bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of the AMP‐activated protein kinase by eicosapentaenoic acid improves vascular endothelial function
Authors: Wu Yong , Xu Jian , Song Ping , Wang Shuangxi , Zou Ming‐hui
Published: June 2021

Ω‐3 fatty acids, including eicosapentaenoic acid (EPA), exert beneficial effects on the cardiovascular system, partly through an increase in nitric oxide (NO) bioavailability. AMP‐activated protein kinase (AMPK) is reported to play an important role in protecting endothelial function. The aim of this study is to determine if EPA activate AMPK resulting in increased NO release. Exposure of bovine aortic endothelial cells to EPA (25 μM, 24 h) increased the phosphorylation of AMPK at Thr172 and eNOS at Ser1177. EPA stimulated NO release, and this action was inhibited in cells pre‐treated with compound C and L‐NAME, or pre‐infected with an adenovirus encoding a dominant negative mutant of AMPKα (Ad‐DN‐AMPK). In addition, Ad‐DN‐AMPK abrogated EPA‐mediated eNOS phosphorylation and NO release. Concomitantly, EPA increased the expression of uncoupling protein 2 (UCP2). Transfection of UCP2 siRNA abolished EPA‐induced AMPK activation. Further, exposure of aortas isolated from apoE−/− mice to AICAR or EPA for 24 h increased endothelium‐dependent vasodilation, which was abolished by compound C. Administration of EPA (500 mg/kg/d) for 4 months increased the phosphorylation of AMPK and UCP2 expression in the aortas of C57BL/6 mice. EPA increased eNOS phosphorylation in aortas from wild type and AMPKα2−/− mice but not α1−/− mice, suggesting EPA‐enhanced eNOS phosphorylation was mediated mainly by AMPKα1. In conclusion, EPA activates AMPK by increasing UCP2 expression. AMPK activation might exert a vasoprotective effect by AMPK 1‐dependent eNOS phosphorylation in vivo.
Functional Variants in the Glutathione Peroxidase‐1 (Gpx‐1) Gene Are Associated With an Increased Risk of Keshan Disease
Authors: Lei Cong , Niu Xiaolin , Wei Jin , Zhu Jianhong , Zhu Yi
Published: June 2021

Keshan disease (KD) is an endemic fatal cardiomyopathy in China. Its etiology is unclear, but might associate with the deficiency in selenium and lower activity of glutathione peroxidase‐1 (Gpx‐1), a selenoprotein. In this study, we screened 71 KD patients and 290 controls, and found that levels of blood selenium and Gpx‐1 activity in KD patients were significantly lower. Two variants (Ala5/6/7 and Pro198Leu) were identified, in which Pro198Leu was statistically differently distributed between the KD patients and the external controls, while the frequency of Ala5/6/7 allele was extremely low. The level of selenium and Gpx‐1 activity in group with Pro/Leu or Leu/Leu on codon 198 polymorphisms were significantly lower than in group with Pro/Pro. The prevalence of variants inversely increased along with the decrease of selenium level, suggesting that low selenium may increase in susceptibility of genomic mutation. Further analysis revealed a synergistic‐multiplicative interaction between Gpx‐1 codon198 alleles and low blood selenium. An in vitro study in neonatal rat myocytes showed that overexpression of Gpx‐1(198Leu) caused a 30% decrease in enzyme activity and less responsive to increasing concentrations of selenium, when compared with Gpx‐1(198Pro). Thus, low blood selenium in carriers with 198Leu susceptible genotype is associated with lower enzyme activity, which may, in turn, increase the risk of KD.
Prenatal Exposure to Maternal Hyperhomocysteinemia and Allele‐Specific Methylation and Expression of H19/Igf2
Authors: Devlin Angela M , Wade Rachel E , Singh Ranji
Published: June 2021

Epigenetic processes may play a role in the metabolic programming associated with prenatal exposure to maternal dietary factors. The goal of this study was to determine the effect of prenatal exposure to maternal hyperhomocysteinemia (HHcy) on allele‐specific methylation and expression of genomically imprinted H19 and Igf2. The H19 gene is located in close proximity to Igf2 with expression of H19 and Igf2 regulated by the methylation status of a differentially methylated domain (DMD). Female C57BL/6J mice with (Cbs +/−) and without (Cbs +/+) heterozygous targeted disruption of the gene for cystathionine‐β‐synthase were mated with male Cast/EiJ mice and fed a control or high methionine/low folate diet to induce HHcy (HH) during pregnancy. The F1 hybrid offspring mice were then challenged with the HH diet or control diet from weaning for 1 month. Mice exposed in utero to maternal HHcy and challenged with the HH diet post weaning had higher plasma total homocysteine than mice not exposed in utero to maternal HHcy but challenged with the HH diet post weaning (12.66±2.7 μM vs 6.99±0.9). This was accompanied by lower (p<0.05) methylation of liver H19 DMD on both the maternal and paternal alleles and higher H19 and lower Igf2 transcript levels in liver. These findings show that prenatal exposure to maternal HHcy results in tissue‐specific changes in H19/Igf2 methylation and expression.
Choline deficiency influences the interaction between REST, chromatin methylation and altered fetal neurogenesis
Authors: Mehedint Mihai George , Niculescu Mihai , Craciunescu Corneliu , Zeisel Steven
Published: June 2021

