18 results on '"Saria EA"'
Search Results
2. Modeling vitamin D insufficiency and moderate deficiency in adult mice via dietary cholecalciferol restriction.
- Author
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Mallya SM, Corrado KR, Saria EA, Yuan FF, Tran HQ, Saucier K, Atti E, Tetradis S, and Arnold A
- Subjects
- Animals, Cholecalciferol administration & dosage, Diet, Disease Models, Animal, Female, Male, Mice, Calcifediol blood, Cholecalciferol pharmacology, Vitamin D Deficiency blood, Vitamins metabolism
- Abstract
Purpose: We sought to develop and characterize a model of human vitamin D nutritional insufficiency/deficiency in the adult mouse, which could have broad utility in examining health consequences of this common condition., Methods: Adult mice were fed diets containing cholecalciferol contents of 0.05 IU/g, 0.25 IU/g, 0.5 IU/g or 1.5 IU/g for four months. We studied induction of steady-state vitamin D insufficiency, and its consequences on primary cholecalciferol metabolite levels, calcium homeostasis, parathyroid physiology, and bone morphology., Results: All diets were well tolerated, without adverse effects on body weight. Diets containing 0.05 IU/g and 0.25 IU/g cholecalciferol significantly lowered serum 25-hydroxyvitamin D levels (median 25OHD, 10.5 ng/ml, and 21.6 ng/ml, respectively), starting as early as one month following initiation of the diets, maintained through the four-month experimental period. The 0.05 IU/g diet significantly decreased 1,25-dihydroxyvitamin D (1,25OH
2 D) levels (median, 78 pg/ml). Despite these decreased 25OHD and 1,25OH2 D levels, the diets did not alter parathyroid gland morphology or parathyroid cell proliferation. There were no statistical differences in the serum total calcium and serum PTH levels among the various dietary groups. Furthermore, the 0.05 IU/g diet did not cause any alterations in the cortical and trabecular bone morphology, as determined by microCT., Conclusions: The dietary manipulations yielded states of vitamin D insufficiency or modest deficiency in adult mice, with no overtly detectable impact on parathyroid and bone physiology, and calcium homeostasis. This model system may be of value to study health effects of vitamin D insufficiency/deficiency especially on extraskeletal phenotypes such as cancer susceptibility or immune function.- Published
- 2016
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3. Regulation of Osteoblast Migration Involving Receptor Activator of Nuclear Factor-kappa B (RANK) Signaling.
- Author
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Golden D, Saria EA, and Hansen MF
- Subjects
- Bone Neoplasms pathology, Bone Remodeling, Cell Line, Tumor, Extracellular Signal-Regulated MAP Kinases metabolism, Humans, Mesenchymal Stem Cells metabolism, Mitogen-Activated Protein Kinase Kinases metabolism, Osteoblasts pathology, Osteosarcoma pathology, Phosphatidylinositol 3-Kinase metabolism, Phosphorylation, Proto-Oncogene Proteins c-akt metabolism, Transcription Factor RelA metabolism, Bone Neoplasms metabolism, Chemotaxis, Osteoblasts metabolism, Osteosarcoma metabolism, Receptor Activator of Nuclear Factor-kappa B metabolism, Signal Transduction
- Abstract
Bone remodeling requires osteoclast activation, resorption, and reversal, prior to osteoblast migration into the bone pit. The Receptor Activator of NF-κB (RANK) signaling pathway plays an important role in bone remodeling. Two components of the RANK signaling pathway, RANK Ligand (RANKL) and the decoy receptor Osteoprotegerin (OPG), are expressed predominantly on the surface of osteoblasts, while RANK is principally expressed on the surface of osteoclasts. However, RANK has also been reported to be expressed on the surface of osteoblasts and osteosarcoma tumor cells. Treatment with soluble RANKL (sRANKL) of both normal osteoblasts and osteosarcoma tumor cells activated phosphorylation of ERK, p38(MAPK) , Akt, and p65(NF-κB). However, modified Boyden chamber assays and wound repair assays showed differential response to sRANKL-induced chemotactic migration in normal osteoblasts and osteosarcoma tumor cells. In contrast to previously published results, both normal osteoblasts and osteosarcoma tumor cells responded to sRANKL-induced chemotactic migration but the normal osteoblasts did so only in the presence of an ERK pathway inhibitor. For both normal and tumor cells, the chemotactic response could be blocked by inhibiting the PI3K/Akt or p65(NF-κB) pathway. Response to sRANKL in normal and tumor cells suggests a role for RANK/ERK-mediated signaling in normal osteoblasts chemotactic migration during bone remodeling that is altered or lost during osteosarcoma tumorigenesis., (© 2015 Wiley Periodicals, Inc.)
- Published
- 2015
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4. Kinase-independent role of cyclin D1 in chromosomal instability and mammary tumorigenesis.
