bluuebunny/crossref_metadata_2025_split huggingface dataset:
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 alsoResults: 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,
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.
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
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.MethodsRabbits (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.ResultsSerial 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.Conclusion3D 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.
Mechanical Stimulation Differentially Controls Proliferation, Morphology, and Vascular Gene and Protein Expression in Mesenchymal Progenitor Cells
Authors: Chew Douglas W. , Maul Timothy M. , Neiponice Alejandro , Vorp David A. 
Published: June 2021

Introduction: Cyclic strain can regulate smooth muscle cell (SMC) gene and protein expression in mesenchymal progenitor cells (MPCs). We tested the hypothesis that the differentiation of BMPCs to endothelial cells (ECs) and SMCs may be controlled by the appropriate type, magnitude, and frequency of mechanical stimulation.Methods: Bone marrow MPCs were seeded onto collagen I substrates and exposed to three stimuli – cyclic strain (CS), cyclic pressure (CP), steady shear stress (SS) – at 3 magnitudes and 2 frequencies for 5 days. Morphology and proliferation were analyzed by Scion Image. Protein expression was examined via immunohistochemistry. RT‐PCR was used to quantify genes relevant to various cell phenotypes and matrix proteins.Results: MPCs responded differentially via changes in morphology, proliferation, gene, and protein expression. Cell size correlated negatively with SS and positively with CP. Proliferation was enhanced by CP, but decreased in for SS. CS caused a sequential increase in the number of SMC proteins. Although gene expression for EC and SMC genes was elevated in CP and SS, respectively, no protein expression was found. Most ECM genes were up‐regulated under CP, while only collagen I was increased in SS. Increases in TGFβ gene expression correlated with SMC and collagen I gene expression.Conclusions: We conclude that the differentiation of MPCs towards SMCs and ECs may be regulated, in part, by the specific mechanical stimulus applied.This work was funded by the NIH BRP grant # R01 HL069368, NIH training grant # T32 EB001026, Ruth L. Kirschstein predoctoral fellowship #F31 EB004791, and American Heart Association predoctoral fellowship #0415437U.
Copper deficiency induces a connective tissue disorder state in a porcine native abdominal aortic aneurysm model
Authors: Sadek Mikel , Hynecek Robert L. , Poblete Honesto , Levin Elizabeth , Shin Hyun‐Joo , Lei Xingen , Faries Peter L. 
Published: June 2021

Objective: Copper deficiency (CD) has been linked to impaired collagen synthesis and the development of abdominal aortic aneurysms (AAA) in swine. This study sought to evaluate the effect of a CD diet on a porcine AAA model.Methods: A CD diet was fed to Yorkshire swine for 3mths. In CD and regular‐fed (RF) swine the infrarenal aorta was balloon dilated and infused with collagenase/elastase (16000U/1000U) (Fig 1). At 2wks, control (suprarenal aorta) and AAA tissues were frozen in liquid nitrogen and pulverized for RNA extraction. RNA from 3 CD and 3 RF samples was biotin‐labeled and hybridized to porcine arrays (20,201 genes). Relative gene intensities equaled log2(AAA/control). A 2‐class unpaired Significance Analysis of Microarray (Fig 2) test was conducted, false discovery rate (FDR) < 2.2%.Results: 55 genes were downregulated and 20 genes were upregulated in CD compared to RF swine (Fig 3). Diminished expression of heme oxygenase and thymidilate synthetase was consistent with vascular smooth muscle and endothelial dysregulation. Differential expression of MHC classes I/II was consistent with an autoimmune etiology for AAA. Inflammatory pathology was supported by altered expressions of cytochrome P450, oligoadenylate synthetase and sphingosine kinase.Conclusion: The addition of a CD diet to the porcine AAA model introduced novel links to known AAA connective tissue biology.Support: NIH‐K08HL073313
The anti‐cataleptic effects of 8‐OH‐DPAT: role of pre‐synaptic 5‐HT1A receptors, and possible implications for combined treatment with antidepressants and antipsychotics
Authors: Koek Wouter , Hidalgo Jose A. , Daws Lynette C. 
Published: June 2021

5‐HT1A agonists attenuate antipsychotic‐induced catalepsy, a model of extrapyramidal symptoms (EPS). To explore the role of pre‐synaptic 5‐HT1A receptors, we examined these effects in mice expressing 50% fewer serotonin transporters (5‐HTT) in which pre‐synaptic 5‐HT1A receptors are down‐regulated, but post‐synaptic 5‐HT1A receptors are not.Mice were pretreated with a 5‐HT1A agonist (ip), treated with haloperidol (cumulative dosing), and examined for catalepsy (bar test). In C57 mice, the 5‐HT1A partial agonist buspirone did not alter the ED50 of haloperidol (0.92 mg/kg). The 5‐HT1A agonist 8‐OH‐DPAT shifted the haloperidol dose‐effect curve to the right, with 10 mg/kg producing a maximum 2.7‐fold shift. Buspirone (32 mg/kg) blocked these effects of 8‐OH‐DPAT. In 5‐HTT(+/+) mice, 10 mg/kg 8‐OH‐DPAT shifted the haloperidol dose‐effect curve 3.5‐fold to the right. In contrast, 8‐OH‐DPAT did not alter haloperidol's ED50 in 5‐HTT(+/−) mice.The finding that 8‐OH‐DPAT had anti‐cataleptic effects in C57 and in 5‐HTT(+/+) but not in 5‐HTT(+/−) mice suggests that pre‐synaptic 5‐HT1A receptors mediate these effects. In as much as 5‐HTT(+/−) mice are a model of chronic treatment with SSRI's, the lack of anti‐cataleptic effects of 8‐OH‐DPAT in these mice suggests that SSRI's may unmask and enhance EPS of antipsychotics with 5‐HT1A activity.Supported by NARSAD (WK) and USPHS Grant MH64489 (LCD)
Redox Regulation of G protein βγ Subunits
Authors: Dessal Axel , Smrcka Alan V 
Published: June 2021

