) groups; With the Ca: P ratio increased, the serum P levels declined significantly(P<0.05); Serum alkaline phosphatase activity was not influenced by dietary Ca: P ratio(P>0.05). In experiment 2. 25 growing pigs were randomly assigned in five replicates to estimate the utilization of Ca,P and microelements(Mg,Cu,Fe,Zn,Mn) in five different dietary Ca: P ratios by the balance experiment. When the Ca: P ratio increased from 0.89 to 1.82, the apparent digestibility of Ca,P,N,Mg,Cu,Fe,Zn and Mn was no difference(P>0.05), but when the Ca: P ratio increased to 2.29, the apparent digestibility of Ca,P,Mg,Cu,Fe,Zn and Mn declined significantly(P<0.05). The results suggested that when the true digestible Ca: P ratio was 1.82, the growth performance and the utilization of Ca,P,Mg,Cu,Fe,Zn,Mn were optimal in 20–50Kg growing pigs fed corn‐rough‐soybean meals.
Molecular Profiling of Generic Versions of the Low Molecular Weight Heparin Enoxaparin and Their Digestion by Heparinase‐I.
Authors: Clark Melanie , Zhu He , Hoppensteadt Debra , Fareed Jawed
Published: June 2021

Generic versions of the low molecular weight heparin (LMWH) enoxaparin (Lovenox, Sanofi‐Aventis, Bridgewater NY) are available in Southeast Asia and South America. The purpose of this study was to compare Lovenox with two generic versions, Lupenox (co. name) and Loparin (co. name). The molecular weight (MW) profile of these agents and unfractionated heparin (UFH) was determined using HPLC prior to and after heparinase‐I digestion. The MW of the generic versions of LMWH ranged from 3.9 to 4.7 kDa. Lupenox exhibited the lowest mean MW (3.9 kDa), whereas Loparin was 4.7 kDa. UFH had a MW of 17.1 kDa. Heparinase‐I digestion resulted in marked decrease in the MW of all agents studied. UFH was the most susceptible to the actions of heparinase‐I digestion (> 50% reduction in MW) whereas the LMWHs exhibited a 20–30% reduction. Loparin was more susceptible to heparinase‐I digestion (30%) compared to Lovenox and Lupenox. The molecular components with a MW > 7.5 kDa ranged from 3–10% for the LMWHs and >90% for UFH. The molecular components with a MW < 7.5 kDa ranged from 90 to 97% for LMWHs and 9% for UFH. These results suggest that the generic versions of LMWHs exhibit differences in their molecular profile and digestion by heparinase‐I. Moreover the studies indicate that relative to UFH, both generic and branded enoxaparins are less susceptible to heparinase‐I digestion.
Connecting the Genome to the Cytoplasm
Authors: Silver Pamela
Published: June 2021

The movement of mRNAs through the nucleoplasm and out of the nucleus to cytoplasmic locations is an elaborately orchestrated and regulated process. The pathway of mRNA export includes: proper processing, packaging into protein‐RNA complexes, targeting and movement through the nuclear pore complex and release into the cytoplasm for translation. In some cases, mRNAs are further localized within the cytoplasm. The presentation will focus on the spatial organization of the genome with respect to its interactions with the nuclear pore complex, nuclear export factors, RNA binding proteins, and the processing and movement of mRNAs out of the nucleus.

We have taken a systems‐wide approach to delineating the complex relationship between mRNA production, processing and transport. In particular, we have used genome‐wide analysis to determine the differential interactions between proteins involved in all stages of the process and the entire genome of both yeast and human cells. In doing so, we have identified the action of new factors in metazoan cells and made the surprising finding that there are distinct ribosomes with specialized functions.
Low Expression of Claudin‐5, a flow‐sensitive endothelial tight junction molecule, correlates with sites vulnerable to atherosclerosis in vivo
Authors: Karamanian Armen A , Davies Peter F
Published: June 2021

In large arteries atherosclerosis‐susceptible sites are associated with disturbed flow (DF) while athero‐protected sites are associated with undisturbed (UF) flow. Increased endothelial permeability to macromolecules, a prominent feature in DF regions, is considered to contribute to this susceptibility. The main determinant of endothelial paracellular permeability is the tight junction, a group of proteins that seal the intercellular space. However, little is known about the tight junction responses to different flow profiles. Endothelial cells were isolated from two DF (aortic arch and renal branch) and UF (descending thoracic aorta and renal artery) regions from normal adult pigs and probed for expression of tight junction molecules. Western blots revealed a 5‐fold lower expression of Claudin‐5 (CLDN5) in DF compared to UF regions whereas expressions of Zonula Occludens‐1, Occludin and VE‐cadherin were not significantly different. In situ immunostaining of CLDN5 protein was weak in DF and prominent in UF regions. CLDN5 is a flow sensitive molecule and its low expression in DF regions of heightened permeability may influence spatial susceptibility to atherosclerosis.