Dietary manipulation of choline during pregnancy alters mitosis, differentiation and apoptosis in the developing fetal hippocampus. In the current study, we asked if varying the availability of choline to neural precursor cells (NPCs) results in alterations of histone methylation and subsequent changes in hippocampal neurogenesis. Timed pregnant C57BL/6 mice were exposed to choline deficiency on gestation days 12–17 (E12 – E17). In fetuses from deficient mothers, the level of mono methyl lysine 9 Histone 3 (1meK9H3) in the ventricular and subventricular zones of E17 hippocampus was decreased (56± 20% of control, P<0.01) as was the level of di‐methyl lysine 9 Histone 3 (2meK9H3) in the pyramidal layer (34±12 % of control, P<0.05). E14 mouse NPCs exposed in vitro to low choline medium exhibited similar changes. In addition, there was increase in the level of calbindin (Calb) relative gene expression (2.34 ± 0.4 folds, P<0.001), at 72 hours, in the low choline cells. REST inhibitor of neuronal genes is a major regulatory mechanism of Calb gene expression. CHIP assays showed that REST bound to chromatin is decreased in the low choline cells (6.1+/−0.3, P<0.01) relative to input DNA, which correlates with Calb mRNA level. At the same sites we report significant changes in the 1meK9H3 (2±0.3, P<0.05), 2meK9H3 (2.5±0.2, P<0.01) and 2meK4H3 (2.3 ±0.4, P<0.05) levels.

This study is supported by the NIH grant AG‐09525.
Changes in global DNA methylation in response to chronic consumption and withdrawal of folic acid is dependent on the MTHFR 677C T polymorphism
Authors: Crider Krista , Quinlivan Eoin , Berry R J , Hao Ling , Li Zhu , Maneval David , Yang Thomas P , Zhu Jiang‐Hui , Rasmussen Sonja , Yang Quan‐He , Bailey Lynn B
Published: June 2021

We evaluated the effect of the MTHFR C→T polymorphism on global DNA methylation from a double‐blind randomized controlled trial in which Chinese women of reproductive age took supplements of folic acid (100, 400, 4000 μg/d). DNA methylation was expressed as the % of methyl‐C/C (measured by a novel LC‐MS/MS assay) at 0, 1, 3, 6 months of supplementation, and 3 months after withdraw, stratified by MTHFR genotype (CC, CT, TT) (n=135; 15 subjects/genotype x 3 treatment groups). Baseline methylation levels (4.4 +/− 0.12) were identical (p>0.05) across the MTFHR genotypes. Folic acid supplementation resulted in a 13% (p< 0.0001) decrease in global methylation after 1 month of exposure, independently of genotype and dose. After 6 months of supplementation, DNA methylation had returned to baseline in the CT‐ and TT‐subjects taking 100 or 400 μg/d folic acid, but not in CC‐subjects, or in any subject receiving 4 mg/d. During the 3 month washout, DNA methylation decreased further in CC‐ and CT‐, but not in the TT‐subjects. This post‐supplementation decrease in methylation was both genotype and dose dependent, being greatest (compared to baseline) in CC‐subjects taking the 100 μg/d dose (42%, p=0.0001). In conclusion, global DNA methylation response to folic acid supplementation and withdrawal varied by MTHFR genotype with a complex genotype‐dose interaction. Support: Collaborative CDC agreement and GCRC Grant # MO1‐RR00082.
A weight loss formula, WL2, alters gene expressions in endocrine organs of DIO rats
Authors: Zhang Lijuan , Gao Ling , Yao Yisang , Zheng Taoye , Zhou Yanjiao , Zhu Jia‐Shi
Published: June 2021

We reported altered gene expressions in liver, muscle and adipose tissues, improving glucose, lipid and energy metabolisms, in response to WL2 therapy (FASEB J 21:A1352 & 21:A329, 2007). We further explored via microarray analysis the effects of WL2 (1.5g/kg for 10wks; n=20) in gene expressions in adrenal gland (AG), thyroid (T), hypothalamus (HT) and pituitary (P) of DIO rats. Along with significantly less gain of body weight, gene expressions were altered in many metabolic pathways in WL2 treated rats, as compared to controls (all p<0.05): (1) up‐regulated: a 3‐fold increase in gene expressions of calcitonin/calcitonin related polypeptide (Calca) in P, and a 68% increase in gene expressions of somatostatin receptor 2 (Sstr2) in AG; (2) down‐regulated: cannabinoid receptor family members (Cnr1, Cnr2) in AG, HT and P, all pro‐inflammation factors in endocrine organs, and thyroid hormone receptor in HT (−30%). Our data suggest that the weight loss by WL2 is a multi‐target outcome through alterations of multiple metabolic pathways, including regulating endocrine functions towards optimization of glucose, lipid, and energy metabolisms and anti‐inflammation.
Two discrete zinc transporter‐2 (ZnT2) variants integrate zinc secretion and efflux in mammary epithelial cells
Authors: Lopez Veronica , Kelleher Shannon L
Published: June 2021

The lactating mammary gland secretes an extraordinary amount of zinc (Zn) (1‐2 mg Zn) into milk each day. Thus, the mammary gland has developed specialized mechanisms to regulate the transfer of such a large amount of Zn. Zinc transport is regulated by Zn‐specific transporters. We previously determined that women with an amino acid substitution in ZnT2 had significantly lower milk Zn concentration verifying that ZnT2 plays a major role in mammary gland Zn secretion. The primary objective of this study was to further characterize the role of ZnT2 in mammary gland Zn secretion. Using a cultured mammary epithelial cell model (HC11), ZnT2 was detected in intracellular vesicles and at the plasma membrane using confocal microscopy. Two ZnT2 protein variants with molecular masses of ~42 and ~32 kDa were identified. Expression of the 42 kDa variant was higher in secreting relative to non‐secreting cells while abundance of the 32 kDa variant was not affected. Over‐expression of both variants resulted in increased Zn transport relative to mock‐transfected cells indicating both proteins are functional Zn transporters. The 42 kDa variant was localized to late endosomal/secretory vesicles in HEK293 cells expressing a 42 kDa ZnT2‐HA fusion protein. In contrast, the 32 kDa variant was detected at the plasma membrane by cell surface biotinylation and confirmed in HEK293 cells expressing a 32 kDa ZnT2‐HA fusion protein. In summary, expression of two ZnT2 variants in mammary cells likely integrates hormonally stimulated Zn secretion and constitutive Zn efflux to provide optimal Zn for secretion into milk. Supported by UCD intramural grants and PSU to SLK.
Zinc regulation of bone‐specific transcription factor Runx2 and bone marker gene expression within 24 hours in osteoblastic MC3T3‐E1 cells
Authors: Cho Young‐Eun , Lomeda Ria‐Ann R , Seo Hyun‐Ju , Shin Hong‐In , Choi Je‐Young , Beattie John H , Kwun In‐Sook
Published: June 2021