- Author
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Casimiro MC, Di Sante G, Crosariol M, Loro E, Dampier W, Ertel A, Yu Z, Saria EA, Papanikolaou A, Li Z, Wang C, Addya S, Lisanti MP, Fortina P, Cardiff RD, Tozeren A, Knudsen ES, Arnold A, and Pestell RG
- Subjects
- Amino Acid Substitution, Aneuploidy, Animals, Catalytic Domain genetics, Cell Transformation, Neoplastic genetics, Cells, Cultured, Centrosome ultrastructure, Chromosomal Instability genetics, Cyclin D1 deficiency, Cyclin D1 genetics, Female, Fibroblasts, Genes, bcl-1, Humans, Mammary Tumor Virus, Mouse physiology, Mice, Mice, Knockout, Mice, Transgenic, Mutation, Piperazines pharmacology, Pyridines pharmacology, Recombinant Fusion Proteins metabolism, Spindle Apparatus ultrastructure, Transduction, Genetic, Adenocarcinoma genetics, Cyclin D1 physiology, Mammary Neoplasms, Experimental genetics
- Abstract
Cyclin D1 is an important molecular driver of human breast cancer but better understanding of its oncogenic mechanisms is needed, especially to enhance efforts in targeted therapeutics. Currently, pharmaceutical initiatives to inhibit cyclin D1 are focused on the catalytic component since the transforming capacity is thought to reside in the cyclin D1/CDK activity. We initiated the following study to directly test the oncogenic potential of catalytically inactive cyclin D1 in an in vivo mouse model that is relevant to breast cancer. Herein, transduction of cyclin D1(-/-) mouse embryonic fibroblasts (MEFs) with the kinase dead KE mutant of cyclin D1 led to aneuploidy, abnormalities in mitotic spindle formation, autosome amplification, and chromosomal instability (CIN) by gene expression profiling. Acute transgenic expression of either cyclin D1(WT) or cyclin D1(KE) in the mammary gland was sufficient to induce a high CIN score within 7 days. Sustained expression of cyclin D1(KE) induced mammary adenocarcinoma with similar kinetics to that of the wild-type cyclin D1. ChIP-Seq studies demonstrated recruitment of cyclin D1(WT) and cyclin D1(KE) to the genes governing CIN. We conclude that the CDK-activating function of cyclin D1 is not necessary to induce either chromosomal instability or mammary tumorigenesis.
- Published
- 2015
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5. ChIP sequencing of cyclin D1 reveals a transcriptional role in chromosomal instability in mice.
- Author
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Casimiro MC, Crosariol M, Loro E, Ertel A, Yu Z, Dampier W, Saria EA, Papanikolaou A, Stanek TJ, Li Z, Wang C, Fortina P, Addya S, Tozeren A, Knudsen ES, Arnold A, and Pestell RG
- Subjects
- Animals, Binding Sites, Breast Neoplasms genetics, Cell Line, Tumor, Chromatin Immunoprecipitation, Chromosome Aberrations, Female, Fibroblasts metabolism, Gene Expression Regulation, Neoplastic, Genome-Wide Association Study, Humans, Karyotyping, Mice, Mice, Transgenic, Transcription, Genetic, Chromosomal Instability, Cyclin D1 genetics
- Abstract
Chromosomal instability (CIN) in tumors is characterized by chromosomal abnormalities and an altered gene expression signature; however, the mechanism of CIN is poorly understood. CCND1 (which encodes cyclin D1) is overexpressed in human malignancies and has been shown to play a direct role in transcriptional regulation. Here, we used genome-wide ChIP sequencing and found that the DNA-bound form of cyclin D1 occupied the regulatory region of genes governing chromosomal integrity and mitochondrial biogenesis. Adding cyclin D1 back to Ccnd1(-/-) mouse embryonic fibroblasts resulted in CIN gene regulatory region occupancy by the DNA-bound form of cyclin D1 and induction of CIN gene expression. Furthermore, increased chromosomal aberrations, aneuploidy, and centrosome abnormalities were observed in the cyclin D1-rescued cells by spectral karyotyping and immunofluorescence. To assess cyclin D1 effects in vivo, we generated transgenic mice with acute and continuous mammary gland-targeted cyclin D1 expression. These transgenic mice presented with increased tumor prevalence and signature CIN gene profiles. Additionally, interrogation of gene expression from 2,254 human breast tumors revealed that cyclin D1 expression correlated with CIN in luminal B breast cancer. These data suggest that cyclin D1 contributes to CIN and tumorigenesis by directly regulating a transcriptional program that governs chromosomal stability.
- Published
- 2012
- Full Text
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6. Tissue-specific regulatory regions of the PTH gene localized by novel chromosome 11 rearrangement breakpoints in a parathyroid adenoma.
