Joe Harford - Academia.edu (original) (raw)
Papers by Joe Harford
Journal of Neurochemistry, Jun 1, 1979
White matter membrane preparations from pig brain catalyze the transfer of [14C]mannose from exog... more White matter membrane preparations from pig brain catalyze the transfer of [14C]mannose from exogenous [14C]mannosylphosphoryldolichol into an endogenous oligosaccharide lipid. Under the same incubation conditions label is also incorporated into endogenous membrane glycoproteins. The enzymatic labeling of both classes of endogenous acceptors is stimulated by the addition of Ca2+. Several enzymatic properties of the mannosyltransferase activity responsible for the transfer of mannose from mannosylphosphoryldolichol into the oligosaccharide lipid intermediate have been examined. The [Man-14C] oligosaccharide lipid synthesized by this in vitro system has the solubility, hydrolytic and chromatographic characteristics of a pyrophosphate-linked oligosaccharide derivative of dolichol. The free [Man-14C]oligosaccharide liberated from the carrier lipid by mild acid treatment is estimated to contain 8 glycose units. All of the [14C]mannosyl units in the [Man-14C]oligosaccharide derived from exogenous [14C]mannosylphosphoryldolichol are released as free [14C]mannose by an α-mannosi-dase. No [14C]mannose is released during incubation with a β-mannosidase. The presence of an N,N′-diacetylchitobiose unit at the reducing end of the lipid-bound [Man-14C]oligosaccharide is indicated by its susceptibility to digestion by endo-β-N-acetylglucosaminidase H. Pronase digestion of the enzymatically labeled [Man-14C]glycoprotein yields a single [Man-14C]gly-copeptide fraction on Bio-Gel P-6 that appears to be slightly larger than the free [Man-14C]oligosac-charide released from the carrier lipid by mild acid hydrolysis. The [Man-14C]glycopeptide is cleaved by endo-β-N-acetylglucosaminidase H, and the neutral [Man-14C]oligosaccharide product appears to be identical to the product formed when the lipid-bound [Man-14C]oligosaccharide is degraded by the endoglycosidase. The glycopeptide linkage in the [Man-14C]glycoprotein is stable to mild alkali treatment. These results are consistent with the dolichol-linked [Man-14C]oligosaccharide, mannosy-lated via exogenous [14C]mannosylphosphoryldoiichol, being subsequently transferred en bloc from dolichyl pyrophosphate to asparagine residues in endogenous membrane polypeptide acceptors. SDS-polyacrylamide gel electrophoresis of the [Man-14C]glycoprotein, labeled when white matter membranes are incubated with [14C]mannosylphosphoryldolichol. revealed a major labeled polypeptide with an apparent mol wt of 24,000. A minor labeled membrane glycoprotein is also seen, having an apparent mol wt of 105,000.
International Journal of Cancer, Aug 5, 2011
Cervical screening for carcinogenic human papillomavirus (HPV) infection is being considered for ... more Cervical screening for carcinogenic human papillomavirus (HPV) infection is being considered for low-income countries. Effectiveness requires targeted screening in older women in whom prevalent infections are more likely to be persistent and predictive of precancer. Some studies in West Africa have found unusually high HPV prevalences across all adult ages, which may reduce the positive predictive value (PPV) of HPV-based screening, if positivity in older women does not sufficiently predict elevated risk. We conducted a population-based study in rural Nigeria to identify HPV prevalence and associated cervical abnormalities. Using stratified random sampling, we enrolled women age 151. Nonvirgins had a cervical exam including liquid-based cytology and PCR HPV DNA testing from residual cytology specimens. Two-thirds of invited women participated, and 14.7% had detectable carcinogenic HPV, a proportion that did not decline with age (p-trend 5 0.36) and showed slight peaks in the 15-29 and 60-69 age groups. Among women of the age typically considered for screen-and-treat programs (30-49 years), 12.8% were HPV positive, and the PPV for high-grade or worse cytology was 16.4%. Comparatively, women age < 30 were more likely to be HPV positive (18.9%, p 5 0.03) with a lower PPV (4.2% p 5 0.05). Among women age 501 (typically excluded from screening in resource-poor settings because inexpensive treatment is not available), HPV positivity was 14.2% with a PPV of 13.9%. In Irun and similar settings where HPV does not decline with age, HPV-based screen-and-treat programs might be feasible for mid-adult women because prevalence is sufficiently low and positivity predicts elevated risk of more easily treated precancer.
Proceedings of the National Academy of Sciences of the United States of America, 1982
Journal of Biological Chemistry, Mar 1, 1983
Cell, Mar 1, 1991
1. Cell. 1991 Mar 8;64(5):881-3. Structural relationship between an iron-regulated RNA-binding pr... more 1. Cell. 1991 Mar 8;64(5):881-3. Structural relationship between an iron-regulated RNA-binding protein (IRE-BP) and aconitase: functional implications. Rouault TA, Stout CD, Kaptain S, Harford JB, Klausner RD. PMID: 2001588 [PubMed - indexed for MEDLINE]. ...
