Leptospira Cultures Maintenance

Leptospira Cultures MaintenanceRESULTSThe Leptospira serovars maintained in the Department of Veterinary Microbiology were used in the present study. For maintenance, EMJH medium (Difco) with ovalbumin supplement was employed and subcultured at seven day intervals and incubation was carried out at 28-30C. In addition, the stock cultures were maintained in semi-solid medium with subculturing at angiotensin converting enzyme month interval.IDENTIFICATION OF LEPTOSPIRESUnder dark field microscope, the live leptospiral organisms were found tightly coiled and actively motile. The motility notice was of both spinning and bending. In highly conpennyrated cultures, the organisms formed entangled masses. No contaminants were observed in most of the time when streaked on blood agar plates for purity checking of the cultures. In case of contamination they were purified by filtration.RECOMBINANT PROTEIN PRODUCTIONPreparation of template DNA from LeptospiraThe genomic DNA isolated from Leptos pira interrogans serovar Australis had a DNA concentration ranging from 40- 60g/ml. The purity of the extracted DNA was checked by measuring the ratio of absorbance (O.D of DNA preparation at 260 and 280 nm). The value of the ratio obtained was found to be in the range of 1.8 to 1.93, indicating that the preparations were nigh free of proteins.(in mat and methods).Amplification of lipl21 gene and lipl32 genesThe amplification of lipl21and lipl32 genes were carried out and observed amplicons of 507 bp and 767 bp, respectively. (Fig 1)Cloning of the lipl21 and lipl32 geneThe colonies of E.coli Dh5 cells transformed with lipl21 and lipl32 genes was picked up separately and tested for the presence of the 2 genes. It was observed that lipl21 clones yielded an amplicon size of 507 bp and lipl32 with 767 bp. These confirmed clones were preserved for further studies (Fig 2)Induction of recombinant proteinThe above clones were subcultured in LB broth containing Ampicillin (100g/ml) and exp ression was optimized with 2 mM IPTG for LipL21 and 1mM IPTG for LipL32. The induced recombinant cells were harvested after six hours and five hours for LipL21 and LipL32, respectively. Uninduced controls were set for each. The cells were then pelleted and lysed. The expression of recombinant 21 kDa (rLipL21) and 32 kDa outer membrane proteins (rLipL32) were confirmed in comparison with that of the uninduced cells where in that respect was no significant protein expression (Fig 3)Purification of recombinant lipl21 and lipl32 proteinsThe rLipL21 and rLipL32 proteins were purified by Nickel chelating affinity chromatography without any contamination. The protein concentrations were estimated to be 0.69 mg/ml and 2.07mg/ml for rLipL21 and rLipL32, respectively.Immunoreactivity of the proteinsThe immunogenicity of both rLipL21 and rLipL32 proteins were tested using bland autocratic dogtooth sera and observed that both the proteins were reacting. Further the protein did not react wh en blotted with hyper immune serum raised against the different bacteria namely E.coli, staphylococcus aureus and Pasteurella multocida.DIAGNOSISMicroscopic Agglutination riseA total of 124 canine serum samples from Leptospira suspected dogs were tested using snap and among this 22 (17.74 per cent) were found to be positive for leptospirosis. Serum samples having a titre of 1400 and above were considered as positive. The infecting serovars identified with canine leptospirosis are depicted in Table 3Enzyme Linked Immunosorbent AssayIndirect ELISA was done in separate microtitre plates employing rLipL21 and rLipL32 as antigens and the results were compared with that of mire.Checker board compendUsing checker board analysis the optimum concentration of antigen, antibody and conjugate were estimated. The optimum concentration of antigen was found to be 50 ng/well and 150 ng/well for rLipL21 and rLipL32, respectively. The rabbit anti-canine immunoglobulin G HRP conjugate concentrati on estimated was 12500 and 12000 for rLipL21 and rLipL32, respectively. A 150 dilution of test serum was used as optimum working dilution in both ELISAs.Determination of cut off valuesIn IgG ELISA, the mean OD and standard deviation for the negative sera samples (n=44) was 0.49 and 0.11 for rLipL21 and 0.59 and 0.09 for rLipL32, respectively. The cut off value obtained was 0.82 for rLipL21 and 0.86 for rLipL32.Test properThe results of rLipL21 and rLipL32 based IgG ELISA are given in Table 4. Among 47 positive samples obtained by rLipL21 ELISA, only 20 found positive with MAT. In case of rLipL32 ELISA, 40 samples were recorded positive out of which 18 found positive with MAT.Comparison of MAT and rLipL21IgG ELISAThe results of rLipL21 IgG ELISA were compared with that of MAT.Among 124 canine sera examined, 47 (37.90 per cent) showed OD more than the cut off value i.e. 0.82 and were considered positive for leptospirosis by rLipL21 IgG ELISA. The sensitivity, specificity and accuracy of rLipL21 IgG ELISA as relative to MAT was measured to be 90.90 per cent, 73.52 per cent and 76.61 per cent, respectively (Table 5).On statistical analysis, it was found that there exists a significant difference between the two tests, ie, rLipL21 ELISA and MAT even at 1 per cent level of significance.Comparison of MAT and rLipL32 IgG ELISAThe results of the IgG ELISA using rLipL32 as the antigen were compared with that of MAT.Among 124 canine sera examined, 40 (32.25 per cent) showed OD more than the cut off value i.e. 0.86 and were considered positive for leptospirosis by IgG ELISA. The sensitivity, specificity and accuracy of rLipL32 IgG ELISA as relative to MAT was calculated to be 81.81per cent, 78.43 per cent and 79.03 per cent, respectively (Table 6).Statistical analysis revealed that there exists a significant difference between the two tests, ie, rLipL32 ELISA and MAT at 1per cent level of significance.Comparison of rLipL21 and rLipL32 IgG ELISAOn statistical analysis usi ng Cochrans Q test, at 1 per cent level of significance it was observed that there exists no significant difference between rLipL21 and rLipL32 ELISA, as the P value was found to be greater than 0.01.Table 3 Infecting serovars identified with MATTable 4 Results of MAT, rLipL21 ELISA and rLipL32 ELISATable 5 Comparison between rLipL21 ELISA and MATSensitivity = a/a+c = 90.90%Specificity = d/b+d = 73.52%Accuracy =a+d/a+b+c+d = 76.61%

Low MicroRNA-21 Expression in HPV-Induced Carcinogenesis

Low Microribonucleic acid-21 Expression in HPV-Induced CarcinogenesisLow microRNA-21 port in the emergence of a favorable microenvironment for HPV-induced carcinogenesisIntroductionHuman papillomaviruses (HPVs) are the most common sexually transmitted agents. gamey risk types of Human Papillomavirus, as HPV16, are the sternutative agents of virtually all cases of cervical cancer and a significant proportion of other anogenital cancers, as well as some head and neck cancers 1-3.The K14-HPV16 transgenic mice mildew is specially utilize to study the HPV-associated squamous cells cancers. In this model, the normal of auriclely region genes (E2-E8) of HPV-16 is driven by the keratin 14 promoter/enhancer 4. The K14-HPV16 transgenic mice develop epidermic hyperplastic lesions that progress to dysplastic lesions and ultimately to invasive cancer. The nerve of HPV oncogenes E6 and E7 induces epithelial carcinogenesis through well-defined premalignant stages before de novo carcinom a tuition 5. The basal cells are mitotically active and thus may develop further mutations in response to a proliferative stimulus, and the expression of K14 has been shown to persist in well-differentiated squamous carcinomas 6. This model of multistep epithelial neoplasia will facilitate the study of both the epi inherited and the genetic factors that regulate neoplastic progression and coordinate malignant conversion.MicroRNAs (miRNA) are small, noncoding RNAs that regulate gene expression by base pairing with informational RNAs, leading to the inhibition of mRNA translation or its degradation. In normal cells, miRNAs control numerous processes including proliferation, differentiation and apoptosis 7. Furthermore, these molecules are described as key regulators in many diseases including, neurological dis orderings, cardiovascular diseases, viral infections and cancer. Some miRNAs are lost during tumorigenesis whereas other miRNAs are upregulated. Previous data indicates that miRNAs are important to distinguish subtypes of cancers, where the histological diagnosis is involved and difficult. Furthermore, it also may be a useful tool to diagnose cancers of unknown origin and to study cancer predisposition 8.MicroRNA-21 (miR-21) has been implicated in divers(a) aspects of carcinogenesis. In most solid tumors, miR-21 is overexpressed and thus bows cell proliferation, differentiation and apoptosis 9-11. However, the interplay in the midst of miRNAs, human papillomavirus (HPV) genes and how these interactions contribute to HPV-associated cancers remain elusive and not well understood.The tumor microenvironment associated to miRNAs plays an increasingly appreciated role in cancer (ref whiteside) however, the microenvironment prior to tumorigenesis can influence the carcinogenesis process. In this study we investigated the expression profile of microRNA-21 in K14-HPV16 transgenic mice, using ear and vanity skin samples. Thus, we evaluate the importance of the microenvironment associated with the miR-21 expression and predisposition to HPV-induced carcinogenesis.Material and methodsTransgenic miceK14-HPV16 mice on a FVB/N background were generously donated by Drs. Jeffrey Arbeit and Douglas Hanahan, from the University of California, through the ground forces National malignant neoplastic disease Institute Mouse Repository. Generation of K14-HPV mice has been previously reported 12. After one week quarantine, the animals were kept as breeder pairs in consent with National (Portaria 1005/92 dated October the 23rd) and European (EU Directive 2010/63/EU) legislation, under controlled conditions of temperature (232C), light-dark cycle (12h light/12h dark) and relative humidity (5010%), using hardwood bedding. A standard diet (Global Diet 2014, Harlan, Barcelona) and water were provided ad libitum. Health checks were performed daily.Genotyping of HPV-E6 and E2 (referencias genotipagem Hugo?)15 offspring females from consecutive litters w ere genotyped at weaning, using one-quarter tip samples. To genotyping were used tail tips of mice of the strain FVB, wild-type or hemizygous. Tissue lysis was performed using MAGNAPure DNA Tissue Lysis Buffer and Proteinase K for 17 h at 65 C. Nucleic acids were extracted by the High Pure Viral Nucleic Acid Kit following the manufacturers instructions. To test the efficacy of the method of DNA extraction was investigated the presence of mouse--globin gene. The presence of integrated HPV was assessed by increase of HPV-E6 and HPV-E2 genes by polymerase chain reaction methodology (PCR) in-house. The resulting genotypes were confirmed to the respective phenotypes. After genotype determination, all animals were sacrificed at 22 to 26 weeks-old and completely necropsied.Ear and chest skin samples were collected into TRIzol (Invitrogen) for miRNA abridgment and matched samples were collected into 10% neutral buffered formalin for histological processing. For these procedures, 13 skin samples (ear and chest) of 8 hemizygous females (+ / -) were collected. As control were used 14 skin samples of 7 wild-type females (- / -).HistologySkin samples were fixated in 10% neutral buffered formalin for 48 hours, routinely processed and paraffin-embedded. Histological sections (2m-thick) were obtained and stained with haematoxylin and eosin (HE) for examination on light microscopy. Samples and their lesions were classified as normal skin, cuticular hyperplasia and epidermal in situ carcinoma by two independent, blind researchers (CL and RGC).miRNA expression analysisTo study miRNA-21 expression, the skin samples were macerated with the TRIzol reagent (Invitrogen) for RNA preservation. The extraction of total RNA was performed using aHigh Pure RNA Isolation Kit(Roche apply Science), according to manufacturers instructions. RNA quality was assessed by beat the absorbance at 260 nm and its purity was evaluated by the ratio of absorbance at 260/280 nm.cDNA synthesisThe conv ersion of miRNA to cDNA was performed using TaqMan MicroRNA Reverse Transcription Kit ( PN 4366596 , Applied Biosystems, Foster CA, USA ), using sequence-specific stem-loop primers from each miRNA (miR-21 and snoRNA-202). The amplification conditions were as follows 30 min at 15C, 52 min at 42C and finally 10 min at 85C.miRNA-21 relative quantificationWe used qPCR technique to measure the relative expression of miR-21 (StepOne Real-time PCR Systems final volume 20 uL, with1 TaqMan Universal Master Mix II Applied Biosystems, Foster City, California USA 1x MicroRNA Assay (Applied Biosystems, Foster City, California USA) and 2 uL cDNA. As endogenous control, we used snoRNA-202.Statistical AnalysisData analysis was performed by the computer software IBMSPSS Statistics for Windows (Version 20.0). The 2Ct method, along with Students t-test was used in order to evaluate any statistical differences in the normalized expression of the miR-21.To analyze the normalized relative expression (-C t) of the different groups, we considered the results corresponding to 99% representation of the population (2). ResultsGenotyping and histological analysisWe observed the presence of integrated HPV DNA (E6 and E2 ORF) in 53% of mice. All cases with HPV-E6 expression also presented HPV-E2 expression. All mice with integrated HPV DNA demonstrated, phenotypically, various degrees of persistent epidermal and squamous mucosal hyperplasia, characteristic lesions associated to HPV infection, previously described in Arbeit et al. study 12. After histologic evaluation we observed that, in all cases with integrated HPV DNA, the ear tissues presented epidermal in situ carcinomas while the chest tissues showed epidermal hyperplasia. In wild-type mice we observed normal histology.MiRNA-21 expression profile in tissue of transgenic versus wild-type miceTo investigate a possible cover influence of HPV16 on the miR-21 expression profile, we analyzed the relative expression between the all tissues (ear and chest) of transgenic mice and the tissues of the control group. We did not found statistical difference in expression levels between both groups (p=0,615). When we compare the miRNA-21 expression in ear and chest samples, singly, we also did not found statistical difference in expression levels between transgenic and wild type mice (Fig.2).MiRNA-21 expression profile in normal tissueTo study the miR-21 normal expression profile in tissues, we quantified the expression of miR-21 in the ear and chest skin samples of the control group. We observed that the ear tissues have lower expression levels when compared to chest tissue (p = 0.036) (Fig.3 a)).Mir-21 expression profile in tissues of transgenic miceTo compare the parity between miR-21 expression and lesion type, we analyzed the histology of ear and chest samples from transgenic mice and we quantified the expression of miR-21 in the same. We detected that