text:
- Research article
- Open Access
The use of whole genome amplification to study chromosomal changes in prostate cancer: insights into genome-wide signature of preneoplasia associated with cancer progression
BMC Genomics volume 7, Article number: 65 (2006)
Prostate cancer (CaP) is a disease with multifactorial etiology that includes both genetic and environmental components. The knowledge of the genetic basis of CaP has increased over the past years, mainly in the pathways that underlie tumourigenesis, progression and drug resistance. The vast majority of cases of CaP are adenocarcinomas that likely develop through a pre-malignant lesion and high-grade prostatic intraepithelial neoplasia (HPIN). Histologically, CaP is a heterogeneous disease consisting of multiple, discrete foci of invasive carcinoma and HPIN that are commonly interspersed with benign glands and stroma. This admixture with benign tissue can complicate genomic analyses in CaP. Specifically, when DNA is bulk-extracted the genetic information obtained represents an average for all of the cells within the sample.
To minimize this problem, we obtained DNA from individual foci of HPIN and CaP by laser capture microdissection (LCM). The small quantities of DNA thus obtained were then amplified by means of multiple-displacement amplification (MDA), for use in genomic DNA array comparative genomic hybridisation (gaCGH). Recurrent chromosome copy number abnormalities (CNAs) were observed in both HPIN and CaP. In HPIN, chromosomal imbalances involving chromosome 8 where common, whilst in CaP additional chromosomal changes involving chromosomes 6, 10, 13 and 16 where also frequently observed.
An overall increase in chromosomal changes was seen in CaP compared to HPIN, suggesting a universal breakdown in chromosomal stability. The accumulation of CNAs, which occurs during this process is non-random and may indicate chromosomal regions important in tumourigenesis. It is therefore likely that the alterations in copy number are part of a programmed cycle of events that promote tumour development, progression and survival. The combination of LCM, MDA and gaCGH is ideally suited for the identification of CNAs from small cell clusters and may assist in the discovery of potential genomic markers for early diagnosis, or identify the location of tumour suppressor genes (TSG) or oncogenes previously unreported in HPIN and CaP.
The worldwide occurrence CaP is increasing and it has now overtaken lung cancer as the most commonly diagnosed malignancy in men in the Western World. In spite of significant progress in its clinical management, comparatively little is known about the aetiology of the disease and predicting outcome is still a challenge for most clinicians. Histopathologically, CaP displays considerable heterogeneity and can contain a substantial admixture of pre-malignant HPIN glands within cancerous foci. HPIN is currently considered to be the most likely precursor to invasive CaP . However, most of these foci are latent and rarely develop into clinically detectable cancer. Unfortunately, a significant number (3%) do progress and can become aggressive and lethal . The major difficulty facing clinicians is the identification of patients presenting with early stage CaP, who are likely to develop life-threatening disease. As a result, intensive research is currently underway to identify the key alterations that may prove to be important for both classification and prognosis prediction.
The advent and development of CGH, which started in the early 1990's , has revolutionised cytogenetics and allowed for genome-wide screening of numerous cancer types and the identification of genomic CNA, which may contribute to cancer development and progression. The accumulation of genetic changes that occurs during the stepwise evolution from normal tissue to metastasis, although likely due to increased genetic instability, may indicate the chromosomal locations of TSG or oncogenes that are important in tumourigenesis. When first introduced, CGH used metaphase chromosomes targets to identify CNAs. However, recent advances have substituted these with arrayed DNA sequences that provide higher resolution, 1 Mb versus 10 Mb, and greater ease of analysis.
Whole genome scans of CaP patients and cell lines have identified consistent, chromosomal alterations, which include recurrent loss of chromosomal regions from 5p, 6q, 8p, 10q, 13, 16q and 17p, in addition to gain on 1q, 3q 7p, 7q, 8q 11p, 17q and Xpq [4–6]. Nonetheless, one potential limitation with current CGH methodology is its requirement for microgram quantities of genomic DNA. When studying cell lines this is not necessarily a matter for concern, as it is relatively straightforward to produce a sufficient quantity of clonal cells to obtain many micrograms of DNA. However, when studying patient tissue if DNA is "bulk extracted" from a heterogeneous non-clonal tumour mass the data obtained will represent an average value for all cells within the specimen and any clinically informative genetic changes, restricted to small cell clusters, may be masked. As a result, when studying a heterogeneous tissue, such as prostate, it is essential to obtain pure cell populations via methods such as macrodissection or LCM. The DNA extracted from the dissected material can then be amplified using a whole genome amplification (WGA) technique , such as MDA [8–11]. The combination of LCM and WGA can generate sufficient amounts of DNA for extensive genome analyses. Several large studies have demonstrated the ability of MDA to accurately amplify human genomic DNA. When used in SNP genotyping [12, 13] an estimated 99.82% of the genome is covered with a reported concordance, to non-amplified DNA, of greater than 99% . In addition, the sequence error rate is only 7.6 × 106  and previous results obtained from gaCGH, using amplified DNA, are almost indistinguishable from those obtained from non-amplified DNA [10, 15].
An understanding of the molecular mechanisms behind cancer development will enable identification of molecular and cytogenetic biomarker(s) that may be useful in predicting early transformation into an aggressive phenotype, as well as, providing fundamental insights into the regulatory pathways of genome integrity that may lead to multistep field cancerization and HPIN and CaP. In the present study, we describe the use of LCM, MDA and gaCGH, using a 1 Mb array (Spectral Genomics, USA) to investigate copy number changes occurring in prostate cancer by the analysis of 7 prostatic samples containing HPIN and 8 CaP specimens.
To determine whether whole genomic amplification exhibited bias or a distortion of imbalance assignments, control CGH experiments comparing the results obtained before and after amplification (normal male versus normal male, normal male versus normal female and DNA from a cancer cell line versus normal control DNA) from the same original DNA samples were performed. These are presented in Additional Files 1, 2, 3 and it is evident that no major distortion of imbalance was introduced by amplification of genomic DNA.
A summary of the chromosomal CNAs detected by CGH for the 7 HPIN and 8 CaP samples is shown in Table 1 and 2 and displayed graphically in Figure 1 (for HPIN blue = loss, yellow = gain; for CaP red = loss, green = gain). Figure 2 shows the frequency of CNAs detected at the level of each chromosomes arm for HPIN and CaP. The general pattern of loss or gain was very similar in the HPIN and CaP samples, however CaP samples possessed significantly more aberrations (90 CNAs) than HPIN samples (41 CNAs) (P<0.001, chi square test). The average number of CNAs for the 7 HPIN samples was 5.8 (range 3 to 8, median 6). For the 8 CaP samples an average of 11.25 (range 5 to 18, median 11.5) amplifications or deletions were observed. When studied at the 850 cytoband level (minus X and Y chromosome cytobands), HPIN samples had the total of 65 affected bands (average 9, range 4 to 14) and CaP samples 143 affected breakpoints bands (average 18, range 8 to 27). This difference is statistically significant (P<0.001, chi square test). In addition, for both the HPIN and CaP samples there were a significantly greater number of cytobands lost than gained (P<0.01, chi squared test) (Table 1).
In the 7 HPIN samples recurrent chromosomal changes, detected in greater than 25% of cases, were losses found on 8p (7 out of 7 cases, 100%), 6q, 13q, 16q (2 out of 7 cases, 28.5%, each), whereas gains were found on 8q and 16p (3 out of 7 cases, 43%, each), 1p, 7p and 20q (2 out of 7 cases, 28.5%, each). For the 8 CaP samples recurrent chromosomal changes, detected in greater than 25% of tumours, were losses on 8p (8 out of 8 cases, 100%), 10q (6 out of 8 cases, 75%), 13q (5 out of 8 cases, 62.5%), 6q (4 out of 8 cases, 50%), 1p, 16q (3 our of 8 cases, 37.5%), 5q, 12p, 18q (2 out of 8 cases, 25%, each) and gains on 7q (6 out of 8 cases, 75%), 7p, 8q (5 out of 8 cases, 62.5%, each), 4p, 16p, 19p (2 out of 8 cases, 25%).
Small consensus regions of consistent CNA were observed for both HPIN and CaP samples (Table 3), in >25% of samples. For example, 8p region was consistently lost in HPIN (100%), but within the cytoband 8p11.23-p23.2 that was commonly lost in >50% of the HPIN samples. Similarly, 8p was consistently lost in CaP (100%), but within the cytoband 8p11.22-p23.2 that was commonly lost in >60% of the CaP samples. A list of the genes present within these consensus or commonly lost regions is displayed in Table 3.
Chromosome copy number abnormalities (CNAs) are common in most cancers, with specific regions of amplification or deletion being associated with specific tumour types, stages or outcomes [16–18]. The introduction of metaphase CGH  to study these CNAs has eliminated the requirement to obtain metaphase spreads from the tumour samples, which was often challenging due to technical difficulties in culturing certain tissues. Though metaphase CGH allows for more samples to be examined, it provides a relatively limited resolution and still requires substantial cytogenetic experience to analyse the results. The recent development of array CGH has opened up the field of CGH research and permitted more laboratories to study an ever-increasing number of tumour types and stages. The application of BAC microarrays to analyse human samples is straightforward and their high-throughput nature makes them the method of choice for rapid detection of genetic alterations.
When analysing heterogeneous tumour samples, the gaCGH results obtained from "bulk extracted" DNA are likely to be inaccurate. However, with the use of LCM and MDA we have been to able obtain highly purified HPIN and CaP DNA and thus identify the particular chromosomal changes associated with the two disease stages.
In this study, we have used a 2,400-element BAC microarray with a resolution of ~1 Mb to study CNAs in a set of 15 patient samples comprised of 7 HPIN cases and 8 CaP cases. For the 7 HPIN cases, 41 genomic alterations (20 gains, 21 losses) were identified, in contrast to the 90 genomic alterations (38 gains, 52 losses) seen for the 8 CaP cases. As with other cancers, CaP development and progression is likely to be the outcome of a series of stepwise genetic changes. The accumulation of CNAs, which occurs during this process, although likely due to increased genetic instability, is non-random and may indicate chromosomal regions important in tumourigenesis. It is suggested that failure in the fidelity of homologous recombination within the repetitive sequences, that comprise the kinetochore complex, could lead to recurrent loss of 8p and gain of 8q by rearrangement of chromosome 8-specific alphoid centromeric sequences. Thus, the high fidelity process of homologous recombination can be the major DNA repair pathway, which is indispensable for the maintenance of genetic stability.