- Author
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Mallya SM, Wu HI, Saria EA, Corrado KR, and Arnold A
- Subjects
- Alleles, Binding Sites, Conserved Sequence genetics, Cyclin D1 genetics, Cyclin D1 metabolism, Gene Expression Regulation, Neoplastic, Genetic Loci genetics, Humans, In Situ Hybridization, Fluorescence, Interphase, Male, Organ Specificity genetics, Reverse Transcriptase Polymerase Chain Reaction, Species Specificity, Transcription Factors, Chromosome Breakpoints, Chromosomes, Human, Pair 11 genetics, Gene Rearrangement genetics, Parathyroid Hormone genetics, Parathyroid Neoplasms genetics, Regulatory Sequences, Nucleic Acid genetics
- Abstract
Parathyroid adenomas can contain clonal rearrangements of chromosome 11 that activate the cyclin D1 oncogene through juxtaposition with the PTH gene. Here we describe such a chromosomal rearrangement whose novel features provide clues to locating elusive cis-regulatory elements in the PTH gene and also expand the physical spectrum of pathogenetic breakpoints in the cyclin D1 gene region. Southern blot analyses of the parathyroid adenoma revealed rearrangement in the PTH gene locus. Analysis of rearranged DNA clones that contained the breakpoint, obtained by screening a tumor genomic library, pinpointed the breakpoint in the PTH locus 3.3 kb upstream of the first exon. Accordingly, highly conserved distal elements of the PTH 5' regulatory region were rearranged at the breakpoint approximately 450 kb upstream of the cyclin D1 oncogene, resulting in overexpression of cyclin D1 mRNA. Thus, PTH-cyclin D1 gene rearrangement breakpoints in parathyroid tumors can be located far from those previously recognized. In addition to expanding the molecular spectrum of pathogenetic chromosomal lesions in this disease, features of this specific rearrangement reinforce the existence of one or more novel cis-enhancer/regulatory elements for PTH gene expression and narrow their location to a 1.7-kb DNA segment in the distal PTH promoter., (Copyright © 2010 American Society for Bone and Mineral Research.)
- Published
- 2010
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7. Comprehensive mutational analysis and mRNA isoform quantification of TP63 in normal and neoplastic human prostate cells.
- Author
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Parsons JK, Saria EA, Nakayama M, Vessella RL, Sawyers CL, Isaacs WB, Faith DA, Bova GS, Samathanam CA, Mitchell R, and De Marzo AM
- Subjects
- Adenocarcinoma pathology, Cell Line, Tumor, Exons genetics, Humans, Male, Prostate pathology, Prostatic Neoplasms pathology, Protein Isoforms metabolism, Transcription Factors, Transplantation, Heterologous, Adenocarcinoma metabolism, DNA Mutational Analysis, Prostate metabolism, Prostatic Neoplasms metabolism, RNA, Messenger metabolism, Trans-Activators metabolism, Tumor Suppressor Proteins metabolism
- Abstract
Background: The role of TP63 in cancer remains controversial since both oncogenic and tumor suppressive actions have been reported. p63 protein is found in the nuclei of basal cells of the normal prostate, yet it is absent in the vast majority of prostate cancer nuclei. Since a complex array of TP63 mRNA transcripts encode polypeptides with distinct functional properties, it is important to determine which forms are expressed in normal and prostate cancer tissue., Methods: We used real-time RT-PCR to distinguish TP63 mRNA isoforms in prostate cancer cell lines (n = 7), and samples from prostate cancer patients. We sequenced all TP63 exons from 20 primary tumors, 20 metastases, 28 tumor xenografts, and 7 prostate cancer cell lines., Results: TP63 mRNA isoforms were present in all tumors, albeit at levels lower than in normal prostate. The most abundant N-terminal variant was DeltaN; the most abundant C-terminal variant was the alpha form. The prostate tumor cell line CWR22Rv1 contained a single G to T substitution in exon 8 that is identical to a dominant-negative DNA binding inactivation mutation occurring in patients with a congenital TP63 deficiency syndrome. One patient tumor contained a somatic mutation in exon 11., Conclusions: The pattern of TP63 mRNA expression in normal prostate tissue is retained in reduced amounts in prostate cancer, and a potentially functional TP63 mutation was identified in one prostate tumor. Thus, if TP63 is a prostate cancer gene it likely functions as a tumor suppressor. Further study of the role of TP63 isoforms in regulating stem cell functions of normal and neoplastic prostate epithelial cells is needed. Prostate 69:559-569, 2009. (c) 2009 Wiley-Liss, Inc.
- Published
- 2009
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8. Somatic mutations in SQSTM1 detected in affected tissues from patients with sporadic Paget's disease of bone.
