EFFECT OF ACTIVE COMPOUNDS ISOLATED FROM PTERIS SEMIPINNATA L ON DNA TOPOISOMERASES AND TYROSINE PROTEIN KINASE AND EXPRESSION OF C-MYC IN LUNG ADENOCARCINOMA CELLS

Objective: To study the effect of active compound 6F and A from Pteris semipinnata L.(PsL) on the activities of DNA topoisomerase (TOPO) I and II, activities of cytosolic and membrane TPK, and expression of oncogene c-myc in lung adenocarcinoma cells. Methods: The effect of compound 6F and A on activities of cytosolic and membrane TPK was measured by scintillation counting; the effect of compound A on expression of oncogene c-myc was determined by flow cytometry indirect fluorimetry. Results: compound 6F and A could inhibit the activities of TOPO I, and they strongly inhibited the TOPO II in 0.01 mg/L and 10.0 mg/L respectively. Compound A slightly inhibited the activities of membrane TPK, but not the cytosolic one. Compound A could inhibit the expression of oncogene c-myc. Conclusion: Topoisomerases are target of compound 6F and A. Compound A could slightly inhibit the activities of TPK, and showed an inhibitory effect on the expression of oncogene c-myc.

Cytotoxicities, Telomerase and Topoisomerases I Inhibitory Activities and Interactions of Terpyridine Derivatives with DNAs

A novel compound 4-methyl-7-{[4-（2,2＇：6＇,2＂-terpyridin-4＇-yl）benzyl]amino}-2H-chromen-2-one（1） was synthesized, and its DNA-binding properties, cytotoxicity, and telomerase and Topo I inhibitory activities were evaluated. For comparison, the anti-proliferative and Topo I inhibitory activities of another two analogues 2 and 3 were also investigated. Compound 1 is able to stabilize the structures of human telomere（h-tert） and promoter（c-myc and c-kit2） G-quadruplexes and h-tert i-motif. The association constants（Kb） are about 106 L/mol for h-tert G-quadruplex and/-motif, while the values are about l0s L/mol for both promoter G-qaudruplexes and calf thymus DNA（ct-DNA）. The binding of compound 1 induces the change of h-tert G-quadruplex from hybrid to antiparallel structure and exhibits 88.7% inhibition of telomerase activity at 8 μmol/L. Both compounds 1 and 3 inhibit signifi- cantly Topo I-mediated relaxation of pBR322 DNA. Compounds 1 and 2 show a high inhibitory efficacy on HepG2 and MCF-7 cancer cell lines with IC50 values of about 10^-6 mol/L. The three compounds also induce a delay of cell cycle progression. The coumarin group is vital for improving the biological activity ofterpyridine derivatives.

Identification of key genes and biological pathways in lung adenocarcinoma by integrated bioinformatics analysis

BACKGROUND The objectives of this study were to identify hub genes and biological pathways involved in lung adenocarcinoma(LUAD)via bioinformatics analysis,and investigate potential therapeutic targets.AIM To determine reliable prognostic biomarkers for early diagnosis and treatment of LUAD.METHODS To identify potential therapeutic targets for LUAD,two microarray datasets derived from the Gene Expression Omnibus(GEO)database were analyzed,GSE3116959 and GSE118370.Differentially expressed genes(DEGs)in LUAD and normal tissues were identified using the GEO2R tool.The Hiplot database was then used to generate a volcanic map of the DEGs.Weighted gene co-expression network analysis was conducted to cluster the genes in GSE116959 and GSE-118370 into different modules,and identify immune genes shared between them.A protein-protein interaction network was established using the Search Tool for the Retrieval of Interacting Genes database,then the CytoNCA and CytoHubba components of Cytoscape software were used to visualize the genes.Hub genes with high scores and co-expression were identified,and the Database for Annotation,Visualization and Integrated Discovery was used to perform enrichment analysis of these genes.The diagnostic and prognostic values of the hub genes were calculated using receiver operating characteristic curves and Kaplan-Meier survival analysis,and gene-set enrichment analysis was conducted.The University of Alabama at Birmingham Cancer data analysis portal was used to analyze relationships between the hub genes and normal specimens,as well as their expression during tumor progression.Lastly,validation of protein expression was conducted on the identified hub genes via the Human Protein Atlas database.RESULTS Three hub genes with high connectivity were identified;cellular retinoic acid binding protein 2(CRABP2),matrix metallopeptidase 12(MMP12),and DNA topoisomerase II alpha(TOP2A).High expression of these genes was associated with a poor LUAD prognosis,and the genes exhibited high diagnostic value.CONCLUSION Expression levels of CRABP2,MMP12,and TOP2A in LUAD were higher than those in normal lung tissue.This observation has diagnostic value,and is linked to poor LUAD prognosis.These genes may be biomarkers and therapeutic targets in LUAD,but further research is warranted to investigate their usefulness in these respects.