lower expression levels of miR-21 are associated with cancerous les ions as in situ carcinomas (ear) compared with hyperplastic lesions (chest) (p=0,043) (Fig.3 b)). handlingRecent studies have associated miR-21 to the pathogenesis of various diseases, including cancer. (ref)MiR-21 overexpression is observed in the majority of carcinomas and hematological malignancies. However, understanding of the potential role of miRNA-21 in previous microenvironment to the development of HPV-associated lesions remains elusive.In cervical cancer, it is accepted that HPV infection is the most important factor for transition from normal cervical epithelium to cervical pre-neoplastic intraepithelial neoplasia and subsequently to invasive cervical cancer. However, the influence of others factors including the microenvironment are poorly investigated. Microenvironment associated to miR-21 may be key factor to the predisposition of cancer. We studied that the expression of miR-21 in normal tissues could be important in development of HPV-associated tumors.Our results d emonstrate that, in transgenic mice, all ear tissues presented epidermal in situ carcinomas and chest tissues showed epidermal hyperplasia. Thus, we hypothesize that these interesting facts could be related to different miR-21 profile expression of both tissues.Our results indicate that there is no statistically significant difference between the miR-21 expression in HPV-positive samples and controls, concluding therefore that the presence of HPV does not straight off influence the expression of this microRNA. This result is consistent with the hypothesis that differences in the miR-21 expression existing in the normal tissue microenvironment are an important determinant of the HPV-induced carcinogenesis process.several(prenominal) studies have expanded the concept that inflammation is a critical component of tumour progression. Many cancers arise from sites of infection, chronic irritation and inflammation 13.Thomas X. Lu et al., identified an IL-12/IFN-gdependent pathway as the m ost prominent upregulated pathway in the lungs of OVAchallenged miR-21-/- mice compared with wild-type littermate controls, providing substantial evidence that this is the major pathway dysregulated in the miR-21deficient mice 14. As IL-12 is a major cytokine that regulates Th1 versus Th2 decisions primarily by inducing T cells to produce the Th1 cytokine IFN- 15, may be related to inflammation responsible for tumor progression.These facts that relate the miR-21 downregulation with change magnitude inflammation may explain our findings that tissues with lower miR-21 expression are more likely to develop a carcinogenic pathway.Pten has been verified as a miR-21 direct in pancreatic cancer, hepatocellular cancer and squamous cell carcinoma 16, 17. ()References1.Walboomers, J.M., et al., Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol, 1999. 189(1) p. 12-9.2.Watson, M., et al., Using population-based cancer register data to assess the burden of human papillomavirus-associated cancers in the United States overview of methods. Cancer, 2008. 113(10 Suppl) p. 2841-54.3.Major, T., et al., The characteristics of human papillomavirus DNA in head and neck cancers and papillomas. J Clin Pathol, 2005. 58(1) p. 51-5.4.Coussens, L.M., D. Hanahan, and J.M. Arbeit, Genetic predisposition and parameters of malignant progression in K14-HPV16 transgenic mice. Am J Pathol, 1996. 149(6) p. 1899-917.5.Masset, A., et al., Unimpeded skin carcinogenesis in K14-HPV16 transgenic mice deficient for plasminogen activator inhibitor. Int J Cancer, 2011. 128(2) p. 283-93.6.Stoler, A., et al., Use of monospecific antisera and cRNA probes to localize the major changes in keratin expression during normal and abnormal epidermal differentiation. J Cell Biol, 1988. 107(2) p. 427-46.7.Chen, C.Z., MicroRNAs as oncogenes and tumor suppressors. N Engl J Med, 2005. 353(17) p. 1768-71.8.Paranjape, T., F.J. Slack, and J.B. Weidhaas, MicroRNAs tools for cancer d iagnostics. Gut, 2009. 58(11) p. 1546-54.9.Chan, J.A., A.M. Krichevsky, and K.S. Kosik, MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells. Cancer Res, 2005. 65(14) p. 6029-33.10.Iyevleva, A.G., et al., High level of miR-21, miR-10b, and miR-31 expression in bilateral vs. unilateral breast carcinomas. Breast Cancer Res Treat, 2012. 131(3) p. 1049-59.11.Kulda, V., et al., Relevance of miR-21 and miR-143 expression in tissue samples of colorectal carcinoma and its liver metastases. Cancer Genet Cytogenet, 2010. 200(2) p. 154-60.12.Arbeit, J.M., et al., Progressive squamous epithelial neoplasia in K14-human papillomavirus type 16 transgenic mice. J Virol, 1994. 68(7) p. 4358-68.13.Coussens, L.M. and Z. Werb, Inflammation and cancer. Nature, 2002. 420(6917) p. 860-7.14.Lu, T.X., et al., MicroRNA-21 limits in vivo immune response-mediated activation of the IL-12/IFN-gamma pathway, Th1 polarization, and the severity of delayed-type hypersensitivity. J Immunol, 2011. 187(6) p. 3362-73.15.Gately, M.K., et al., The interleukin-12/interleukin-12-receptor system role in normal and pathologic immune responses. Annu Rev Immunol, 1998. 16 p. 495-521.16.Darido, C., et al., Targeting of the tumor suppressor GRHL3 by a miR-21-dependent proto-oncogenic network results in PTEN way out and tumorigenesis. Cancer Cell, 2011. 20(5) p. 635-48.17.Meng, F., et al., MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology, 2007. 133(2) p. 647-58.Fig. 1 Mice genotyping. The presence of integrated HPV was assessed by amplification of HPV-E2 (b) and HPV-E6 (c) genes by polymerase chain reaction methodology (PCR) in-house. Mouse--globin gene was used as endogenous control (a).Fig. 2 Normalized relative expression of miR-21 in transgenic mice (HPV+) and wild-type mice (HPV-), in ear (a) and chest tissue (b).1