- Author
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Merchant A, Smielewska M, Patel N, Akunowicz JD, Saria EA, Delaney JD, Leach RJ, Seton M, and Hansen MF
- Subjects
- Adolescent, Alleles, Base Sequence, Bone and Bones metabolism, Case-Control Studies, DNA Mutational Analysis, Exons genetics, Gene Expression Regulation, Neoplastic, Humans, Microdissection, Molecular Sequence Data, Osteitis Deformans blood, Osteitis Deformans complications, Osteitis Deformans pathology, Osteosarcoma complications, Osteosarcoma genetics, Osteosarcoma pathology, Sequestosome-1 Protein, Viral Proteins, Adaptor Proteins, Signal Transducing genetics, Bone and Bones pathology, Mutation genetics, Osteitis Deformans genetics
- Abstract
Paget's disease of bone (PDB) is a focal disorder of bone remodeling that leads to overgrowth of affected bone, with rare progression to osteosarcoma. Extensive studies of familial PDB showed that a majority of cases harbor germline mutations in the Sequestosome1 gene (SQSTM1). In contrast, little is known about the mutational status of SQSTM1 in sporadic PDB. We hypothesized that somatic SQSTM1 mutations might occur in the affected tissues of sporadic PDB and pagetic osteosarcoma. We used laser capture microdissection to capture homogeneous populations of cells from the affected bone or tumor of patients with sporadic PDB or pagetic osteosarcoma, respectively. DNA from these samples and appropriate controls was used for sequence analysis and allelic discrimination analysis. Two of five patients with sporadic PDB had SQSTM1(C1215T) mutations detected in their affected bone but not in their blood samples, indicating a somatic origin of the mutations. Samples from three of five sporadic pagetic osteosarcoma patients had the SQSTM1(C1215T) mutation, whereas the normal adjacent tissue from two of these tumors clearly lacked the mutation, again indicating an occurrence of somatic events. No SQSTM1 mutations were found in primary adolescent osteosarcomas. The discovery of somatic SQSTM1 mutations in sporadic PDB and pagetic osteosarcoma shows a role for SQSTM1 in both sporadic and inherited PDB. The discovery of somatically acquired mutations in both the diseased bone and tumor samples suggests a paradigm shift in our understanding of this disease.
- Published
- 2009
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9. PHC3, a component of the hPRC-H complex, associates with E2F6 during G0 and is lost in osteosarcoma tumors.
- Author
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Deshpande AM, Akunowicz JD, Reveles XT, Patel BB, Saria EA, Gorlick RG, Naylor SL, Leach RJ, and Hansen MF
- Subjects
- Base Sequence, DNA, DNA-Binding Proteins genetics, Humans, Loss of Heterozygosity, Nuclear Proteins, Polycomb Repressive Complex 1, Protein Binding, Tumor Cells, Cultured, DNA-Binding Proteins metabolism, E2F6 Transcription Factor metabolism, Osteosarcoma metabolism, Resting Phase, Cell Cycle
- Abstract
Polyhomeotic-like 3 (PHC3) is a ubiquitously expressed member of the polycomb gene family and part of the human polycomb complex hPRC-H. We found that in normal cells PHC3 associated with both hPRC-H complex components and with the transcription factor E2F6. In differentiating and confluent cells, PHC3 and E2F6 showed nuclear colocalization in a punctate pattern that resembled the binding of polycomb bodies to heterochromatin. This punctate pattern was not seen in proliferating cells suggesting that PHC3 may be part of an E2F6-polycomb complex that has been shown to occupy and silence target promoters in G(0). Previous loss of heterozygosity (LoH) analyses had shown that the region containing PHC3 underwent frequent LoH in primary human osteosarcoma tumors. When we examined normal bone and human osteosarcoma tumors, we found loss of PHC3 expression in 36 of 56 osteosarcoma tumors. Sequence analysis revealed that PHC3 was mutated in nine of 15 primary osteosarcoma tumors. These findings suggest that loss of PHC3 may favor tumorigenesis by potentially disrupting the ability of cells to remain in G(0).
- Published
- 2007
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10. Expression of secretory phospholipase A2 in colon tumor cells potentiates tumor growth.