Runx2, a bone‐specific transcription factor, increases bone formation through the stimulation of bone‐marker gene transcription in osteoblasts. In our previous study, Runx2 protein expression decreased under Zn deficiency at 24 h with the delayed gene/protein expression pattern, which implies the optimal Runx2 expression would be within 24 h. In the present study, we investigated whether Runx2 protein expression is delayed by Zn deficiency within 24 h and whether the delayed Runx2 protein expression decreased bone marker gene transcription level. Osteoblastic MC3T3‐E1 cells were treated with Zn− (1 μM as ZnCl2) & Zn+ (15 μM as ZnCl2) along with a control (normal osteogenic medium) up to 24 h and collected in every 4 h. Runx2 protein expression was delayed in Zn− having the highest expression at 24 h, while at 16h in Zn+. Protein expression of BMP‐2, as upstream regulator of Runx2, protein expression was also delayed in Zn‐, which is consistent to the Runx2 protein expression pattern. Transcription of bone‐related genes (Pro COLI, ALP, OP, OC & PTHR) showed a delaying pattern, as being consistent to Runx2 protein expression pattern. The study results suggested that Zn deficiency delayed Runx2 protein expression which may be regulated by BMP‐2 protein and again the delayed Runx2 protein expression delayed bone marker gene transcription in 24 h in osteoblasts. (Supported by KOSEF, F01‐2006‐000‐10172‐0)
Effects of iron chelators on iron export across the basolateral membrane of human intestinal Caco‐2 cells
Authors: Kim Eun‐Young , Han Okhee
Published: June 2021

Very little is known about the process of iron transfer from the enterocyte into circulation. Recently, it was proposed that ferrous ion is transferred across the basolateral membrane via the iron exporter ferroportin, and then oxidized to ferric ion by the ceruloplasmin (Cp) homologue hephaestin on the basolateral surface prior to release into transferrin in the circulation. Although the specific mechanism has not been defined, it has been suggested that iron transfer across the basolateral membrane of the enterocyte into circulation may be the rate‐limiting step in iron absorption during various patho‐physiological conditions. To study the cellular mechanism of iron release across the basolateral membrane of enterocyte into circulation, differentiated human intestinal Caco‐2 cells grown on microporous membrane inserts were used. The differentiated Caco‐2 cells exported approximately 6.5 pmol Fe/mg protein during 1 h incubation when 10 μM 55Fe(NTA)2 was added to the apical compartment. The export of 55Fe was enhanced by 3 – 4 fold when deferoxamine, an Fe(III) chelator was present in the basolateral chamber. The cellular levels of 55Fe were decreased by 30–40% when deferoxamine was added in the basolateral chamber during iron release study. In contrast, bathophenanthroline disulfonate, an Fe(II) chelator and apo‐transferrin had minimum effects on iron release. Similarly, DFO also stimulated iron release from hepatocytes.
Evaluation of automated methods for quantifying serum 25‐hydroxyvitamin D
Authors: Courtney‐Hanwell Heather Elaine , Wagner Dennis , Vieth Reinhold
Published: June 2021

Overview: The accepted biomarker of vitamin D nutritional status is serum 25‐hydroxyvitamin D [25(OH)D]. However, a clear understanding of vitamin D research is confounded by the variability among assays used for measuring 25(OH)D.

Objectives: To compare two new automated assays with the well‐established DiaSorin radioimmunoassay (RIA) for 25(OH)D.

Methodology: The 25(OH)D from human sera (n=160) was quantified using the RIA and two automated non‐radioactive immunoassays: DiaSorin Liaison â Total 25(OH)Dâ and Roche â Elecsys 25‐OHâ. Correlation was assessed with the Pearson test and agreement with the Bland‐Altman method.

Results: The RIA and Liaison correlated well (r = 0.919) and with negligible bias (bias +/â 95% limits of agreement = â 0.72 +/â 16.01nmol/L). The Roche correlated similarly with both the RIA and Liaison but with higher bias versus the RIA (r = 0.871; bias = â 2.55 +/â 21.06 nmol/L) than the Liaison (r = 0.861; bias = â1.72 +/â 21.34 nmol/L). Imprecision (CV%) within‐run for the RIA, Liaison and Roche was 9.66%, 6.29%, and 4.52%, and between‐run was 10.8%, 6.67%, and 9.13%, respectively.

Conclusion: For quantification of serum 25(OH)D, the Liaison â Total 25(OH)Dâ demonstrated a stronger correlation and better agreement with the reference method (RIA) than the Roche â Elecsys 25‐OHâ.
Absence of diabetes indicators in a selenium‐supplementation trial
Authors: Combs Gerald F , Watts Jennifer C , Johnson LuAnn K , Canfield Wesley K , Davis Cindy D , Milner John A
Published: June 2021

