bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibitory constituents of aldose reductase in the fruiting body of Phellinus linteus
Authors: Kim Hye Mi , Lee Yeon Sil , Kang Young‐Hee , Kang Il‐Jun , Jung Ju‐Young , Shin Hyun‐Kyung , Lim Soon Sung
Published: June 2021

Aldose reductase (alditol/NADP+ oxidoreductase, AR), the first enzyme in the polyol pathway, catalyzes the reduction of the aldehyde form of D‐glucose to D‐sorbitol with concomitant conversion of NADPH to NADP+. It is generally accepted that this polyol pathway plays an important role in the development of some degenerative complications of diabetes such as retinopathy, neuropathy, and nephropathy. In an effort to characterize active principles for diabetic complication from medicinal mushroom, AR inhibitors were isolated from the fruiting body of Phellinus linteus and identified as hispidin, phelligridimer A, davallialactone, methyldavallialactone, hypholomine B, interfungins A, and inoscavin A (11), together with protocatechuic acid (1), protocatechualdehyde, caffeic acid, and ellagic acid. Their structures were elucidated by spectroscopic analyses. Among them, davallialactone, hypholomine B, and ellagic acid exhibited potent rat lens AR and human recombinant AR inhibitory activity with IC50 values of 0.33, 0.82, 0.63 ¥ìM and 0.56, 1.28, 1.37 ¥ìM, respectively. Supported by Brain Korea 21.
Protective effects of quercetin against oxidized LDL‐induced apoptosis through p38 MAP kinase‐dependent and JAK‐responsive pathways in human endothelial cells
Authors: Choi Jung Suk , Li Jing , Kang Young Hee
Published: June 2021

OxLDL is present in atherosclerotic lesions and has been proposed to play an important role in atherogenesis. The present study elucidated multiple MAPK‐responsive death/survival signaling pathways, through which quercetin exerted anti‐apoptosis in endothelial cells exposed to oxLDL. Quercetin substantially diminished the oxLDL‐induced cytotoxicity and DCF staining, suggesting the inhibition of accumulation of oxLDL‐triggered ROS and consequent apoptosis. Additionally, oxLDL induced p53 activation for the Bax translocation and the luciferase activity of p53, which induction was strikingly down‐regulated by quercetin. The Western blot data identified that quercetin at ≤ 10 μM promptly attenuated phosphorylation of ASK‐1, p38MAPK but not JNK, and its downstream c‐myc evoked by oxLDL. Furthermore, this study also attempted to explore JAK2/STAT3 activation in oxLDL‐induced endothelial apoptosis. It was found that quercetin rapidly attenuated the phosphorylation of JAK2 and STAT3 within 30 min. Our findings suggest that the interplay between p38MAPK and is involved in dietary quercetin protection against oxLDL through hampering p38 MAPK‐dependent pathways involving the activation of JAK2. Thus, quercetin may act as anti‐atherogenic agents blocking oxLDL‐induced endothelial apoptosis.

Grant Funding Source: KRF‐2003‐041‐C20338 from Korea Research Foundation and Brain Korea 21.
Regulation of HIFs and PGK1 by Oxidative Stress and Antioxidants in Human Colon Cancer Cells
Authors: Kim Jong‐Sang , Lee InAE , Quintos Lesley , Ha Young Ran , Seo Ji Yeon , Lim Ji Sun , Sung Mi Kyung , Lee Sung Joon
Published: June 2021

Our previous study attempted to identify proteins differentially expressed in human colon tumor cells treated with hydrogen peroxide as prooxidant, delphinidin as antioxidant, and both. Phosphoglycerate kinase 1 (PGK1) was the protein only upregulated selectively by hydrogen peroxide and its ROS‐induced induction is efficiently suppressed by delphinidin, one of antioxidants. In this study we attempted to confirm the previous observations using various antioxidants including alpha‐tocopherol, trolox, BHT in both p53‐positive (HCT116) and ‐negative (HT29) colon cancer cells. Treatment with hydrogen peroxide induced PGK1 in both HT29 and HCT116 cells in a dose‐dependent manner. Antioxidants (alpha‐tocopherol, trolox, BHT, delphinidin) consistently decreased the expression of PGK1 and suppressed the increase of PGK1 expression induced by hydrogen peroxide. However, the expression of hypoxia‐inducible factors (HIFs) known as a major upstream regulator of glycolytic enzymes including PGK1 were not affected by hydrogen peroxide but downregulated by antioxidants. The data suggest that intracellular level of PGK1 might be a biomarker for oxidative status and partly regulated by the levels of HIFs.
Antioxidant status and lipid peroxidation in pediatric patients with acute lymphoblastic leukemia
Authors: Phoonlapdacha Phanphen , Chongviriyaphan Nalinee , Pakakasama Samart , Kalpravidh Rachaneekorn W. , Suthutvoravut Umaporn
Published: June 2021

Oxidative stress is associated with cancers via many mechanisms including lipid peroxidation. Antioxidants play roles in protection against cancers. This study was conducted to determine the status of antioxidant enzymes and vitamins in pediatric patients with acute lymphoblastic leukemia (ALL).

Ten patients aged 1–12 years, newly diagnosed with ALL and 10 age‐matched healthy controls were included in the study. The activities of erythrocyte superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were assessed. Levels of erythrocyte reduced glutathione (GSH), plasma vitamin C (VC), vitamin E (VE), and malondialdehyde (MDA, a marker of lipid peroxidation) were evaluated.

We found that patients with ALL had higher erythrocyte SOD and GPx activities as well as plasma MDA level than controls (3062.9 ± 243.4 vs. 2305.4 ± 71.1, 37.4 ± 5.5 vs. 21.0 ± 0.8 U/g Hb; p < 0.02, and 0.40 ± 0.04 vs. 0.19 ± 0.02 uM; p = 0.006, respectively). But they had lower levels of GSH, VC, and VE than controls (49.9 ± 2.3 vs. 65.4 ± 2.0, 0.7 ± 0.1 vs. 1.3 ± 0.1 mg/dL, and 799.8 ± 49.1 vs. 1180.4 ± 50.2 μg/dL, respectively; p < 0.05).