- Author
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Belinsky GS, Rajan TV, Saria EA, Giardina C, and Rosenberg DW
- Subjects
- Arachidonic Acid pharmacology, Base Sequence, Butyric Acid pharmacology, Cell Line, Tumor, Colonic Neoplasms enzymology, DNA Primers, Group II Phospholipases A2, Humans, Phospholipases A2, RNA, Messenger genetics, Tumor Necrosis Factor-alpha pharmacology, Cell Division drug effects, Colonic Neoplasms pathology, Phospholipases A metabolism
- Abstract
Secretory phospholipase A2 (sPLA2-IIA) has been shown to attenuate intestinal tumorigenesis in Apc(Min) mice, demonstrating that it is a tumor modifier. To further explore the actions of sPLA2-IIA in tumorigenesis, sPLA2-IIA was overexpressed in two cell lines where it is normally absent, the murine colon tumor cell line AJ02nm0, and human colon carcinoma cell line HCT-116. Two allelic variants of sPLA2-IIA were tested in this study; sPLA2-IIA(AKR) and sPLA2-IIA(SWR), which are derived from AKR/J and SWR/J mice, respectively, and differ by a single amino acid at position 63 in the calcium- and receptor-binding domain. There was no change in cell-doubling time for either allele when compared to vector controls. Furthermore, sodium butyrate and arachidonic acid (AA)-induced cell death were unchanged in control and transfected cells. Addition of the sPLA2 substrate, palmitoyl-arachidonoyl-phosphatidic acid (PAPA), to AJ02nm0 cells resulted in a modest (12%-24%), but significant (P < 0.01), inhibition of growth that was dependent on sPLA2-IIA expression. However, when AJ02nm0 and HCT-116 cells were injected subcutaneously (sc) into nude mice, Pla2g2a expression resulted in a 2.5-fold increase in tumor size. In addition, sPLA2-IIA expressing HCT-116 tumors were found to be more infiltrative than controls. We conclude that the ability of sPLA2-IIA to slow tumor cell growth is dependent upon the availability of substrate, and that in some instances sPLA2-IIA may actually enhance tumor growth. Mechanisms that may account for differences between the tumor explant model versus the Apc(Min) model of intestinal cancer are discussed., (2006 Wiley-Liss, Inc.)
- Published
- 2007
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11. Cyclooxygenase-2 is up-regulated in proliferative inflammatory atrophy of the prostate, but not in prostate carcinoma.
- Author
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Zha S, Gage WR, Sauvageot J, Saria EA, Putzi MJ, Ewing CM, Faith DA, Nelson WG, De Marzo AM, and Isaacs WB
- Subjects
- Atrophy enzymology, Blotting, Western, Cyclooxygenase 2, Disease Progression, Epithelium enzymology, Epithelium pathology, Humans, Immunohistochemistry, Isoenzymes genetics, Male, Membrane Proteins, Prostaglandin-Endoperoxide Synthases genetics, Prostatic Neoplasms genetics, Prostatic Neoplasms pathology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Reverse Transcriptase Polymerase Chain Reaction, Stromal Cells enzymology, Stromal Cells pathology, Tumor Cells, Cultured, Up-Regulation, Isoenzymes biosynthesis, Prostaglandin-Endoperoxide Synthases biosynthesis, Prostate pathology, Prostatic Neoplasms enzymology
- Abstract
Cyclooxygenase-2 (COX-2) is the inducible isoform of the rate-limiting enzymes that convert arachidonic acid to proinflammatory prostaglandins as well as a primary target for nonsteroidal anti-inflammatory drugs. Accumulating evidence suggests that up-regulation of COX-2 is associated with carcinogenesis in multiple organ systems including the large bowel, lung, breast, and prostate. In this report, we examine the expression of COX-2 protein and mRNA in prostate tissue containing various lesions and in prostate cancer cell lines. In the cell lines, LNCaP, DU145, PC-3, and TSU, COX-2 protein expression was undetectable under basal conditions but could be induced transiently by phorbol ester treatment in PC-3 and TSU cells, but not in DU145 and LNCaP cells. Immunohistochemical analysis of 144 human prostate cancer cases suggested that, in contrast to several previous reports, there was no consistent overexpression of COX-2 in established prostate cancer or high-grade prostatic intraepithelial neoplasia, as compared with adjacent normal prostate tissue. Positive staining was seen only in scattered cells (<1%) in both tumor and normal tissue regions but was much more consistently observed in areas of proliferative inflammatory atrophy, lesions that have been implicated in prostatic carcinogenesis. Staining was also seen at times in macrophages. Western blotting and quantitative RT-PCR analyses confirmed these patterns of expression. These results suggest that if nonsteroidal anti-inflammatory drugs are indeed chemopreventive and/or chemotherapeutic for prostate cancer, their effects are likely to be mediated by modulating COX-2 activity in non-PCa cells (either inflammatory cells or atrophic epithelial cells) or by affecting a COX-2-independent pathway.
- Published
- 2001
12. Intrauterine epidermal necrosis.
- Author
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Allee JE, Saria EA, Rosenblum D, Prose NS, Prieto VG, and Shea CR
- Subjects
- Fatal Outcome, Female, Herpes Simplex complications, Humans, Infant, Newborn, Necrosis, Skin Diseases complications, Epidermis pathology, Infant, Premature, Diseases pathology, Skin Diseases pathology
- Abstract
Background: Epidermal necrosis in a neonate is an uncommon event with a variety of potential cases., Result: We report a case of intrauterine epidermal necrosis in a preterm infant, with death occurring soon after birth. The histopathology of the denuded skin revealed full-thickness epidermal necrosis and calcification within both the epidermis and follicular structures., Conclusion: We believe this represents the fourth reported case of lethal intrauterine epidermal necrosis and follicular calcification.