We conducted a yr‐long intervention with selenium (Se) to characterize dose‐response relationships of biomarkers of Se status. Volunteers from Grand Forks, ND, were screened by interview, medical history and clinical biochemistry; individuals with liver or renal dysfunction, uncontrolled hypertension, unmanaged diabetes, or BMI>40 were ineligible. Of those eligible, 261 (106 men, 155 women) were randomized to Se supplements (0, 50, 100 or 200 ug Se [as L‐selenomethionine]/day). Baseline plasma Se levels were relatively high (141.5±23.7 ng/ml), with >50% in the upper quintile of the NHANES III cohort, and 6% in the range achieved only by Se supplementation in the NPC trial. As secondary analyses of those studies have suggested associations of plasma Se level and diabetes risk, we evaluated that relationship in our cohort. At baseline, 15 subjects reported having diabetes and/or using diabetic medications; they showed higher (P<0.05) fasting glucose levels (120±25 mg/dl) than subjects without apparent diabetes (88±9 mg/dl), but similar plasma Se levels (132.4±18.3 ng/ml vs. 142.6±23.7 ng/ml) with cases similarly distributed over the tertiles of baseline plasma Se. Subjects without apparent diabetes showed similar fasting glucose levels across those tertiles (low: 86.4±9.0 mg/dl; mid: 87.5±8.3 mg/dl; high: 90.2±10.1 mg/ml). These results do not support a relationship of Se status and diabetes risk.
Modeling zinc (Zn) absorption (AZ) from single test meals (SM) as a function of dietary zinc (DZ) and phytate (DP)
Authors: Hambidge K Michael , Miller Leland V , Westcott Jamie E , Krebs Nancy F
Published: June 2021

Background: Previous modeling of Zn absorption as a function of DZ and DP has focused on total daily diets (TD) (Miller et al, J Nutr, Jan 2007). Numerous studies, however, have been limited to SM.

The objectives of this study were to compare the data from published SM to our model for TD studies and to fit the model to the SM data.

Methods: For the first objective, AZ as a function of DZ and DP was plotted along with the surface of the TD model and the residuals were examined. Then the model was fit to the AZ data using nonlinear regression analysis and fitting statistics were evaluated.

Results: SM data were clustered at the low end of the TD Zn and phytate ranges and AZ were consistently lower than that predicted by the TD model. This appears to indicate that, if the contents of the single meals were ingested over an entire day, the resulting Zn absorption would be notably higher. The AZ data ran parallel to the TD model in the DP axis, but deviated with increasing Zn in the DZ axis. Modeling of the AZ data produced an adequately determined phytate parameter with a value similar to the TD model, but values for the parameters relating AZ to DZ were very high and ill‐determined, indicating the absence of a saturation effect with increasing DZ.

Conclusion: Though modeling data from SM can provide some insight into AZ as a function of DZ and DP, this relationship is better examined using TD data.

Supported by IAEA and MRDDR Center Grant P30 HD004024.
Effects of vegetable‐derived phytochemicals on differentiation‐associated factors in 3T3‐L1 preadipocytes
Authors: CHANG HSIAO‐PEI , CHEN YUE‐HWA
Published: June 2021

The study was aimed at investigating the effect of various vegetable‐derived phytochemicals, including indole‐3‐carbinol (I3C), phenethyl isothiocyanate (PEITC) and benzyl isothiocyanate (BEITC) from cruciferous vegetable, diallyl sulfide (DAS) from garlic, as well as capsaicin from red pepper, on the various factors related to adipocyte differentiation in 3T3‐L1 preadipocytes. The results indicated that these phytochemicals not only dose‐dependently suppressed tumor necrosis factor‐£\ (TNF‐£\)‐interferon£^‐lipopolysaccharide‐stimulated NO production in the undifferentiated 3T3‐L1 preadipocytes, but also inhibited intracellular lipid accumulation and triglyceride contents in the differentiated 3T3‐L1 adipocytes. Additionally, the secretion of IL‐6 and adiponectin by differentiated adipocytes was also modulated by the treatment of these phytochemicals. In conclusion, phytochemicals from cruciferous vegetable, garlic and red pepper are able to dysregulate preadipocyte differentiation by inhibiting lipid and triglyceride accumulation, and such suppression is associated with various adipocyte differentiation mediators, including NO, IL‐6, and adiponectin,
The hexane/ethanol extract of licorice induces apoptosis and cell cycle arrest in DU145 human prostate cancer cells
Authors: Seon Mira , Choi Hyun Ju , Lim Soon Sung , Kim Jong‐Sang , Park Jung Han Yoon
Published: June 2021

Licorice is frequently used in traditional oriental medicine and reported to have anti‐carcinogenic effects. This study examined the effects of the hexane/ethanol extract of licorice (HEL) on apoptosis and cell cycle progression of DU145 cells. HPLC analysis revealed that HEL did not contain glycyrrhizin that is known to induce hypertension. Cells were cultured in DMEM/F12 supplemented with 1% charcoal stripped FBS with 0–7.5 μg/mL HEL. HEL dose‐dependently decreased viable cell numbers and DNA synthesis and induced apoptosis in DU145 cells. HEL increased the number of cells with depolarized mitochondrial membranes and induced the release of cytochrome c from the mitochondria to the cytosol. Western blot analysis of total cell lysates revealed that HEL increased the levels of Fas and truncated Bid. HEL dose‐dependently increased the levels of cleaved caspase‐9, ‐8, ‐7, ‐3 and poly(ADP‐ribose) polymerase. In addition, HEL induced a G1 phase arrest and decreased the protein levels of cyclin‐dependent kinases 2 and 4 and Cyclin A and D1. These results indicate that HEL inhibits DU145 cell growth by the induction of apoptosis and G1 phase arrest and HEL has potential as a cancer chemopreventive agent.
Licorice extract inhibits migration, invasion, and adhesion of DU145 human prostate cancer cells
Authors: Park So Young , Sung Lim Soon , Kim Jin‐Kyung , Kang Il‐Jun , Kang Young‐Hee , Yoon Park Jung Han
Published: June 2021