Journal of Research in Medical Sciences, 2014
Background: Cancer incidence trends use by health officials in order to program evaluations and d... more Background: Cancer incidence trends use by health officials in order to program evaluations and development of cancer control strategies. The trends of cancer incidence have used to evaluate programs and develop the cancer control strategies. The aim of this study is to analyze changes of breast cancer incidence trends in Isfahan city using joinpoint regression models. Materials and Methods: The study was based on all cases of breast cancer reported among women residing in the city of Isfahan for the period 2001-2010. Age-standardized rates were calculated for each tumor characteristics, using the direct method. Joinpoint regression was used to provide estimated annual percentage change. Results: A plot of the age-specific rates of breast cancer showed an increase in all age groups from 30 to 69 years and sharp increase in the incidence of breast cancer confined to estrogen receptor-positive and progesterone receptor-positive tumors and the significant change (2003) by progesterone receptor − tumors. The analysis by tumor size and grade, incidence rates decreased for tumors >5 cm by 10.6% since 2006.7 and for poorly differentiated tumor by 26.1% since 2007.8. No decrease in incidence was observed for group of proportion of positive lymph nodes to lymph node surgery ≥25%. The proportion of positive lymph node to surgery node ≤25% (nonsignificant) was upward. Conclusion: The trend of incidence rates with tumor size ≤2, well-differentiated tumor grade, moderately differentiated tumor grade, positive estrogen and progesterone hormone receptors was upward. The pattern of breast cancer can help to cancer prevention and prognosis, selecting the best type of surgery.
Cold Spring Harbor Monograph Archive, 1998
The expression of two proteins that participate in cellular iron homeostasis, ferritin and the tr... more The expression of two proteins that participate in cellular iron homeostasis, ferritin and the transferrin receptor (TfR), are coordinately regulated at the posttranscriptional level by the intracellular iron (for background information on cellular iron metabolism, see Brock et al. 1994). When iron is abundant, synthesis of ferritin, the protein responsible for iron sequestration, increases, whereas expression of the TfR falls, resulting in less iron acquisition from the cell’s exterior. When the cell is deprived of iron, the opposite occurs—TfR synthesis increases and ferritin synthesis decreases. The iron-dependent regulation of these two proteins is mediated by structurally related RNA elements found in their mRNAs (see Fig. 1) (Klausner et al. 1993; Harford et al. 1994; Theil 1994; Hentze and Kuhn 1996). These RNA elements, termed iron-responsive elements (IREs), are moderately stable stem-loop structures of approximately 30 nucleotides (see Fig. 2). In ferritin transcripts, a single IRE is found in the 5′-untranslated region (UTR), whereas in the TfR transcript, five IREs are found in the 3′ UTR. The IREs of both transcripts interact with cytosolic IRE-binding proteins referred to as IRP1 and IRP2 (for i ron r egulatory p roteins). The interaction(s) between IREs and IRPs is responsible for the posttranscriptional regulation of the expression of the genes encoding ferritin and the TfR (Fig. 1). In the case of ferritin regulation, the interaction of an IRP with the transcript inhibits protein synthesis through an inhibition of translation initiation. In the case of TfR regulation, interaction of one or more IRPs with IREs in the...
Cold Spring Harbor Monograph Archive, 2000
REGULATION OF IRON HOMEOSTASIS Iron is an important nutrient in mammalian cells, and the processe... more REGULATION OF IRON HOMEOSTASIS Iron is an important nutrient in mammalian cells, and the processes of uptake and sequestration of iron are highly regulated to ensure that supplies are adequate and toxicity is controlled. A major mediator of iron uptake is the transferrin receptor, a plasma membrane protein that binds serum diferric transferrin at the cell surface. Upon transferrin binding, the transferrin receptor– diferric transferrin complex is internalized in endosomes; within endosomes, iron is released from transferrin and transported into the cytosol and other compartments of the cell such as the mitochondria. When iron levels in the cell are high, toxic products generated by the reaction of iron with oxygen species can oxidize and damage cellular proteins, lipids, and nucleic acids (Halliwell and Gutteridge 1992). To protect against iron toxicity, cells synthesize ferritin, a 24-subunit spherical protein that sequesters as many as 4,500 iron atoms within precipitates in its interior (Theil 1987). Although ferritin serves a useful purpose in cells with high cytosolic iron levels, excess production of ferritin can be deleterious in iron-depleted cells because it may compete with other proteins for the limited amounts of available iron (Picard et al. 1998). Translational regulation of ferritin expression is the mechanism utilized by mammalian cells to ensure that ferritin is expressed primarily when it is needed for sequestration of excess iron (Fig. 1). Binding of cytosolic iron-sensing proteins known as iron regulatory proteins (IRPs) to a conserved RNA stem-loop element in the ferritin transcript interferes with ferritin translation in iron-depleted...