In conclusion, pediatric patients with ALL have higher oxidative stress but lower levels of antioxidants than healthy counterparts. An intervention to increase antioxidant status in patients with ALL should be considered.

Funded by the Faculty of Medicine Ramathibodi Hospital Research Grant
Effects of soy isoflavones on aberrant crypt foci and DNA damage in colon of rats treated with dimethylhydrazine
Authors: Choi Young‐Sun , Min Won‐Kyung
Published: June 2021

Soy isoflavones have received considerable attention for the potential role in reducing the risk of cancers. We previously observed significant reduction in the number of colonic aberrant crypt foci and expression of cyclooxygenase‐2 in rats which were fed soy isoflavones after injection of dimethylhydrazine (DMH) for 12 weeks. In this study, we investigated whether feeding soy isoflavones before and during treatment with DMH affects aberrant crypt foci formation and DNA damage in colon. Four weeks old Sprague Dawley male rats were fed AIN93‐based experimental diets supplemented with 0 (control diet), 50 or 500 mg soy isoflavones per kg diet for 8 weeks, and received subcutaneous injections of DMH (20mg/kg body weight) twice per week for 4 weeks while maintaining experimental diets. Then all the groups were given the control diet for another 6 weeks. Colon cells were isolated and subjected to the alkaline single cell gel electrophoresis. DNA damage was determined by measuring tail lengths of the cells with fluorescence microscope. There was no significant difference in weight gains and food efficiencies among groups. Numbers of total aberrant crypts and aberrant crypt foci of colon tended to be lower in the rats pre‐fed soy isoflavones as compared with the control group. DNA damages of colonic cells were insignificant among the groups. Being considered that rats were sacrificed at only 6 weeks after DMH treatment, the results indicate that intake of soy isoflavones before and during DMH treatment may reduce formation of aberrant crypt foci in colon, which was irrelevant to colonic cell DNA damage. This research was supported by RIC program of MCIE.
Soy protein diet protects against liver steatosis using DMBA‐Induced mammary tumors in an ovariectomized obese zucker rat model
Authors: Hakkak Reza , Saeid Shaaf , Talley Lori , Fuchs George , Korourian Soheila
Published: June 2021

Human and animal studies have suggested that obesity can play important role in promotion of fatty liver. Recently, we have reported that short‐term soy protein diet can reduce fatty liver. The main objective of this experiment was to investigate the long‐term effects of obesity and soy diet on liver steatosis using 7,12‐dimethylbenz(a)anthracene (DMBA)‐Induced mammary tumors in an ovariectomized obese zucker rat model. treatment. Ovariectomized lean (n=56) and Obese (n=60) Zucker rats were maintained on either casein or soy protein diet from one week before and until 155 days after DMBA treatment. All rats were orally gavaged at age 50 days with 65 mg/kg DMBA. Our results demonstrated obesity caused a significant increased on liver weight (P<0.05) and soy diet reduce liver weight in lean and obese group (P<0.05). Obesity caused a significant increase in fatty liver steatosis with rats fed casein diet had marked steatosis with small foci of mononuclear infiltration (P<0.05) but soy‐fed lean (1.00±0.0) and obese (3.20±0.26) rats significantly had lower steatosis than casein‐fed lean (1.96±.16) and obese (4.42±.10) rats. Our results suggest that long‐term dietary soy treatment can protects against liver steatosis in lean and obese ovariectomized animals. The mechanisms responsible for increasing fatty liver steatosis in obese rats are under investigation.
Effects of a soy protein‐soluble fiber beverage in subjects with ABCG5/8 polymorphism with mild hypercholesterolemia.
Authors: Guevara‐Cruz Martha , Torres Nimbe , Tovar Armando R
Published: June 2021

The ATP binding cassette G5/8 (ABCG5/8) polymorphisms have been postulated to play a role in response to dietary cholesterol. The objective of this study was to determine whether the combination of soy protein diet and soluble fiber in a low saturated fat diet may reduce total and LDL‐cholesterol, and to determine if the responsiveness to the diet was associated with the presence of the polymorphisms of the ABCG5 (C1950G), or ABCG8 (D19H) genes in subjects with hyperlipidemia. SUBJECTS: 51 subjects with an average age of 43 ± 9 y were studied, with serum total cholesterol concentration of 260 to 330mg/dl. Subjects consumed a low saturated fat diet (LSFD) according to the ATP III program for four weeks. Then, patients started to eat a LSFD with a soy protein‐soluble fiber beverage that provided 25g of soy protein and 15 g of soluble fiber for two months. Each month serum cholesterol (TC), triglycerides (TG), high‐ density lipoprotein‐cholesterol (HDL) and low density lipoprotein‐cholesterol (LDL) were determined. RESULTS: Allele frequencies (wild‐type, variant) were C1950G (0.29, 0.40) and D19H (0.90, 0.09). Both genotypes were distributed according to the Hardy‐Weinberg equilibrium. There was a significantly reduction in TC and TG with the protein‐soluble fiber beverage (p< 0.0001) regardless of the polymorphisms, however patients with the ABCG8 D19H variant had lower serum TC, TG and LDL levels. CONACYT grant C01‐016.
Diets containing soy or rice protein isolate (SPI, RPI) increase insulin sensitivity and improve lipid homeostasis in weanling rats fed high fat, high cholesterol Western diets as a result of activation of PPAR and LXR‐mediated pathways
Authors: Ronis Martin J , Chen Ying , Badeaux Jamie , Shankar Kartik , Badger Thomas M
Published: June 2021