Supported by grants from the NHLBI.
Determination of Rodent Tropoelastin in the Skin by Competitive ELISA
Authors: Barron Adam G. , Warner Roscoe L. , Bhagavathula Narasimharao , Johnson Kent J. , Varani James
Published: June 2021

Current interest in novel compounds with dermal repair potential has led to the development of a rapid and accurate assay to screen large numbers of samples for newly synthesized tropoelastin in the dermis of natural and photo aged skin. Herein we report the development of a competitive enzyme‐linked immunosorbent assay (ELISA) for rodent tropoelastin using commercially available reagents. Utilizing a polyclonal antibody to mouse tropoelastin and rat lung alpha elastin (Elastin Products Company, Inc) we were able to quantify tropoelastin production in the supernatants of cultured skin and lysate of dermal punch biopsies. Briefly, micro titer plates were coated with antibody (10 micro g/ml) in carbonate buffer (pH 9.6) overnight at 4oC, washed and blocked with 1% BSA in PBS for 1 hour to reduce non‐specific binding. Equal volumes of samples or elastin (standards) and biotinylated elastin were simultaneously added to wells and incubated for 2 hours at room temperature. Plates were washed to remove unbound antigen and Streptavidin‐HRP followed by OPD substrate was used for detection. The range of detection of elastin standard curve was 0.1–200 micro g/ml with a coefficient of variation of < 3%. This assay will prove a valuable tool in determining the ability of novel compounds to increase dermal tropoelastin production and subsequently increase dermal elasticity. Work supported by NIH RO1 HL07097
Chronic hypoxia impaired tolerance to ischemia: attenuation by aeration and phosphodiesterase‐5 inhibition
Authors: Milano Giuseppina , Morel Sandrine , Vassalli Giuseppe , Corno Antonio F , von Segesser Ludwig Karl , Samaja Michele
Published: June 2021

Aim. We investigated the effects of sildenafil and aeration on myocardial tolerance to infarct in a model of chronically hypoxic rat with pulmonary hypertension and right ventricular hypertrophy.

Methods. Rats were exposed for 2 weeks to chronic hypoxia (CH, 10% O2) and divided into three groups: CH alone, treatment with sildenafil (CH+S, 1.4 mg/kg/day, i.p.) or repeatedly aerated (CH+A, 1 h/day). Normoxic animals served as control. At the end of the observation, hearts were subjected to 30‐min regional ischemia followed by 3 h reperfusion, or to right/left ventricle pressure measurement, or to freeze‐clamp to measure proteins of interest.

Results. CH increased right ventricle hypertrophy, right systolic pressure and the infarct size. In addition, the expression level of P‐eNOS and cGMP was reduced, while increasing plasma nitrates+nitrites and pro‐apoptotic caspase‐3. These changes were reversed in CH+S and CH+A, with reduced right ventricle hypertrophy and right systolic pressure, restored expression levels of P‐eNOS, cGMP and caspase‐3, and enhanced tolerance to ischemia‐reperfusion.

Conclusions. Either inhibition of phosphodiesterase 5 by sildenafil during CH, or repeated aerations provided an efficient strategy for the treatment of pulmonary artery hypertension and cardiac hypertrophy, and improved myocardial tolerance to coronary artery occlusion.
Dynamin 2 is involved in Angiotensin II‐induced NADPH oxidase activity mediated through cortactin in vascular smooth muscle cells
Authors: Si Shuyi , Zhou Jianzhong , Xiao Chunling , Su Lianfang , Jiang Wei , He Xiaobo , Zuo Lian
Published: June 2021

Although the actin cytoskeleton has been implicated in the control of NADPH oxidase activation induced by angiotensin II (Ang II), very little is known about the detailed molecular mechanisms. The dynamin family of large GTPases has been suggested in the formation of nascent vesicles in endocytic pathways. However, there is no report about the role of the large GTPase dynamin 2 (Dyn2) on the activity of NADPH oxidase induced by Ang II in vascular smooth muscle cells (VSMC). In this study we found that Dyn2 was colocalized with cortactin, the actin‐binding protein, by immunofluorencence. By performing immunoprecipitation, Ang II stimulation enhanced the association between Dyn2 and cortactin. Importantly, both cortactin siRNA and Dyn2 siRNA significantly inhibited Ang II‐induced ROS production, whereas cytochalasin D and latrunculin A suppressed the ROS formation by Ang II. Furthermore, Dyn2 siRNA significantly inhibited Ang II‐stimulated redox‐sensitive Akt phosphorylation without affecting redox‐insensitive ERK1/2 phosphorylation. These findings provide the first demonstration that Dyn2 is involved in Ang II‐stimulation NADPH oxidase activation mediated through cortactin, which may play an important role in temple‐spatial role in ROS‐dependent Ang II signaling in vascular smooth muscle cells.
Maintenance of glutathione homeostasis by NRF2‐dependent regulation of glutathione reductase during cigarette smoke‐induced oxidative stress
Authors: Ling Guoyu , Harvey Christopher J , Thimmulappa Rajesh K , Singh Anju , Biswal Shyam
Published: June 2021