Examination of our array results indicates that aberrations involving parts or all of 1p, 6q, 7p, 7q, 8p, 8q, 10q, 13q 16p and 16q are most common, which is concordant with previous metaphase [19, 20] and array CGH [4, 21] results. It is therefore likely that the alterations in copy number are part of a programmed cycle of events that promote tumour development and progression as well having an impact on disease-specific survival.
A comparison of the CNAs present in the HPIN and CaP samples identified a significant increase in copy number for 7q (P<0.01, chi square test) and a significantly increased frequency of loss for 10q (P<0.01, chi square test) and 13q (P<0.0001, chi square test). In genotype/phenotype correlations, gain of chromosome 7q  and loss of 13q  have been associated with advancing tumour stage and aggressiveness, which is in agreement with the results presented here. However, gain of 8q  and loss of 16q  have also been linked to tumour progression, but our data do not show any significant difference for these CNAs in our HPIN and CaP samples. This would suggest that 8q and 16q CNA's are likely to be early events in tumourigenesis. In addition, they may also identify HPIN and CaP samples that are likely to progress.
Apart from the commonly reported CNAs, additional alterations that have been less frequently reported in earlier CGH studies have also been identified. These include gains on 12q (HPIN) and loss of 4q and 10q (HPIN and CaP). Whether the identification of 12q and 4q regions will provide additional insight into CaP progression is not yet clear. Further analysis using a platform such as tissue microarrays, which permits the screening of different disease stages from large patient cohorts, will help better identify their frequency and also the potential use of genomic imbalance in diagnosis of CaP. The value of genomic analysis in CaP was recently demonstrated by the discovery of a high frequency of chromosomal translocations leading to rearrangement of the fusion oncoproteins ERG or ETV1 with TMPRSS2 .
A common problem when analysing archival tissue is the availability of a suitable quality and quantity of RNA to study expression changes. Though RNA amplification techniques  can generate a sufficient quantity, obtaining RNA of the required quality is often challenging. Previous reports have demonstrated a relationship between alterations in chromosomal copy number and alterations in gene expression [28, 29]. The ability to distinguish these regions will point to genes, which may either directly (tumour suppressor gene loss or oncogene gain) or indirectly contribute to tumour development and progression. As a result, CGH can be used as a surrogate for gene identification. Candidate genes, which have previously been implicated in CaP have been highlighted in bold in Table 3. The roles of MYC [30–32], PSCA [33–35], and MDM2 [36–38] have all been well reported and alterations in gene dosage correlate well with their change in expression. However, other candidate genes have been less well studied. For example, EBAG9, whose increased expression in CaP is a negative prognostic indicator, has a potential role in progression by enabling cancer cells to evade the immune response . NBS1, which has been identified as a founder mutation causing an increased susceptibility to prostate cancer , is involved in processing/repair of DNA double strand breaks and in cell cycle checkpoints, thus its deregulation will likely contribute to chromosomal instability. FKHR, which is a member of the FOXO forkhead transcription factor family, is thought to play a regulatory role in several cellular functions including cell proliferation and survival . Loss of FKHR expression, as observed in CaP cell lines, is likely to abrogate this control leading to tumour cell growth. Though these genes have previously been implicated in CaP there are additional genes, including both oncogenes and tumour suppressor genes, which reside within all of the affected regions that may play an important role in the aetiology of the disease. Further analysis of these other candidates may identify their potential as molecular targets for diagnosis and treatment.
Several of the genes present within altered regions of the genome are associated with genetic pathways, indicating that these pathways are likely to be important in prostate tumourigenesis.
The combination of techniques used in this study has allowed for the identification of consistent regions of copy number change, ranging from specific cytobands to whole chromosomes, starting with as little as 5–10 ng of DNA. Through our use of LCM, we have also identified several lower frequency CNAs which may not have been detected when using bulk extracted DNA due to the heterogeneous nature of prostate tissue. In addition, several interesting candidate genes have been observed within the altered chromosomal regions. Although some have already been extensively studied others, which have not, may prove to be of clinical importance. Genetic screening strategies that combine FISH and immunohistochemistry for the detection of various combinations of chromosomal gains and/or losses and altered gene expression are likely to be of great use in diagnosis and prognosis prediction.
Fresh prostate tissue was obtained from radical prostatectomies performed at The University Health Network (UHN), Toronto, with the informed patient consent and approval of the institutional research ethics board. The tissue was embedded in OCT frozen section medium (Stephens Scientific, Riverdale NJ, USA) and stored at -80°C until required. For all samples, the presence of HPIN and CaP was verified by histologic assessment of frozen sections by a urological pathologist (A.E.).
Patient samples for gaCGH analysis were selected based on two criteria; first, the presence of clearly identifiable regions of HPIN and/or CaP and second, the availability of good quality high molecular weight DNA. DNA quality was assessed by DNA extraction from a single tissue section and visualisation by agarose gel electrophoresis. A cohort consisting of 15 cases (7 HPIN and 8 CaP derived from different patients) were selected based on these criteria.
Laser capture microdissection
The selected fresh-frozen prostate tissue samples were cut onto microscope slides (8 μm thickness) and stained using the HistoGene LCM frozen section staining kit (Arcturus, USA). Foci of HPIN and CaP were identified from stained prostate tissue sections and a minimum of 1000 cells were removed from these regions by LCM using the Cell Robotics LaserScissors system (Cell Robotics Inc., USA).
DNA was extracted using the QIAamp DNA Micro Kit (Qiagen, USA) and the DNA concentration was determined using the PicoGreen dsDNA Quantitation kit (Molecular Probes Inc., USA). Both procedures were performed following manufacturers instructions.
Strand displacement amplification
DNA was amplified using the GenomiPhi Amplification Kit (Amersham Biosciences, USA) according to manufacturer's instructions. Briefly, amplification was carried out in two individual steps. The step 1 reaction mixture contained 5–10 ng (1000 to 2000 cell equivalents) of DNA (from diluted reference or test DNA) in 1 μl of sterile water and 9 μl of Sample Buffer. This mixture was heated at 95°C for 3 minutes and then chilled on ice. The step 2 reaction (amplification) mixture contained 9 μl of Reaction Buffer, 1 μl of Enzyme Mix and the 10 μl from Step 1. The amplification reaction was incubated at 30°C for 16–18 hours. The enzyme was inactivated by heating at 65°C for 10 minutes, followed by cooling to 4°C. This method of WGA consistently produced in excess of 5 μg of DNA from a starting concentration of 5–10 ng.
Assessment of DNA quality and strand displacement amplification
Five microlitres of each amplification reaction was electrophoresed through a 1% agarose gel and stained with ethidium bromide in order to assess product yield and product length. All amplification products were purified by phenol-chloroform extraction and DNA concentration and purity were determined by measuring absorbance at A260 and A280.
Spectral genomic BAC arrays
The genomic DNA arrays used in these experiments were obtained from Spectral Genomics Inc. and consist of approximately 2400 unique BAC and PAC clones, which provide an average genomic resolution of 1 Mb. CGH experiments were performed using the amplified patient DNA as the "test" and amplified female placental DNA as the "normal". In addition, control experiments were performed, corresponding to DNA before and after amplification (normal non-neoplastic prostate epithelial DNA versus normal non-neoplastic prostate epithelial DNA, normal non-neoplastic prostate epithelial DNA versus normal female placental DNA and DNA from a cancer cell line versus normal control DNA).
Reference DNA and test DNA were first digested overnight at 37°C using 10 units of RsaI (Invitrogen) in a 10 μl reaction. The digested DNA was labeled using the protocol optimized by Spectral Genomics, with separate labeling reactions for Cy3 and Cy5 being set up for both reference and test DNA. Briefly, labeling reaction were set up containing 2 μg of DNA, 20 μl of 2.5× random primer/reaction buffer mix (Invitrogen) and sterile water up to a final volume of 45 μl. The reaction mix was boiled for 5 minutes prior to cooling on ice and the addition of 2.5 μl of Spectral labeling buffer (Spectral Genomics, Houston, U.S.A), 1.5 μl of either Cy3-dCTP (1mM, Applied Biosystems, Foster City, U.S.A) or Cy5-dCTP (1 mM, Applied Biosystems) and 1 μl of Klenow Fragment (BioPrime labeling kit, Invitrogen). The reaction was mixed gently and then incubated for 2 hours at 37°C. Following incubation the reaction was stopped by the addition of 5 μl 0.5 M EDTA (pH8) and heating at 72°C for 10 minutes.
The Cy3 labeled test DNA was combined with the Cy5 labeled normal reference DNA and vice versa. Each combined probe was mixed with 45 μl of Spectral Hybridisation Buffer (Spectral Genomics), 11.3 μl of 5 M NaCl and 110 μl of room temperature isopropanol. The samples were incubated in the dark at room temperature for 10–15 minutes and centrifuged at 16,000 x g for 10 minutes and the supernatant discarded. The pellets were then washed with 500 μl of 70% ethanol. The supernatant was carefully removed and the pellets air-dried at room temperature in the dark. For hybridisation, the pellets were first resuspended in 10 μl of sterile water prior to being mixed with 30 μl of Spectral Hybridisation Buffer II (Spectral Genomics) by pipetting. The reconstituted probes were then incubated at 72°C for 10 minutes, placed on ice for 5 minutes and then incubate for 30 minutes at 37°C. The probes were hybridised to BAC arrays, covered with a 22 × 60 mm coverslip and incubated for 12–16 hours at 37°C in a humidified chamber.
The wash buffers, with the exception of Wash I, were pre-warmed to 50°C. The slides were gently dipped into and out of Wash I (2× SSC, 0.5% SDS) until the coverslip detached from the slide. The slides were then washed once in Wash II (2 × SSC, 50% deionized Formamide, pH 7.5) for 20 minutes, followed by successive washes in Wash III (2× SSC, 0.1% NP-40, pH 7.5) for 20 minutes and Wash IV (0.2× SSC, pH 7.5) for 10 minutes. All washes were performed at 50°C, with the exception of Wash I. The slides were briefly submerged in distilled deionized water for 5–10 seconds and centrifuged for 5 minutes at 750 rpm to dry.