Male Sprague‐Dawley rats were placed on AIN‐93G diets containing casein (CAS)on PND24 or onto CAS, SPI or RPI‐based Western diets (Harlan Teklad TD88137) made with high fat and 0.5% cholesterol. Prior to sacrifice on PND50, an oral glucose tolerance test was performed. High fat/high cholesterol feeding impaired glucose disposal (p < 0.05), suggesting peripheral insulin resistance. This was prevented by feeding SPI‐ or RPI‐containing diets (p < 0.05). Increased serum and hepatic triglyceride and cholesterol levels occurred with high fat/cholesterol feeding. Both SPI and RPI reduced serum and hepatic cholesterol (p < 0.05) and hepatic triglyceride levels (p < 0.05). Expression of hepatic PPAR alpha‐, PPAR gamma‐ and LXR‐regulated genes were induced (p < 0.05) by feeding SPI‐ and RPI‐containing diets relative those in CAS‐fed rats, as determined by real time RT‐PCR. This was accompanied by increased binding of the PPARs and LXR to their response elements on the ACO, glucokinase and CYP7A1 gene promoters as measured in EMSA and ChIP assays. Increased expression of PPARs and LXR mRNA and protein were observed (p < 0.05). These data suggest that SPI and RPI elevated insulin sensitivity via increased PPAR gamma signaling, and improved lipid homeostasis as a result of signaling via PPAR alpha and LXR. Supported in part by ARS CRIS #6251‐51000‐005‐00D and the Solae Company (St. Louis, MO).
Soybean snack does not inhibit post‐prandial fat oxidation
Authors: Iwashita Soh , Sakurai Masao , Suyama Toru , Hayase Hideki , Morimoto Taketoshi , Hamada Koichiro
Published: June 2021

Soybean (Soy) intake after overnight fasting increased post‐prandial energy expenditure (EE) compared to wheat intake. We assessed the metabolic consequences of Soy or wheat snacks consumption. Ten healthy male volunteers (29.2 yrs, BMI= 23.7) attend 3 sessions (non‐snack: Non, Soy, Wheat) in a random order with at least one week of wash out period. Soy and Wheat trials were blinded. In each testing day, subjects ate regular breakfast at 0730 and lunch at 1200, and test snack (17 kJ/kg) at 1515 in Soy and Wheat sessions. Expired gas and blood samples were collected every an hour from 1100 until 2115 to determine EE and substrate oxidation (OX) and glucose, insulin and lipid profiles in the blood. Plasma glucose and serum insulin levels were markedly lower in Soy than in Wheat (P<0.05). And those levels in Soy were only slightly higher compared to Non (P<0.05). EE after snack was not different between three sessions. However, change in fat OX was higher and carbohydrate OX was lower in Soy and Non than in Wheat (P<0.05). In addition, serum triglyceride level was higher in Soy than in Wheat and Non (P<0.05). Serum free fatty acid level was higher in Non than in Soy, and further lower in Wheat (P<0.05). These findings suggested that soybean snack does not inhibit acceleration of post‐prandial fat OX, but wheat snack does, possibly by insulin's action of fat storage.
Dietary micronutrients are associated with higher cognitive function gains among primary school children in rural Kenya
Authors: Gewa Constance Awuor , Weiss Robert , Bwibo Nimrod , Sigman Marian , Whaley Shannon , Suzanne Murphy , Harrison Gail , Neumann Charlotte
Published: June 2021

With the exception of iodine and iron, there is still very limited information on the effect of micronutrients on cognitive function, especially among school‐age children. This analysis evaluates the relationship between dietary iron, zinc and B‐vitamins and, gains in cognitive test scores among school children in rural Kenya. Data for this study were obtained from The Child Nutrition Kenya Project, a 2‐year longitudinal. Dietary nutrient values were based on monthly and bimonthly 24‐hour recall data collected during the study period. In longitudinal regression analyses, available iron, available zinc, vitamin B‐12 and riboflavin showed significant relationships with improved cognitive test scores, after controlling for confounders such as energy intake, school, socio‐economic status and morbidity. This analysis demonstrates the influence of improved dietary micronutrient status on school children's cognitive function. Support: GLCRSP
The prevalence and severity of anaemia and iron deficiency after marriage and before childbearing among adolescent and adult females in rural Bangladesh
Authors: Khambalia Amina Zehra , Chowdhury Nuzhat , Hyder Ziauddin , Zlotkin Stanley
Published: June 2021

Background: Anemia and iron deficiency (ID) during pregnancy are associated with adverse maternal and infant outcomes. An estimated 300–500 mg of storage Fe is required for pregnancy.

Objective: To examine iron (Fe) status of nulliparious, married females.

Methods: Baseline data was from a RCT on periconceptional Fe supplementation in Kalingonj, Bangladesh.

Results: Of 272 females, 37% were anaemic (Hb<120 g/L), 16% were ID (SF<12 ìg/L or TfR>4.4 mg/L) and 11% had ID and anaemia (ID with Hb<120 g/L). Median storage Fe was 328.1 (171.9–388.8). 20% of females had <300 mg of storage Fe and all females had <500 mg of Fe.Adolescents (<19 yrs) had significantly lower SF (32.6 vs 42.7, p<0.0007) and storage Fe values(321.2 vs 332.6, p=0.0035).

Conclusions: A significant proportion of females have compromised Fe status and at least one fifth will enter pregnancy with insufficient Fe stores. Adolescent females represent a high‐risk group. Efforts around periconceptional Fe supplementation, education on dietary intake of iron‐rich foods and delayed childbearing are needed.
Expression of Jejunal Apical Excitatory Amino Acid Carrier 1 (EAAC1) is Maintained in Piglets with Bowel Inflammation
Authors: Yang Chengbo , Lackeyram Dale , Archbold Tania , Mine Yoshinori , Fan Ming
Published: June 2021