Synthesis, SAR, and in Silico ADME Screening Studies of Some 9-Amino-3-Phenylacridone Derivatives as Topoisomerase II Inhibitors

Cancer is a leading cause of death globally, claiming about 9.6 million lives and approximately 420 million new cases of cancer will be diagnosed in the world by the year 2025. The aim of this study was to synthesize and computationally evaluate pharmacological potential of some derivatives of 9-amino-3-phenylacridone, as topoisomerase II (Topo II) inhibitors. In this study, 10 derivatives of 3-phenyl-9-aminoacridone were chemically synthesized and characterized, and the potential pharmacological indications of these compounds were computationally predicted by methods such as ADMET prediction, molecular target prediction and molecular docking. The results showed that two derivatives (58e and 58j) were non-permeant of blood-brain barrier, and this property was found similar to that of amsacrine and etoposide. The results of molecular docking of the ten derivatives of 3-phenyl-9-aminoacridone that were synthesized in this work showed that the synthetic compounds (58a-j) and the standard drugs have overall best binding affinities for human acetylcholine esterase than butyrylcholinesterase, and overall best binding affinities for human topo IIα than human topo IIβ. Overall, the results of this study suggest that the synthetic compounds 58a, 58c, 58f, 58g, and 58i could probably inhibit topo IIα by catalytic inhibition as seen with amsacrine, but only 58b and 58e possessed DNA non-intercalation properties as seen with etoposide, serving as topo II poison. In conclusion, this study showed that 3-phenyl-9-aminoacridone derivatives are potential inhibitor of topo IIα/β both by catalytic inhibition and poison as non-intercalator of DNA.

应用组织芯片探讨宫颈癌前病变及宫颈癌中DNA拓扑异构酶Ⅱα的表达及其临床意义

目的：探讨DNA拓扑异构酶Ⅱα（TopoⅡα）蛋白在宫颈癌前病变及宫颈癌中的表达及其在宫颈癌发生发展中的作用。方法选取2007-2008年在山西省肿瘤医院行宫颈病变诊治的患者，其中包括慢性宫颈炎，低级别和高级别宫颈上皮内瘤变（CIN）和宫颈浸润性鳞癌（SCC）的患者分别21、33、30、73例，制作组织芯片，应用免疫组织化学法（SP）检测宫颈组织中TopoⅡα蛋白的表达情况。结果结果TopoⅡα蛋白在慢性宫颈炎上皮组织、CINⅠ、CINⅡ～Ⅲ、SCC中的表达率分别是5％，12％，90％，97％，表达率逐渐增加，高级别CIN和浸润性鳞癌的表达显著高于低级别CIN和正常宫颈组织，差异有统计学意义；而高级别CIN与浸润性癌之间、低级别CIN与正常宫颈上皮之间的表达差异均无统计学意义。结论 TopoⅡα蛋白表达随着宫颈癌前病变的严重程度的加重而增加，因此T o poⅡα蛋白可能参与了宫颈上皮的异常增生和恶性转变。