- Published
- 2001
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13. Prognostic value of MIB-1 in advanced ovarian carcinoma as determined using automated immunohistochemistry and quantitative image analysis.
- Author
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Layfield LJ, Saria EA, Berchuck A, Dodge RK, Thompson JK, Conlon DH, and Kerns BJ
- Subjects
- Antibodies, Monoclonal, Antigens, Nuclear, Cell Cycle, DNA, Neoplasm analysis, DNA, Neoplasm genetics, Female, Humans, Immunohistochemistry, Ki-67 Antigen, Middle Aged, Ovarian Neoplasms mortality, Ovarian Neoplasms pathology, Paraffin Embedding, Ploidies, Prognosis, Survival Rate, Image Processing, Computer-Assisted, Nuclear Proteins analysis, Ovarian Neoplasms diagnosis
- Abstract
Background and Objectives: The monoclonal antibody MIB-1 is an immunohistochemical marker reacting most strongly with cells in late S phase, G2, and M portions of the cell cycle. This antibody, reactive in formalin-fixed, paraffin-embedded tissue, allows the quantitation of a proliferation index (PI) in both current clinical cases and archival material using a computerized image analyzer (CIA)., Methods: Since many laboratories make use of automated immunohistochemistry (AIH), this study was performed to explore the technical feasibility of using AIH (Ventana ES 320) in combination with CIA (CAS 200) to evaluate MIB-1 PI as a prognostic marker as assessed by overall survival in 50 archival (formalin-fixed, paraffin-embedded), advanced stage primary ovarian carcinomas., Results: Exploratory methods confirmed that 15% was a cutpoint that could dichotomize these 50 patients into two prognostic groups based on overall survival. The median survival of patients whose carcinoma had a high MIB-1 expression (> or = 15%) was 16 months compared with 30 months in the patients whose tumors demonstrated low MIB-1 expression (< 15%, P = 0.01). After adjustment for age, MIB-1 retained its prognostic significance (P = 0.02). Patients 60 years and older had shorter survival than younger patients (P = 0.06), but these two groups did not differ with respect to PI (P = 0.76). Those patients with a negative second look laparotomy had a longer median survival of 70 months compared with 18.5 months in patients with a positive second look (P < 0.001); the median PIs were 17% and 27%, respectively (P = 0.36). There were no significant relationships between clinical stage, nuclear grade, DNA ploidy, p53, and either survival or PI., Conclusions: In this study, we concluded that the combination of AIH and CIA yielded a reliable quantitation of MIB-1 proliferative index and that this proliferative marker had prognostic significance in late stage ovarian carcinoma. Further studies in a larger group of patients are needed to confirm the relationship between proliferation index and other known clinicopathologic and genetic variables.
- Published
- 1997
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14. Estrogen and progesterone receptor status determined by the Ventana ES 320 automated immunohistochemical stainer and the CAS 200 image analyzer in 236 early-stage breast carcinomas: prognostic significance.
- Author
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Layfield LJ, Saria EA, Conlon DH, and Kerns BJ
- Subjects
- Aged, Automation, Biopsy, Needle, Carcinoma, Ductal, Breast pathology, Charcoal, Coloring Agents, Dextrans, Female, Fixatives, Formaldehyde, Humans, Middle Aged, Neoplasm Staging, Paraffin Embedding, Prognosis, Survival Rate, Breast Neoplasms pathology, Carcinoma pathology, Image Processing, Computer-Assisted instrumentation, Immunohistochemistry instrumentation, Receptors, Estrogen analysis, Receptors, Progesterone analysis
- Abstract
The quantitation of estrogen and progesterone receptors (ER and PgR) has become the standard of care in the evaluation of patients with primary breast carcinoma. It has been demonstrated that ER and PgR detected by immunohistochemical methods in formalin-fixed paraffin-embedded tissue can be quantified by computerized image analysis. In this study, ER and PgR levels were determined by using an automated immunochemistry stainer (Ventana ES 320) and an image analyzer (CAS 200) in a series of 236 patients with stage I/II carcinoma of the breast. The degree of correlation of the ER and PgR levels determined by the dextran-coated charcoal method (DCC) with image analysis quantitation was high (r=0.75). The agreement between both methods was 77% for ER and 73% for PgR. Hormone receptor levels were correlated with prognosis as determined by overall survival. An ER level of 30 fmol/mg as determined by image analysis was established to stratify the patient population most effectively into favorable and unfavorable prognostic groups (P=0.003). An ER level of 20 fmol/mg for prognostic stratification reached statistical significance (P=0.03). The DCC method was not able to stratify the patients into prognostic groups at the traditionally accepted cutpoint of 10 fmol/mg (P=0.52). We conclude that when used in combination, automated immunohistochemistry and quantitative image analysis offer a favorable alternative to the DCC method in assessment of ER and PgR status in human mammary carcinoma. In addition, quantitative immunocytochemistry techniques may prove superior to the DCC method in specimens in which there is limited tumor volume (including fine-needle aspirates), stroma-rich tumors, and early-stage lesions including intraductal carcinoma.