Intravascular papillary endothelial hyperplasia

Intravascular papillose endothelial hyperplasiaIntravascular papillary endothelial hyperplasia, regarding a subjectINTRODUCTIONIntravascular papillary endothelial hyperplasia (IPEH) was first exposit as a malignant lesion by Pierre Masson in 1923(1), warning his histological similarity to angiosarcoma. Later, Henschen(2) described an intravascular endothelial proliferation that he interpreted like a reagent process due to inflammation and stasis vascular.We present a case of a patient diagnosed with intravascular papillary endothelial hyperplasia in nephritic vein.CASE REPORT61 years old male referred to our department with symptoms compatible with right renal colic. He has previous accounting of hypertension, obstructive sleep apnea syndrome and benign prostatic hyperplasia with alpha-blocker therapy.The patient relates intense pain in the right flank, sometimes radiating to ipsilateral inguinal region. On physical examination revealed pain with percussion in the right flank. Blood and urine analysis, abdominal radiography and ultrasound are requested, all of them normal. Due to the perseveration of symptoms, CT scan with contrast was requested (Fig. 1), in which a solid mass of 3.5 x 3 x 4 cm, lobulated and with manifold necrotic totality located at the right renal hilum infiltrates renal vein is seen . This mass is adjacent to lower portion of adrenal gland, kidney, and posterior portion of the duodenum, not clearly identifying dependency. To try to understand the dependence of this mass MRI is requested (Fig. 2), which describes hypointense on T1 and hyperintense on T2 tumor, with probable adrenal gland dependence.Suspecting adrenal injury, hormonal and metabolous study is carried on in blood and urine of 24 hours, all was normal. Scintigraphy was also performed with MIBG finding no enhancing lesions.Given previous findings, and the likelihood of adrenal malignance (nonfunctional), surgery is decided. By a subcostal laparotomy, a tumor about 4 cm , firmly adhered to the renal hilum is observed. Right radical nephrectomy is performed due to impossibility the tumor excision alone.Postoperative course without incidences and the patient was discharged on the fourth day.Microscopic examination of the surgical specimen revealed kidney and adrenal gland unaltered. Near of the renal hilum and, at least partially, contained within a dilated vein, thrombosis and endothelial reactive area proliferation (capillary and papillary), compatible with papillary endothelial hyperplasia intravascular (Fig 3).Currently the patient is reviewed e actually sestet months, with analytical tests and CT normal.DISCUSSIONEnzinger and Clearkin(3) suggested several morphological features that served in the differential diagnosis between IPEH and angiosarcoma, including intraluminal location of the lesion, absence of necrotic tissue and presence of thrombotic material, and proposed the intravascular papillary endothelial hyperplasia term.The etiology of I PEH is still unknown. Trauma has been proposed as the main etiological factor, but the traumatic history is exceptional. Several authors agree with the view of Clearkin and Salyer who believe it is due to an alteration in the thrombosis process, consisting of an unusual and peculiar way of organizing thrombus.( 3,6 )IPEH can occur at any age, more frequently in female. Most of the cases are localize to skin vessels, in head and neck, where they appear as small hard mass of bluish red coloration to the skin. Although there have also been little frequent locations as jejunum, central nervous system, liver, and lungs (4,5).Three forms have been described Primary, on dilated vascular lakes secondary or mixed, with preexisting vascular lesion as hemangioma , arteriovenous malformations or pyogenic granuloma and the third and less frequent, extravascular, resulting in a hematoma (7).The finding of IPEH in the renal vein is rare, there are very few cases reflected in the literature. The symptoms are variable, ranging from an incidental finding, asymptomatic, to colic pain and hematuria.In imaging, with the CT scan with contrast we can find a solid lesion with heterogeneous contrast enhancement, and in MRI a lesion hypointense on T1 and hyperintense on T2 , which does not exclude malignancy(8, 9).In a case reported(10), preoperative diagnosis was performed with