乳腺浸润性导管癌中拓扑异构酶Ⅱ等相关因素表达与腋窝淋巴结转移的关系研究

目的分析拓扑异构酶Ⅱ(TopoisomeraseⅡ)、nm23基因、雌激素受体(ER)、孕激素受体(PR)和C-erbB-2基因等在乳腺浸润性导管癌中的表达与腋窝淋巴结转移的关系,并探讨其临床意义,为进一步治疗、准确判断患者预后提供数据支持。方法采用免疫组织化学-SP法检测101例乳腺浸润性导管癌患者病理组织中的Topoi-someraseⅡ、nm23基因、ER、PR和C-erbB-2基因等,分析其与腋窝淋巴结转移的关系。结果 C-erbB-2(-)、(+)、(++)、(+++)4组的腋窝淋巴结转移发生率依次升高,分别为18.2%(2/11)、69.7%(23/33)、70.0%(21/30)、81.5%(22/27),差异有统计学意义(χ2=14.718,P=0.002)。TopoisomeraseⅡ(-)组腋窝淋巴结转移发生率(45.2%)低于TopoisomeraseⅡ(+)组(77.1%),差异有统计学意义(χ2=9.990,P=0.002)。而nm23(-)组腋窝淋巴结转移发生率(85.0%)高于nm23(+)组(55.7%),差异有统计学意义(χ2=9.404,P=0.002)。Logistic回归分析结果显示,TopoisomeraseⅡ(OR=8.870,P=0.001)、C-erbB-2(OR=1.848,P=0.041)是腋窝淋巴结转移的危险因素;而nm23(OR=0.151,P=0.008)是保护因素。结论 C-erbB-2基因和TopoisomeraseⅡ的阳性表达与腋窝淋巴结转移有关,nm23基因低表达患者腋窝淋巴结转移的危险较高。

CD44V6、P-gp、Top-Ⅱ在结直肠癌中的表达与临床病理的关系及其临床意义

目的:探讨CD44拼接变异体V6(CD44variantV6,CD44V6)、P-糖蛋白(P-glycoprotein,P-gp)、拓扑异构酶Ⅱ(TopoisomeraseⅡ,Top-Ⅱ)在结直肠中表达及其临床意义。方法:采用HE方法验证结直肠癌组织和正常大肠黏膜组织。采用免疫组织化学法MaxVision二步法检测80例结直肠癌组织及46例正常大肠黏膜组织中CD44V6、P-gp、Top-Ⅱ蛋白的表达情况,将其与性别,临床Dukes分期,浸润深度,淋巴转移等临床病理学资料进行统计学分析。结果:CD44V6、P-gp、Top-Ⅱ蛋白在结直肠癌组织中的表达显著高于正常大肠黏膜组织。CD44V6在结直肠癌中的表达强度与肿瘤的临床Dukes分期(rs=0.328,P<0.05),浸润深度(rs=0.356,P<0.01),淋巴转移(rs=0.406,P<0.01)成显著正相关性,与性别无相关性;P-gp在结直肠癌中的表达强度与性别,临床Dukes分期,肿瘤的浸润深度,淋巴转移无相关性。Top-Ⅱ在结直肠癌中的表达强度与性别,临床Dukes分期,肿瘤的浸润深度无相关性,但与淋巴转移有相关性(rs=0.245,P<0.05)。结论:P-gp、Top-Ⅱ、CD44V6在结直肠癌中均高表达。CD44V6在结直肠癌中的表达与临床Dukes分期,肿瘤的浸润深度及淋巴转移情况存在正相关性。