NRF2, a transcription factor, protects from oxidative stress through maintenance of the redox status which is largely mediated through changes in intracellular GSH levels. Oxidative stress leads to oxidation of GSH, which is then reduced back to GSH by glutathione reductase (GSR). The present study was designed to investigate the role of NRF2 in GSR induction and GSH homeostasis during smoke‐induced oxidative stress. Wild type and NRF2 deficient (NRF2−/−) mice and their embryonic fibroblasts were exposed to acute cigarette smoke (CS) and CS extract (CSE) respectively. Induction of GSR, proliferation, and oxidized/reduced glutathione was measured. Exposure to either CS or CSE resulted in increased GSR mRNA and protein levels in mice and in cell culture. Induction of GSR was dependent on NRF2. In mouse embryonic fibroblasts, inhibition of GSH synthesis, by buthionine sulfoxide (BSO), an inhibitor of glutathione synthetase, did not affect cell proliferation. However, inhibition of GSR, using 1,3‐bis(2‐chloroethyl)‐1‐nitroso‐urea (BCNU), reduced cell proliferation and resulted in accumulation of oxidized glutathione. Transient transfection studies, using reporter constructs revealed one functional antioxidant response element in mouse GSR promoter. We conclude that Nrf2 dependent regeneration of glutathione by GSR plays a critical role in protection against cigarette smoke induced oxidative stress.
Capture and enrichment of CD34+ hematopoietic stem cells from blood circulation using P‐selectin in an implantable device
Authors: Wojciechowski Joel Christopher , Narasipura Srinivas D , Charles Nichola , Mickelsen Deanne , Blair Martha L , King Michael R
Published: June 2021

Clinical isolation and infusion of hematopoietic stem cells (HSCs) is a vital step for restoration of hematopoietic function in many cancer patients. Previously, we have demonstrated an ability to capture HSCs using monolayers of P‐selectin in parallel plate flow chambers. The aim of the current study was to determine if HSCs could be captured directly from circulating blood in vivo. Biocompatible micro‐tubes, coated internally with P‐selectin, were inserted into the femoral artery of rats mobilized with granulocyte colony stimulating factor. Blood flow through the device was verified and wall shear stress was measured to be 4–6 dynes/cm2. In devices removed after 1‐hr blood perfusion, immunofluorescence microscopy and flow cytometric analysis revealed successful capture of mononuclear cells positive for the HSC‐surface marker CD34. Purity of captured CD34+ cells represented a 7‐fold enrichment over levels found in whole blood, with an average purity of 28%. Robust cell capture and HSC enrichment were also demonstrated in P‐selectin coated devices implanted in a closed‐loop arterial‐venous shunt conformation and perfused for 2 hrs. Furthermore, adherent cells were viable in culture and differentiated into burst forming units. The study demonstrates an ability to mimic the physiologic arrest of HSCs from blood in an implantable device and may represent a practical alternative for HSC capture and enrichment.
Endocrine Pancreas Changes in Experimental DMH Administration
Authors: Farinha Rodrigo , Varanda Nuno , Rafael Ana I.V. , Castro Ricardo F. , Cabrita A.M. Silvério
Published: June 2021

Dimetilhydrazine (DMH) is a carcinogen currently used to induce bowel neoplasms in rat and mice. In this study we evaluate changes in the endocrine pancreas of animals submitted to the administration of DMH in a protocol of colon cancer induction.

A total of 40 wistar male rats, four weeks old were divided into two groups of twenty animals each. The group I was a control group kept with no manipulation. The group II was submitted to the administration of 15mg DMH/Kg twice a week during eight weeks. The animals were sacrificed by the end of the 8th week and a necropsy was performed. The pancreas was collected and processed for histology, stained with haematoxylin and eosin. The section was used for histopathologic evaluation and morphometric analysis of the islets of Langerhans’ areas (La), using SigmaScan Pro®.

In the control group (group I) La is 30804.36 ± 13109.6 px and varies between 29020.62 px and 59824.98 px for a 95% confidence interval. The distribution of the La in this group is: VL 0%; L 20,75%; M 58,49%; H 20,75% and VH 0%.

In the test group (group II) La is 27367.64 ± 15053.85 px and varies between 25529.84 px and 29205.44 px for a 95% confidence interval. The La distribution in this group is: VL 0%; L 30.93%; M 54.68%; H 7.91% and VH 6.47%.