The βγ‐subunit of G protein (Gβγ) dissociates from the α‐subunit upon activation of G protein‐coupled receptors. Once free, Gβγ is able to regulate many target proteins, including phospholipase C β2 (PLCβ2), phosphoinositide 3‐kinase γ (PI3Kγ) and G protein‐coupled receptor kinase 2 (GRK2). Our group has used a combination of virtual and manual screening to find small molecules that inhibit the interaction between Gβγ and the aforementioned proteins with affinities in the high nM range. Recently, we discovered another set of compounds (some with heavy metals, Hg2+, Cu2+ and Ag+ in their structure) that binding of a Gβγ‐binding peptide (SIGK) in an enzyme‐linked immunosorbent assay (ELISA) at concentrations in the low nM range. The effect of these small molecules persists after washing them away, and it is both prevented and reversed by dithiothreitol (DTT). This suggests that these compounds act directly at Gβγ by a reversible redox mechanism. Investigation of the mechanism of action of these compounds may lead to uncovering mechanisms for Gβγ regulation by both natural and synthetic redox active molecules.This work was funded by National Institutes of Health GM060286
Platelet growth factor induces smooth muscle migration through an integrin linked kinase‐dependent pathway
Authors: Esfandiarei Mitra , Yazdi Sahar Abdoli , van Breemen Cornelis 
Published: June 2021

Integrin linked kinase (ILK) is a cytoplasmic interactor of integrin receptors regulating cell survival, spreading, and contraction. In this study we aimed to evaluate the role of ILK in regulating smooth muscle cell (SMCs) migration, an important event during the development of atherosclerotic plaque and coronary artery disease. Primary mouse SMCs were cultured in the presence or absence of platelet growth factor (PDGF) and various specific inhibitors. Cell lysates were used for kinase and Western blot assays. Cell migration was measured using colorimetric migration or wound healing assays utilizing fluorescent imaging. Our study showed that PDGF could activate ILK, Akt, and Erk in SMCs without any effect on focal adhesion kinase and α‐paxillin. ILK inhibition had no effect on PDGF‐induced Akt and Erk activation, indicating that their activation is through an ILK‐independent pathway. Inhibition of ILK, PI3K/Akt, and Erk1/2 decreased SMCs migration, signifying the cooperation between these signaling pathways in regulating SMC migration. To our knowledge, this is the first report of a regulatory role for ILK in SMCs migration, which is independent from Akt and Erk1/2 pathways, and conveys new insights in our efforts to establish a novel therapeutic target for treatment of coronary artery occlusion.This work was supported by the Canadian Diabetes Association & the Michael Smith Foundation for Health Research.
Increased vascular contractile responses to phenylephrine in Doca‐salt mice is normalized by Pyk2 blockade
Authors: Giachini Fernanda R. C. , Carneiro Fernando Silva , Lima Victor Vitorino , Webb R. Clinton , Tostes Rita Cassia 
Published: June 2021

Background: The calcium‐dependent proline‐rich tyrosine kinase (Pyk2), a nonreceptor protein activated by tyrosine phosphorylation, links G‐protein‐coupled receptors to vascular responses. DOCA‐salt hypertension is characterized by enhanced vascular reactivity. We tested the hypothesis that changes in vascular responses in DOCA‐salt mice are due to increased activation of Pyk2.Methods and results: Aorta and small mesenteric arteries from DOCA‐salt and uninephrectomized (UNI) male C57bl/6 mice were incubated with vehicle or Tyrphostin A‐9 (0.1μM, Pyk2 inhibitor). Systolic blood pressure (mmHg) was higher in DOCA (126±3) vs UNI (100±4). Vascular responses to PE (1nM to 10mM) were greater in vessels from DOCA‐salt than UNI, but treatment with Tyrphostin A‐9 abolished the difference among the groups (Figure 1). Pyk2 levels, as well as Pyk2Tyr402 phosphorylation, paxillin and paxillinTyr118 phosphorylation were increased in DOCA‐salt aorta (Figure2). Incubation with Tyrphostin A‐9 restored phosphorylation of Pyk2 and paxillin.Conclusion: Increased Pyk2 expression and activity are involved in increased vascular contractile‐responses in Doca‐salt mice.Financial Support: NIH (HL‐74167, RCW); FAPESP and CAPES, Brazil.