Cold Spring Harbor Monograph Archive, 1996
The translation of messenger RNAs encoding ferritin is highly regulated by iron. A single copy of... more The translation of messenger RNAs encoding ferritin is highly regulated by iron. A single copy of an RNA motif known as an iron-responsive element or IRE (see Fig. 1) is found within the 5′-untranslated region (5′ UTR) of the mRNAs that encode all known vertebrate ferritins. The control of translation of ferritin mRNA by iron is mediated by regulation of an interaction between the IRE and a cytosolic protein. This protein, which binds to the IRE with high affinity and specificity when cells are iron-depleted, has been called the IRE-binding protein (IRE-BP) (Leibold and Munro 1988; Rouault et al. 1988), iron regulatory factor (IRF) (Mullner et al. 1989), ferritin repressor protein (FRP) (Walden et al. 1989), and P90 (Harrell et al. 1991). Recently, a consensus has emerged that favors the term iron regulatory protein (IRP), and we adhere to this convention here. There are now two relatively well-characterized iron regulatory proteins that are described in this chapter. The protein originally called IRE-BP is henceforth referred to as IRP1, and a second IRP that is less abundant in the majority of tissues tested is referred to as IRP2. In addition to the role of the IRE in mediating the regulation of ferritin biosynthesis, an IRE that functions to produce IRP1-mediated translational regulation is present in the 5′ UTR of the mRNA for the erythroid form of δ-aminolevulinic acid (ALA) synthase, the rate-limiting step in the heme biosynthetic pathway (Dierks 1990; Cox et al. 1991; Dandekar et al. 1991; Bhasker et al....
Molecular and Cellular Biology, 1986
Publisher Summary A specific receptor exists in the plasma membrane of mammalian hepatocytes that... more Publisher Summary A specific receptor exists in the plasma membrane of mammalian hepatocytes that mediates endocytosis of desialylated glycoproteins. The receptor has been isolated from Triton X-100 extracts of rat liver by affinity chromatography on ligand-Sepharose. The binding activity appears as a high MW entity by gel filtration, but sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) reveals a major band corresponding to 42,000 daltons and two less-prominent bands of slightly higher MW. This chapter describes an experiment where spleen cells from BALB/C mice immunized with a soluble receptor preparation were fused with P3X63–Ag8653 myeloma cells. Media from the resultant hybridomas were screened for their ability to bind a radioiodinated receptor. After subcloning, positive ascites was produced in BALB/C mice by i.p. injection of 106 cells. A clone was identified that produces antibody directed toward a determinant that is related to receptor function; it blocked the binding of ligand by plasma membranes. The SDS-PAGE pattern of the radiolabeled receptor preparation bound by this antibody was indistinguishable from the Coomassie-blue-staining pattern of the preparation used as antigen. However, because of the high apparent MW of the Triton X-100 extract, this result was viewed inconclusive. No reaction could be detected with the individual polypeptides recovered by a preparative SDS-PAGE using the gel system of Laemmli. Accordingly, a modified preparative SDS-PAGE system was devised that proved to be less destructive of antigenic reactivity. These results indicated that the recognized determinant is shared, at least in part, by the three polypeptides.
The endocytosis leading to degradation of 125I- labeled asialo-orosomucoid specifically bound to ... more The endocytosis leading to degradation of 125I- labeled asialo-orosomucoid specifically bound to the surface of freshly isolated hepatocytes was monitored as a function of time at 370C. Experimental values were determined for the rates of internalization, dissociation of the receptor-ligand complex, and degradation of the labeled ligand. Compartmental analysis and computer modeling revealed that the data were compatible with dissociation of ligand from receptor preceding ligand degradation. The rate coefficient for internalization was calculated to be an order of magnitude greater than that for receptor-ligand disso- ciation. 'Ligand internalization did not result in concomitant de- pletion in the total number of cell surface receptors. Our data are taken to indicate that ligand remains associated with the receptor after internalization, that the complex is dissociated,prior to deg- radation, and that new, unoccupied receptors are promptly re- turned to the cell surface from an internal pool.
Ejc Supplements, Apr 1, 2008
I n the early 1700s, Leeuwenhoek noted that avian and amphibian blood cells contained a “clear ar... more I n the early 1700s, Leeuwenhoek noted that avian and amphibian blood cells contained a “clear area,” almost certainly corresponding to the structure we now know as the nucleus. This represents probably the earliest recognition that eukaryotic cells are not simply sacs of protoplasm, but instead contain subcellular structures. Although the concept of the nucleated cell became firmly established in the 18th and 19th centuries, the real beginning of subcellular fractionation had to await the emergence in the mid-1940s of the art form known as electron microscopy. The electron microscope revealed much more complexity than Leeuwenhoek could have imagined. Concurrent with improvements in the techniques of electron microscopy was the development of methodologies for subcellular fractionation. Through the 1950s, these parallel approaches resulted in the discovery and isolation of the major organelles that comprise the eukaryotic cell. Finally, as biochemical functions were associated with specific subcellular compartments, a much clearer picture of the eukaryotic cell began to emerge, and with it the field of modern cell biology.