The objective of this study was to examine jejunal excitatory amino acid carrier 1 (EAAC1) expression in response to bowel inflammation induced with dextran sodium sulfate (DSS) in the piglet model. Piglets at the age of d 5 were fed a commercial source of liquid formula, fitted with an intra‐gastric catheter and administrated with either 2.5 g of DSS/kg body weight (DSS, n = 5) in saline or saline (control, n = 6) in two injections daily for a period of 10 d prior to jejunal tissue harvesting. Small and large bowel inflammations were confirmed with representative pro‐inflammatory cytokine profiling. Predominant EAAC1 immuno‐reactive bands of 57 and 73 kDa were detected in the jejunal homogenate and the apical membrane (AM), respectively, consistent with the nonglycosylated pre‐mature and the glycosylated mature forms of the EAAC1 protein. Expression of the 57‐kDa pre‐mature EAAC1 protein in the jejunal homogenate was lower (P < 0.05) in the DSS group compared with the control. However, there was no difference (P > 0.05) in abundance of the 73‐kDa mature EAAC1 in AM between the two groups. Furthermore, there was no difference (P > 0.05) in EAAC1 mRNA abundance between the DSS and the control groups. These results have suggested that expression of 73‐kDa mature EAAC1 protein in the gut apical membrane is maintained in piglets with during bowel inflammation. Supported by AMFNET of NSERC.
Massive small intestinal adaptation occurs in a neonatal miniature piglet model of short bowel syndrome
Authors: Dodge Elaine , Bertolo Robert , Brunton Janet
Published: June 2021

Our objective was to characterize small intestinal (SI) adaptation in a novel piglet model. Piglets (n = 27, 12 d old) were randomized to 1) 80% SI resection with total parenteral nutrition (TPN) feeding (R‐TPN), 2) 80% SI resection with TPN + enteral feeding (R‐EN) or 3) sham SI transection with TPN + enteral feeding (Sham‐EN). In the R pigs, the distal 100 cm of ileum (residual SI) and 30 cm of proximal SI was left intact. All pigs received TPN after surgery. R‐EN and Sham‐EN piglets also received continuous gastric infusions of elemental diet starting on d 3 (40:60 TPN:EN). Piglets were kept for 4, 6 or 10 d post‐surgery. The weights of residual SI and mucosa were greater in R‐EN compared to other pigs at 4, 6 and 10 d. By d 10, weight per length of residual SI in R‐EN animals was more than 25% greater than other groups; R‐EN mucosal mass was 65% higher. Rate of residual SI length growth in the R‐TPN and R‐EN was twice that of the Sham‐EN, indicating massive adaptation responsive to enteral feeding. A rapid decline in plasma arginine and glutamate occurred in enterally fed groups compared to R‐TPN pigs (p<0.05). Ornithine decarboxylase activity in mucosa of R‐EN pigs at d 10 was 35% higher than Sham‐EN and 60% higher than R‐TPN. Route of amino acid delivery may have affected whole body availability, as the gut may have extracted significant amounts of enteral arginine and glutamate to support intestinal adaptation growth. (SickKids Foundation)
Platelet Sensitivity is Increased by Acrolein
Authors: Sithu Srinivas D , Srivastava Sanjay , Conklin Daniel , O'Toole Timothy E , Bhatnagar Aruni , D'Souza Stanley E
Published: June 2021

Acrolein is a toxic environmental aldehyde. It is also produced during lipid peroxidation. Since acrolein exposure is associated with cardiovascular distress, we studied its effect on mouse platelets. In a platelet aggregation assay, acrolein (10–25μM) added to platelets‐rich plasma (PRP) or fibrinogen (fg) (200nM) added washed platelets, 30s prior to 10μM ADP or 250μM arachidonic acid, significantly augmented aggregation (15–20%) when compared with untreated platelets. To study in vivo effects, blood from mice fed with 1–5 mg/kg/day acrolein for 1–3 days was pooled. ADP‐induced platelet aggregation was augmented (10–20%) in a dose and time dependent manner in acrolein treated animals, both in the PRP and washed platelet system. In acrolein treated mice, plasma levels of platelet factor‐4 and P‐selectin were significantly increased, platelet‐monocyte aggregates in blood increased by 15% and the tail bleeding time was shortened by 25% while plasma fg levels were unchanged. Mice exposed to acrolein inhalation for 6h also showed augmented ADP‐induced platelet aggregation. These data suggest that acrolein exposure at environmentally relevant concentrations could increase platelet adhesion in vivo and such effects may be responsible in part for the increased thrombogenicity of acrolein containing pollutants such as automobile exhaust and cigarette smoke. The work was funded by NIEHS grant ES11860.
Developmental sensitivity to environmentally‐relevant levels of cadmium in the zebrafish, Danio rerio
Authors: Warren Kerri Staroscik , LeMasters Elizabeth H , Wilcox Kenton W , Goss Ashley J , O'Shea Stephen
Published: June 2021

We find that embryonic zebrafish possess a differential sensitivity to cadmium ion that is stage‐, duration‐ and concentration‐dependent. After characterizing cadmium response, we developed a bioassay that includes a general indicator of cadmium‐induced lethality (swim bladder inflation failure) and a specific cardiac response to cadmium (cardiac conduction block). Using these endpoints, we determined that doses as low as 0.5 μM CdCl2 are lethal when administered after 24 hours of development, but not when chronically provided from early stages forward. ICP‐MS analysis of cadmium in embryos subjected to early and late treatments showed that cadmium accumulation was similar in 4 day old larvae of both treatments even though the prognosis for survival differed greatly. Preliminary examination of the role of environment on the biological impact of cadmium ion suggested that entry and accumulation is sensitive to the nature of the cadmium‐ligand complex. Exposure to cadmium doses above 10μM is associated with bradycardia and a reduced incidence of cardiac conduction block. Supporting the hypothesis that heart rate reduction masks the ventricular disorder by eliminating chamber rate mismatch, we found that genetic reduction of heart rate protected slow mo mutant zebrafish larvae from cadmium‐induced atrio‐ventricular delay.

RI‐INBRE NIH P20 RR016457
Some biochemical parameters and histological findings in the kidneys of Hybro chickens treated with chromium (VI) and vitamin C
Authors: Gashi Agim Maliq , Bajraktari Ismet Dervish , Halili Fetah Murtez , Bislimi Bislimi , Bislimi Kemalj , Alija Avdulla Jashar , Fetahu Shukri , Sopa Ramadan
Published: June 2021

In this study we have investigated the effect of ingestion of chromium (VI) as well as coeffect of chromium (VI) and vitamin C on biochemical parameters of the serum (uric acid, aspartate aminotransferase ‐ AST, lactatdehydrogenase ‐ LDH, and alkaline phosphatase ‐ AP), and as well as histopathological findings in kidneys. The chickens were divided in five groups (I–V), each consisting of 10 chickens, and were treated orally for six months with different dosage levels of chromium (VI) and vitamin C.