excision of the lesion and kidney preservation. simply in most cases, it was impossible to rule out malignancy, and the radical surgery is usual, either for technic impossibility by proximity to the renal vessels, or the suspicion of malignancy. No metastases or malignant degeneration has been reported.Preoperative diagnosis of IPEH is difficult as there are no characteristic symptoms or an imaging test that allows decorous differential diagnosis.There are several neoplastic and non-neoplastic lesions that can be found in the renal hilum. Among them renal carcinoma, angiomyolipoma , schwannoma , myelolipoma , hemangiopericytoma , lymphoma, cysts, Castleman disease or lipomas.Therefore, this condition, although rare, must add up within the diagnostic possibilities, especially if their dependency or proximity to vessels is detected.CONFLICT OF INTERESTThe authors declare no conflict of interestREFERENCESMasson P. Hemangioendothelioma vegetant intra-vasculaire. Bull Soc Anat Paris 19239351723.Henschen F. Lendovasculite proliferante thrombopoietique dans la lesion vasculaire locale. Ann anat Pathol 19329113-21.Clearkin KP, Enzinger FM. Intravascular papillary endotelial hiperplasia. ARch Pathol Lab MEd 197610441-4.Johraku A, Miyanaga N, Sekido N, et al. A case of Intravascular Papillary endothelial Hyperplasia Arising from Renal sinus.Jpn J clin Oncol 1997 27(6) 433-36.Pelosi G, Sonzogni A, VIale G. Intravascular Papillary Endothelial Hyperplasia of the renal vein. Int J Surg Pathol 19(4) 518-20Salyer WR, Salyer DC. Intravascular angiomatosis development and distinction from agniosarcoma . Cancer. 1975 36 995-1001Hashimoto H, Daimaru Y, Enjoji M. Intravascular papillary endothelial hyperplasia. A clinicopathological study of 91 cases. AM J Dermatopathol. 1983 5539-46Kuo T, Sayers CP, Rosai J. Massons Vegetant intravascular hemangioendothelioma a lesion often mistaken for angiosarcoma. Cancer 1976 38 1227-36.Van den bogaert S, Boel K, Van Poppel H, et al. Massons tumour of the kidney. Cancer Imaging. 2002 2 116-9.Akhtar M, Aslam MAL-Mana H, et al. Intravascular Papillary endothelial Hyperpasia of Renal Vein. disgusting Pathol Lab med. 2005129 516-520LEGENDS TO FIGURESFig.1CT heterogeneous mass with necrotic core located in the right renal hilum.Fig. 2 T1-weighted MR image hypointense mass on the right renal hilum.Fig. 3 Histological specimen. Close to the renal vascular pole and partially including a dilated vein, an intravascular thrombus and a reactive endothelial proliferation zone (capillary and papillaroid) are observed.Fig. 4 great magnification of the prev ious image, which can be seen proliferation of papillary structures that tend to anastomose that are lined by a row of endothelial cells, centered on an axis of collagen and fibrin. No images of necrosis, atypia or mitosis (not characteristic of malignancy) were observed.

Halfway There :: essays papers

Halfway TherePeople. Droves of them, hustling off to their appointed provide with seventeen suitcases strapped to themselves alike(p) pack mules. All scowling, furrowing their brows. Hoping to get to where they want to go, and with all seventeen suitcases they came with. Me? I only had two bags, but one of them was large enough to be a body bag. Beside me was my cousin, a tall 16-year old, the jock type, with broad shoulders and pimple covered cheeks. He, of course, got stuck carrying my oversized bag. As we made our way past the ticket counter the automatonlike doors whooshed open, nearly sweeping us away in a blast of icy air. It was December in Vermont, which means one thing glacial. The kind of cold that hurt the skin, just breathing made people cough. As we zigged and zagged our way through the seething maze of bodies, we kept looking down feather at the rush information in my hands. Gate B-17, Im sure of it I said, none too convincingly apparently, for he kept r eading aloud the gates and their destinations. We reached a fairly quiet section of the airport, and all the sounds became subdued. It had the tone of voice of a library to it old, peaceful, and undisturbed. Is that our gate? I asked. He looked up at the monitor and said, escapism 182 to Pittsburgh, I think thats us. We stepped up to the woman behind the counter and handed her our tickets. She looked up at us, crows feet at the edges of her eyes, soft blond hair, and slightly handsome hands, a very attractive middle-aged woman. She had a soft voice, meek and unassuming. Right this way please, she said. We followed her down the steep incline to the plane. The closer we got the louder the noise became, leaden to deafen us. I could see the pilots huddled over the glowing panels in the cockpit, pressing a button here, turning a knob there, and making me feel secure just by looking busy. We stepped into the cabin and the sound became suddenly muffled, like someone thr ew a wool blanket over us. As we sidestepped down the aisle, I kept glancing over my shoulder into the cockpit, maybe out of curiosity as to what all the flashing buttons did, or maybe to continually remind myself that the pilots knew what they were doing.