Data collection and analysis
The slides were scanned using an Axon GenePix 4000 A confocal scanner, each fluorescence signal was collected separately and quantified with the GenePix Pro 3.0 software (Axon Instruments, U.S.A). The data was normalised and analysed using Normalise Suite v2.4 , all regions of loss or gain were determined as those that were 2 standard deviations above the mean baseline for each separate sample.
van der Kwast TH, Labrie F, Tetu B: Persistence of high-grade prostatic intra-epithelial neoplasia under combined androgen blockade therapy. Hum Pathol. 1999, 30: 1503-1507. 10.1016/S0046-8177(99)90174-0.
Al-Maghrabi J, Vorobyova L, Toi A, Chapman W, Zielenska M, Squire JA: Identification of numerical chromosomal changes detected by interphase fluorescence in situ hybridization in high-grade prostate intraepithelial neoplasia as a predictor of carcinoma. Arch Pathol Lab Med. 2002, 126: 165-169.
Kallioniemi A, Kallioniemi OP, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D: Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science. 1992, 258: 818-821.
van Dekken H, Paris PL, Albertson DG, Alers JC, Andaya A, Kowbel D, van der Kwast TH, Pinkel D, Schroder FH, Vissers KJ, Wildhagen MF, Collins C: Evaluation of genetic patterns in different tumor areas of intermediate-grade prostatic adenocarcinomas by high-resolution genomic array analysis. Genes Chromosomes Cancer. 2004, 39: 249-256. 10.1002/gcc.20001.
Wolf M, Mousses S, Hautaniemi S, Karhu R, Huusko P, Allinen M, Elkahloun A, Monni O, Chen Y, Kallioniemi A, Kallioniemi OP: High-resolution analysis of gene copy number alterations in human prostate cancer using CGH on cDNA microarrays: impact of copy number on gene expression. Neoplasia. 2004, 6: 240-247.
Clark J, Edwards S, Feber A, Flohr P, John M, Giddings I, Crossland S, Stratton MR, Wooster R, Campbell C, Cooper CS: Genome-wide screening for complete genetic loss in prostate cancer by comparative hybridization onto cDNA microarrays. Oncogene. 2003, 22: 1247-1252. 10.1038/sj.onc.1206247.
Hughes S, Arneson N, Done S, Squire J: The use of whole genome amplification in the study of human disease. Prog Biophys Mol Biol. 2005, 88: 173-189. 10.1016/j.pbiomolbio.2004.01.007.
Dean FB, Hosono S, Fang L, Wu X, Faruqi AF, Bray-Ward P, Sun Z, Zong Q, Du Y, Du J, Driscoll M, Song W, Kingsmore SF, Egholm M, Lasken RS: Comprehensive human genome amplification using multiple displacement amplification. Proc Natl Acad Sci U S A. 2002, 99: 5261-5266. 10.1073/pnas.082089499.
Dean FB, Nelson JR, Giesler TL, Lasken RS: Rapid amplification of plasmid and phage DNA using Phi 29 DNA polymerase and multiply-primed rolling circle amplification. Genome Res. 2001, 11: 1095-1099. 10.1101/gr.180501.
Lage JM, Leamon JH, Pejovic T, Hamann S, Lacey M, Dillon D, Segraves R, Vossbrinck B, Gonzalez A, Pinkel D, Albertson DG, Costa J, Lizardi PM: Whole genome analysis of genetic alterations in small DNA samples using hyperbranched strand displacement amplification and array-CGH. Genome Res. 2003, 13: 294-307. 10.1101/gr.377203.
Lasken R: Multiple displacement amplification of genomic DNA. Whole Genome Amplification: Methods Express. Edited by: Hughes SLR. 2005, Oxford, Scion Publishing Ltd., 99-118.
Barker DL, Hansen MS, Faruqi AF, Giannola D, Irsula OR, Lasken RS, Latterich M, Makarov V, Oliphant A, Pinter JH, Shen R, Sleptsova I, Ziehler W, Lai E: Two methods of whole-genome amplification enable accurate genotyping across a 2320-SNP linkage panel. Genome Res. 2004, 14: 901-907. 10.1101/gr.1949704.
Paez JG, Lin M, Beroukhim R, Lee JC, Zhao X, Richter DJ, Gabriel S, Herman P, Sasaki H, Altshuler D, Li C, Meyerson M, Sellers WR: Genome coverage and sequence fidelity of phi29 polymerase-based multiple strand displacement whole genome amplification. Nucleic Acids Res. 2004, 32: e71-10.1093/nar/gnh069.
Tzvetkov MV, Becker C, Kulle B, Nurnberg P, Brockmoller J, Wojnowski L: Genome-wide single-nucleotide polymorphism arrays demonstrate high fidelity of multiple displacement-based whole-genome amplification. Electrophoresis. 2005, 26: 710-715. 10.1002/elps.200410121.
Hughes S, Lim G, Beheshti B, Bayani J, Marrano P, Huang A, Squire JA: Use of whole genome amplification and comparative genomic hybridisation to detect chromosomal copy number alterations in cell line material and tumour tissue. Cytogenet Genome Res. 2004, 105: 18-24. 10.1159/000078004.
Hughes S, Damato BE, Giddings I, Hiscott PS, Humphreys J, Houlston RS: Microarray comparative genomic hybridisation analysis of intraocular uveal melanomas identifies distinctive imbalances associated with loss of chromosome 3. Br J Cancer. 2005, 93: 1191-1196. 10.1038/sj.bjc.6602834.
Cleton-Jansen AM, Buerger H, Haar N, Philippo K, van de Vijver MJ, Boecker W, Smit VT, Cornelisse CJ: Different mechanisms of chromosome 16 loss of heterozygosity in well- versus poorly differentiated ductal breast cancer. Genes Chromosomes Cancer. 2004, 41: 109-116. 10.1002/gcc.20070.
Kimura Y, Noguchi T, Kawahara K, Kashima K, Daa T, Yokoyama S: Genetic alterations in 102 primary gastric cancers by comparative genomic hybridization: gain of 20q and loss of 18q are associated with tumor progression. Mod Pathol. 2004, 17: 1328-1337. 10.1038/modpathol.3800180.
Joos S, Bergerheim US, Pan Y, Matsuyama H, Bentz M, du Manoir S, Lichter P: Mapping of chromosomal gains and losses in prostate cancer by comparative genomic hybridization. Genes Chromosomes Cancer. 1995, 14: 267-276.
Visakorpi T, Kallioniemi AH, Syvanen AC, Hyytinen ER, Karhu R, Tammela T, Isola JJ, Kallioniemi OP: Genetic changes in primary and recurrent prostate cancer by comparative genomic hybridization. Cancer Res. 1995, 55: 342-347.
Yano S, Matsuyama H, Matsuda K, Matsumoto H, Yoshihiro S, Naito K: Accuracy of an array comparative genomic hybridization (CGH) technique in detecting DNA copy number aberrations: comparison with conventional CGH and loss of heterozygosity analysis in prostate cancer. Cancer Genet Cytogenet. 2004, 150: 122-127. 10.1016/j.cancergencyto.2003.09.004.
Paiss T, Worner S, Kurtz F, Haeussler J, Hautmann RE, Gschwend JE, Herkommer K, Vogel W: Linkage of aggressive prostate cancer to chromosome 7q31-33 in German prostate cancer families. Eur J Hum Genet. 2003, 11: 17-22. 10.1038/sj.ejhg.5200898.
Dong JT, Boyd JC, Frierson HFJ: Loss of heterozygosity at 13q14 and 13q21 in high grade, high stage prostate cancer. Prostate. 2001, 49: 166-171. 10.1002/pros.1131.
Alers JC, Rochat J, Krijtenburg PJ, Hop WC, Kranse R, Rosenberg C, Tanke HJ, Schroder FH, van Dekken H: Identification of genetic markers for prostatic cancer progression. Lab Invest. 2000, 80: 931-942.
Matsuyama H, Pan Y, Yoshihiro S, Kudren D, Naito K, Bergerheim US, Ekman P: Clinical significance of chromosome 8p, 10q, and 16q deletions in prostate cancer. Prostate. 2003, 54: 103-111. 10.1002/pros.10173.
Tomlins SA, Rhodes DR, Perner S, Dhanasekaran SM, Mehra R, Sun XW, Varambally S, Cao X, Tchinda J, Kuefer R, Lee C, Montie JE, Shah RB, Pienta KJ, Rubin MA, Chinnaiyan AM: Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science. 2005, 310: 644-648. 10.1126/science.1117679.
Zhao H, Hastie T, Whitfield ML, Borresen-Dale AL, Jeffrey SS: Optimization and evaluation of T7 based RNA linear amplification protocols for cDNA microarray analysis. BMC Genomics. 2002, 3: 31-10.1186/1471-2164-3-31.
Collins C, Rommens JM, Kowbel D, Godfrey T, Tanner M, Hwang SI, Polikoff D, Nonet G, Cochran J, Myambo K, Jay KE, Froula J, Cloutier T, Kuo WL, Yaswen P, Dairkee S, Giovanola J, Hutchinson GB, Isola J, Kallioniemi OP, Palazzolo M, Martin C, Ericsson C, Pinkel D, Albertson D, Li WB, Gray JW: Positional cloning of ZNF217 and NABC1: genes amplified at 20q13.2 and overexpressed in breast carcinoma. Proc Natl Acad Sci U S A. 1998, 95: 8703-8708. 10.1073/pnas.95.15.8703.
Shayesteh L, Lu Y, Kuo WL, Baldocchi R, Godfrey T, Collins C, Pinkel D, Powell B, Mills GB, Gray JW: PIK3CA is implicated as an oncogene in ovarian cancer. Nat Genet. 1999, 21: 99-102. 10.1038/5042.
Savinainen KJ, Linja MJ, Saramaki OR, Tammela TL, Chang GT, Brinkmann AO, Visakorpi T: Expression and copy number analysis of TRPS1, EIF3S3 and MYC genes in breast and prostate cancer. Br J Cancer. 2004, 90: 1041-1046. 10.1038/sj.bjc.6601648.
Cassinelli G, Supino R, Zuco V, Lanzi C, Scovassi AI, Semple SC, Zunino F: Role of c-myc protein in hormone refractory prostate carcinoma: cellular response to paclitaxel. Biochem Pharmacol. 2004, 68: 923-931. 10.1016/j.bcp.2004.06.002.
Bernard D, Pourtier-Manzanedo A, Gil J, Beach DH: Myc confers androgen-independent prostate cancer cell growth. J Clin Invest. 2003, 112: 1724-1731. 10.1172/JCI200319035.
Zhigang Z, Wenlv S: Prostate stem cell antigen (PSCA) expression in human prostate cancer tissues and its potential role in prostate carcinogenesis and progression of prostate cancer. World J Surg Oncol. 2004, 2: 13-10.1186/1477-7819-2-13.