Methods in Enzymology, 1985
Publisher Summary Endocytosis is the process by which cells engulf extracellular materials within... more Publisher Summary Endocytosis is the process by which cells engulf extracellular materials within the infoldings of their plasma membrane that pinch off to form cytoplasmic vesicles. This general process is utilized by cells in the uptake of particulates and for the nonselective internalization of extracellular fluid. The mechanism of uptake and receptor reutilization follows, in many cells, a common pattern with only minor and subtle alterations designed to meet specific metabolic requirements. The chapter explains that one such system, the hepatic receptor for asialoglycoproteins, has been extensively examined in a number of laboratories. The chapter presents various methodologies employed and the conclusions drawn. It discusses that a critical evaluation of the techniques involved in receptor reutilization does not lend itself readily to the more conventional format for describing well-established methodologies.
Cancer Epidemiology, Biomarkers & Prevention, Oct 25, 2011
Background: Inflammatory breast cancer (IBC) is a rare and highly aggressive form of primary brea... more Background: Inflammatory breast cancer (IBC) is a rare and highly aggressive form of primary breast cancer. Little is known about the risk factors for IBC, specifically the association with socioeconomic position (SEP). Methods: The association between breast cancer type (IBC vs. non-IBC) with county-level SEP in the Surveillance, Epidemiology, and End Results database for cases diagnosed from 2000 to 2007 was examined. County-level SEP characteristics included metropolitan versus non-metropolitan residence, percentage below the poverty level, percentage less than high-school graduate, and an index combining the poverty and high-school variables. IBC and non-IBC age-adjusted incidence rates were calculated, stratified on SEP and race/ethnicity. The odds of IBC versus non-IBC given a particular SEP characteristic, adjusting for age and race/ethnicity, was examined through fitting of hierarchical logistic regression models (HLM). Results: Incidence rates for IBC generally increased as SEP decreased, whereas the opposite was found for non-IBC. HLM results showed that low SEP is associated with higher odds of IBC: highest (!20%) versus lowest (<10%) persons below the poverty level [OR (95% confidence interval, CI) ¼ 1.25 (1.09-1.43)]; highest (>28.76%) versus lowest (15.99%) persons less than high-school graduate [OR (95% CI) ¼ 1.25 (1.10-1.42)]; and low SEP as measured by poverty-high school index versus high SEP [OR (95% CI)¼ 1.26 (1.11-1.44)]. Conclusion: Overall breast cancer has been found to be positively associated with SEP, whereas in this analysis, IBC was associated with decreasing SEP. Impact: Studies focused on understanding the disparity in IBC incidence, as well as interventions to eliminate these differences are needed. Cancer Epidemiol Biomarkers Prev; 21(1); 155-65. Ó2011 AACR.
Journal of Clinical Oncology
4139 Background: Nearly all stage IV pancreatic adenocarcinoma (PAC) patients progress after firs... more 4139 Background: Nearly all stage IV pancreatic adenocarcinoma (PAC) patients progress after first-line treatment, and second-line options are limited. SGT-53 is an investigational product for tumor-targeted TP53 gene therapy that has completed phase Ia/Ib trials [Senser et al (2013), Mol Ther 21:1096; Pirollo et al (2016) Mol Ther 24:1697]. Methods: Here we provide an interim analysis of a Phase II trial (SGT53-02-1; NCT02340117) combining SGT-53 with gemcitabine/nab-paclitaxel (GEM/ABX). Eligible were first-line patients or those who had progressed after FOLFIRINOX (FFX) and/or gemcitabine-based therapy (second-line). In a 7-week treatment cycle, SGT-53 (3.6 mg DNA) was given once or twice weekly with GEM/ABX (1000 mg/m2/wk and 125 mg/m2/wk, respectively, for 3 of 4 weeks). Progression-free survival (PFS) and objective response rate (ORR) are primary endpoints.Overall survival (OS) and PFS are estimated by Kaplan-Meier analysis. Results: Of all evaluable patients (n=20), best resp...
Journal of Biological Chemistry, 1982
The major polypeptide (43,000 daltons) of the rat liver receptor for asialoglycoproteins was isol... more The major polypeptide (43,000 daltons) of the rat liver receptor for asialoglycoproteins was isolated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Antibodies raised in a goat against this SDStreated polypeptide exhibited marked cross-reactivity toward the SDS-denatured forms of the two other prominent polypeptides (54,000 and 64,000 daltons) of the receptor preparation. Monoclonal antibodies directed against the receptor were prepared using the spleen cells of mice immunized with the soluble, active receptor purified by affinity chromatography. The most extensively characterized of the monoclonal antibodies, designated D3-5D3, recognized the solubilized receptor and bound to the exterior surface of isolated rat hepatocytes. The binding of D3-5D3 to hepatocytes prevented subsequent binding of the ligand, '251-asia10-orosomucoid. Conversely, occupation of the receptor with ligand inhibited binding of "'I-IgG prepared from D3-5D3 ascites fluid. The secondary structure of the receptor appears to be critical for recognition by D3-5D3, since denaturation of the receptor with 1% SDS, 5% /3mercaptoethanol at 100 "C abolished antibody binding. Under less denaturing conditions (0.1% SDS, 25 "C), antigenic reactivity was retained by the receptor. Preparative electrophoresis using the latter conditions permitted the demonstration that D3-5D3 recognized a unique determinant that is present in each of the three polypeptides.