In comparison with control chicken, the serum of chickens which were treated for 180 days with different dosage levels of chromium (VI) and vitamin C, shows that levels of concentration of uric acid and the activity of AST and LDH had increased, whereas the activity of AP had decreased. Considerable histopathological changes of tubules, interstitial areas and some glomerulus of kidneys of these chickens were observed as well. No protective effect of the vitamin C in this concentration was observed. Moreover, in the case of kidneys vitamin C amplifies a toxic effect of chromium (VI). The effects of chromium (VI) depend on the dosage administered, its duration of action and biochemical parameter.
Pathological Changes In Melanoma Induced by Multiple Nanosecond Pulsed Electric Fields Treatments And Possible Mechanism Analysis
Authors: chen xinhua , Swanson Robbert James , Schoenbach Karl H , Zheng Shusen
Published: June 2021

Objective : Nanosecond pulsed electric fields (nsPEFs) can affect the intracellular structures of cells in vitro. This study is to reveal the pathological characteristics of B16‐F10 melanoma on SKH‐1 mice exposed to nsPEFs and analysis of the possible underlying mechanism.

Methods: Melanoma model was set up by injecting B16‐F10 on 120 female SKH‐1 mice. After the multiple treatments nsPEFs ( intensity as 100 kV/cm and duration as 300 nanosecond ), morphologic changes of melanoma was detected and Caspase‐3,6,7 and histone H2AX were also

Results: After multiple nsPEFs, the melanoma blood supply was blocked and melanoma shrink 90% compared to the control group. Histological exams revealed the features of increased metabolic activity and apoptosis body in the treated melanomas. Moreover, TUNEL showed an increase in the apoptotic activity in treated melanoma. These alterations were associated with caspase3,6,7 and histone H2AX increased significantly after pulse as revealed by immunohistochemistry.

Conclusion: nsPEFs has potential applications in cancer therapy,
A2B adenosine receptors promote tumor growth
Authors: Ryzhov Sergey , Novitskiy Sergey V. , Zaynagetdinov Rinat , Goldstein Anna E. , Biaggioni Italo , Dikov Mikhail M. , Feoktistov Igor
Published: June 2021

Due to rapid growth, solid tumors experience severe hypoxia and produce high levels of extracellular adenosine. We have previously shown that A2B adenosine receptors (AR) upregulate VEGF and other angiogenic factors. To test the hypothesis that A2BAR contribute to tumor growth, we used an isograft model of Lewis lung carcinoma (LLC) in A2BAR knockout (A2BKO) and control mice. We found that genetic ablation of A2BAR in the host mice significantly increased median animal survival from 35 to 40 days. On day 14, tumor volume, weight, vascular density and VEGF protein levels were significantly lower in A2BKO (by 48, 38, 30 and 41%, respectively), compared to control mice. Host‐derived CD45+ cells represented approximately 30% of total tumor cell populations in both A2BKO and control mice. After MACS separation, tumor associated CD45+ cells isolated from A2BKO mice produced significantly lower levels of VEGF (37±5 vs 149±8 pg/106 cells in control mice cells) when stimulated with the stable adenosine analog NECA (10 μM). In contrast, we found no difference in VEGF production between LLC CD45‐negative cells isolated from A2BKO and control mice. We conclude that A2BAR expressed on host CD45+ cells infiltrating LLC promote tumor vascularisation and growth, at least in part, by stimulation of VEGF production. Supported by NIH grants.
Basal‐like breast cancers: More than just triple negatives
Authors: Beyer Ginine , Yeh I‐Tien
Published: June 2021

Basal‐like breast carcinomas are aggressive tumors that are typically ER‐, PR‐ and HER2‐ (known as “triple‐negative”, TN), and EGFR+. This study compares breast cancer expression of basal keratins and the TN phenotype.

Methods:

We reviewed immunomarkers on invasive breast carcinoma biopsies from 9/06 to 10/07, including ER, PR, and HER2. Immunohistochemistry (IHC) for CK5/6 was performed on all breast cancers. CK14 and EGFR were performed on CK5/6 + cases and TN cases. Proportion and intensity of staining for CK5/6 and CK14 were evaluated with the Allred score. EGFR staining was graded as 0–3+.

Results:

24 basal‐like breast carcinomas were identified by IHC, representing 16% of the total cases of invasive breast cancer. H&Es demonstrated tumors to be high grade with poor tubule formation, prominent necrosis and high mitotic count. Cases included 19 TNs and 5 that marked with CK5/6 but were not TN. Of the TNs, only 11 were CK5+. All cases were + with CK14. CK14 scores were on average higher than CK5/6 scores (5.4 vs. 3.9). EGFR was + in all but 3 cases, 2 of which were CK5/6 +.

Conclusions:

Not all breast cancers expressing basal keratins are TN. CK14 marked all TN breast cancers and all CK5/6 positive cases. EGFR marked the vast majority of basal‐like breast cancers. Keratin identifies more basal‐like breast cancers than the TN profile.
The effect of RNA inteference‐medicated inhibition of VASP expression on migration of MCF‐7 cells
Authors: zhou xuan , Wei Lei , Chen Huaqi , Wang Jing , Wei Yun
Published: June 2021