Fuessel S, Sickert D, Meye A, Klenk U, Schmidt U, Schmitz M, Rost AK, Weigle B, Kiessling A, Wirth MP: Multiple tumor marker analyses (PSA, hK2, PSCA, trp-p8) in primary prostate cancers using quantitative RT-PCR. Int J Oncol. 2003, 23: 221-228.
Reiter RE, Gu Z, Watabe T, Thomas G, Szigeti K, Davis E, Wahl M, Nisitani S, Yamashiro J, Le Beau MM, Loda M, Witte ON: Prostate stem cell antigen: a cell surface marker overexpressed in prostate cancer. Proc Natl Acad Sci U S A. 1998, 95: 1735-1740. 10.1073/pnas.95.4.1735.
Chang CJ, Freeman DJ, Wu H: PTEN regulates Mdm2 expression through the P1 promoter. J Biol Chem. 2004, 279: 29841-29848. 10.1074/jbc.M401488200.
Mu Z, Hachem P, Agrawal S, Pollack A: Antisense MDM2 oligonucleotides restore the apoptotic response of prostate cancer cells to androgen deprivation. Prostate. 2004, 60: 187-196. 10.1002/pros.20044.
Leite KR, Franco MF, Srougi M, Nesrallah LJ, Nesrallah A, Bevilacqua RG, Darini E, Carvalho CM, Meirelles MI, Santana I, Camara-Lopes LH: Abnormal expression of MDM2 in prostate carcinoma. Mod Pathol. 2001, 14: 428-436. 10.1038/modpathol.3880330.
Takahashi S, Urano T, Tsuchiya F, Fujimura T, Kitamura T, Ouchi Y, Muramatsu M, Inoue S: EBAG9/RCAS1 expression and its prognostic significance in prostatic cancer. Int J Cancer. 2003, 106: 310-315. 10.1002/ijc.11205.
Cybulski C, Gorski B, Debniak T, Gliniewicz B, Mierzejewski M, Masojc B, Jakubowska A, Matyjasik J, Zlowocka E, Sikorski A, Narod SA, Lubinski J: NBS1 is a prostate cancer susceptibility gene. Cancer Res. 2004, 64: 1215-1219. 10.1158/0008-5472.CAN-03-2502.
Huang H, Muddiman DC, Tindall DJ: Androgens negatively regulate forkhead transcription factor FKHR (FOXO1) through a proteolytic mechanism in prostate cancer cells. J Biol Chem. 2004, 279: 13866-13877. 10.1074/jbc.M314143200.
Beheshti B, Braude I, Marrano P, Thorner P, Zielenska M, Squire JA: Chromosomal localization of DNA amplifications in neuroblastoma tumors using cDNA microarray comparative genomic hybridization. Neoplasia. 2003, 5: 53-62.
We would like to thank the researchers at Princess Margaret Hospital tissue procurement facility for their help and technical assistance in securing good quality samples for this study. We also like to thank Roger Lasken for his advice. This work was supported by the National Cancer Institute of Canada with funds from the Canadian Cancer Society and by awards to S.H. and M.Y. from the United States Army Medical Research and Materiel Command [DAMD17-03-1-0154].
SH and MY designed and carried out the study, in addition to preparing the final manuscript for publication. BB helped with the analysis of the data. AE helped in the selection of cases and worked closely with JS in the study design and coordination.
Electronic supplementary material
Additional File 1: Genomic comparison of gaCGH results obtained from non-amplified male DNA versus non-amplified male DNA hybridisation and amplified male DNA versus amplified male DNA hybridisation. Blue line indicates the chromosome position plotting of amplified DNA data. Black line indicates the chromosome position plotting of non-amplified DNA data. (PNG 9 KB)
Additional File 2: Genomic comparison of gaCGH results obtained from non-amplified male versus non-amplified female DNA hybridisation and amplified male DNA versus female DNA hybridisation. Blue line indicates the chromosome position plotting of amplified DNA data. Black line indicates the chromosome position plotting of non-amplified DNA data. (PNG 12 KB)
Additional File 3: Genomic comparison of gaCGH results obtained from non-amplified DNA from the colorectal cell line DLD1 versus non-amplified control DNA male hybridisation and amplified DNA from the colorectal cell line DLD1 versus amplified control DNA hybridisation. Blue line indicates the chromosome position plotting of amplified DNA data. Black line indicates the chromosome position plotting of non-amplified DNA data. Detection of gain and loss is shown by cyan and pink bars, corresponding to amplified and non-amplified DNA, respectively. (PNG 9 KB)
Additional File 4: H&E sections show an example of LCM. Dissection of HPIN. picture represents the tissue before dissection, middle picture is after dissection, and bottom picture is the cap tissue. (TIFF 703 KB)
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Cite this article
Hughes, S., Yoshimoto, M., Beheshti, B. et al. The use of whole genome amplification to study chromosomal changes in prostate cancer: insights into genome-wide signature of preneoplasia associated with cancer progression. BMC Genomics 7, 65 (2006). https://doi.org/10.1186/1471-2164-7-65
- Laser Capture Microdissection
- Copy Number Change
- Whole Genome Amplification
- Promote Tumour Development
- Cell Robotic

domain: health


text:
Hands Off the People of Iran chair Yassamine Mather assesses the current situation in Iran at Communist University 2010.
The attempt by the two wings of the Iranian regime to shelve their differences is unlikely to defuse the mass movement, writes Yassamine Mather
More than two weeks after the demonstrations of December 27 2009, the political repercussions of these events, and the reaction to the anger and radicalism of the protesters, continue. Clearly now no-one, from the government to the ‘reformists’, to the revolutionary opposition, has any doubt that the current protests are no longer about who should be the ‘president’ of the Islamic Republic, but represent a serious challenge to the very existence of the religious state.
Ashura is a day of mourning for Shia Muslims, as they commemorate the martyrdom of Hussein, a grandson of the prophet Mohammed in 680AD. In December 2009 it coincided with the seventh day following the death of a clerical critic of the regime, ayatollah Montazeri. Throughout Iran hundreds of thousands of protesters took to the streets with slogans against the supreme leader, Ali Khamenei, and calling for the overthrow of the Islamic Republic. When security forces attacked, the crowds fought back. Tehran was “covered in thick smoke from fires and tear gas” and there was “hand-to-hand combat between security forces and the protesters,” with reports of street battles in other major cities.1 For the first time in the last 30 years, many women came out into the streets to join the demonstrations wearing no headscarves or hijabs.
At a number of locations in Tehran security forces were forced to retreat, as demonstrators burnt police vehicles and bassij posts and erected barricades. There are videos showing instances where police and bassij were captured and detained by demonstrators and three police stations in Tehran were briefly occupied. Demonstrators also attacked Bank Saderat in central Tehran, setting it on fire.
The government’s reaction was predictable. Since December 27 bassij and pasdaran (revolutionary guards) have been unleashed to impose further repression. Hundreds of people have been incarcerated. The summary arrest of leftwing and worker activists, the death sentences issued against left political prisoners, the sacking of workers already in prison are part of a deliberate attempt by the regime to impose an atmosphere of terror.
Ultra-conservative clerics have also called for the arrest and execution of ‘reformist’ leaders. In a speech on January 9 the supreme leader told government security forces and the judiciary to act decisively against “rioters and anti-government demonstrators”.
Despite the bravado of Khamenei, there are clear signs that the demonstrations of December 27 have divided the conservatives further on how to respond to the protests. While supporters of president Mahmoud Ahmadinejad openly call for more arrests and even the execution of political opponents, the ‘principlist’ faction2 within parliament is preaching caution.
On January 9, a parliamentary committee publicly blamed Tehran’s former prosecutor, Saeed Mortazavi, a close ally of Ahmadinejad, for the death of three prisoners arrested during anti-government protests in June 2009. The committee found that Mortazavi had authorised the imprisonment of 147 opposition supporters and 30 criminals in a cell measuring only 70 square metres in Kahrizak detention centre. The inmates were frequently beaten and spent days without food or water during the summer.
Ali Motahhari, a prominent fundamentalist parliamentarian, told the weekly magazine Iran Dokht: “Under the current circumstances, moderates should be in charge of the country’s affairs.” He suggested Ahmadinejad should also be held accountable for the deaths in Kahrizak and for fuelling the post-election turmoil. Iranian state television is broadcasting debates between ‘radical’ and ‘moderate’ conservatives, in which Ahmadinejad is blamed by some for causing the crisis.
There are two reasons for this dramatic change in line:
1. The December 27 demonstrations were a turning point, in that both conservatives and ‘reformists’ came to realise how the anger and frustration of ordinary Iranians with the political and economic situation is taking revolutionary forms.
2. The principlists are responding to a number of ‘proposals’ by leading ‘reformists’ as a last attempt to save the Islamic Republic. Fearful of revolution, ‘reformist’ leaders from the June 2009 presidential candidate Mir-Hossein Moussavi to former president Mohammad Khatami have made conciliatory statements, and the moderate conservatives have responded positively to these approaches.
In a clear sign that ‘reformists’ have heard the cry of the revolution, Moussavi’s initial response to the Ashura demonstrations was to distance himself from the protests, emphasising that neither he nor Mehdi Karroubi had called for protests on that day. His statement on January 1 entitled ‘Five stages to resolution’ (of the crisis) was a signal to both his supporters and opponents that this was truly the last chance to save the Islamic regime from collapse.
Western reportage of the statement concentrated on his comment, “I am ready to sacrifice my life for reform.” Of course, Iranians are well known for their love of ‘martyrdom’, from Ashura itself to the Fedayeen Islam in 1946,3 to the Marxist Fedayeen (1970s-80s). Iranians have been mesmerised by the Shia concept of martyrdom, inherited from Sassanide ideals, a yearning to put their lives at risk for what they see as a ‘revolutionary cause’. But Moussavi will no doubt go down in history as the first Iranian who is putting his life on the line for the cause of ‘reform’ and compromise!
His five-point plan is seen as a compromise because it does not challenge the legitimacy of the current president and “presents a way out of the current impasse” in order to save the Islamic Republic, basically demanding more freedom for the Islamic ‘reformist’ politicians, activists and press, as well as accountability of government forces, while reaffirming his allegiance to the constitution of the Islamic regime, as well as the existing “judicial and executive powers”. The preamble to the proposal explains very well Moussavi’s message to the supreme leader and the conservative faction: it is not too late to save the regime, but this could be our last chance.