Journal of Neurochemistry, Jun 1, 1979
White matter membrane preparations from pig brain catalyze the transfer of [14C]mannose from exog... more White matter membrane preparations from pig brain catalyze the transfer of [14C]mannose from exogenous [14C]mannosylphosphoryldolichol into an endogenous oligosaccharide lipid. Under the same incubation conditions label is also incorporated into endogenous membrane glycoproteins. The enzymatic labeling of both classes of endogenous acceptors is stimulated by the addition of Ca2+. Several enzymatic properties of the mannosyltransferase activity responsible for the transfer of mannose from mannosylphosphoryldolichol into the oligosaccharide lipid intermediate have been examined. The [Man-14C] oligosaccharide lipid synthesized by this in vitro system has the solubility, hydrolytic and chromatographic characteristics of a pyrophosphate-linked oligosaccharide derivative of dolichol. The free [Man-14C]oligosaccharide liberated from the carrier lipid by mild acid treatment is estimated to contain 8 glycose units. All of the [14C]mannosyl units in the [Man-14C]oligosaccharide derived from exogenous [14C]mannosylphosphoryldolichol are released as free [14C]mannose by an α-mannosi-dase. No [14C]mannose is released during incubation with a β-mannosidase. The presence of an N,N′-diacetylchitobiose unit at the reducing end of the lipid-bound [Man-14C]oligosaccharide is indicated by its susceptibility to digestion by endo-β-N-acetylglucosaminidase H. Pronase digestion of the enzymatically labeled [Man-14C]glycoprotein yields a single [Man-14C]gly-copeptide fraction on Bio-Gel P-6 that appears to be slightly larger than the free [Man-14C]oligosac-charide released from the carrier lipid by mild acid hydrolysis. The [Man-14C]glycopeptide is cleaved by endo-β-N-acetylglucosaminidase H, and the neutral [Man-14C]oligosaccharide product appears to be identical to the product formed when the lipid-bound [Man-14C]oligosaccharide is degraded by the endoglycosidase. The glycopeptide linkage in the [Man-14C]glycoprotein is stable to mild alkali treatment. These results are consistent with the dolichol-linked [Man-14C]oligosaccharide, mannosy-lated via exogenous [14C]mannosylphosphoryldoiichol, being subsequently transferred en bloc from dolichyl pyrophosphate to asparagine residues in endogenous membrane polypeptide acceptors. SDS-polyacrylamide gel electrophoresis of the [Man-14C]glycoprotein, labeled when white matter membranes are incubated with [14C]mannosylphosphoryldolichol. revealed a major labeled polypeptide with an apparent mol wt of 24,000. A minor labeled membrane glycoprotein is also seen, having an apparent mol wt of 105,000.
International Journal of Cancer, Aug 5, 2011
Cervical screening for carcinogenic human papillomavirus (HPV) infection is being considered for ... more Cervical screening for carcinogenic human papillomavirus (HPV) infection is being considered for low-income countries. Effectiveness requires targeted screening in older women in whom prevalent infections are more likely to be persistent and predictive of precancer. Some studies in West Africa have found unusually high HPV prevalences across all adult ages, which may reduce the positive predictive value (PPV) of HPV-based screening, if positivity in older women does not sufficiently predict elevated risk. We conducted a population-based study in rural Nigeria to identify HPV prevalence and associated cervical abnormalities. Using stratified random sampling, we enrolled women age 151. Nonvirgins had a cervical exam including liquid-based cytology and PCR HPV DNA testing from residual cytology specimens. Two-thirds of invited women participated, and 14.7% had detectable carcinogenic HPV, a proportion that did not decline with age (p-trend 5 0.36) and showed slight peaks in the 15-29 and 60-69 age groups. Among women of the age typically considered for screen-and-treat programs (30-49 years), 12.8% were HPV positive, and the PPV for high-grade or worse cytology was 16.4%. Comparatively, women age < 30 were more likely to be HPV positive (18.9%, p 5 0.03) with a lower PPV (4.2% p 5 0.05). Among women age 501 (typically excluded from screening in resource-poor settings because inexpensive treatment is not available), HPV positivity was 14.2% with a PPV of 13.9%. In Irun and similar settings where HPV does not decline with age, HPV-based screen-and-treat programs might be feasible for mid-adult women because prevalence is sufficiently low and positivity predicts elevated risk of more easily treated precancer.
Proceedings of the National Academy of Sciences of the United States of America, 1982
Journal of Biological Chemistry, Mar 1, 1983
Cell, Mar 1, 1991
1. Cell. 1991 Mar 8;64(5):881-3. Structural relationship between an iron-regulated RNA-binding pr... more 1. Cell. 1991 Mar 8;64(5):881-3. Structural relationship between an iron-regulated RNA-binding protein (IRE-BP) and aconitase: functional implications. Rouault TA, Stout CD, Kaptain S, Harford JB, Klausner RD. PMID: 2001588 [PubMed - indexed for MEDLINE]. ...