According to these results the endocrine pancreas is a target of DHM and there is a significant decrease in the islets of Langerhans’ area in the sections.
To what extent are free testosterone (FT) values reproducible between the two Washingtons, and when can calculated FT be used in lieu of direct measurements?
Authors: Abadie Jude , DeVan Michale , Bankson Daniel
Published: June 2021

Accurate assessment of free testosterone (FT) by equilibrium dialysis and liquid chromatography tandem mass spectroscopy (LCMS/MS) is the gold standard in evaluating many endocrine patients. FT levels can be calculated using measured total testosterone, albumin, and sex hormone binding globulin (SHBG). We hypothesized that a calculated FT value could substitute for measured values. This study compares FT results reported by Walter Reed Army Medical Center (WRAMC) in Washington, DC to results reported by the Seattle Veterans Affairs (VA) Health Care System in Seattle, WA. Calculated FT values are compared to gold standard measurements. Groups 1 and 2 were composed of 54 patients evaluated at the Seattle VA (Group 1) and 94 patients evaluated at a primary care clinic in Alaska (Group 2), whose care resulted in ordering a FT. Group 3 was composed of 64 patients evaluated by the WRAMC endocrine clinic. FT levels were measured by gold standard technology and compared to calculated values. Calculated FT values between the two facilities demonstrated a strong correlation (R2 = 0.98). When comparing calculated FT values to measured levels, Group 3 demonstrated an R2 = 0.93. However, samples from this group with FT values < 50pg/mL demonstrated a poorer correlation (R2 = 0.45). Calculated FT values may be substituted in the clinical setting for gold standard technology when levels are > 50pg/mL.
Simvastatin (40mg/day) Reduced the Activity of Circulating Plasminogen Activator Inhibitor 1 in Volunteers with Metabolic Syndrome
Authors: Wang Long , Rockwood Jason , Zak Danielle , Devaraj Sridevi , Jialal Ishwarlal
Published: June 2021

The Metabolic Syndrome (MS) confers an increased risk for diabetes and cardiovascular disease. We previously showed that Simvastatin has concomitant benefits in reducing LDL‐cholesterol and inflammation in MS subjects. Plasminogen Activator Inhibitor 1(PAI‐1) levels are increased in MS. The current study was to investigate the effects of simvastatin on sP‐selectin, sCD40 ligand, and PAI‐1 which play important roles in the development of atherosclerosis. Fifty subjects with MS were randomized into either placebo or simvastatin (40 mg/day) group for 8 weeks. Blood samples were obtained at 0 and 8 weeks. PAI‐1, sP‐selectin and sCD40 ligand values were obtained by ELISA. There was no baseline differences in any of the parameters studied. Compared to baseline, simvastatin significantly (P<0.05) reduced the circulating PAI‐1 activity (24.3 ± 5.2 U/mL at baseline vs. 21.4 ± 3.9 U/mL after 8 weeks of treatment). Simvistatin did not alter (P>0.05) the levels of sP‐selectin (111.4 ± 35.9 ng/mL at baseline vs. 118.5 ± 71.2 ng/mL after 8 weeks) or sCD40 (2.0 ± 1.6 μg/L at baseline vs. 1.5 ± 1.0 μg/L after 8 weeks). Our data indicate that simvastatin therapy has significant effects on the fibronolytic system in MS subjects as evidenced in a reduction in PAI‐1.
PLCG1 (Phospholipase C, gamma 1) polymorphism and the risk of hepatocellular carcinoma in a Korean population.
Authors: Oh Ah‐Reum , Lee Seung Ku , Kang Hyun Jun , Kim Min ho , Kim Hyoun Geun , Kim Kyoung‐Yeon , Kim Jeong‐Geun , Cheong Jae Youn , Cho Sung Won , Kang Jason J. , Yang Song‐Ju , Kwack KyuBum
Published: June 2021

PLCG1 (Phospholipase C, gamma 1) as an important component of signal transduction processes involved in several cellular processes, including mitogenesis, differentiation, transformation, and tumor migration. We have examined association between polymorphisms of the PLCG1 gene and the risk of transformation from chronic liver diseases (CLD) to hepatocellular carcinoma (HCC). Total of 314 Korean subjects, 96 patients with HCC and 218 patients with CLD were included in this case‐control study. Single nucleotide polymorphisms (SNPs) of PLCG1 gene were searched for and genotyped using the GoldenGate genotyping assay. Only one SNP among the six SNPs of PLCG1 tested was significantly associated. This is a non‐synonymous SNP in exon 21 coded for SH3 domain of PLCG1. The risk of HCC was significantly lower in patients with the G/G genotypes than in those G/A and A/A genotypes. The result from our study provides a novel evidence for an association of the PLCG1 polymorphism with HCC and a genetic susceptibility marker in Korean population.
Localization and Expression of Cytosolic Sulfotransferases (SULTs) in Normal Human Brain
Authors: Salman Emily Deanna , Falany Charles N
Published: June 2021