Cell migration is critical to many pathophysiological processes, such as tumor invasion and metastasis. It is a complicated process that is regulated by cell‐matrix adhesions and actin cytoskeleton. We report here that Vasodilator‐stimulated phosphoprotein (VASP), which has complex effects on cytoskeletal organization and cell motility. Small interfering RNAs(siRNAs) mediate RNA interference(RNAi), providing a powerful new tool for gene silencing. Herein, we explore the effect of VASP‐siRNAs on migration of MCF‐7 cells. Three pairs of VASP‐siRNAs (942, 1002, 1024) were designed, which were transfected into MCF‐7 cells via Lipofectamine 2000. To evaluate the inhibition activity of VASP‐siRNAs, VASP expression were examined with RT‐PCR and Western blotting. In MCF‐7 cells, we observed that VASP‐942 siRNA could effectively inhibit VASP expression at mRNA and protein level with a concomitant decrease of migration significantly by a wound‐healing assay. Meanwhile, the activation of Rac1 pathway by fMLP could promote the migration of MCF‐7 cells via elevating the VASP expression level. Thus, our study showed that the knockdown of VASP contributes to the reduction of migration in MCF‐7 cells. It provided that the important role of VASP in breast tumour metastasis might be regulated through Rac1 pathway. (This work was supported by National Natural Sciences Foundation of China under Grant No.30570751)
Basal‐like breast cancers express a hypermethylation defect
Authors: Risher J. Devon , Parker Joel S. , Rivenbark Ashley G. , Jones Wendell D. , Carey Lisa , Livasy Chad A. , Coleman William B.
Published: June 2021

Breast cancer cell lines of the hypermethylator phenotype are characterized by the methylation‐dependent silencing of six genes (CEACAM6, CDH1, ESR1, SCNN1A, LCN2, CST6), and exhibit elevated DNMT activity related to overexpression of DNMT3b protein. Examination of the UNC Microarray Database reveals that 19/88 (22%) primary breast tumors lack expression of these genes. Of these putative hypermethylator tumors, 100% are of the basal‐like subtype. These putative hypermethylator tumors represent 83% (19/23) of basal‐like tumors in the dataset. Analysis of a second microarray dataset (from MD Anderson Cancer Center, n=133) identified 26/32 (81%) basal‐like tumors to be putative hypermethylators. Methylation‐specific PCR confirmed that lack of normal gene expression among basal‐like tumors was due to promoter hypermethylation: 7/10 (70%) basal‐like tumors exhibit methylation of 5–6 indicator genes and bisulfite sequencing showed extensive promoter hypermethylation. These findings strongly suggest that basal‐like breast cancers express a hypermethylation defect and that the resulting molecular signature contributes to the clinical behavior and poor prognosis associated with these neoplasms. Further, recognition of this fundamental biological property of the basal‐like breast cancers presents new molecular targets for development of novel treatment strategies. Support: NIH CA78343.
CD6, a novel cell surface marker for prostate cancer progression?
Authors: Crisan Mihaela , Dadzan Elham , Parwani Anil , Logar Alison , Peault Bruno , Croisille Laure
Published: June 2021

CD6 is involved in cell–cell interactions via binding with CD166. Over‐expression of CD166 is a marker for disease progression in prostate, colon, and breast cancer, whereas the expression of CD6 by epithelial tumor cells has not yet been described.

We studied CD6 and CD166 expression, by immunohistochemistry, in 14 prostate primary tumors of Gleason Scores (GS) 6 to 9, 3 metastatic lymph nodes (LNs), and, by flow cytometry in 2 prostate cancer cell lines (PC3 and DU145).

We show that prostate tumor cells express CD166, with a stronger expression in high GS. 6/9 specimens of GS 6 and 7 did not show any tumor cells expressing CD6. Few CD166+CD6+ cells were respectively detected in 1/3 and 1/6 specimens of GS 6 and 7. In contrast, large areas with numerous CD166+CD6+ tumor cells were detected in all (5/5) specimens of GS 8 and 9, and 1/6 GS7. Infiltration by CD6 expressing tumor cells was detected in all 3 invaded LNs. CD6 was strongly expressed by 15% of the PC3 and DU145 cells, while most of these cells weakly expressed CD6, and a few were CD6‐. Interestingly all MUC‐1 positive PC3 cells strongly expressed CD6.

Our study shows for the first time CD6 expression in prostate cancer, in high GS specimens and LNs, and suggests that CD6/CD166 might be involved in cancer progression. Further studies are necessary to assess the functional phenotype of tumor cells subsets depending on CD6 and CD166 expression.
PREDICTIVE FACTORS OF DYSPLASIA ON COLONIC POLYPS
Authors: Ramos Rui , Duarte Patricia , Pereira Marta , Vicente Celia , Medeiros Jose , Cabrita Antonio , Casteleiro Carlos
Published: June 2021

Introduction: The clinical importance of colonic polyps resides in the risk of adenoma progression to colon cancer (the “adenoma‐carcinoma” sequence).

Objectives: To evaluate the presence of dysplasia on colon polyps and identify factors which are predictive of high‐grade changes.

Material and methods: Histopathologic studies were carried out on 357 polyps, excised from 334 patients, out of 1423 colonoscopies performed in 2006.

Results: Most (77.5%) polyps were distal to the splenic flexure, and the majority was found in the sigmoid colon; 71.8% were tubular adenomas. Low‐grade changes were found in 22.5% of the adenomas, while 77.5% showed high‐grade dysplasia. Only 6.4% of polyps were larger than 2 cm, while 60.1% measured less than 1 cm; 11.2% of the latter had high‐grade dysplasia.

Conclusions:

1 ‐ All polyps should be excised, regardless of their size, since high‐grade dysplasia can be found in more than 10% of small polyps;

2 – High‐grade dysplasia was more likely in polyps larger than 1 cm, and especially those larger than 2 cm;

3‐ High‐grade dysplasia was also more frequent in distal, villous and stalked polyps.
Nur 77 Is a Poor Prognostic Factor in Diffuse Large B‐Cell Lymphoma
Authors: Almiski Muhamad Sadek , Kurrer Michael , Levi Edi
Published: June 2021

Background: Nur77 is a pro‐apoptotic member of the orphan nuclear receptor family. It has been implicated in the pathogenesis of some T‐cell lymphomas. Its main mechanism of action is conversion of bcl2 to a proapoptotic molecule on the mitochondrial membrane. In this study we evaluated the possible prognostic significance of Nur77 expression in diffuse large B‐cell lymphomas.