Journal of Research in Medical Sciences, 2014
Background: Cancer incidence trends use by health officials in order to program evaluations and d... more Background: Cancer incidence trends use by health officials in order to program evaluations and development of cancer control strategies. The trends of cancer incidence have used to evaluate programs and develop the cancer control strategies. The aim of this study is to analyze changes of breast cancer incidence trends in Isfahan city using joinpoint regression models. Materials and Methods: The study was based on all cases of breast cancer reported among women residing in the city of Isfahan for the period 2001-2010. Age-standardized rates were calculated for each tumor characteristics, using the direct method. Joinpoint regression was used to provide estimated annual percentage change. Results: A plot of the age-specific rates of breast cancer showed an increase in all age groups from 30 to 69 years and sharp increase in the incidence of breast cancer confined to estrogen receptor-positive and progesterone receptor-positive tumors and the significant change (2003) by progesterone receptor − tumors. The analysis by tumor size and grade, incidence rates decreased for tumors >5 cm by 10.6% since 2006.7 and for poorly differentiated tumor by 26.1% since 2007.8. No decrease in incidence was observed for group of proportion of positive lymph nodes to lymph node surgery ≥25%. The proportion of positive lymph node to surgery node ≤25% (nonsignificant) was upward. Conclusion: The trend of incidence rates with tumor size ≤2, well-differentiated tumor grade, moderately differentiated tumor grade, positive estrogen and progesterone hormone receptors was upward. The pattern of breast cancer can help to cancer prevention and prognosis, selecting the best type of surgery.
Cold Spring Harbor Monograph Archive, 1998
The expression of two proteins that participate in cellular iron homeostasis, ferritin and the tr... more The expression of two proteins that participate in cellular iron homeostasis, ferritin and the transferrin receptor (TfR), are coordinately regulated at the posttranscriptional level by the intracellular iron (for background information on cellular iron metabolism, see Brock et al. 1994). When iron is abundant, synthesis of ferritin, the protein responsible for iron sequestration, increases, whereas expression of the TfR falls, resulting in less iron acquisition from the cell’s exterior. When the cell is deprived of iron, the opposite occurs—TfR synthesis increases and ferritin synthesis decreases. The iron-dependent regulation of these two proteins is mediated by structurally related RNA elements found in their mRNAs (see Fig. 1) (Klausner et al. 1993; Harford et al. 1994; Theil 1994; Hentze and Kuhn 1996). These RNA elements, termed iron-responsive elements (IREs), are moderately stable stem-loop structures of approximately 30 nucleotides (see Fig. 2). In ferritin transcripts, a single IRE is found in the 5′-untranslated region (UTR), whereas in the TfR transcript, five IREs are found in the 3′ UTR. The IREs of both transcripts interact with cytosolic IRE-binding proteins referred to as IRP1 and IRP2 (for i ron r egulatory p roteins). The interaction(s) between IREs and IRPs is responsible for the posttranscriptional regulation of the expression of the genes encoding ferritin and the TfR (Fig. 1). In the case of ferritin regulation, the interaction of an IRP with the transcript inhibits protein synthesis through an inhibition of translation initiation. In the case of TfR regulation, interaction of one or more IRPs with IREs in the...
Cold Spring Harbor Monograph Archive, 2000
REGULATION OF IRON HOMEOSTASIS Iron is an important nutrient in mammalian cells, and the processe... more REGULATION OF IRON HOMEOSTASIS Iron is an important nutrient in mammalian cells, and the processes of uptake and sequestration of iron are highly regulated to ensure that supplies are adequate and toxicity is controlled. A major mediator of iron uptake is the transferrin receptor, a plasma membrane protein that binds serum diferric transferrin at the cell surface. Upon transferrin binding, the transferrin receptor– diferric transferrin complex is internalized in endosomes; within endosomes, iron is released from transferrin and transported into the cytosol and other compartments of the cell such as the mitochondria. When iron levels in the cell are high, toxic products generated by the reaction of iron with oxygen species can oxidize and damage cellular proteins, lipids, and nucleic acids (Halliwell and Gutteridge 1992). To protect against iron toxicity, cells synthesize ferritin, a 24-subunit spherical protein that sequesters as many as 4,500 iron atoms within precipitates in its interior (Theil 1987). Although ferritin serves a useful purpose in cells with high cytosolic iron levels, excess production of ferritin can be deleterious in iron-depleted cells because it may compete with other proteins for the limited amounts of available iron (Picard et al. 1998). Translational regulation of ferritin expression is the mechanism utilized by mammalian cells to ensure that ferritin is expressed primarily when it is needed for sequestration of excess iron (Fig. 1). Binding of cytosolic iron-sensing proteins known as iron regulatory proteins (IRPs) to a conserved RNA stem-loop element in the ferritin transcript interferes with ferritin translation in iron-depleted...