Cytosolic SULTs are a family of Phase II enzymes that catalyze the transfer of a sulfonate group (SO3−) from 3′‐phosphoadensoine 5′‐phoshosulphate (PAPS) to endogenous and xenobiotic compounds. Message for several of the SULT isoforms has been detected in brain; however, protein expression patterns have not been extensively characterized. Immunoblot analysis was performed on cytosolic fractions isolated from superior temporal gyrus, hippocampus, cerebellum, occipital pole, frontal pole and temporal pole regions of normal brains. SULT1A1 expression is highest in cerebellum and occipital pole, while SULT1A3 expression is highest in frontal and temporal poles. SULT2B1b expression was higher in cerebellum, frontal and temporal poles. SULT1B1 expression was also identified in all brain regions examined, whereas SULT1E1 and 2A1 expression has not been detected. Immunohistochemistry was performed on hippocampus, cerebellum, frontal cortex, temporal lobe and pituitary paraffin sections. SULT1A1, 1A3, and 2B1b localize primarily to neurons whereas SULT1B1 also localizes to non‐neuronal cells. The differences in expression pattern and cellular localization of the SULT isoforms may be associated with selective substrate conjugation and functions in the different brain regions. Research supported by PHS grant GM38953‐22 to CNF.
Effect of Zanthoxylum schinifolium on TNF‐α‐induced vascular inflammation in human umbilical vein endothelial cells
Authors: Cao Li Hua , Lee Yun Jung , Kang Dae Gill , Moon Mi Kyoung , Hwang Sun Mi , Kim Jin Sook , Lee Ho Sub
Published: June 2021

Pro‐inflammatory cytokines induced the injury of endothelial cells caused by increases of adhesion molecules, leading to vascular inflammation and the development of atherosclerosis. In this study, we investigated whether an ethanol extract of Zanthoxylum schinifolium (EZS) inhibited the expression level of cellular adhesion molecules in human umbilical vein endothelial cells (HUVEC). EZS suppressed the cell adhesions between HUVECs and HL‐60 cells induced by TNF‐α in a dose dependent manner. EZS also suppressed the expression levels of adhesion molecules such as intracellular cell adhesion molecule‐1 (ICAM‐1) and vascular cell adhesion molecule‐1 (VCAM‐1) induced by TNF‐α. TNF‐α‐induced increases of MCP‐1 and IL‐8 mRNA expression were also attenuated by pretreatment with EZS. In addition, treatment of HUVEC with EZS suppressed TNF‐α‐induced reactive oxygen species (ROS) production. Furthermore, EZS inhibited NF‐κB activation and IκB‐αphosphorylation induced by TNF‐α, subsequent degradation of IκB‐α. These results demonstrated that EZS could suppress vascular inflammatory process, which may be closely related with the inhibition of ROS and NF‐κB activation in HUVEC.
Effects of hawthorn, a herbal medicine, on arterial blood pressure in anaesthetized rats
Authors: Leung Susan W.S. , Man Ricky Y.K.
Published: June 2021

Hawthorn is a herbal medicine that has been used to treat various cardiovascular disorders. It belongs to a member of the Rosaceae family and its major components include flavonoids and oligomeric procyanidin. The present study examined the cardiovascular effects of a commercial available hawthorn extract (WS 1442) in anaesthetized rats. The carotid artery was cannulated in anaesthetized rats for blood pressure and heart rate measurement. After bolus intravenous injections of WS 1442 (3.125, 6.25, 12.5 and 25 mg/kg), mean arterial blood pressure was reduced transiently in a dose‐dependent manner. A greater effect on the diastolic than on the systolic blood pressure was observed. Heart rate was not significantly affected by all doses of WS 1442. Phenylephrine (1, 3 and 10 μg/kg) dose‐dependently increased arterial blood pressure. Twenty minutes after the administration of the hawthorn extract (10 and 28 mg/kg/min for 7 min), the hypertensive actions of phenylephrine was significantly reduced when compared to rats without exposure to WS 1442. From these findings, it appears that the hawthorn extract possesses hypotensive action. Pre‐exposure to the hawthorn extract also resulted in an impaired response to phenylephrine, suggesting that hawthorn may have a modulatory effect on blood pressure regulation. (Supported by a CRCG grant of HKU)
Treatment with Glucagon‐like Peptide‐1 During Reperfusion Decreases Ischemia‐Reperfusion Injury in Type 2 Diabetic Rat Hearts
Authors: Dokken Betsy B , La Bonte Laura R , Davis‐Gorman Grace , McDonagh Paul F
Published: June 2021