Licorice root extracts have been used in traditional Oriental medicine for the treatment of inflammatory processes. In the present study, we investigated the effects of licorice extract (LE) treatment on the migration and invasion characteristics of DU145 cells. Because we previously observed that epidermal growth factor (EGF) stimulated migration of DU145 cells, the cells were treated with LE in the absence or presence of EGF. LE inhibited the basal and EGF‐induced cell migration, invasion and adhesion in dose‐dependent manners. Gelatin zymography and Western blot analysis exhibited that LE significantly down‐regulated secretion of matrix metalloproteinase (MMP)‐9 and MMP‐2. LE decreased the protein levels of tissue inhibitor of metalloproteinase (TIMP)‐1, intercellular adhesion molecule, and vascular cell adhesion molecule but increased TIMP‐2 levels. These results suggest that LE is a potent antimetastatic agent, which can markedly inhibit the metastatic and invasive capacity of malignant prostate cancer cells. The decreases in MMP‐9, MMP‐2, and adhesion molecules and increase in TIMP‐2 expression may be a part of mechanisms by which LE inhibits prostate cancer cell migration and adhesion.
Identification of the active compound from the Driselase‐treated fraction of rice bran that suppresses hypertension, hyperlipidemia, and hyperglycemia in stroke‐prone spontaneously hypertensive rats
Authors: Ardiansyah Ardiansyah , Shirakawa Hitoshi , Koseki Takuya , Komai Michio
Published: June 2021

We previously demonstrated that a fraction of rice bran treated with Driselase—a mixture of plant cell wall‐degrading enzymes including cellulases, xylanases, and laminarinases—can reduce high blood pressure (BP) and improve plasma lipid parameters. In this experiment, we isolated and identified the active compound from the Driselase‐treated fraction (DF) of rice bran. The DF was fractionated by silica gel column chromatography and HPLC with an ODS column, and the obtained fractions were orally administered to stroke‐prone spontaneously hypertensive rats (SHRSP). By NMR analyses of the active fraction, the active compound was identified as adenosine, and it improved hypertension and the plasma triglyceride, glucose, and nitric oxide (NO) levels at 2 h after administration. Subsequently, the effects of chronic adenosine administration in SHRSP were studied. SHRSP aged 10 weeks were divided into a control and an adenosine group that were administered water or water with adenosine, respectively, for 3 weeks. The development of high BP was suppressed in the SHRSP from the adenosine group compared to those from the control group.　The plasma lipid, glucose, and NO levels and glucose tolerance were significantly improved in the adenosine‐treated SHRSP compared to the untreated SHRSP. In conclusion, adenosine appears to be beneficial for the suppression of hypertension, hyperlipidemia, and hyperglycemia.
Apoptosis induced by piplartine on PC‐3 cells
Authors: KONG Eun Hee , An Soon Cheol , Meydani Mohsen
Published: June 2021

Prostate cancer treatment is androgen ablation of frequently option but this treatment has limited scope, especially, for hormone‐refractory cancer. Piplartine, a main component of Piper species have known anti‐proliferation and anti‐cancer activity. We investigated the effects of piplartine on PC‐3 cells by using the MTT assay, DAPI staining, PI staining assay, Annexin V binding assay, FACS analysis and Western bloting. The apoptosis in the PC‐3 cells is induced by piplartine and the caspase‐3‐mediated PARP cleavage is involved in the apoptosis process. In addition, the treated cells arrest at a G2 point in the cell cycle might be involved in apoptosis induced by piplartine. In conclusion, these results provide the molecular basis for this potent leading inhibitors to act as anti‐tumor agents in chemotherapy for human prostate cancer and piplartine may be a lead candidate compound to explore anticancer drug potential.
Detection of cranberry juice flavonols in the plasma of type 2 diabetics.
Authors: Singh Ajay P , Wilson Ted , Franco Paul , Vorsa Nicholi
Published: June 2021

Cranberry flavonol absorption and metabolism studies have primarily focused on the aglycone quercetin and the glycoside quercetin‐3‐glucoside that is considered to be more bioavailable than the aglycone, as well as other sugar conjugates. This study of type 2 diabetics characterized the pharmacokinetics of cranberry juice (CBJ) flavonol glycosides following consumption of: unsweetened low calorie CBJ (19 Cal/240 ml), normal calorie CBJ (120 Cal/240 ml), isocaloric low calorie sugar water, or isocaloric normal calorie sugar water. LC‐MS‐ESI and HPLC‐PDA techniques were developed to detect flavonols and metabolites. Quercetin glycosides were indicated by the molecular ion m/z 463, 479, 447 to the specific quercetin fragment of m/z301, enabling the monitoring and quantification of flavonol glycosides in the pre‐ and postprandial plasma of type 2 diabetic patients. Flavonols were not observed following administration of control treatments (isocaloric low calorie sugar water) and could be observed in 50% of individuals following administration of the CBJ treatments, respectively. This study validated LC‐MS and HPLC methods for characterizing plasma glycosylated flavonols. There was no evidence of proanthocyanidin absorption and significant variation among individuals was apparent regarding flavonol pharmacokinetics.
Absorption of anthocyanins through intestinal epithelial cells. Effect of ethanol.
Authors: Faria Ana , Pestana Diogo , Azevedo Joana , De Freitas Victor , Azevedo Isabel , Mateus Nuno , Calhau Conceição
Published: June 2021

Anthocyanins are phenolic compounds that are consumed from many dietary sources such as red fruit, vegetables and red wine. They present a wide range of biological activities including antioxidant, anti‐inflammatory and anticarcinogenic activities. Bioavailability is a major issue regarding the biological effects of these compounds in vivo and remains unclear, with few data available on this matter.

This work aimed to evaluate the absorption of anthocyanins on intestine using Caco‐2 cells. Anthocyanin extract, rich in malvidin‐3‐glucoside, was obtained from grape skins. Anthocyanins were tested on Caco‐2 cells cultured on transwell inserts for 25 days. The transported anthocyanins were detected by HPLC.