Design: A tissue array containing 92 cases of diffuse large B‐cell lymphoma, followed up at University Hospital, Zurich was utilized. Three random, representative cores were obtained from each case and inserted in a grid pattern into a recipient paraffin block using a tissue arrayer. Sections were then cut from each TMA and stained with antibodies to CD10, bcl‐6, MUM1, BCL2, and Nur77 (Santa Cruz). Each core was evaluated for the percentage of tumor cells staining by visual estimation and recorded in 3 grades (up to 10%, 10–25%, >25% of cells ). Data was analyzed by Kaplan‐Meier and Log Rank tests as well as Spearmanrank correlation.

Result: In normal lymphoid tissues Nur 77 was expressed mainly in the germinal centers. Among the 92 diffuse large B‐cell lymphomas, 33 cases stained up to 10% of tumor cells, 30 cases stained 10–25 %, and 29 cases stained >30% of tumor cells. Expression of Nur77 was a bad prognostic factor in both univariate and multivariate analyses (p= 0.0058). Expression of Nur77 had no correlation with CD10, bcl‐6, or bcl‐2.

Conclusion: Nur 77 expression correlates with poor overall survival in diffuse large B‐cell lymphoma. Nuclear functions of Nur77 other than apoptosis induction are more likely to be in play in diffuse large B‐cell lymphomas.
Prenatal Exposure to Tungstate/Arsenite May Increase Susceptibility to Childhood Leukemia
Authors: Fastje Cynthia D. , Vargas Jason E. , Wong Simon S. , Witten Mark L.
Published: June 2021

Two concurrent, childhood leukemia clusters have been identified in the Southwestern U.S. at Fallon, NV and Sierra Vista, AZ. Using advances in medical geology, we have identified tungsten (W) and arsenic (As) as potentially relevant to leukemogenesis and utilized Affymetrix 430A 2.0 mouse microarray and real time RT‐PCR of the gene, Deleted in Malignant Brain Tumors 1 (DMBT1), to ascertain transcriptome‐expression in spleen tissue collected from 4‐week old C57BL/6 mouse pups (N=6–8/group/gender) exposed in utero to W, As, W/As and longitudinal controls at 20% of the normalized exposure a human mother would receive during gestation at mean environmental concentrations. Significantly altered pathways for in utero exposure to W/As includes Mek/Erk/Wnt pathways, antigen presentation and processing pathways, and cytokine interactions involving lymphocytic signaling. Additionally, significantly elevated genes associated with spermatogenesis were noted. Prenatal exposure to W was found to be associated with a 37 ± 1.2‐fold (p=0.012) decrease in DMBT1 expression in mice expressing DMBT1 at high levels. Histological examination of testicular tissue from sires exposed to W during spematogenesis showed a 50% vacancy rate of the seminiferous tubules. W/As prenatal‐exposure may be leukemogenic through increased susceptibility to viruses. Exposure to W may also be associated with reproductive disease.

Supported by OAFSR/EPA
Atypical B‐cell infiltrate in a pediatric patient: diagnostic and pathogenetic considerations
Authors: Florea Alina Emilia Dulau , Abbott Ira Richmond , Ratech Howard , Llena Josefina , Lui Yvonne , Farinhas Joaquim , Weidenheim Karen
Published: June 2021

Pediatric primary central nervous system lymphoma (PCNSL) represents1–1.5% of all PCNSL. Pediatric PCNSL are usually high grade B‐cell lymphoma (30%) or anaplastic large cell lymphomas. Herein we report a 15 year‐old male with a well‐circumscribed, enhancing mass centered on the right thalamus with mass effect on the 3rd ventricle and obstructive hydrocephalus. Endoscopic biopsy produced only hemorrhage; repeat biopsy three months later produced multiple fragments of pink‐tan soft tissue including pineal gland, grey matter, granulation tissue, choroid plexus, and a diffuse mononuclear cell infiltrate composed of a mixture of small round and large atypical lymphoid cells with irregular vesicular nuclei, 2 to 3 nucleoli attached to the nuclear membrane, and moderately abundant eosinophilic cytoplasm. The large lymphoid cells showed immunoreactivity for CD20, PAX‐5, and CD79a (B‐cell markers) but were negative for epithelial and germ cell markers; small T lymphocytes were also present. The lymphocytic process infiltrated brain tissue containing atypical astrocytes and large binucleated neurons. Immunoglobulin heavy chain (Ig‐VDJ; B‐cell) and T‐cell receptor (TCR) beta and gamma gene rearrangement studies showed a monoclonal B‐cell population. This atypical B‐cell infiltrate with features of lymphoma may have arising in an inflammatory infiltrate accompanying a pre‐existing glio‐neuronal tumor.
Role of CD36‐HRGP‐TSP‐1 Pathway in Tumor Biology
Authors: Hale James , Febbraio Maria , Silverstein Roy
Published: June 2021

Tumor growth and metastasis requires angiogenesis. Previous studies have demonstrated the anti‐angiogenic activity of Thrombospondin‐1 (TSP), a protein contained primarily in platelet alpha granules, was dependent upon signaling mediated by endothelial CD36 receptor binding. Additionally, it was shown that Histidine‐rich glycoprotein (HRGP), a circulating plasma protein, may block this interaction by sequestering TSP. We hypothesize that the TSP‐CD36‐HRGP pathway regulates angiogenesis in the context of tumor growth and metastasis. Murine B16 melanoma tumor volumes were assessed 10 and 17 days following subcutaneous injection into HRGP or CD36 knock out (KO) and wild type (WT) mice. Over 17 days average tumor volume was greater in CD36 KO mice compared to WT. Immunofluorescence (IF) and hematoxylin and eosin (H&E) staining showed larger vessels in tumors from CD36 KO mice. Initially, tumors from HRGP KO mice had larger average volume compared to WT, with no difference by day 17. Tumors from HRGP KO mice displayed increased necrosis with loss in overall cellularity compared to WT, as evidenced by H&E staining. IF showed fewer vessels in tumors from HRGP KO mice compared to WT. These data suggest that TSP‐CD36 interaction leads to inhibition of angiogenesis and subsequent reduction in tumor growth. Additionally, HRGP may have a time dependent or rate limiting function in the modulation of this interaction.
3D MR neovascular mapping of rabbit Vx2 tumor response to Avastin (bevacizumab)
Authors: Keene Jeffery , Schmieder Anne , Williams Todd , Allen John , Scott Michael , Caruthers Shelton , Wickline Samuel , Lanza Gregory
Published: June 2021

The objective of the present study was to characterize angiogenesis and growth response to bevacizumab in the Vx2 rabbit tumor model using MR molecular imaging and 3D neovascular mapping.