It reads: “Today the situation of the country is like an immense roaring river, where massive floods and various events have led to its rising and then caused it to become silted. The solution to calm down this great river and clear its water is not possible in a quick and swift action. Thinking of these kinds of solutions that some should repent and some should make deals and there should be some give and take to solve this great problem is in practice going off the track … I also believe that it is still not too late and our establishment has the power to accomplish this important task, should it have insight and a respectful and kind view toward all of the nation and its layers.”
This statement was followed on January 4 by a ‘10-point proposal’ from the self-appointed ‘ideologues’ in exile of Iran’s Islamic ‘reformist’ movement: the former Pasdar, Akbar Ganji (nowadays introduced on BBC and CNN as a “human rights activist!”), Abdolkarim Soroush, Mohsen Kadivar, Abdolali Bazargan and Ataollah Mohajerani.4
Fearful that the Moussavi plan will be seen by many as too much of a compromise, the group of five call for the resignation of Ahmadinejad and fresh elections under the supervision of a newly established independent election commission to replace that of the Guardian Council. In the last few days both Khatami and another former president, Ali Akbar Rafsanjani, have publicly declared their support for the compromise, while condemning “radicals and rioters”. Khatami went further than most, insulting demonstrators who called for the overthrow of the Islamic regime.
All in all, it has been a busy two and a half weeks for Iran’s ‘reformists’, terrified by the radicalism of the demonstrators and desperate to save the clerical regime at all cost. Inevitably the reformist left, led by the Fedayeen Majority, is tailing the Moussavi-Khatami line. However, inside Iran there are signs that the leadership of the green movement is facing a serious crisis.
None of the proposals addresses the most basic democratic demand of the Iranian people: separation of state and religion. A widely distributed leaflet and web post inside Iran entitled ‘Who is the leader of the current protest movement in Iran?’ refers to comments made by ayatollah Taleghani 31 years ago,5 at the height of the revolutionary movement. Taleghani, faced with a similar question, replied that it was the shah who led the protest movement because the repression he imposed and his inability to compromise caused it to move forward day by day. The leaflet concludes that the current force leading the movement is supreme leader Khamenei, who by his words and actions is fuelling the revolutionary fervour.
Working class response
In every event Iranians see real and imaginary parallels with the 1978-79 uprising that led to the shah’s downfall. Last week the publication of Khamenei’s alleged escape plans and the revelations that senior clerics had arranged to send their fortunes abroad to avoid sanctions and the consequences of an uprising reminded Iranians of January 1979, when the shah and his entourage were busy making similar arrangements.
The Iranian left is not immune to such nostalgia. Arguments about the ‘principal contradiction’ and ‘stages of revolution’ seem to dominate current debates. While some Maoists argue in favour of a ‘democratic stage’ of the revolution, citing the relative weakness of the organised working class, the Coordinating Committee for the Setting Up of Workers’ Organisations (Comite Hahamhangi) points out that the dominant contradiction in Iran, a country where 70% of the population lives in urban areas, is between labour and capital. They point out that the level and depth of workers’ struggles show radicalism and levels of organisation and that the Iranian working class is the only force capable of delivering radical democracy.
Leftwing organisations and their supporters are also discussing the lessons to be learnt from the Ashura demonstrations. Clearly sections of the police and soldiers are refusing to shoot at demonstrators and the issue of organising radical conscripts in order to divide and reduce the power of the state’s repressive forces must be addressed. In some working class districts around Tehran and other major cities the organisation of neighbourhood shoras (councils) has started.
The current debates within the ruling circles have had no impact on the level of protests undertaken by workers and students. There are reports of strikes and demonstrations in one of Iran’s largest industrial complexes, Isfahan’s steel plant, where privatisation and contract employment have led to action by the workers. Leftwing oil workers/employees are reporting disillusionment with Moussavi and the ‘reformist’ camp amongst fellow workers and believe there is an opportunity to radicalise protests in this industry despite the fact that close control and repression has intensified over the recent period.
Last week a number of prominent labour activists, including Vahed bus worker Mansour Ossanlou, who are currently in prison (some incarcerated for over a year) were sacked from their jobs for ‘failing to turn up at work’, which prompted protests in Vahed depots and the Haft Tapeh sugar cane plant. In late December workers at the Lastic Alborz factory went on strike demanding payment of unpaid wages. This week workers have been holding protests at dozens of workplaces, including the Arak industrial complex, the Mazandaran textile factory, at the Polsadr metro construction and in Tonkabon.
Over the next few weeks Iranian workers will face major challenges. Even if the two main factions of the regime achieve a compromise, it will be unlikely to defuse the movement. In fact the conciliatory line of Moussavi and Khatami is certain to further reduce their influence amongst protesters. However, if the religious state is able to reunite, it will be more difficult to attend demonstrations, call strikes and hold sit-ins, etc.
Whatever happens, Iranian workers will need our solidarity more than ever. That is why Hands Off the People of Iran is currently planning a week of solidarity and fundraising actions in February – check the Hopi website for more details (www.hopoi.org).
1. New York Times December 29 2009.
2. One of the groups in the conservative faction of the Iranian parliament.
3. Fedayeen Islam was one of the first truly Islamic fundamentalist organisations in the Muslim world. It was founded in Iran by Navab Safavi in 1946 for the purpose of demanding strict application of the sharia and assassinating those it believed to be apostates and enemies of Islam.
5. See http://en.wikipedia.org/wiki/Mahmoud_Taleghani.
On December 30 two students were critically wounded and scores injured by knife wielding members of Ansar-e Hezbollah and Basij militia, up to 500 thugs were brought in to attack students at Mashhad University after they staged anti-regime protests during Ashura. One of the students professors was also attacked and sustained knife wounds, whilst a young female student was badly injured after being struck repeatedly over the head with a piece of wood. Students at the university were holding silent mourning ceremonies for the Ashura were they opposed the repression of popular protests. The police aided the Basij and Hezbollah by blocking the roads leading up to the University and attacking crowds of students with tear gas and batons. Around 210 students and youth were arrested by the state-repressive forces throughout the recent Ashura protests. Below is the video of the brutal attack by Basij and Hezbollah on students:
The day after over 4000 students and professors staged protests against the attacks and arrests at Ferdowsi University of Mashhad and Azad University of Mashhad but were laid siege by security forces and militia. Students, professors and parents have trie dto find out information on the arrested and injured. The local judiciary is refusing to release information on the two seriously injured students, leading to speculation that the student have died and the authorities do not want there funerals to become a focus of further anti-regime protests. Students sent a delegation of representatives from the Islamic Organisation of the university to a meeting with university officials where they were arrested. Amongst those arrested is Seyed Sadra Mirada a relative of Khamanei. Below is a video of the gathering of students and youth on December 31:
Below is a brief report of the moves the regime has been taking against known leftwing activists and the threats that leading officialis have been giving on state television. This report was sent to us by Anahita Hosseini of the ‘Independent Leftist Students’ who represent an anti-imperialist socialist tendency within the student movement in Iran
After the mass protests of Sunday December 17 the regime is showing its fear of people uprising by going to well known activists homes one by one and arresting them. This morning armed plain cloths forces went to Mahin Fahimis home who is a member of the organization of: mothers for peace and arrested her and her son Omid Montazeri who is a known leftist student activist. Omid is Hamid Montazeriz son a known communist activist who was executed by the regime during the mass murders of the leftists and Mujahadeen in prison in 1988.
Ardavan Tarakameh another leftist student activist who was staying in Omids home this morning was arrested, afterwards the plain cloths forces went to Ardavan’s parents home and searched it all and took some books and notes, and told his mother she is not allowed to ask any questions about what they are doing or where her son is. Zohreh Takaboni one of the mothers for peace whose husband was also executed as a leftist in 1988 has also been arrested.
The regime has started a new scenario since this morning on all of their TV channels they are talking about what happened in the 80s they are talking about the leftist opposition of Iran in those days and how the regime killed them! because of their activities, they are frankly threatening people that they are not afraid of repeating the history.
After yesterdays uprising it became more obvious that no one is of the illusion of re-running the elections. The slogans are aimed at the regime and Khamenei himself, radicalization of the movement has made the regime fearful of the effect of the lefftist and the other radical activists on the current uprising. They are threatening to bring back the black and the bloody decade of 80’s in which they mass murdered thousands of the bravest, purest and the true believers of freedom and equality especially in 1988 when they executed thousands (possibly 30 thousand) leftists and Mujahadeen and buried them in the mass graves. Now they are threatening their children and all the other activists and all people who are yelling their anger against them, in their official news today they said: the rebels have crossed the red lines by having slogans against Khamenei and they will all pay back for it. what is obvious is that they will not be able to repeat the bloody years of 80s because they cant mass murder a nation. But we should take the threat serious on the level that we know this regime has nothing to lose and before its final collapse they may do anything for revenge. They may try to limit the number of activists against them, the threats they have started against people is important on these levels, and it is our responsibility to fight until the release of each and every political prisoner in Iran alongside supporting the peoples uprising. Underestimating the threats of the dictator regime in taking revenge on the protesters can end in a catastrophe.
We will fight until all of our classmates, comrades and friends are released. We wont let the Islamic Republic take the revenge of its inevitable collapse on activists.
Unity – Struggle – Victory
December 27 was the bloodiest and most violent convulsion in Iran since the June elections. Millions of ordinary Iranians came out onto the streets to use the Ashura ceremonies and mourning as a focal point of opposition protests. In every part of Iran security forces backed up by Basij militia and the revolutionary guard (Pasdaran) resorted to ever intensifying violence as swarms of protestors over-ran state-repressive forces. It is unclear how many have been killed and arrested at this time, the regime say that only 4 have been killed, whilst student websites and news feeds from Iran put the number around 15. ‘Reformist’ leader Mir-Hossein Mousavi’s nephew is among the dead. The official reason for these deaths have been accidents and murder by ‘uknown assailants’. The regime has admitted to arresting over 300 protestors yesterday, this number will undoubtedly be much greater.
Clashes took place in Shiraz, Isfahan, Ardebil, Arababad, Mashhad. Whilst Marshal Law was declared in Najaf-Abad, at least four have been killed in the city of Tabriz and the house of recently deceased Ayatollah Montazeri was the scene of heavy fighting in Qom.