Anthocyanins (200 μg/ml) were tested in Hanks medium, in absence or presence of 1% ethanol, for different incubation times. Our results showed that this transport was significantly increased in the presence of ethanol especially after 60 min of incubation. In addition, cells that were pretreated for 4 days with anthocyanins (200 μg/ml) showed an increase in their own transport. Cells viability was assayed and was not altered in these conditions. This result could indicate an interference of these compounds with expression/activity of the transporters involved on their own transport.

Supported by Fundação para a Ciência e Tecnologia (POCI, FEDER, Programa Comunitário de Apoio, SFRH/BD/28160/2006).
Chlorogenic acid, a coffee polyphenol and antioxidant, hastens clinical onset of disease but prolongs life span in the G93A mouse, an animal model of ALS, as compared with caffeine
Authors: Hamadeh Mazen J , Tarnopolsky Mark A
Published: June 2021

The G93A mouse, an animal model of amyotrophic lateral sclerosis, exhibits elevated markers of oxidative stress (malondialdehyde, protein carbonyls) and a compensatory upregulation of antioxidant enzyme activity (Mn‐SOD, catalase) in the gastrocnemius, compared with wildtype control. We aimed to study the effects of chlorogenic acid (CHLA, an antioxidant) and caffeine (CAFN), both ingredients in coffee, on functional and disease outcomes. We hypothesized that CHLA would prolong life span as compared with CAFN. Starting at age 40 d, 20 female and 15 male separately‐caged G93A mice were randomly divided into two groups: CHLA (9 females; 7 males) and CAFN (11 females; 8 males). CHLA and CAFN were provided in the food mix, equivalent to amounts found in 5–10 cups of coffee/day (mg/g body weight). Differences in disease outcomes were evident in females, but not males. Clinical onset was attained 11 d sooner in the CHLA (97 ± 5 d) vs. CAFN (108 ± 2 d) group, a 10% difference (P = 0.036). Disease progression was 20 d longer in the CHLA (39 ± 9 d) vs. CAFN (19 ± 2 d) group, a 2 fold difference (P = 0.037). Life span was 9 d longer in the CHLA (136 ± 5 d) vs. CAFN (127 ± 2 d) group, a 7% difference (P = 0.048). Compared with CAFN, CHLA hastens onset of disease but prolongs life span in female G93A mice. (Supported by the Hamilton Health Sciences Foundation and NSERC).
Antioxidant effects of Panax Ginseng extracts on the LPO production in the hypercholesterolemic apo E KO mice
Authors: Jang Soojeong , Park Miyoung , Kim Kkotchbyul , Shin Eunjung , Lee Myoungsook*
Published: June 2021

Ginsenosides, the principal active components of ginseng, have been shown to possess a variety of beneficial effects on human health, including anti‐inflammatory, antioxidant and anticancer effects. There are two major classes of gensenosides, namely protopanaxadiol(PD;Rb1,Rb2,Rc,Rd,Rg3,Rh2) and protopanaxatriol(PT;Rg1,Rg2,Re,Rf,Rh1). We hypothesized that the manipulated ginsenosides PD:PT ratio(1or 2) have the different effects on the lipid profiles and lipid peroxidation production in the atherogenic animal model. We treated the different PD:PT ratio with total ginseng extracts,100mg/kg/day, intraperitoneally to apo E KO mice 4 weeks after high atherogenic cholesterol diet (cholesterol 1.25%, cocoa butter 7.5% ) was induced atherogenesis for 8 weeks. Both of PD:PT=1 and PD:PT=2 decreased TG&TC in plasma, heart and liver tissues, and increased plasma HDL level. In case of LPO production, it was decreased by PD:PT=1 rather than PD:PT=2 in plasma and liver corresponding to the reduction of markers for liver damages, AST and ALT. Both of the gensenosides treatments decreased LPO production in heart tissues. We found that adiponectin was accumulated in adipocytes in the both treated PD:PT=1 and PD:PT=2 groups even though this tendency was not consistent with plasma adiponectin level. We concluded that the antioxidant effects of ginseng extract with various compounds of gensenosides on the lipid profiles, LPO production and cytokine releasing was affected by the types of tissues, such as the plasma, liver and heart.[This study was supported by Rural Development Administration]
Unusual malignant glioma with extensive fibrosis in an adult
Authors: Weidenheim Karen Marie , Shifteh Keivan , LaSala Patrick A
Published: June 2021

Collagen production is seen in gliosarcomas, desmoplastic infantile astrocytomas and gangliogliomas, and gliomas that are infarcted or treated with radiotherapy. The term “gliofibroma” has been suggested for very rare astrocytomas with extensive collagen production (Acta Neuropathol 1993;85:349; JNEN 1978;38:300‐313) by the glial cells. An unusual malignant glioma with extensive desmoplasia arose in a 63 year old male with a 3 month history of seizures. Imaging studies showed a focally enhancing lesion in the right motor cortex. MR spectroscopy demonstrated an increased choline/creatine ratio and decreased NAA metabolite. A well‐defined neoplasm 3 millimeters below the pial surface had a matrix rich in collagen and reticulin, and several patterns. Less cellular, more fibrotic regions contained anaplastic polygonal to spindly cells . More cellular regions had sheets of pleomorphic oval cells with apoptotic bodies and frequent mitoses. Rounded nodules of clear, uniform tumor cells were present. Tumor cells expressed S100, vimentin and GFAP but were negative for epithelial, mesenchymal and neuronal markers. There was no necrosis, nor endothelial proliferation, but MIB‐1 proliferation index was approximately 30%. The term “malignant gliofibroma” may be useful for rare tumors of glial origin, which have produced a collagen‐ and reticulin‐rich matrix without treatment, infarct or associated sarcoma.
Tumor‐to‐meningioma metastasis: Report of two cases
Authors: Takei Hidehiro , Powell Suzanne Z
Published: June 2021

Background: Tumor‐to‐tumor metastasis is a well recognized phenomenon. Meningioma is the most common intracranial host tumor, with the most common primary sites being reportedly the lung and breast. We report two such cases of metastasis with the primary tumor being lung adenocarcinoma and malignant melanoma (MM) of vulva, respectively. The latter is the second reported case of MM metastasized to meningioma.