Cold Spring Harbor Monograph Archive, 1996
The translation of messenger RNAs encoding ferritin is highly regulated by iron. A single copy of... more The translation of messenger RNAs encoding ferritin is highly regulated by iron. A single copy of an RNA motif known as an iron-responsive element or IRE (see Fig. 1) is found within the 5′-untranslated region (5′ UTR) of the mRNAs that encode all known vertebrate ferritins. The control of translation of ferritin mRNA by iron is mediated by regulation of an interaction between the IRE and a cytosolic protein. This protein, which binds to the IRE with high affinity and specificity when cells are iron-depleted, has been called the IRE-binding protein (IRE-BP) (Leibold and Munro 1988; Rouault et al. 1988), iron regulatory factor (IRF) (Mullner et al. 1989), ferritin repressor protein (FRP) (Walden et al. 1989), and P90 (Harrell et al. 1991). Recently, a consensus has emerged that favors the term iron regulatory protein (IRP), and we adhere to this convention here. There are now two relatively well-characterized iron regulatory proteins that are described in this chapter. The protein originally called IRE-BP is henceforth referred to as IRP1, and a second IRP that is less abundant in the majority of tissues tested is referred to as IRP2. In addition to the role of the IRE in mediating the regulation of ferritin biosynthesis, an IRE that functions to produce IRP1-mediated translational regulation is present in the 5′ UTR of the mRNA for the erythroid form of δ-aminolevulinic acid (ALA) synthase, the rate-limiting step in the heme biosynthetic pathway (Dierks 1990; Cox et al. 1991; Dandekar et al. 1991; Bhasker et al....
Molecular and Cellular Biology, 1986
Publisher Summary A specific receptor exists in the plasma membrane of mammalian hepatocytes that... more Publisher Summary A specific receptor exists in the plasma membrane of mammalian hepatocytes that mediates endocytosis of desialylated glycoproteins. The receptor has been isolated from Triton X-100 extracts of rat liver by affinity chromatography on ligand-Sepharose. The binding activity appears as a high MW entity by gel filtration, but sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) reveals a major band corresponding to 42,000 daltons and two less-prominent bands of slightly higher MW. This chapter describes an experiment where spleen cells from BALB/C mice immunized with a soluble receptor preparation were fused with P3X63–Ag8653 myeloma cells. Media from the resultant hybridomas were screened for their ability to bind a radioiodinated receptor. After subcloning, positive ascites was produced in BALB/C mice by i.p. injection of 106 cells. A clone was identified that produces antibody directed toward a determinant that is related to receptor function; it blocked the binding of ligand by plasma membranes. The SDS-PAGE pattern of the radiolabeled receptor preparation bound by this antibody was indistinguishable from the Coomassie-blue-staining pattern of the preparation used as antigen. However, because of the high apparent MW of the Triton X-100 extract, this result was viewed inconclusive. No reaction could be detected with the individual polypeptides recovered by a preparative SDS-PAGE using the gel system of Laemmli. Accordingly, a modified preparative SDS-PAGE system was devised that proved to be less destructive of antigenic reactivity. These results indicated that the recognized determinant is shared, at least in part, by the three polypeptides.
The endocytosis leading to degradation of 125I- labeled asialo-orosomucoid specifically bound to ... more The endocytosis leading to degradation of 125I- labeled asialo-orosomucoid specifically bound to the surface of freshly isolated hepatocytes was monitored as a function of time at 370C. Experimental values were determined for the rates of internalization, dissociation of the receptor-ligand complex, and degradation of the labeled ligand. Compartmental analysis and computer modeling revealed that the data were compatible with dissociation of ligand from receptor preceding ligand degradation. The rate coefficient for internalization was calculated to be an order of magnitude greater than that for receptor-ligand disso- ciation. 'Ligand internalization did not result in concomitant de- pletion in the total number of cell surface receptors. Our data are taken to indicate that ligand remains associated with the receptor after internalization, that the complex is dissociated,prior to deg- radation, and that new, unoccupied receptors are promptly re- turned to the cell surface from an internal pool.
Ejc Supplements, Apr 1, 2008
I n the early 1700s, Leeuwenhoek noted that avian and amphibian blood cells contained a “clear ar... more I n the early 1700s, Leeuwenhoek noted that avian and amphibian blood cells contained a “clear area,” almost certainly corresponding to the structure we now know as the nucleus. This represents probably the earliest recognition that eukaryotic cells are not simply sacs of protoplasm, but instead contain subcellular structures. Although the concept of the nucleated cell became firmly established in the 18th and 19th centuries, the real beginning of subcellular fractionation had to await the emergence in the mid-1940s of the art form known as electron microscopy. The electron microscope revealed much more complexity than Leeuwenhoek could have imagined. Concurrent with improvements in the techniques of electron microscopy was the development of methodologies for subcellular fractionation. Through the 1950s, these parallel approaches resulted in the discovery and isolation of the major organelles that comprise the eukaryotic cell. Finally, as biochemical functions were associated with specific subcellular compartments, a much clearer picture of the eukaryotic cell began to emerge, and with it the field of modern cell biology.