Protests began in the morning around 10 am with heavy security presence on major streets, squares and transport links. In Tehran the supreme leader’s residence was surrounded by massed ranks of Pasdaran and police. Throughout the day chants against Khamanei, such as ‘this month is a month of blood!- Khamanei will be toppled’. A clear indication of how far the protest movement has come since June, not only is the regime fearful of a re-run of the election, but are now considerably worried that a revolution is underway. In Tehran clashes erupted at many religious sites as soon as people started to gather for the planned opposition protests. The fighting was intense, with security forces taking several defeats as demonstrators burnt police vehicles, stations, Basij posts and erected barricades. In a couple of instances police and Basij were arrested and detained by the people and three police stations in Tehran were briefly occupied by protestors.. Demonstrators also attacked the Saderat Bank in central Tehran, setting it on fire.
As the day wore on the security forces began to crack, the first division of the special forces refused orders to shoot protestors. There are many pictures and videos that show police retreating or being beaten back by protestors (some are in this report). There is also unconfirmed statements from sections of the army declaring that they will not be used to put down popular unrest. During the evening clashes erupted outside the IRIB headquarters with security forces firing tear gas and bullets into the crowds who responded with rocks and burning barricades. Later on there was fighting in and around Hospitals in central Tehran.
Following the protests several aides to opposition leaders have been arrested whilst injured protestors have been interviewed, beaten and arrested whilst in hospital, the many injured have had to endure interrogation with painful injuries. In response to this it has been reported that medical staff have been patching people up instead of admitting them to the already overcrowded wards. In many parts of Tehran residents opened their doors to the injured and exhausted demonstrators.
The Ashura protests saw a qualitative change in the protests, the people of Iran attacked and won street battles in Tehran, attacked a set fire to police stations and security forces vehicles, demonstrators arrested and detained many riot police and Basij throughout the day. Possibly more importantly the regime has undermined its own religious credibility by making martyrs on Ashura day. Neither side of the regime can now back down, and through this split the mass movement is breaking down the Islamic Republic. Many calls have come not just for the end of Khamanei’s rule, or Ahmadinejad’s government but for the end of the Islamic Republic itself. On the streets protestors have begun chanting ‘Independence, freedom, Iranian Republic’, a slogan which has been condemned by ‘reformist’ leader Mousavi as too radical. The Ashura protests have further underlined that the Islamic Republic is facing the greatest existential threat since its inception and the Iraq-Iran war.
Below are some videos of the protests:
Iranian demonstrations have given a real boost to working class opponents of the regime, writes Yassamine Mather
Every year November 4, the anniversary of the 1979 take-over of the US embassy in Tehran, is marked in Iran with a state-organised demonstration outside the building that used to house the American ambassador and his staff. On that date 30 years ago militant Islamic students stormed the embassy and took 71 hostages. Nineteen were released within weeks, but the remaining 52 were held for 444 days.
The ceremony commemorating the 30th anniversary of the ‘US hostage crisis’ was no different from recent years: a lacklustre ritual addressed by an insignificant minister. However, no-one in Iran will ever forget November 4 2009. It was the day when illegal demonstrations in at least six separate locations in Tehran and 20 cities and university campuses throughout the country overshadowed the state-organised event. As the national broadcasting service was showing live pictures of the gathering outside the former US embassy, shouts of “Death to the dictator” from protesters on neighbouring streets and squares were so loud that it was difficult to hear the minister’s speech. In Tehran the six locations were Enghelab Square, Ferdowsi, Haft Tir, Enghelab Square, Vali Asr and Vanak Square.
Revolutionary guards had issued stern warnings that they would not tolerate any protest demonstrations, and the night before dozens of political activists were arrested. On the morning of November 4 itself, government offices closed their doors at around 10am to stop employees leaving their workplace to join the protests. The ministry of the interior deployed special units of anti-riot police, many on motorbikes, as well as the religious bassij militia, to block main roads, intimidate potential demonstrators and attack any gathering. Yet despite all these measure, by all accounts – including admissions in the pro-Ahmadinejad press – tens of thousands of Iranians joined the protests against the regime.
Highly significant was the absence of any slogans regarding the rigged elections. Four months and 22 days after the June 2009 presidential poll, demonstrators in Iran have clearly moved on. Even the BBC Persian Service, that staunch defender of the ‘green movement’, had to admit in its broadcasts and analyses what most of the left has been saying for some time: as a result of the impasse within the factions of the Islamic regime the protests are no longer about the results of the presidential elections. Protesters are now challenging the very existence of that regime. ‘Reformist’ leaders are tailing the masses.1
The advice of their ‘leaders’ – most of whom, with the exception of presidential candidate Mehdi Karroubi, did not even dare show their face at the demonstrations – was totally ignored. Fellow ‘reformist’ candidate Mir-Hossein Moussavi had spent the previous 10 days warning everyone against “radical” slogans that would only “benefit the enemy”. Yet demonstrators did the exact opposite.
Even the bourgeois media had to admit that the radicalisation of the demonstrations has marked a new phase in the life of the opposition. The main slogans that dominated the day were directed at the supreme leader himself: “Our guardian is a murderer [the supreme leader’s official religious title is ‘guardian of the nation’]. His rule is null and void” (Vali ma ghateleh velayatesh bateleh), plus the usual “Death to Khamenei, death to the Islamic republic”.
The crowds were also at odds with Moussavi over the nuclear issue. In late October he and Karroubi met to discuss the recent US-EU offer to Iran, and made it clear that they considered Ahmadinejad’s response to be a sell-out. Moussavi was quoted by his own website Kalameh as saying: “If the promises given are realised then the hard work of thousands of scientists would be ruined.” Yet for the first time in many years, it looked like the nationalist defenders of a nuclear Iran had no supporters amongst the protesters, whose slogans were very clear: “We don’t want reactors, we don’t want the atomic bomb.”
A week earlier, Moussavi, after a lot of dithering, had called on his supporters to back the November 4 demonstrations, yet on the day he failed to show up at any of the protests. His supporters claimed he was prevented from leaving a cultural centre by the security forces, but witnesses deny this. For all his faults, Karroubi, the 70-year-old cleric, showed more courage. He was prepared to join the demonstrations, even though one of his bodyguards was badly injured and ended up in hospital.
In another qualitative development angry demonstrators tore down posters of Khamenei and trampled all over them in what were unprecedented scenes. The man who is supposed to be god’s representative on earth (for Shia Muslims) was called a murderer and his image defiled by demonstrators wiping their feet on his posters.
Most of all, though, November 4 will be remembered as the day Iranians realised their strength and found the courage to stand up to the regime’s supporters and security forces. A number of bloggers have remarked on how government supporters leaving the official gathering hid memorabilia and photos of the supreme leader that had been dished out at that event when they saw the huge number of protesters in neighbouring streets.
There were many reports and films of the bassij and militia attacking protesters, especially women. However, there were also many incidents where demonstrators confronted those forces and actually got the better of them. In some incidents old women defended young protesters and shamed the security forces into retreating.
Some protesters have also taken up a new chant: “Obama, Obama – either you’re with them or you’re with us.” On the face of it, this does not sound like the most radical of slogans. However, this is a country obsessed with conspiracy theories regarding foreign interference and it was the first time since 1979 that Iranians have directed a slogan at the leader of the hegemon capitalist power in the face of such conspiracy theories. It should be noted that since Irangate2 no-one in Iran takes slogans like “Death to America” and “Death to Israel” shouted at official demonstrations seriously.
A number of foreign reporters were detained, most of whom have now been released, together with an Iranian journalist working for Agence France Presse. The stupid leaders of the regime had thought that by making such arrests they would stop the world hearing about the protests, but the reality is that now Iran has millions of reporters, with their text messages, emails and video footage captured on mobile phones. Perhaps the regime will consider banning all electronic equipment in their desperation to stop the ‘wrong’ news spreading.
The demonstrations have given a real boost to working class opponents of the regime. For the first time in many years they are finding allies in their struggle against the Islamic government. Sections of the left, including Rahe Kargar, have been talking of setting up neighbourhood resistance committees and clearly, given the vicious attacks by security forces on the growing opposition, such committees are necessary. For the first time in many years Iranians are discussing the need for the masses to be armed to confront the state security forces, while maintaining their opposition to ‘militarist’ tactics.
But the regime will not give up easily. More than 200 people were arrested in Tehran and the provinces on or around November 4, while a number of labour activists from the Haft Tapeh sugar cane company have been sent to prison for organising strikes. There are unconfirmed reports that despite many efforts to save the life of Kurdish leader Ehsan Fattahian, he was executed on November 11 in Sanandaj Central Prison. Ehsan’s 10-year prison sentence for membership of an illegal Kurdish organisation was recently changed to execution for no apparent reason.
Hundreds of protesters remain in prison and we must do all we can to support and defend them. Let us step up our solidarity with the working class and democratic opposition.
- See BBC Newsnight report, including interview with BBC Persian Service presenter: www.youtube.com/watch?v=wPgi2LUNdqI
- See en.wikipedia.org/wiki/Iran%E2%80%93Contra_affair for a summary of the ‘Iran Contra affair’, also known as ‘Irangate’.
From Weekly Worker 793
Ahmadinejad uses the ‘enemy without’ to justify increased repression, arrests and the torture of the ‘enemy within’, writes Yassamine Mather
The dramatic statements by Obama, Brown and Sarkozy about Iran’s undisclosed nuclear enrichment plant, made in a ‘breaking news’-style press conference on the first day of the G20 gathering in Pittsburgh, were clearly intended to prepare the world for a new conflict in the Middle East. The presentation of the ‘news’ and the language used in delivering the threats were reminiscent of the warnings about Iraq’s ‘45-minute’ strike capability.
According to Obama, “Iran is on notice that when we meet with them on October 1 they are going to have to come clean, and they will have to make a choice.” The alternative to sticking to ‘international rules’ on Iran’s nuclear development, would be “a path that is going to lead to confrontation”.
Yet in some ways the existence of a second uranium enrichment plant is old news. By all accounts US and UK secret services had known about this plant for at least three years – Israel and France also knew about it for some time and had delivered their finding to the International Atomic Energy Agency earlier this year. The ‘dramatic’ disclosures came at a time when Russia was already on board regarding further sanctions. Given its billion-dollar trade with Iran, China – one of Iran’s major commercial partners – is unlikely to change its opposition to further sanctions.
So what was the main purpose of the Obama-Sarkozy-Brown show on September 25? Could it be it was directed mainly to audiences in the US, UK and France, to convince them that, at a time of economic uncertainty, western leaders have to deal with a ‘major external threat’ posed by Iran’s nuclear development?