Myocardial infarctions are much more prevalent and severe in diabetic patients, yet the mechanisms underlying the excess injury are not well understood. Glucagon‐like peptide‐1 (GLP‐1) is an incretin with pleiotrophic effects, and is deficient in type 2 diabetes. GLP‐1 has been found to have cardioprotective properties in non‐diabetic hearts undergoing ischemia and reperfusion. We tested the hypothesis that GLP‐1, given in‐ vivo during the reperfusion period after myocardial ischemia, decreases infarct size in diabetic rats. Male Zucker Diabetic Fatty (ZDF) rats, 11–13 weeks of age, (n = 5), were anesthetized and subjected to 30 minutes of myocardial ischemia (LAD occlusion). Recombinant human GLP‐1 (30 pmol/kg/min; Sigma‐Aldrich) was infused IV for 2 hours of reperfusion. Age‐matched ZDF rats (n = 7) were subjected to the same experimental protocol, but treated with vehicle. Left ventricular infarct size (stained with TTC) within the risk zone (stained with Trypan blue), was determined and expressed as a percentage of the area at risk (I/AAR%). GLP‐1 treatment caused a significant decrease in LV infarct size in the ZDF animals (Untreated: 41% ± 7.9%; Treated: 9.6% ± 4.4%, p< 0.05). These findings suggest that treatment with GLP‐1 provides myocardial protection from I/R injury, even when given after the ischemic event. Supported by the University of Arizona Sarver Heart Center, NIH HL58859 and NIH HL07249.
O‐GlcNAc signaling attenuates mitochondrial permeability transition
Authors: Ngoh Gladys Afor , Jones Steven P
Published: June 2021

We have shown that glycosylation of proteins with O‐linked β‐N‐acetylglucosamine (O‐GlcNAc) preserves mitochondrial membrane potential, attenuates post‐hypoxic injury, and reduces myocardial infarct size in vivo. Based on our recent identification of voltage dependent anion channel (VDAC) as an O‐GlcNAc modified protein, we hypothesized that O‐GlcNAc signaling mediates cardioprotection by attenuating sensitivity to mPTP formation. Here, isolated cardiac mitochondria from Vehicle‐ (normal O‐GlcNAc), PUGNAc‐ (high O‐GlcNAc; 50mg/kg, 18hours), or TT04‐treated (low O‐GlcNAc; 10mg/kg, 18hours) C57BL/6J mice showed no difference in total VDAC levels (n>/=3/group). for O‐GlcNAc and immunoblot for VDAC (n>/=3/group), and vice versa, revealed a significant increase in O‐ GlcNAc modified VDAC for cardiac mitochondria from high O‐GlcNAc mice with Vehicle, while a significant reduction of O‐GlcNAc‐modified VDAC was observed with cardiac mitochondria from low O‐GlcNAc mice compared to Vehicle. To assess functional effects of changes in O‐GlcNAc modification of VDAC on mPTP formation, cardiac mitochondria from high, normal, and low O‐GlcNAc mice were challenged with 100μM CaCl2 and the decrease in spectrohotometric absorbance recorded as an index of mitochondrial swelling. Mitochondria from high O‐GlcNAc mice were significantly less sensitive to calcium‐induced swelling, those from low O‐GlcNAc mice were more sensitive to calcium‐induced swelling compared with Vehicle. We that prevention of mPTP formation, potentially via O‐GlcNAc modification of VDAC, represents a novel mechanism of cardioprotection. NIH (HL0833220), AHA (0535270N) and AHA (0715493B).
KATP Channels are Important Downstream Targets of FGF2‐Initiated Cardioprotective Signaling in Mice
Authors: Pietras Daniel Anthony , Newman Gilbert , Schultz Jo El Jean
Published: June 2021

Our laboratory demonstrated that fibroblast growth factor‐2 (FGF2), a molecule whose vascular actions involve opening of surface adenosine triphosphate‐sensitive potassium (KATP) channels, confers resistance to ischemia‐reperfusion (I‐R) injury, a phenomenon termed cardioprotection. Other evidence indicates that KATP channel opening, sarcolemmal or mitochondrial, is necessary for several modes of cardioprotection. Thus, our lab hypothesized that KATP activation plays a major role in FGF2‐induced cardioprotection. Mouse hearts expressing a cardiospecific FGF2 transgene (Tg) and their nontransgenic (NTg) cohort were subjected to ex vivo global low‐flow I‐R. Hearts were treated before ischemia (isch) or before reperfusion (rep) with 10 μM sarcKATP inhibitor (HMR1098) or before rep with 100 μM putative mitoKATP inhibitor (5‐HD). Cell damage was assessed by creatine kinase (CK) release and infarct size was determined by triphenyltetrazolium chloride (TTC) staining. HMR1098 before isch or 5‐HD before rep demonstrated reduction of recovery in treated FGF2 Tg hearts vs. vehicle‐treated cohorts (p<0.05). 5‐HD also attenuated the FGF2‐induced decrease of infarct size and increased CK release from all hearts. Thus, sarcKATP and mitoKATP each participate in FGF2‐induced cardioprotection, but other effectors may also be involved.
Gain of NRF2 function in Non small cell lung cancer promotes tumorigenecity and confers therapeutic resistance
Authors: Biswal Shyam , Singh Anju
Published: June 2021