Ki-67及TopoisomeraseⅡ在脑胶质母细胞瘤组织中的表达及其生物学意义

目的探讨人胶质母细胞瘤中Ki-67及TopoisomeraseⅡ（TopoⅡ）之间的相关性及其与胶质母细胞瘤病人预后的关系。方法收集O6-甲基鸟嘌呤-DNA甲基转移酶（MGMT）低表达的胶质母细胞瘤标本60例,应用免疫组化法检测Ki-67和TopoⅡ的表达,研究其相关性。结果在MGMT低表达的胶质母细胞瘤中,Ki-67与TopoⅡ表达呈正相关（γ=0.83,P〈0.05）。对本组标本来源的60例胶质母细胞瘤病人随访12~23个月,其中死亡38例,肿瘤复发46例。结论 Ki-67和TopoⅡ在胶质母细胞瘤中的表达强度相对一致;其表达能客观反映肿瘤细胞的增殖活性和恶性程度,且可能影响胶质母细胞瘤病人的预后。

免疫组化检测大肠癌GST-π、TopoisomeraseⅡ-α表达的临床意义

目的 探讨GST -π、TopoisomeraseⅡ -α蛋白与大肠癌病理学特征之间的关系。方法 分别用鼠抗人TopoisomeraseⅡ -α单克隆抗体和兔抗人GST -π抗体对 6 0例大肠癌石蜡标本进行免疫组化研究。结果 1.GST -π、TopoisomeraseⅡ -α蛋白的表达和肿瘤大小、部位及类型无关 (P >0 .0 5 ) ;但与肿瘤的分化程度、临床分期、淋巴结转移有统计学差异 (P <0 .0 0 1)。 2 .GST -π在癌灶及癌旁组织中的表达意义不同 ,在肿瘤组织中低分化组高表达而在癌旁组织中则低表达 ,二者有统计学差异 (P <0 .0 0 1) ;TopoisomeraseⅡ -α在高分化组的表达高于低分化组 (P <0 .0 0 1)。结论 1.GST -π在瘤组织中的表达强度及在癌旁组织中的表达均与肿瘤的分化及临床分期有关 ,可作为肿瘤预后的指标。 2 .Topoi someraseⅡ -α在肿瘤组织中的表达和其分化、淋巴结转移有关 。

TopoisomeraseⅡ-α在大肠癌中的表达和临床意义

目的 探讨TopoisomeraseⅡ α蛋白与大肠癌病理学特征之间 ,耐药之间的关系。 方法 分别用鼠抗人TopoisomeraseⅡ α单克隆抗体对 60例大肠癌石蜡标本进行免疫组化研究。 结果 TopoisomeraseⅡ α蛋白及癌旁组织中的表达和性别、肿瘤大小、部位及类型无关 (P >0 .0 5 ) ;但与肿瘤的分化程度、Dukes分期、TMN分期、临床分期、淋巴结转移有统计学差异 (P <0 .0 0 1)。TopoisomeraseⅡ α在高分化组的表达高于低分化组 (P <0 .0 0 1) ,淋巴结转移组表达则低于无淋巴结转移组 (p <0 .0 0 1)。结论 TopoisomeraseⅡ α在肿瘤组织中的表达 ,强度和肠癌生物学特性密切相关 ,与肿瘤的分化及临床分期、淋巴结转移有关 ,可做为衡量恶性程度 ,耐药性及预后的指标之一。

Surveillance of Fluoroquinolone-Resistant Clinical Isolates of <i>Pseudomonas aeruginosa</i>