Case 1: A 57‐year‐old male smoker presented with a headache and was found to have a left parietal contrast‐enhancing mass on MRI. Histologically, biopsy specimen revealed moderately differentiated adenocarcinoma within meningothelial meningioma. Immunohistochemically, the tumor cells were positive for cytokeratin 7 and negative for TTF‐1. Subsequent CT examination revealed a mass in his lung. Case 2: A 75‐year‐old woman with a history of radical vulvectomy for MM died of widespread systemic metastasis. At autopsy, a 2.5×2×2 cm, firm nodule attached to the falx was incidentally found, with focal black discoloration at the periphery of the mass. Histologic examination showed fibroblastic meningioma with foci of metastatic MM.

Discussion: Meningeal lesions in patients with malignant tumors may reflect a metastasis, a meningioma, or a metastasis in a meningioma, and the histological examination is crucial. Metastasis in meningioma may be an initial presentation of the primary tumor, as seen in case 1.
Intracranial chondroma of the falx in a child; Case report & review of the literature
Authors: Aljerian Khaldoon , Lach Boleslaw , Chorneyko Kathy , Nguyen Clara , Hollenberg Robert D.
Published: June 2021

Intracranial chondromas (ICC) comprise 0.2 – 0.3% of intracranial tumours. They are usually located at the skull base and thought to arise from foci of synchondrosis. Extra‐skeletal ICC are less common; only 50 cases have been reported. These tumors have no bone attachment and their histogenesis isn't known. We present the immunohistochemical (IHC) and electron microscopy (EM) characteristics of a falx chondroma in a 15 year old child.

The tumor was lobulated, 3.5 × 3.0 × 3.0 cm in diameter, pearly‐white externally and gray, with multiple gelatinous islands and small foci of ossification on the cut surface. The paraffin embedded tissue was used for routine and IHC stains. EM was performed on glutaraldehyde fixed material.

Microscopic examination revealed a well differentiated chondroma with foci of ossification and islands of vacuolated cells (VC) encircling thin‐walled capillaries. The majority of VC was represented by PAS, Oil‐red‐O, Vimentin (VIM), CD68, CD31, trypsin‐anti‐chymotrypsin (TACT) and ferritin‐ positive macrophages. A subpopulation of spindle VC displayed reaction for S100, ferritin, TACT and VIM and on EM showed transitional morphological features suggestive of transformation to chondrocytes. There was no evidence of transitional forms to arachnoid cell.

In conclusion, our IHC and EM studies suggest origin of extra‐skeletal ICC from a perivascular multipotential mesenchymal cell.
Aggressive pediatric meningioma with soft tissue and lymph node metastases: a case report
Authors: Doxtader Erika , Fuller Christine
Published: June 2021

Metastatic meningioma is extremely rare, occurring in an estimated 0.1% of cases. We report a case of pediatric meningioma metastatic to cervical soft tissue and lymph nodes. An 8‐year‐old boy presented with headaches, dizziness, involuntary eye flickering, and 3‐year history of cervical lymphadenopathy. MRI revealed a 7.5 cm parasagittal dural‐based mass with venous sinus encasement. Therapeutic embolization was followed by bilateral craniotomy, achieving subtotal resection. Histopathologic examination revealed an atypical meningioma (WHO grade II) with regions of hypercellularity, small cell morphology, sheeting architecture, increased mitoses, and brain invasion. Deletions of NF2 and DAL1 (protein 4.1B) were documented by fluorescence in situ hybridization. Subsequent surveillance MRI studies showed growth of residual intracranial tumor, large extra‐axial and subgaleal CSF collections, and enlarging cervical soft tissue masses with posterior triangle lymphadenopathy. Gamma knife and surgical resection were employed for the intracranial tumor burden; resection of the soft tissue masses revealed metastatic meningioma, with diffuse soft tissue infiltration, extensive lymphatic invasion, and metastasis to 8 lymph nodes. This case demonstrates the aggressive biologic potential of pediatric meningiomas, with potential for distant spread via CSF leakage and lymphatic invasion.
Retrospective survival comparison of patients with cerebral versus cerebellar glioblastomas
Authors: Rivera Andreana L. , Pelloski Christopher E. , Giannini Caterina , Aldape Kenneth D.
Published: June 2021

Glioblastoma (GM) is one of the most common malignant parenchymal tumors in adults with the majority being supratentorial. Cerebellar GMs are far less common, comprising less than 1% of all intracranial GMs. To our knowledge, no large scale studies have been done to compare the survival outcome of cerebellar to cerebral GMs.

Clinical characteristics of 42 patients with newly diagnosed cerebellar GMs from the files of M.D. Anderson Cancer Center between 1985 and 2005 were reviewed, and compared with 649 cerebral GMs. The median age of patients with cerebellar GM was 38.7, which was significantly younger than for cerebral GM (56.7, p<0.001). Survival outcome between the two patient populations was also compared. The median survival for the cerebellar GM group was 61.6 weeks, not significantly different from the cerebral GM group (81.3 weeks, p=0.156). However, long‐term survivorship, as defined by survival at 2 years was 17.5% and 28.6% for cerebral and cerebellar GMs, respectively (p<0.001).