Rationale: Loss of function mutations in the Nuclear factor erythroid‐2 related factor‐2 (NRF2) inhibitor, Kelch‐like ECH‐associated protein (KEAP1), in non‐small‐cell lung cancer (NSCLC) results in gain of NRF2 function. This study was designed to investigate the dysregulated NRF2‐KEAP1 pathway as a novel determinant of tumorigenecity and therapeutic resistance. Methods: Global gene expression profiling of A549 cells transfected with siRNA. Generation of A549 and H460 cells stably expressing NRF2 shRNA to (a) to evaluate drug cytotoxicity using cell viability assays. (b) growth of these cells using in vitro and in vivo assays. Results: siRNA mediated lowering of NRF2 levels in NSCLC (A549 and H460) cell lines results in global decrease in cellular thiol levels and the drug efflux proteins involved in counteracting electrophiles and detoxification of a broad spectrum of drugs. Cancer cells expressing NRF2 shRNA demonstrated enhanced drug accumulation and sensitivity to commonly used chemotherapeutic drugs. NRF2 shRNA abrogated neoplastic cellular proliferation in vitro and in vivo. These results demonstrate that gain of NRF2 function in NSCLC cells plays an important role in promoting tumor growth and confers resistance against drug induced cell death. Conclusion: Targeting NRF2 activity in lung cancer could be a promising strategy suppress tumor growth and circumvent therapeutic resistance.
Synergistic Anticancer Potential of the Combination of the Novel Curcumin Analog EF24 and p38 MAPK Inhibitors
Authors: Thomas Shala , Khuri Fadlo , Snyder James , Liotta Dennis , Fu Haian
Published: June 2021

Curcumin has been demonstrated to be an effective agent against human cancer cells. This anticancer activity has been in part attributed to the inhibition of the nuclear factor of kappa B (NFkB), a transcription factor that controls the expression of genes involved in cellular survival and the inflammatory response. This knowledge has been exploited to develop curcumin‐derived analogs, including the fluorinated EF24 compound, with higher potency in in vitro and in vivo models. In order to determine the mechanism of this analog, we examined the contribution of major mitogen‐activated protein kinases (MAPKs) in EF24 induced signaling pathways and revealed upregulated ERK, JNK, and p38 MAPK. We utilized pharmacological MAPK inhibitors to determine pathways that are important for EF24‐induced cell death. Interestingly, EF24 in combination with pyridinyl‐imidazole p38 MAPK inhibitors elicited cytotoxic synergy based on combination index (CI) analysis and colony formation assays. The synergy was associated with the induction of apoptosis. These studies suggest that p38 MAPK has a role in the survival response of lung cancer cells and is possibly one of the major survival pathways induced by EF24. Our finding suggests a potential therapeutic approach by combining a curcumin analog, such as EF24, with a p38 MAPK inhibitor for the effective treatment of lung cancer.
Cytotoxic effect of a tepary bean (Phaseolus acutifolius) lectin on human cancer cell lines
Authors: López‐Martinez Josue , Castañeda‐Cuevas Ana Luisa , Yllescas‐Gasca Lorena , Mendiola‐Olaya Elizabeth , Blanco‐Labra Alejandro , Garcia‐Gasca Teresa
Published: June 2021

Lectins can specifically recognize carbohydrates of cell membranes and some of them exhibit anticancer properties. In a previous work, we identified a cytotoxic lectin from tepary bean with differential antiproliferative effect on normal and transformed fibroblasts. The goal of the present work was to evaluate the effect of a concentrated tepary bean lectin fraction (TBLF) on human cancer cell lines growth. TBLF was obtained by molecular weight exclusion and ion exchange chromatography and it was tested on breast cancer cells (MCF‐7 and ZR‐75), cervix cancer cells (HeLa, SiHa and C33A) and colon cancer cells (CaCo2). All cell lines showed a cytotoxic effect, after 72 h of treatment. Inhibitory (IC50) and lethal (LC50) concentrations were calculated for each cell line. The most sensible cells were colon cancer cells with IC50 of 0.119 and LC50 of 3.0 μg protein/mL. CaCo2 and normal intestinal cells from rat illeon (IEC‐18), included as normal counterpart, were also tested at 24 h, LC50 did not change for CaCo2 but IC50 was of 0.563 μg protein/mL; for IEC‐18, IC50 and LC50 were of 0.993 and 7.95 μg protein/mL, respectively. Our results show that TBLF is able to recognize selectively different types of cancer cells, more over between normal and cancer cells. Current studies are focusing on TBLF mechanism action and on in vivo studies against colon cancer.
Catch‐up growth in males affected by the Union Carbide disaster of 1984 in Bhopal, India
Authors: Varma Daya R , Pal Ritesh , Katgara Diana , Sarangi Satinath , Zaidi Tasneem , Holleran Steven , Ramakrishnan Rajashekhar , Mulay Shree
Published: June 2021