But the elephant in that press conference room was the Israeli nuclear programme. While Iran might be approaching nuclear military capability by 2010-15 (no-one is claiming it has such capability now), another ‘religious’ state in the Middle East is exempt from IAEA regulations and possesses between 100 and 200 nuclear warheads (this according to US estimates), yet it maintains a policy of ‘deliberate ambiguity’ on whether it has nuclear weapons.
Former IAEA director general Mohamed El Baradei regarded Israel as a state possessing nuclear weapons, but there has been no IAEA inspection, hence the ambiguity over the number of warheads it possesses. Strictly speaking, as a beneficiary of the largest cumulative recipient of US foreign assistance since World War II, Israel is not supposed to have any. Yet every year the US congress approves billions of dollars of US military aid to Israel. For the fiscal year 2010, Obama is requesting $2.775 billion.
The Symington and Glenn amendments to foreign aid law specifically prohibit US aid to nuclear states outside the Nuclear Non-proliferation Treaty (NPT). Iran has signed the NPT. Israel has not.
Of course, none of this justifies the Iranian rulers’ obsession with reaching a stage where they can produce nuclear weapons. Unlike middle class nationalist Iranians, who even in their opposition to the regime, favour the government’s nuclear programme, the Iranian working class has been clear on this issue, as shown by placards on recent demonstrations: “We don’t want nuclear power – we don’t want huge salaries. We work so that we can live – we don’t live to work.”
Millions of Iranian workers have not been paid for months, while capitalists and the religious government keep telling them of Iran’s economic crisis and shortfalls in both the state and private funds, yet the Islamic regime seems to have sufficient funds to equip one more nuclear enrichment plant, paying billions – presumably to dubious sources – for black market equipment. The current escalation of the conflict also exposes the stupidity of the Iranian rulers who only admitted to the existence of this ‘secret’ plant after its existence was ‘exposed’.
Of course, Iranians have become so used to hearing total lies from the leaders of all factions of the Islamic regime that the revelation of the existence of this facility, hidden not far from the capital, did not come as a surprise. After all, this is the same government that used Photoshop to pretend a failed rocket did succesfully launch, the same government that cheated in the presidential elections, then lied about the number of people killed in the subsequent protests, and the same government whose president claims to have seen a white light descending from another world while he was addressing the UN assembly in 2007.
Further sanctions will bring more poverty for Iranian workers and it will be the Iranian people who will pay the price for the foolishness of the very leaders they have been protesting against for over two months. The US is keen on sanctions against companies exporting refined oil to Iran (which imports 60% of its requirements). It now looks like France and Germany are sceptical about such sanctions. They refer to the Iraq experience and the ease with which petrol can be smuggled across land borders.
The Iranian government has already indicated that it will cut petrol subsidies. It is blaming the west and hopes such a move will unite the country against the ‘foreign enemy’. Contrary to the pessimism of sections of the Iranian left in exile who ‘despair’ of the growth of the ‘Green’ movement or who have joined the bandwagon behind ‘reformist’ presidential candidate Mir-Hossein Moussavi, workers in oil refineries in Iran are well aware of the historic role of their class in the current situation and there have been discussions regarding strikes in this industry for the last few weeks. These workers have two valid concerns: (1) that their strike should not benefit Moussavi (he is hated by these workers, some of whom remember his time in power); and (2) that their strike should not help US efforts for regime change from above.
Western countries are also considering options including an embargo on investment in Iran’s oil and gas sector, an end to loan guarantees to all companies investing in Iran, a ban on Iranian businesses trading in euros, and a ban on foreign companies insuring Iranian shipping and air transport. All of these measures will target the Iranian people, the majority of whom hate the clerical state.
If the Iranian government lied about its nuclear installations, Ahmadinejad’s speech last week at the UN was also full of deceit. His holocaust-denial comments, repeated in every interview he gave while in the US, were a deliberate attempt to divert attention from mass protests at home and to heighten the tension with the rest of the world. This regime and this president rely on foreign crises to survive – he desperately needs enemies abroad to divert attention from problems at home, and the Obama-Brown-Sarkozy trio gave him that.
However, his speech contained other lies. The man who has printed money in an attempt to solve Iran’s economic problems told the world: “It is no longer possible to inject thousands of billions of dollars of unreal wealth into the world economy simply by printing worthless paper assets, or transfer inflation as well as social and economic problems to others through creating severe budget deficits.” He also criticised “liberal capitalism” (as opposed to clerical capitalism?). After all, this is the president of a government that is busy privatising every industry in Iran, from services in the oil industry to car plants and Iran’s national telecommunications. The telecom company was privatised and sold to the ‘revolutionary guards’ in the last week of September, although Iran’s ‘monopoly regulatory commission’ is now said to be investigating this.
However, such actions by Iran’s Sepah Pasdaran (Revolutionary Guards) do not imply that the country is under military capitalist rule: they are controlled by the most conservative sections of Iran’s clerical elite. The Pasdaran ownership of the telecommunication services is only another success for supreme leader Ali Khamenei, his son and the clerics around him, as this ideological military force has no life and no significance without clerical rule.
The few delegates in the UN assembly hall who heard Ahmadinejad condemn the excesses of “liberal capitalism” might have thought Iran is an egalitarian religious society. Nothing could be further from reality. After 30 years in power Iran’s Islamic regime has created one of the most unequal, corrupt societies of the region, where the gap between the rich and the poor is amongst the highest in the world. As Ahmadinejad was speaking, Iran’s car workers (amongst the best paid sections of the working class) were protesting at long shifts causing ill health and workers throughout Iran were on strike or demonstrating against non-payment of wages. While factory closures due to privatisation continue, Aryaman Motors, a Tehran-based company specialising in reproducing classic cars, launched a new series of replica vehicles based on the original design of the earliest Rolls Royce models at $120,000 each – wealthy Iranians have already pre-paid for the first models that will be finished later this year.
In his speech Ahmadinejad also referred to the disastrous wars in Iraq and Afghanistan, failing to mention Iran’s role in support of US aggression in both – as leaders of the Campaign Against Sanctions and Military Intervention in Iran keep reminding us! The Iranian president then referred to breaches of human rights in Guantanamo and Abu Ghraib. Of course, it is inevitable that abuse of human rights by the ‘torch holders’ of liberal democracy in the US and the UK will be used by every tinpot leader in Africa, the Middle East and elsewhere to justify the torture and execution of opponents. The Iranian president is the leader of a government that has killed at least 72 civilians and tortured hundreds in the last two months alone, yet the actions of western governments allow him to stand up in New York and give moral lectures about ‘human right abuses’. We truly live in irrational times.
Protests and divisions
The first days of the new university term in Iran saw major protests on campuses throughout the country – the largest being at Tehran University on September 27-28. Students shouted “Death to the dictator” and booed the new minister of higher education. Security forces retreated from the campus. On Tuesday September 29 students protested at Sharif University, once more causing the minister for higher education to abandon plans to speak. Meanwhile, security forces are warning football crowds not to chant political slogans at the Tehran derby between Esteghlal and Persepolis on October 2.
As former president and leading ‘reformist’ Ali Akbar Rafsanjani continues his efforts to find a compromise between the regime’s warring factions, the first signs of a rift amongst ‘reformists’ has appeared. In an open letter addressed to Rafsanjani, another ‘reformist’ presidential candidate, Mehdi Karoubi, writes: “What is your answer to the people who, under dangerous conditions, question the actions of the Assembly of Experts under your leadership? … By what measure have you preserved the ideals of the revolution in your role as chair of the Assembly of Experts, whose first duty is fighting injustice?”
Moussavi’s latest statement on September 28 is also predictably uninspiring. Its repeated references to the “wisdom” of Iran’s first supreme leader, Ruhollah Khomeini, confirmed his continued allegiance to the ‘imam’s line’. But this will not gain him much support amongst young Iranians, who will not accept any solution short of the overthrow of the entire regime. Moussavi’s call on his supporters to “avoid any radical measures which could damage the achievements so far made by the opposition” expose once more his fear of radical change and his determination to save the religious state.
All this is very good news for the revolutionary forces. However, the threat of sanctions and war only strengthens Khamenei and Ahmadinejad. In the words of UN weapons inspector Hans Blix, any “rush to punitive sanctions – tightened to the point where ordinary Iranians, already suffering the effects of chronic unemployment, had to endure petrol shortages or big fuel price hikes – could backfire spectacularly”.
Hands Off the People of Iran has always condemned sanctions and threats of war against Iran. We oppose them not only because we want to see imperialism defeated, but because they increase patriotism and nationalism, thus helping the reactionary regime. The government will use the ‘threat of the enemy without’ to increase repression, to arrest and torture its ‘enemy within’. Sanctions disorganise the working class, as people are forced to squander their fighting energies on day-to-day struggles to keep their jobs and feed their families – Iranian oil workers are right to be concerned about going on strike at a time when sanctions will also target ‘imported refined oil’.
The proposed US-European sanctions dramatically degrade the ability of the working people to struggle collectively on their own account, to organise and to fight. In other words, for the sake of Iranian working class we must continue our opposition to war, sanctions and regime change from above, while increasing our solidarity with the revolutionary movement inside Iran.
More videos after the break…

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Barely a few days after a group called ‘Northern Elders’ protested President Tinubu’s order for the CBN to relocate some of its key departments to Lagos, the Federal Government has again directed FAAN to move its national head office from Abuja to Lagos!
The question is, what is President Tinubu’s government trying to achieve by moving key Government parastatals from the FCT to Lagos? From all indications, Lagos is congested and cannot absorb more federal institutions. On the other hand, Abuja is still sparsely populated and needs more such federal institutions to accord it the very essence of a federal capital territory.
President Tinubu’s attempt to relocate some of these federal agencies from Abuja to Lagos would amount to a waste of time and money. No federal agency wants to be far from the seat of Government. Secondly, most of their meetings and operational engagements are done in Abuja, not Lagos.
Read Also: Tinubu Dey Carry Us Go Where We No Know!
Therefore, relocating such agencies to Lagos would only increase their costs in terms of overhead, operations, logistics, etc. And lastly, when another northerner becomes president after Tinubu, those agencies would be returned to Abuja again. By then, what has Nigeria gained by such wasteful back and forth?
It’s time to rethink our federal structure!