- Published
- 1996
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15. Evaluation of anti-CD5 ricin A chain immunoconjugate for prevention of acute graft-vs.-host disease after HLA-identical marrow transplantation.
- Author
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Przepiorka D, LeMaistre CF, Huh YO, Luna M, Saria EA, Brown CT, and Champlin RE
- Subjects
- Acute Disease, Adult, Antibodies, Monoclonal adverse effects, CD5 Antigens immunology, Female, Humans, Immunotoxins adverse effects, Male, Middle Aged, Pilot Projects, Ricin adverse effects, T-Lymphocyte Subsets immunology, Antibodies, Monoclonal therapeutic use, Bone Marrow Transplantation methods, Graft vs Host Disease prevention & control, Immunotoxins therapeutic use, Ricin therapeutic use
- Abstract
Anti-CD5 ricin A chain immunoconjugate (XZ-CD5) is an immunotoxin that inhibits proliferative and cytotoxic responses to alloantigen in vitro and has activity in the treatment of acute graft-vs.-host disease (GVHD). To determine if XZ-CD5 could be used to prevent acute GVHD, 11 adult recipients of HLA-identical allogeneic marrow received XZ-CD5 0.1 mg kg-1 day-1 intravenously with high-dose methyl-prednisolone for 10, 14 or 17 doses early post-transplant. Six additional patients received 17 doses of XZ-CD5 and cyclosporine (CSA). All patients engrafted. Severe capillary leak syndrome was the most common serious toxicity and occurred more frequently in patients receiving CSA (5/5 vs. 3/11, P = 0.03). All evaluable patients developed acute GVHD; 88% had grade II-IV GVHD. Flow cytometric analysis demonstrated a substantial number of circulating CD5+ and CD3+ lymphocytes during and early after administration of XZ-CD5. These results suggest that the immunotoxin did not eliminate alloreactive T cells in this setting.
- Published
- 1994
16. Effects of an anti-CD5 immunoconjugate (CD5-plus) in recent onset type I diabetes mellitus: a preliminary investigation. The CD5 Diabetes Project Team.
- Author
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Skyler JS, Lorenz TJ, Schwartz S, Eisenbarth GS, Einhorn D, Palmer JP, Marks JB, Greenbaum C, Saria EA, and Byers V
- Subjects
- Adolescent, Adult, Antibodies, Monoclonal, Antigens, CD blood, C-Peptide blood, C-Peptide metabolism, CD5 Antigens, Diabetes Mellitus, Type 1 blood, Diabetes Mellitus, Type 1 immunology, Female, Humans, Immunotoxins therapeutic use, Insulin metabolism, Insulin Secretion, Lymphocyte Depletion, Male, Regression Analysis, Ricin therapeutic use, T-Lymphocytes immunology, Antigens, CD immunology, Diabetes Mellitus, Type 1 therapy, Immunotoxins toxicity, Ricin toxicity
- Abstract
Type-I (insulin-dependent) diabetes mellitus is an immunologically mediated disease that results in destruction of the insulin secreting beta cells of the pancreas. T cells have been implicated in the pathogenesis of this disease. One novel form of anti-T-cell therapy is the immunoconjugate CD5-Plus. This agent is composed of the murine IgG1 monoclonal antibody H65, which is directed toward the CD5+ antigen; and ricin A chain, a ribosomal inhibitor protein. We performed a pilot study to evaluate the safety of the immunoconjugate in subjects with type-I diabetes mellitus. We conducted a dose-escalation study using CD5-Plus given as an intravenous infusion for 5 consecutive days. Fifteen subjects (12 men and 3 women) with a mean age of 26 years, a mean duration of diabetes of 4.8 months, and a minimum stimulated C peptide of 0.3 pmol/mL were entered. Six subjects each were treated at the 0.1 and 0.2 mg/kg/day dosage levels, and three subjects were treated at the 0.33 mg/kg/day dose. Glycemic control was determined monthly by recording the glycohemoglobin, total daily insulin requirements, and fasting blood glucoses. Beta-cell function was measured by determining the C-peptide response to a mixed formula meal (Sustacal) at baseline and at 1,3,6,9, and 12 months after treatment. The area under the curve (AUC) of the C-peptide response was calculated and, to reduce variability, related to that of the same subject at baseline. An analysis of subjects who retained at least 80% of their baseline beta-cell function as measured by the AUC was performed.(ABSTRACT TRUNCATED AT 250 WORDS)
- Published
- 1993
17. Phase I/II study of murine monoclonal antibody-ricin A chain (XOMAZYME-Mel) immunoconjugate plus cyclosporine A in patients with metastatic melanoma.