Methods in Enzymology, 1985
Publisher Summary Endocytosis is the process by which cells engulf extracellular materials within... more Publisher Summary Endocytosis is the process by which cells engulf extracellular materials within the infoldings of their plasma membrane that pinch off to form cytoplasmic vesicles. This general process is utilized by cells in the uptake of particulates and for the nonselective internalization of extracellular fluid. The mechanism of uptake and receptor reutilization follows, in many cells, a common pattern with only minor and subtle alterations designed to meet specific metabolic requirements. The chapter explains that one such system, the hepatic receptor for asialoglycoproteins, has been extensively examined in a number of laboratories. The chapter presents various methodologies employed and the conclusions drawn. It discusses that a critical evaluation of the techniques involved in receptor reutilization does not lend itself readily to the more conventional format for describing well-established methodologies.
Cancer Epidemiology, Biomarkers & Prevention, Oct 25, 2011
Background: Inflammatory breast cancer (IBC) is a rare and highly aggressive form of primary brea... more Background: Inflammatory breast cancer (IBC) is a rare and highly aggressive form of primary breast cancer. Little is known about the risk factors for IBC, specifically the association with socioeconomic position (SEP). Methods: The association between breast cancer type (IBC vs. non-IBC) with county-level SEP in the Surveillance, Epidemiology, and End Results database for cases diagnosed from 2000 to 2007 was examined. County-level SEP characteristics included metropolitan versus non-metropolitan residence, percentage below the poverty level, percentage less than high-school graduate, and an index combining the poverty and high-school variables. IBC and non-IBC age-adjusted incidence rates were calculated, stratified on SEP and race/ethnicity. The odds of IBC versus non-IBC given a particular SEP characteristic, adjusting for age and race/ethnicity, was examined through fitting of hierarchical logistic regression models (HLM). Results: Incidence rates for IBC generally increased as SEP decreased, whereas the opposite was found for non-IBC. HLM results showed that low SEP is associated with higher odds of IBC: highest (!20%) versus lowest (<10%) persons below the poverty level [OR (95% confidence interval, CI) ¼ 1.25 (1.09-1.43)]; highest (>28.76%) versus lowest (15.99%) persons less than high-school graduate [OR (95% CI) ¼ 1.25 (1.10-1.42)]; and low SEP as measured by poverty-high school index versus high SEP [OR (95% CI)¼ 1.26 (1.11-1.44)]. Conclusion: Overall breast cancer has been found to be positively associated with SEP, whereas in this analysis, IBC was associated with decreasing SEP. Impact: Studies focused on understanding the disparity in IBC incidence, as well as interventions to eliminate these differences are needed. Cancer Epidemiol Biomarkers Prev; 21(1); 155-65. Ó2011 AACR.
Journal of Clinical Oncology
4139 Background: Nearly all stage IV pancreatic adenocarcinoma (PAC) patients progress after firs... more 4139 Background: Nearly all stage IV pancreatic adenocarcinoma (PAC) patients progress after first-line treatment, and second-line options are limited. SGT-53 is an investigational product for tumor-targeted TP53 gene therapy that has completed phase Ia/Ib trials [Senser et al (2013), Mol Ther 21:1096; Pirollo et al (2016) Mol Ther 24:1697]. Methods: Here we provide an interim analysis of a Phase II trial (SGT53-02-1; NCT02340117) combining SGT-53 with gemcitabine/nab-paclitaxel (GEM/ABX). Eligible were first-line patients or those who had progressed after FOLFIRINOX (FFX) and/or gemcitabine-based therapy (second-line). In a 7-week treatment cycle, SGT-53 (3.6 mg DNA) was given once or twice weekly with GEM/ABX (1000 mg/m2/wk and 125 mg/m2/wk, respectively, for 3 of 4 weeks). Progression-free survival (PFS) and objective response rate (ORR) are primary endpoints.Overall survival (OS) and PFS are estimated by Kaplan-Meier analysis. Results: Of all evaluable patients (n=20), best resp...
Journal of Biological Chemistry, 1982
The major polypeptide (43,000 daltons) of the rat liver receptor for asialoglycoproteins was isol... more The major polypeptide (43,000 daltons) of the rat liver receptor for asialoglycoproteins was isolated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Antibodies raised in a goat against this SDStreated polypeptide exhibited marked cross-reactivity toward the SDS-denatured forms of the two other prominent polypeptides (54,000 and 64,000 daltons) of the receptor preparation. Monoclonal antibodies directed against the receptor were prepared using the spleen cells of mice immunized with the soluble, active receptor purified by affinity chromatography. The most extensively characterized of the monoclonal antibodies, designated D3-5D3, recognized the solubilized receptor and bound to the exterior surface of isolated rat hepatocytes. The binding of D3-5D3 to hepatocytes prevented subsequent binding of the ligand, '251-asia10-orosomucoid. Conversely, occupation of the receptor with ligand inhibited binding of "'I-IgG prepared from D3-5D3 ascites fluid. The secondary structure of the receptor appears to be critical for recognition by D3-5D3, since denaturation of the receptor with 1% SDS, 5% /3mercaptoethanol at 100 "C abolished antibody binding. Under less denaturing conditions (0.1% SDS, 25 "C), antigenic reactivity was retained by the receptor. Preparative electrophoresis using the latter conditions permitted the demonstration that D3-5D3 recognized a unique determinant that is present in each of the three polypeptides.