Ciprofloxacin (CPFX) and pazufloxacin (PZFX) have strong antibacterial activity against Pseudomonas aeruginosa. We investigated the sensitivity of P. aeruginosa to CPFX and PZFX in 373 strains isolated from inpatients (321 strains) and outpatients (52 strains) during September 2010 to September 2011 at Toho University Ohashi Medical Center. The percentage of CPFX-non-susceptible (≥3.91 μg/mL) among inpatients was 22.4%, but that among outpatients was 1.9%. As the major resistance mechanism to fluoroquinolones in P. aeruginosa involves modification of type II topoisomerases (DNA gyrase and topoisomerase IV), we examined mutations in the quinolone-resistance-determining regions (QRDRs) of gyrA and parC of P. aeruginosa isolates. Among the 373 isolates, 73 isolates had reduced CPFX-susceptibility and 88 had reduced PZFX-susceptibility. Sequencing of gyrA and parC revealed base substitutions that resulted in amino acid replacements in QRDR of GyrA in 70 P. aeruginosa isolates, while Thr83Ile (in GyrA) and Ser87Leu (in ParC) substitutions were found in 12 strains. These replacements were clearly associated with reduced susceptibility to CPFX and PZFX. However, we also found strains with high MICs to quinolones without mutations in either gyrA or parC. We then investigated the effect of efflux pumps in CPFX-resistance in these isolates. In the presence of an efflux pump inhibitor, MIC values in 12 of 66 strains decreased to 1/23. We also sequenced genes related to overexpression of efflux pumps, viz., mexZ, mexR, and nfxB. Eight of the strains without mutations in QRDRs had a mutation in mexZ, 7 strains had a mutation in mexR, but no mutation was identified in nfxB.

氟喹诺酮类抗菌药在动物医学上的研究进展

在动物医学上研究和应用的氟喹诺酮类抗菌药有恩诺沙星(Enrofloxacin,乙基环丙沙星),达诺沙星(danofloxacin,甲基环丙沙星),诺氟沙星(norfloxacin,氟哌酸)。氟甲喹(flumequine)等。其共同特点是抗菌谱广,除对革兰氏阴性菌有很强的杀菌作用外,对革兰氏阳性需氧菌、支原体和细胞内病原菌及衣原体也有杀灭作用。杀菌机制为抑制细菌DNA回旋酶亚单位A,该酶为Ⅱ型拓扑异构酶(type—Ⅱtopoisomerase)。

Inhibitory effects of lapachol on rat C6 glioma in vitro and in vivo by targeting DNA topoisomerase Ⅰ and topoisomerase Ⅱ

OBJECTIVE The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS First,the model of C6 glioma in Wistar rats was established and verified by hemotoxylin and eosin staining,immunohistochemical staining and magnetic resonance imaging(MRI).Then different doses of lapachol were gavaged and tumor volumes of the C6 glioma were detected by MRI.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phen-azinemethosulfate(PMS)assay,Hoechst33358 staining,AnnexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomeraseⅠ(TOPⅠ)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322 DNA relaxation assay.Molecular docking was used to predict the interaction of lapachol-TOPⅠand lapachol-TOPⅡ.TOP I and TOPⅡexpression levels in C6 cells were determined by Enzymelinked immunosorbent assay kits and real-time polymerase chain reaction(RT-PCR).RESULTS The rat C6 glioma model was successfully established.High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).MTS/PMS assay,Hoechst 33258 staining,AnnexinⅤ-FITC/PI staining,and comet assay showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cells in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠandⅡ.Molecular docking showed that lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡ.Enzyme-linked immunosorbent assay and RT-PCR showed that lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠand TOPⅡactivities,as wel as TOPⅡexpression.

THE ANTICANCER EFFECT AND ANTI－DNA TOPOISOMERASE II EFFECT OF EXTRACTS OF CAMELLIA PTILOPHYLLA CHANG AND CAMELLIA SINENSIS

The cytotoxic effect of extract of camellia ptilophyllachang(ECPC) and extract of camellia sinensis(ECS) onHeLa cell line, poorly differentiated nasopharyngealcarcinoma cell line(CNE2) and gastric cancer cell line(MGC-803 ) in vitro was studied using MIT assay method.The results showed that ECPC and ECS possessed significantcytotoxic effect on above three cell lines. The anticancer testin mice showed that ECPC had marked inhibitory effectagainst Ehrlich solid carcinoma(ESC) with inhibition ratesof 17. 8 48. 3% and with inhibition rates of 28. 3-54. 5% against reticular cell sarcoma(L2), and that ECShad inhibition rates of 31 . 5 -49. 4 % against ESC and 35. 8- 50% against L2. These two extracts had only marginalinhibitory effect against sarcoma- 180. The unknottingactivity of DNA topoisomerase II was inhibited completelyby ECPC and ECS at the concentration of 50 μg/ mlsuggesting that DNA topoisomerase II might be a targetenzyme of these two extracts.