Joseph Conrad :: essays research papers

Joseph ConradJoseph Conrad, born Tedor Josef Konrad Nalecz Korzeniowski, was born December 3, 1857 in a Russian-ruled province of Poland. His parents involvement in the Polish independence movement had them kicked out of northerly Russia in 1863. After his parents deaths, he moved in with relatives where he was often ill and received little schooling. At sixteen years of age, Conrad decided to drive a seaman and he joined the British merchant marines in 1878. His lack of speaking the English language did not discourage him. During his ten years of service, Conrad became a British citizen, traveled the western continents, developed into a Captain and learned the English language. Health problems caused his early retirement of the British merchant marines. In 1894, he started his career as a writer, using his seaman and sailing experience to write. In 1895, Conrads first novel, Almayers Folly, was published, with some of the book macrocosm written in the service. One year after his first novel, on March 24, 1896, Conrad married Jessie George. They had two children, Alfred Borys and John Alexander. In Kent, England, 1924, Joseph Conrad suffered a heart sharpshoot and died.For the rest of his writing career, Conrad would have difficulty being a writer. He found it difficult to write in the English language he conception it was a slow and unbearable torment. His novel Chance was his first financial success. His other novels and short stories that were published in the first ten years of the 20th century are thought of as his well-nigh important works. Throughout his career, Conrad examined the ridiculousness of living by a traditional code of conduct his novels apprize that the complication of the human spirit allows neither absolute loyalty to any ideal nor even to ones conscience. It is presented in all of his novels that failure is a fact of human existence. The novel Nostromo, which deals mainly with revolution, politics and financial manipulation, is b est at portraying failure. This novel is widely recognized as Conrads most ambitious novel.The Secret Agent A Simple Tale begins with Mr. Verlocs being summoned to a certain foreign embassy. Strolling down the street he did not tang like the agent provocateur he was meant to be. He owned an ambiguous little shop where his family lived close by. He often entertained anarchists from London that he had to keep and eye on and hid his actual occupation.

The Tragedy Of Hamlet :: Shakespeare Hamlet Essays

The Tragedy Of crossroadsHardship, unfortunately, is a part of everyones life. It is unavoidable, and in villages case he found out that bad share comes in commodious amounts at a time.Most people see bad luck as getting splashed by a cable car in the rain, or findingout that the idiots at McDonalds forgot the fries in your order. But Hamlet gota quadruple dose of bad luck. First his mystify was unjustly murdered. accordingly theghost of his father comes back and tells him that he is to avenge his death. Totop it all off Hamlet finds out that his mother has just hook up with his latefathers assassin. When Hamlet tries to expose the new king of killing his father,he is exiled to England because the other people thought that he was mentallyill. When Hamlet returns to Denmark he finds his transcendental love Ophelia existenceburied. Hamlet feels that he is living in a world of horror, and by the end ofthis miserably disheartening play, his fathers death is avenged, but at quite acost. Hamlet, Claudius, Gertrude, Ophelia, Laertes, and Polonius are all dead. Iwould have to say that all of the adversity in hamlets life had to have a great regard on his spirit. superstar can not go through life, and Hamlet had a short one,lose all of the people that you love and expect it to not dishearten you alittle. And in Hamlets case it pretty much drove him insane.The human spirit is a very fragile thing, and something as tragic as the deathof a loved one can damage it greatly. As in Hamlets case, when his father wasmurdered, this started a sort of devastating chain reaction of the psyche. Hestarted to go kooky, and it showed. The people around him started noticing thisdrastic change in his personality. But his insanity was most evident during theplay which he set up and called The Mousetrap. Hamlet sat fidgeting in hischair, staring at Caudius with accusing eyes. When his little trap had run itscourse, Claudius had just about been broken. Hamlet watched as Claudius l aborand chewed his nails. When Claudius could take no more he stumbled out of theroom and into the streets where hamlet proceeded to follow, dancing andscreaming like a mad man. Hamlet could take no more of this harrowing life,watching his mother hang off of the man who murdered his father. His spirit hadbeen irreversibly damaged, and insanity was the price.This play is a classic font of one of the greatest, if not the greatestThe Tragedy Of Hamlet Shakespeare Hamlet EssaysThe Tragedy Of