Our 2001 study found that exposure of toddlers and parents in Bhopal to toxic gases in 1984 caused stunting of boys but not of girls (JAMA, 290:1856, 2003); here we report on a follow‐up study of anthropometrics in 2006. Informed consent of all subjects was obtained. Data were analyzed by two‐way ANOVA with the factors exposure (yes, no) and age (born before, in utero, born after), and their interaction, followed by pairwise contrasts of each exposed group with its control. There was a significantly greater increase from 2001 to 2006 in height (mean±SD in cm) of boys exposed to gases as toddlers (5.1± 5.3, n=25, p<0.05) vs controls (1.2±2.6, n=14), or born to exposed parents (15.5±7.8, n=6, p<0.01) vs controls (7.4±4.0, n=7), resulting in nearly complete catch‐up; also, body weight and mid‐arm circumference increased significantly more in exposed than in control males. There was a significant correlation in 2001 for height versus age in boys but not in girls, suggesting that the growth deficit found in exposed boys in 2001 was not present in girls because the girls attained their full height at a younger age. The mechanism underlying early stunting followed by a catch‐up growth is unclear. An important adverse effect of the Bhopal disaster was lasting pulmonary pathology. The catch‐up growth in these boys might be similar to that observed in asthmatic children. (Supported by the Canadian Institutes for Health Research)
A fermented Cordyceps sinensis product Cordymax‐6 improves sexual functions via PDE‐5 Inhibition and central pathways
Authors: Gao Ling , Zhang Yan , Zhao Chunsheng , Yang Jieying , Zhou Yanjiao , Tan Ningzhi , Zhu Jia‐Shi
Published: June 2021

Cordyceps sinensis (Cs) is an anti‐aging medicinal herb with profound therapeutic activities in enhancing endurance and sexual functions (JACM 4:289, and 4:429, 1998). Sexual enhancement activities of CordyMax (CM), a Cs replacement fermented with Paecilomyces hepiali, were proven in humans (CTPM 9:29, 1988). We reported the activities of CM in anti‐fatigue and glucose‐lipid metabolisms. We now investigated CM6 on the improvement of sexual functions in vivo. Male Wistar rats were given CM6 (0.32, 0.8 or 2.0g/kg) or a control Viagra (0.05g/kg) by gavage. Platelets were isolated after animals received 3 doses of treatment. PDE‐5 activity was assayed, showing 23% to 40% inhibitions by CM6 therapy (p<0.01). In the 2nd experiment after received 3 doses of CM6 (0.6, 1.2 or 1.8g/kg), male KM mice in CM6/1.8g/kg group showed shortened mounting latency (p<0.01). In the 3rd experiment, Orchiectomized SD rats were given CM6 (0.6, 1.2 or 1.8g/kg) by gavage. Erection latency (EL) was assessed on Days 3 & 22. CM6 shortened EL in orchiectomized rats in a dose‐dependent manner. Compared to controls, rats in CM6/1.8g/kg group showed decreased EL (p=0.048). Our data indicate sexual enhancement activities of CM6, possibly through PDE‐5 inhibition and central pathways.
Nutrient Sensing Through Metabolic Transcription‐Dependent Acetylation
Authors: Puigserver Pere
Published: June 2021

Fluctuations in nutrients are sensed in cells and trigger a metabolic adaptive response to ensure homeostasis for energy and nutrients. For example, nutrient deprivation increases genes involved in mitochondrial fatty acid oxidation and glucose production. A key control occurs at the transcriptional level and coordinates the expression of metabolic genes. Among the major components of this transcriptional circuitry are the PGC‐1α transcriptional coactivator complexes. Using skeletal muscle cells as a metabolic model system we found a nutrient sensing pathway that control fatty acid mitochondrial oxidative function. This pathway involves a response to low nutrients and glucose that in a cell‐autonomous manner switches from glucose to fatty acid oxidation via an increase in a specific subset of mitochondrial genes. This coordination switch to fatty acid oxidation and full respiration activities is controlled through a mechanism of low glucose stimulated SIRT1‐dependent deacetylation and activation of PGC‐1α. De‐acetylated PGC‐1α is bound to promoters and activates genes involved in oxidation of fatty acids. In this regard, SIRT1 deficient cells lack the ability to undergo this nutrient switch under low glucose concentrations. Conversely, in high glucose concentrations PGC‐1α is acetylated through the GCN5 Acetyl Transferase Complex, leading to decreases in mitochondrial fatty acid oxidation. Mechanistically, acetylation of PGC‐1α results in inactivation of the transcriptional activity and displacement from the active chromatin to a different subnuclear localization. In summary, we have identified a nutrient‐dependent molecular mechanisms that involve regulation of PGC‐1α and mediate control of skeletal muscle mitochondrial nutrient oxidation activities.