Cerebellar GMs occur in a younger patient population than cerebral GMs. The median survival does not appear to differ, but there is an increase in long‐term survivorship in cerebellar tumors. Comparison of molecular signatures, treatment modalities and tumor progression among the patients are under investigation to better stratify the patient populations.
Cell kinetics underlying grading of gliomas
Authors: Alvord Ellsworth C. , Claridge Jonathan , Anderson Alexander , Swanson Kristin R
Published: June 2021

The development of a bio‐mathematical model of the dynamics of glioma growth (Tracqui et al 1995, Swanson 1999, Murray 2003) based on the net rates of proliferation (ρ) and dispersal (D) of glioma cells led to the discovery of the inevitability of “progression” of low‐grade gliomas to death (Pallud et al 2006) and provided a basis for predicting the development of contrast‐enhancement and the duration of survival given the size and velocity of expansion (Alvord & Swanson 2007). The addition of hypoxia‐driven neo‐angiogenesis and necrosis provides a dynamic picture of the continuum of changes in cell concentration, mitoses, vascular proliferation and necrosis, criteria commonly used in grading gliomas. We have applied these criteria to random high‐power fields of each of 15 D's x 15 ρ's = 225 ρ and D combinations (virtual gliomas) and generated a map of probabilities of a particular grade being associated with each combination of D and ; at each point in time/size. The grade progressively changes with time and size, consistent with about 30% of grade II gliomas “progressing” to grade III and essentially all of these “progressing” to grade IV. At the moment measurement of the velocity of radial expansion and calculation of D and ρ requires two MRI observations without intervening treatment, but the accuracy of the prognosis for the individual seems clear.
Neuronal hypertrophy in asymptomatic Alzheimer's Disease in the BLSA
Authors: Iacono Diego , O’Brien Richard , Resnick Susan M , Zonderman Alan , Pletnikova Olga , Rudow Gay , Yang An , West Mark J , Crain Barbara , Troncoso Juan C
Published: June 2021

Objective: To examine the sizes of neurons, nuclei, and nucleoli of asymptomatic Alzheimer's Disease (AD) subjects (ASYMAD) in specific cerebral regions: CA1‐hippocampus (CA1), anterior cingulate gyrus (ACG), posterior cingulate gyrus (PCG) and primary visual cortex (PVC); and to compare them with age‐matched controls (C), mild cognitive impairment (MCI) and definite AD subjects (AD).

Introduction: The pathology of AD may evolve gradually before the disease becomes clinically manifest. Thus, it is common to find substantial amounts of Aβ‐plaques (NP) and neurofibrillary tangles (NFT) in brains of older subjects with normal cognition (ASYMAD).

Methods: We examined and compared 15 autopsy brains per group: C, ASYMAD, MCI, AD. All subjects were from the Baltimore Longitudinal Study of Aging (BLSA). Using designed‐based stereology (nucleator), we measured the volume of neuronal cell bodies, nuclei, and nucleoli in CA1, ACG, PCG and PVC.

Results: We found a significant hypertrophy of the neuronal cell bodies, nuclei, and nucleoli of CA1 and ACG neurons in ASYMAD subjects compared to MCI (p<0.0001) and to controls (p<0.0001). In PCG and PVC, the hypertrophy was limited to the nuclei and nucleoli. The hypertrophy of neurons and their nuclei and nucleoli in ASYMAD may represent an early reaction to the presence of neurotoxic Aβ or Tau, or an adaptive mechanism that prevents the progression of the disease into dementia.
Co‐existence of diffuse multisystem tauopathy and cerebral amyloid angiopathy in an elderly patient with dementia
Authors: Rivera Andreana L. , Powell Suzanne Z. , Ngo Thu , Dickson Dennis W.
Published: June 2021

85 year old man with progressive dementia and history of stroke clinically diagnosed with Alzheimer's disease (AD) came to autopsy. There was diffuse bilateral atrophy of the frontal/temporal lobes and focal cavitations grossly consistent with remote infarcts (confirmed microscopically) were noted in parietal lobe, basal ganglia, and cerebellar hemispheres. Microscopically, numerous tau inclusions including neurofibrillary tangles, globular inclusions, grains and threads with tau immunohistochemical stain were seen. Tau deposits were present throughout the brain with increased numbers in the limbic lobe, basal telencephalon and diencephalon. Leukoencephalopathy was due to atherosclerosis and deposition of amyloid β protein predominantly in the cortical capillaries, with diffuse cortical involvement. Only rare senile plaques were noted with amyloid immunostains. The distribution of tauopathy was unusual and similar to argyrophilic grain disease (AGD), however, the morphology of tau deposits seen are inconsistent with AGD. The presence of the cerebral amyloid angiopathy is unusual due to involvement of parenchymal capillaries outside the primary visual cortex and suggests a genetic component. A low Braak stage and scattered cortical plaques were insufficient for diagnosis of AD. To our knowledge the constellation of findings seen are unique and not previously reported.
Relative neuron loss (NL) in substantia nigra (SN) and locus coeruleus (LC) in neurodegenerative disorders
Authors: Hedreen John C
Published: June 2021

The pattern of pigmented neuron NL in SN and LC varies in advanced cases of different types of neurodegenerative disease. It is well‐known that in Parkinson disease (PD) and diffuse Lewy body disease (DLBD) there is usually moderate to severe NL in both SN and LC, whereas in Alzheimer disease (AD) NL is typically much more severe in LC than in SN. This study investigates relative NL in SN and LC by review of semiquantitative estimates of NL in the author's Neuropathology Reports at the McLean Brain Bank for the last 7 years. The accepted patterns for AD, PD, and DLBD are confirmed. MSA follows the PD pattern. In progressive supranuclear palsy, and frontotemporal degeneration tauopathies and non‐tauopathies, NL is somewhat variable; typically there is moderate to severe NL in SN with none or mild NL in LC. In argyrophilic grain dementia, SN has modest NL and LC little. In familial severe amyloid angiopathy dementia cases, half have an AD‐type pattern and half only mild NL, worse in LC than SN. In severe tauopathy cases with a Guam Parkinson‐dementia tangle distribution pattern, SN NL varies from none to moderate, and all have moderate to severe NL in LC. This study establishes typical patterns of SN and LC NL for different neurodegenerative diseases. No explanation is presently available for these different patterns.