HamletHardship, unfortunately, is a part of everyones life. It is unavoidable, and inHamlets case he found out that bad luck comes in colossal amounts at a time.Most people see bad luck as getting splashed by a car in the rain, or findingout that the idiots at McDonalds forgot the fries in your order. But Hamlet gota quadruple dose of bad luck. First his father was unjustly murdered. Then theghost of his father comes back and tells him that he is to avenge his death. Totop it all off Hamlet find s out that his mother has just married his latefathers assassin. When Hamlet tries to expose the new king of killing his father,he is exiled to England because the other people thought that he was mentallyill. When Hamlet returns to Denmark he finds his secret love Ophelia beingburied. Hamlet feels that he is living in a world of horror, and by the end ofthis miserably disheartening play, his fathers death is avenged, but at quite acost. Hamlet, Claudius, Gertrude, Ophelia, Laertes, and Polonius are all dead. Iwould have to say that all of the adversity in hamlets life had to have a greataffect on his spirit. One can not go through life, and Hamlet had a short one,lose all of the people that you love and expect it to not dishearten you alittle. And in Hamlets case it pretty much drove him insane.The human spirit is a very fragile thing, and something as tragic as the deathof a loved one can damage it greatly. As in Hamlets case, when his father wasmurdered, this started a sort of de vastating chain reaction of the psyche. Hestarted to go nuts, and it showed. The people around him started noticing thisdrastic change in his personality. But his insanity was most evident during theplay which he set up and called The Mousetrap. Hamlet sat fidgeting in hischair, staring at Caudius with accusing eyes. When his little trap had run itscourse, Claudius had just about been broken. Hamlet watched as Claudius sweatand chewed his nails. When Claudius could take no more he stumbled out of theroom and into the streets where hamlet proceeded to follow, dancing andscreaming like a mad man. Hamlet could take no more of this torturous life,watching his mother hang off of the man who murdered his father. His spirit hadbeen irreversibly damaged, and insanity was the price.This play is a classic example of one of the greatest, if not the greatest

Portrait of a Lady - From Novel to Film Essay -- Movie Film Essays

Portrait of a Lady - From Novel to Film Jane Campions most recent exact, Portrait of a Lady (1996), offers a distinct departure from her previous work, The easygoing (1993), with which some critics have found fault. In her 1998 article, for example, while commending Campion for introducing two characters able to desert the gender warfare that characterizes Western culture, Diane Long Hoeveler criticizes Campion for celebrating marriage, the idea that women cannot survive with go forth a man at the center of their lives (Hoeveler 110, 114). Second, she asserts that while Campion toys with feminist issues and images, Piano is Aromantic and escapist, with Adas decision to be reborn with Baines a step hardly worthy of the serious feminist issues that Campion seems to be raising in the film (Hoeveler 114). Finally, she points out that Campion is heavily indebted to a 1920s work, The Story of a New Zealand River by Jane Mander. Partly as a consequence of not acknowledging this deb t, the film has conflicting sources, Campions rather permissive twentieth century script about adultery, superimposed on Manders original, in which the Victorian heroine is not united sexually with her raw sienna until after her husbands death. Enacting a basically contemporary drama in anachronistic costumes and setting, Hoeveler says the film contains gaps, ...fissures we sense while viewing it (Hoeveler 114). For example, how likely is it, she asks, that an 1850s heroine would conduct an adulterous contest? In (Re)Visioning the Gothic (1998), Cyndy Hendershot echoes this view, calling Baines, the films nontraditional male (Harvey Keitel), a deus ex machina, a fairy-tale character, an imaginary resolution to two real problems, on the one extend the castratio... ..., Campion breaks his barrier of reticence about sex, money and behavior and delivers the facts straight. Hardly faithful to him as she is, though, Jane Campions work is itself made possible by the original master, a tomic number 1 James. Sources CitedBluestone, George. Novels Into Film. California UP, 1971.Campion, Jane. The Piano. London Bloomsbury, 1993.Dapkus, Jeanne R. Sloughing off the Burdens. Film literature Quarterly 25.3 (1997) 177-187.Giannetti, Louis. Understanding Movies. New Jersey Prentice-Hall, 1972.Hendershot, Cyndy. (Re)Visioning the Gothic. Film Literature Quarterly 26.2 (1998) 97-108.Hoeveler, Diane Long. Silence, Sex, and Feminism. Film Literature Quarterly 26.2 (1998) 109-116.James, Henry. The Portrait of a Lady. 1881. New York Random House, 1996.Jones, Laura. The Portrait of a Lady. New York Penguin, 1996.