Inhibition of DNA-Topoisomerase I by Acylated Triterpene Saponins from Pittosporum angustifolium Lodd.

Previous phytochemical investigation of the leaves and seeds of Pittosporum angustifolium Lodd.led to the isolation and structural elucidation of polyphenols and triterpene saponins.Evaluation for cytotoxicity of isolated saponins revealed that the predominant structural feature for a cytotoxic activity are acyl substituents at the oleanane aglycon backbone.The present work reports the results of a screening of 10 selected acylated saponins for their potential to inhibit the human DNA-topoisomerase I,giving rise to IC50 values in a range of 2.8-46.5 lM.To clarify the mode of observed cytotoxic action and,moreover,to distinguish from a pure surfactant effect which is commonly accompanied with saponins,these results indicate an involvement of the topoisomerase I and its role as a possible target structure for a cytotoxic activity.In addition,computational predictions of the fitting of saponins to the topoisomerase I-DNA complex,indicate a similar binding mode to that of clinically used topoisomerase I inhibitors.Graphical Abstract Ten acylated triterpene saponins from Pittosporum angustifolium were investigated for their potential to inhibit the human DNA-topoisomerase I and computational predictions of the fitting of saponins to the topoisomerase I-DNA complex were carried out.

Therapy-related myeloid neoplasms - what have we learned so far?

Therapy-related myeloid neoplasms are neoplastic processes arising as a result of chemotherapy, radiation therapy, or a combination of these modalities given for a primary condition. The disease biology varies based on the etiology and treatment modalities patients receive for their primary condition. Topoisomerase Ⅱ inhibitor therapy results in balanced translocations. Alkylating agents, characteristically, give rise to more complex karyotypes and mutations in p53. Other etiologies include radiation therapy, high-dose chemotherapy with autologous stem cell transplantation and telomere dysfunction. Poor-risk cytogenetic abnormalities are more prevalent than they are in de novo leukemias and the prognosis of these patients is uniformly dismal. Outcome varies according to cytogenetic risk group. Treatment recommendations should be based on performance status and karyotype. An in-depth understanding of risk factors that lead to the development of therapyrelated myeloid neoplasms would help developing riskadapted treatment protocols and monitoring patients after treatment for the primary condition, translating into reduced incidence, early detection and timely treatment.

Cu(II) Propionyl-Thiazole Thiosemicarbazone Complexes: Crystal Structure, Inhibition of Human Topoisomerase IIα, and Activity against Breast Cancer Cells

Two new thiosemicarbazone ligands, 2-propionylthiazole ethylthiosemicarbazone (PTZ-ETSC), and 2-propionylthiazole tert-butylthiosemicarbazone (PTZ-tBTSC), along with their two copper(II) complexes, [Cu(PTZ-ETSC)Cl] and [Cu(PTZ-tBTSC)Cl], are reported here for the first time. Once characterized by NMR and MS, these mono-anionic tridentate ligands were reacted with Cu2+ to form the square planar metal complexes [Cu(PTZ-ETSC)Cl] and [Cu(PTZ-tBTSC)Cl]. The x-ray crystal structure of the [Cu(PTZ-tBTSC)Cl] complex shows that the complex adopts a square planar arrangement around the copper(II) ion, but forms a sulfur-bridged dimer in the solid state. Both of the copper complexes displayed strong inhibition of human topoisomerase IIα at activities between 2-4 μM for [Cu(PTZ-ETSC)Cl], and between 8-10 μM for the [Cu(PTZ-tBTSC)Cl] complex. The EC50 values for the MDA-MB-231 breast cancer cell line were 82.6 μM for (PTZ-ETSC), 17.9 μM for [Cu(PTZ- ETSC)Cl], 97.8 μM for (PTZ-tBTSC), and 1.41 μM for [Cu(PTZ-tBTSC)Cl]. The EC50 values for the MCF7 breast cancer cell lines were 9.36 μM for (PTZ-ETSC), 0.13 μM for [Cu(PTZ-ETSC)Cl], 0.333 μM for (PTZ-tBTSC), and 0.093 μM for [Cu(PTZ-tBTSC)Cl].