Methods

Rabbits (n=20) implanted with the Vx2 adenocarcinoma were administered bevacizumab (5mg/kg) or an irrelevant IgG1 antibody IV on days 8, 10, 12, and 14. MRI (3T) and 3D MR neovascular mapping were performed with ανβ3‐targeted nanoparticles on days 8 and 16.

Results

Serial treatment with bevacizumab decreased Vx2 tumor size relative to control (2.1 vs. 1.2 cm3, p<0.05), while angiogenesis was diminished by 16% from baseline (p>0.05). 3D neovascular maps illustrated the “angiogenic switch” between days 8 and 16, and revealed peripheral distributed neovasculature with densely coalesced regions interspersed with reticular network of angiogenic regions (Fig). Incomplete suppression of neovascularity with bevacizumab may reflect an under dosing of the drug, given its lower VEGF binding affinity in rabbits and suggests that further benefit of bevacizumab could be obtained with continued treatment at higher dosages.

Conclusion

3D MR molecular imaging permits unique assessments of tumor neovasculature, which could have an important role in patient stratification and longitudinal management of patients receiving antiangiogenic treatment strategies. Support: NIH/NCI U54 CA 119342
Increased expression of Suppressor of Cytokine Signaling‐1 (SOCS‐1) by dendritic and CD4+ T cells of HIV‐1 transgenic rats: A mechanism for dysregulated T helper‐1 responses
Authors: Yadav Anjana , Reitz Marvin , Reid William C.
Published: June 2021

Antigen‐specific T cell proliferation, maturation or maintenance of memory T cells, T helper‐1 responses and dendritic cell functions are compromised in HIV‐1 infected individuals. To better understand these immune abnormalities,we developed an HIV‐1 transgenic(Tg)rat which was earlier reported to be defective in Th1 cytokine production and generation/maintanence of effector/memory phenotype T cell subsets. We now report defects in IL‐12p40 production which are likely mediated by factors that increase IL‐10 production by bone marrow derived dendritic cells (BMDCs), resulting in the induction of suppressor of cytokine signaling (SOCS‐1) mRNA in BMDCs and CD4+ T cells. Increased basal expression of SOCS‐1 in CD4+ T cells is correlated with a decrease in IFN‐gamma production under neutral conditions but not in presence of exogenous IL‐12. We also demonstrate that IL‐12p40 production following IFN‐gamma and LPS stimulation of Tg rat BMDCs can be increased by inhibiting SOCS‐1 expression using RNA interference.

These results suggest a link between IL‐10, SOCS‐1, IL‐12p40 production and IFN‐gamma and lipopolysaccharide(LPS)signaling and development/maintanence of Th1 memory responses in HIV‐1 Tg rat.
Plasma membrane cholesterol modulates Campylobacter jejuni‐induced translocation of non‐invasive intestinal bacteria
Authors: Kalischuk Lisa D , Inglis G. Douglas , Buret Andre G
Published: June 2021

Bacterial enteritis, caused by pathogens such as Campylobacter jejuni, has been implicated as a factor that exacerbates inflammatory bowel disease (IBD) symptoms. The mechanisms whereby translocation of intestinal bacteria secondary to epithelial barrier dysfunction may contribute to the pathogenesis of IBD, have yet to be elucidated.

Aims: The aims of this study were to determine whether C. jejuni induces translocation of non‐invasive intestinal bacteria and examine the mechanisms by which this occurs.

Results:

Wild‐type C. jejuni 81‐176, but not an invasion‐defective FlaAFlaB mutant, induced translocation of intestinal bacteria to the spleens, livers, and mesenteric lymph nodes of infected mice. In human colonic T84 monolayers, C. jejuni induced internalization and translocation of non‐invasive Escherichia coli HB101 via a transcellular pathway in the absence of increased epithelial permeability to a 3 kDa dextran probe. Translocation was prevented by depletion of plasma membrane cholesterol with MbetaCD/lovastatin, and translocating E. coli co‐localized with lipid raft caveolin.

Conclusion:

C. jejuni may contribute to the pathogenesis of IBD by inducing the internalization and translocation of non‐invasive intestinal bacteria via a mechanism consistent with lipid raft‐mediated endocytosis.

Research supported by NSERC and the Crohn's and Colitis Foundation of Canada.
Proregion anionic amino acids mediate inhibition of mouse pro‐α‐defensin bactericidal activity
Authors: Figueredo Sharel Menezes , Weeks Colby , Young Steven , Ouellette Andre
Published: June 2021

Mouse Paneth cell α‐defensins are activated by proteolysis of inactive precursors, e.g., proCrp4 to bactericidal forms by the convertase MMP‐7. The activating step cleaves 24 residues, including 9 acidic amino acids that collectively block the membrane disruptive behavior of the Crp4 peptide (J Biol. Chem 281:28932–28943 2006). This inhibitory mechanism has been investigated further to identify whether specific cluster(s) of anionic amino acids in proCrp4(20–43) are preferentially responsible for blocking bactericidal activity and membrane disruption. To test this hypothesis, sets of Asp and Glu in proCrp4(20–43) were mutagenized to Gly. The recombinant proCrp4 variant peptides were expressed in E. coli, purified by HPLC to homogeneity and then assayed for differential effects of bactericidal inactivity. Generally, Gly for Glu/Asp replacements produced additive effects with respect to both bactericidal activity and membrane disruption of live E. coli ML35, regardless of their position. These findings support the conclusion that Asp and Glu residues contribute additively via intramolecular interactions to neutralize Crp4 membrane disruptive mechanisms. Supported by NIH grants DK044632 and AI059346