- Author
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Selvaggi K, Saria EA, Schwartz R, Vlock DR, Ackerman S, Wedel N, Kirkwood JM, Jones H, and Ernstoff MS
- Subjects
- Aged, Animals, Antibodies, Monoclonal adverse effects, Antibodies, Monoclonal metabolism, Antibody Formation, Combined Modality Therapy, Female, Humans, Immunotoxins adverse effects, Immunotoxins metabolism, Male, Melanoma metabolism, Melanoma secondary, Mice, Middle Aged, Ricin adverse effects, Ricin metabolism, Antibodies, Monoclonal administration & dosage, Cyclosporine administration & dosage, Immunotoxins administration & dosage, Melanoma therapy, Ricin administration & dosage
- Abstract
XOMAZYME-Mel (XMMME-001-RTA) is an immunoconjugate comprised of ricin A chain conjugated to a murine monoclonal antibody directed against high molecular weight melanoma antigens. Although not necessarily related to increased toxicity or decreased efficacy, the development of anti-immunoconjugate antibodies may limit repetitive dosing with an immunoconjugate. We evaluated the role of cyclosporine A in blocking the antibody response in patients with melanoma treated with XMMME-001-RTA. Patients received cyclosporine in divided daily doses to achieve serum levels by HPLC of 150-200 ng/ml on days 1-22. On day 3, XMMME-001-RTA was begun at dosages 0.2-0.6 mg/kg daily for 5 days. Treatment was repeated every 35 days. Three patients were treated in each dosage tier (0.2, 0.4, 0.6 mg/kg). Nine patients were entered and all nine were evaluable. Patients had histologically confirmed melanoma. Metastatic sites included skin, soft tissue, and lymph nodes (seven), lung (two), liver (one), and spleen (one). There were four men and five women aged 46-75 years. Toxicities included myalgia, arthralgia, hypoalbuminemia, fatigue, elevations in liver function tests, and increased peripheral edema. Four patients received two to five repeated dosages of XMMME-001-RTA. One wheal-and-flare reaction from an immunotoxin test dose of XMMME-001-RTA was noted after five cycles. After a test dose subsequent to one cycle, two patients experienced chest tightness without ECG changes and were removed from the study. All toxicities resolved without sequelae. One patient experienced partial lymph node remission for 9 months. A second patient had stable mediastinal disease for 20 months. XMMME-001-RTA is safe when given repeatedly with cyclosporine.
- Published
- 1993
- Full Text
- View/download PDF
18. Investigation and prevention of artifactual staining in flow cytometric analyses of whole blood samples from patients treated with H65-RTA, an anti-CD5 monoclonal antibody conjugated to ricin A chain.
- Author
-
Fishwild DM and Saria EA
- Subjects
- Animals, Antibodies, Anti-Idiotypic analysis, Antibodies, Monoclonal immunology, Artifacts, CD4 Antigens analysis, CD5 Antigens, CD8 Antigens analysis, Goats, Humans, Staining and Labeling, Antibodies, Monoclonal therapeutic use, Antigens, CD immunology, Flow Cytometry, Immunotoxins therapeutic use, Ricin therapeutic use
- Abstract
To assess the biological effect of therapeutic monoclonal antibodies (mAbs) immunoconjugates, flow cytometric assays are often performed on patients' blood samples. We report here that, using standard protocols, staining whole blood samples with mAb-fluorochromes can result in erroneous immune cell phenotyping. Blood samples were obtained from patients treated with H65-RTA, a murine IgG1 anti-CD5 mAb conjugated to ricin A chain. While a transient decrease in CD4+ and CD8+ cells was observed during the 5 days of therapy, alterations in lymphocyte phenotype were also noted, beginning around days 15-30. The most prominent effect was an apparent loss of CD4+ cells. However, if the cells were separated from plasma prior to staining, the patients' samples demonstrated their pre-treatment lymphocytic phenotypes. Co-incubation of post-treatment (day greater than or equal to 15) patients serum with lymphocytes from normal donors also resulted in artifactual staining. The effects of the post-treatment serum could be correlated with the onset of a human immune response directed against H65-RTA. Moreover, co-incubation of normal PBMC with goat anti-mouse immunoglobulin could replicate the artifactual staining patterns. Even though the human immune response was generated against a murine IgG1 mAb, there were sufficient human antibodies crossreactive with other murine IgG isotypes to induce artifactual staining with all mAb-fluorochromes tested. To minimize artifacts, cells should be separated from plasma prior to flow cytometric analysis as well as for other immunoassays involving murine mAbs.
- Published
- 1991
- Full Text
- View/download PDF
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