SCRIPT CREDIT: Facebook.com / Uzogara Tobechukwu
Disclaimer: “The views/contents expressed in this article are the sole responsibility of Uzogara Tobechukwu and do not necessarily reflect those of The World Satellite (TWS). TWS will not be responsible or liable for any inaccurate or incorrect statements contained in this article.”


Congratulations at last to U.S. Senator Barack Obama. The first bill bearing his name passed the Senate, and was signed into law by President Bush. Obama, while appreciative of the notoriety he has received before and since being elected to the Senate, has joked in his Gridiron speech, “Thanks. When I actually do something, we’ll let you know.”
Well, aside from raise mountains of money for other democrats and, with his wife Michelle, pull in a whopping $1.67 million for his family, today he’s done something for America.
The “Federal Funding Accountability and Transparency Act”, will create a Web-based searchable database of federal government spending. It looked for a while that the bill would suffer defeat, but bloggers from the left and the right united in protest, and the bill passed.
Good work, Senator!


text:
by Owen McNally
CHRISTMAS IN NEW ORLEANS
The Dukes of Dixieland
Hurricane Katrina nearly wiped out this holiday release, which marks the first Christmas album by the New Orleans-based Dukes of Dixieland.
Just four days before katrina, the tapes were Fed Exed from New Orleans to Los Angeles for mastering — lucky timing that may have saved them from being swept away.
Paradoxically, the tapes fared far better than the Dukes themselves. Drummer Richard Taylor lost his home to Katrina. And all six band members became evacuees, scattered across the United States from Sarasota, Fla., to Seattle.
Because these tracks were recorded pre-Katrina, they light up like a Christmas tree with bright, festive spirit.
the band uses holiday fare like "O Tannenbaum" and "Winter Wonderland" as jam session goodies seasoned with a Creole flavor. It trumpets hosannas to the glories of the Big Easy on such pieces as "Christmas Time in New Orleans," "Holiday Time in New Orleans," and "Second Line Santas."
Fellow New Orleanians the Pfister Sisters, bluesman Luther Kent, and Moses Hogan's Gospel Choir join the Dukes in their jubilant celebration of Christmas in the Crescent City.
Since returning, the Dukes, who are old hands at riverboat rambling, have toured on the Mississippi and Ohio Rivers, raising money for the Bush-Clinton Katrina Relief Fund.
As a sign of devotion to its mother city, the combo has embraced a new working name, New Orleans' Own Dukes of Dixieland. It lives by a new, passionate slogan: "We will rebuild!"

domain: geography


text:
Women suffering from bloating are more likely to turn to probiotic yogurts than visit their general practitioner to check for Ovarian Cancer, a study has found.
Persistent bloating is one of the key symptoms of cancer but research by Target Ovarian Cancer found just 34 percent of women in UK would visit the doctors if they had concerns about regular bloating. While there is no figure for India, the estimates could be much lower in India.
Half of the British women would change their diet if they...
Congress President Rahul Gandhi is visiting the election-bound state of Meghalaya. He held several meetings and a road show in Shillong earlier today. In his trademark style, Gandhi took on Prime Minister Narendra Modi. Talking to journalists in Shillong he said that Prime Minister Narendra Modi is not against corruption, but an instrument of corruption.
He said, “One after the other you will see these scams. They are coming out. We have already seen Nirav Modi, we have seen Rotomac, and we have...
Bharatiya Janata Party’s Karnataka chief BS Yeddyurappa on Wednesday targeted the Congress President Rahul Gandhi over his recent visit to poll-bound Karnataka by taking a jibe and called him a kid. Yeddyurappa, the BJP chief ministerial candidate asserted that his party would win over 150 seats.
“By bringing that baccha (Rahul Gandhi) in Karnataka, we now know that we will win more than 150 seats here,” Yeddyurappa said at a BJP social media convention.
He had earlier alleged that the Congress...
Way before the trailer was released, Tiger Shroff and Disha Patani-starrer 'Baaghi 2' has piqued the interest of the masses for various reasons, one being the tactic pairing of leading duo, who reached to unveil the action-packed first glimpse of their film at Mumbai's Mahalaxmi Race Course.
Their unconventional entry in a chopper raised the level of excitement for the fans. Dressed like a true 'rebel', Disha paired a simple white T-shirt with cargo pants and converse whereas Tiger was in action...
Syria’s daily death toll on Tuesday reached its highest point in the last three years. The death toll from two days of bombing by Syrian Government of a rebel-held area Damascus has risen to 250.
The Britain-based Syrian Observatory for Human Rights claimed it was the highest 48-hour death toll in the Syria conflict since a 2013 chemical attack in the area. It is the worst violence in the Eastern Ghouta area near Damascus. The report stated that more than 50 children are among the dead.
Another handsome bachelor of Bollywood is now off the market. 'Raag Desh' actor Mohit Marwah got hitched to his longtime girlfriend Antara Motiwala in a typical Punjabi wedding on Tuesday. The lavish ceremony was hosted at Waldorf Astoria Ras Al Khaimah in UAE.
And he is off ... ????????❤❤❤ #antumoh????
A post shared by Maheep Kapoor (@maheepkapoor) on Feb 20, 2018 at 6:24am PST
Being the member of influential Kapoor family has its own perks which were noted...
Meri Aashiqui Tum Se Hi: Ishaani’s Grand Entry In Ranveer’s House Post Leap[Inside Pics & Video]
NEWS WORLD INDIA | 0
September 16 , 2015 , 16:45 IST
Ranveer Ishaani's Meri Aashiqui Tum Se Hi fans rejoice! Finally, the small screen hit Jodi has been reunited. After all the drama and hurdles, Ishani is back with her lover Ranveer and looks like happiness is finally knocking the doors of Vaghela family.
In the last episode, we saw how Ranveer saved Ishaanni's life by diffusing the bomb that was tied across her waist.
ALSO READ: Meri Aashiqui Tumse Hi: Ranveer Saves Ishaani From Bomb Blast; Shikhar To Exit Show
Also, Ranveer's mom Amba apologised to Ishaani for not trusting her and accepted her as bahu. The show then takes six months leap!
In the upcoming episodes, get ready for the most awaited scene i.e. Ishaani's Grihapravesh in Vaghela house. The episode has already been shot and will soon be aired on television.
ALSO READ: Meri Aashiqui Tum Se Hi: ‘Double Trouble’ In Ranveer-Ishaani’s Love Story!
Isn't it exciting fans? We have heard that Amba has planned a grand entry for her bahurani and brings her inside the house with all rituals.
Watch Ishaani's Grihapravesh:
With this, we have also heard that Ranveer and Ishaani will renew their wedding vows. Makers are planning a grand wedding for IshVeer and till then they will live separately in different rooms!
So get ready for Ranveer and Ishaani's remarriage in the upcoming episodes looks like happiness is FINALLY stepping in their life.


text:
Conventional medicine does not look at the body as a whole, instead viewing it in terms of isolated systems, with a separate doctor for each one. Generally, lupus patients are under the care of a rheumatologist and a doctor who specializes in the area in which they are experiencing symptoms–for example, a nephrologist for your kidneys, and a dermatologist for your skin.
And it’s important to point out that people who are initially diagnosed with systemic lupus (or SLE) can also get lupus rashes. One of the common rashes that occurs in people with systemic lupus is malar rash. It’s alternatively called a butterfly rash, and it spreads across the bridge of the nose and cheeks and is telltale sign of the disease because its appearance is so distinctive, Caricchio says. A malar rash can be flat or raised. While it usually isn’t painful, it can itch and burn. (3) And the rash can show up on other parts of the body, including the arms, legs, and torso.
Arthritis is inflammation of one or more joints and it involves the breakdown of cartilage. Cartilage normally protects a joint, allowing it to move smoothly. Cartilage also absorbs shock when pressure is placed on the joint, such as when you walk. Without the normal amount of cartilage, the bones rub together, causing pain, swelling (inflammation), and stiffness. Usually the joint inflammation goes away after the cause goes away or is treated. Sometimes it does not. When this happens, you have chronic arthritis. Arthritis may occur in men or women. Osteoarthritis is the most common type.
Levels of stress-related illnesses are on the rise, and stress, both emotional and physical, has been shown to trigger and intensify autoimmune disorders. Stress is your body’s response to a threat–a wound, injury, or infection. Acute stress revs up your immune system to help you deal with an immediate crisis, and then calms it back down once the threat is removed. On the other hand, chronic stress (the kind we face in this day and age) leads to long-term inflammation and actually suppresses your immune system. This can trigger or worsen autoimmune conditions, and can lead to the reactivation of latent viruses linked to lupus, perpetuating a vicious cycle.
The Accelerating Medicines Partnership (AMP) is a bold new venture between the NIH, 10 biopharmaceutical companies and several non-profit organizations to transform the current model for developing new diagnostics and treatments by jointly identifying and validating promising biological targets of disease. The ultimate goal is to increase the number of new diagnostics and therapies for patients and reduce the time and cost of developing them.
The panel judged the effect of extended AC as a large benefit, reducing VTD with increase in bleeding risk as a moderate harm. For the comparisons of different AC intensities, the panel decided to use the evidence from observational studies because it judged that it probably better reflects reality given that the randomised controlled trials (RCT) are severely flawed (indirectness of intervention as most patients did not reach the INR >3 goal). They judged the reduction in VTD as a large benefit and the bleeding increase as a large harm. Hence, the panel considered that the balance could favour the intervention only when the risk of VTD recurrence is particularly high.
Moderate use of alcohol is usually not a problem for people with lupus, but alcohol can lower the effectiveness of some medications, cause new health problems, and/or can make existing problems worse. For example, non-steroidal anti-inflammatory drugs -- such as aspirin, ibuprofen (Motrin®), naproxen (Naprosyn®), and celecoxib (Celebrex®) -- can cause ulcers and bleeding in the stomach and intestines at any time during treatment; the chance of developing an ulcer or internal bleeding increases with alcohol use. Also, anticoagulant medicines such as warfarin (Coumadin®) and the chemotherapy drug, methotrexate, may not be as effective if you are drinking alcohol.
For instance, a dermatologist for cutaneous lupus (skin disease), a cardiologist for heart disease, a nephrologist for kidney disease, a neurologist for brain and nervous system disease, or a gastroenterologist for gastrointestinal tract disease. A woman with lupus who is considering a pregnancy needs an obstetrician who specializes in high-risk pregnancies.
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Please Note: The material on this site is provided for informational purposes only and is not medical advice. Always consult your physician before beginning any diet or exercise program.
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domain: medical