Associations with Organ Involvement and Autoantibodies in Systemic Sclerosis: Results from the Canadian Scleroderma Research Group (CSRG)

Objective: Serum from SSc patients was analyzed centrally to determine ANA patterns and extractable nuclear antigens (ENAs) between lcSSc and dcSSc and associations with organ involvement. Methods: 1145 SSc patients had ANA and ENA analyzed by indirect immunofluorescence on HEp-2 substrate at a screening serum dilution of 1/160. Most ENA antibodies [Sm. U1-RNP, Ro52, SS-A/Ro60, topoisomeraseI (Topo1), SS-B/La, chromatin, ribosomal P and Jo1] were measured by laser bead immunoassay;and RNA polymerase III (RNAP) by ELISA. Results: ANA was positive in 95% (same in lcSSc, and dcSSc). Centromere pattern was present in 34%, speckled 22%, nucleolar 18%, homogeneous and speckled (H&S) 16%, multiple nuclear dots 6%. Anti-centromere Ab (ACA) occurred in 46% of lcSSc and 11% of dcSSc (P = 0.0001). ENAs that differed between lcSSc and dcSSc subsets were Topo1 (OR 2.4, P = 0.0001) and RNAP (OR 5.6, P 0.0001) more common in dcSSc. Overall, 15% had positive Topo1;usually with a H&S pattern (67%);Topo1 was associated with ILD on CXR (OR 2.3;95% CI 1.5 - 3.5) and HRCT (OR 3.8;95% CI 1.8 - 8.2). RNAP occurred in 18.5% (35.4% in dcSSc vs. 8.9% in lcSSc). Scleroderma renal crisis (SRC) was 13 times more likely if RNAP positive;P = 0.0001. ACA was only weakly associated with sPAP > 50 mmHg (OR 1.8;95%CI 1.1 - 3.0). Conclusion: ANA homogeneous pattern alone is rare in SSc;ACA was significantly more common in lcSSc. Many ENAs are equal in lcSSc and dcSSc except RNAP and Topo1. RNAP has the highest OR of SRC. Topo1 is less strongly associated with ILD. Abstract word count: 249, Body word count 1246, Figures 2, Tables 2. Key Messages: 1) 95% of SSc has a positive ANA and ANA patterns in SSc include centromere, nucleolar, and homogeneous and speckled together;2) Most ENAs are equal in both dcSSc and lcSSc except anti RNA polymerase III and topoisomerase I;3) RNA polymerase III has the highest association (odds ratio) with scleroderma renal crisis, topoisomerase I is associated with interstitial lung disease;whereas anticentromere was not associated with elevated pulmonary arterial pressures on echocardiogram.

Cu(II) Benzoylpyridine Thiosemicarbazone Complexes: Inhibition of Human Topoisomerase IIα and Activity against Breast Cancer Cells

The focus of this research is on the study of a series of copper (II) benzoylpyridine thiosemicarbazone complexes. Of the six benzoylpyridine thiosemicarbazone ligands used in this study, two are reported for the first time;2-benzoylpyridine tert-butyl thiosemicarbazone (BZP-tBTSC), and 2-benzoylpyridine benzyl thiosemicarbazone (BZP-BzTSC). Once characterized by NMR, melting point, and MS, these mono-anionic tridentate ligands were then reacted with Cu2+ to form the new square planar metal complexes [Cu(BZP-tBTSC)Cl] and [Cu(BZP-BzTSC)Cl]. All of the copper complexes display marked inhibition of human topoisomerase IIα. The [Cu(BZP-tBTSC)Cl] complex shows marked activity against human breast cancer cell lines.