Chuck Logsdon - Academia.edu (original) (raw)

Papers by Chuck Logsdon

Research paper thumbnail of Reminding Us That We Are "Tough and Competent

American Journal of Critical Care

Reminding Us That We Are "Tough and Competent" Thank you for the inspiring (and for me, timely) e... more Reminding Us That We Are "Tough and Competent" Thank you for the inspiring (and for me, timely) editorial, "Tough and Competent." 1 I grew up in awe of space travel and desperately wanted to be a "nurse in space" on the space shuttle. So I read the editorial that recounted the history of NASA with enjoyment and fascination. In this current health care environment, there is a lack of leaders who will shine the light on "carelessness, incapacity, and neglect." They are not inspiring us to be tough and competent, and to never compromise regarding our responsibilities. But this piece spoke to my head and heart when you referred to NASA's operations and the parallels to critical care. A couple of weeks ago I had lost my inspiration when 5 health care-associated infections were sent to me tagged to the units in my division. This occurred after we had several months of 1 or 2 health care-associated infections. I was feeling defeated and worried that all of the work on reducing them was meaningless. The next day I decided to move forward and embrace the current state of events as a continued challenge. You are right about critical care professionals, we are tough and competent. We just need to be reminded of that.

Research paper thumbnail of Down-regulation of the G-proteins G q α and G 11 α by transfected human M3 muscarinic acetylcholine receptors in Chinese hamster ovary cells is independent of receptor down-regulation

Biochemical Journal, 1995

Chinese hamster ovary cells stably transfected with human M3 muscarinic acetylcholine receptors s... more Chinese hamster ovary cells stably transfected with human M3 muscarinic acetylcholine receptors show a 40-50% reduction in the immunoreactive G-proteins Gq alpha and G11 alpha when stimulated with the cholinergic agonist carbachol. This effect is seen after 9 h, is maximal after 24 h, and occurs over a range of carbachol concentrations that activate phosphoinositide hydrolysis in these cells. The effect is specific for Gq alpha family proteins as Gs alpha was slightly increased after carbachol treatment and G13 alpha was unchanged. Using a urea gel system, we were able to resolve Gq alpha and G11 alpha, both of which were down-regulated by carbachol. An M3 receptor mutant, with C-terminal threonines changed to alanines as described previously, binds ligand and activates phosphoinositide hydrolysis normally but is not down-regulated in response to carbachol. This receptor, however, induces Gq alpha/G11 alpha down-regulation similarly to wild-type M3 receptors, indicating that G-prote...

Research paper thumbnail of Psychosocial and functional outcomes in women after coronary artery bypass surgery

Critical care nurse, 1997

In this longitudinal study of women after CABS, the women were coping admirably with good psychos... more In this longitudinal study of women after CABS, the women were coping admirably with good psychosocial and functional outcomes up to 3 months postoperatively. It is suggested that women be instructed how to find and use their support systems and be told that most women do well and return to normal activities of daily living after CABS.

Research paper thumbnail of Bisphosphonates Inhibit Stellate Cell Activity and Enhance Antitumor Effects of Nanoparticle Albumin-Bound Paclitaxel in Pancreatic Ductal Adenocarcinoma

Molecular Cancer Therapeutics, 2014

Research paper thumbnail of Yap Part of the Hippo Pathway is Overexpressed in Pancreatic Cancer

Research paper thumbnail of Increased CDC20 expression is associated with pancreatic ductal adenocarcinoma differentiation and progression

Journal of Hematology & Oncology, 2012

Purpose: Cell division cycle 20 (CDC20) homolog is an anaphase-promoting complex activator that i... more Purpose: Cell division cycle 20 (CDC20) homolog is an anaphase-promoting complex activator that is essential for cell division, but whether its expression in pancreatic ductal adenocarcinoma (PDAC) is significant is unknown. In this retrospective study, we determined whether aberrant CDC20 expression can be used as a biomarker in pancreatic ductal adenocarcinoma (PDAC) tumorigenesis and whether its expression reflects clinical progression. Experimental design: We compared CDC20 expression levels in normal, cancerous, and inflamed pancreatic tissues from stage II PDAC patients with clinical outcomes and determined CDC20 levels in seven PDAC cell lines. CDC20 was identified using a cDNA microarray database containing gene expression profiles for PDAC tissues and cell lines and chronic pancreatitis and normal pancreas tissues. Its expression was confirmed by real-time quantitative reverse-transcriptase-polymerase chain reaction (qRT-PCR). An immunohistochemical analysis of tissue microarrays from resected PDAC tumors and paired benign pancreatic tissues was done and CDC20 levels were correlated with clinical outcome. Results: Fifty-six patients were included in this study. A microarray analysis revealed 5-fold higher CDC20 expression in PDAC tissue than in chronic pancreatitis tissue. A qRT-PCR analysis confirmed a mean 20-fold higher CDC20 level in PDAC tissue than in normal pancreas and pancreatitis tissue. RNA and protein CDC20 expression was detected in several PDAC cell lines. An immunohistochemical analysis revealed higher CDC20 protein expression levels in PDAC tissue than in normal pancreas tissue, and high CDC20 expression was associated with poor differentiation (P = 0.020) and a significantly lower 5-year recurrence-free survival rate (P = 0.039); we also found a trend toward a shorter overall survival duration. Conclusions: Aberrant CDC20 expression may play an important role in PDAC tumorigenesis and progression and may thus be useful as a marker of disease progression and prognosis and as a therapeutic target.

Research paper thumbnail of Phosphatidylinositol 3-kinase and trypsin activation in pancreatitis

Journal of Clinical Investigation, 2001

Research paper thumbnail of The CUL7/F-box and WD Repeat Domain Containing 8 (CUL7/Fbxw8) Ubiquitin Ligase Promotes Degradation of Hematopoietic Progenitor Kinase 1

Journal of Biological Chemistry, 2014

Background: HPK1 is lost in Ͼ95% pancreatic cancer through proteasome-mediated degradation. Resul... more Background: HPK1 is lost in Ͼ95% pancreatic cancer through proteasome-mediated degradation. Results: The ubiquitination and degradation of HPK1 mediated by CUL7/Fbxw8 ubiquitin ligase required HPK1 kinase activity and autophosphorylation. Conclusion: Targeted HPK1 degradation by CUL7/Fbxw8 ubiquitin ligase constitutes a negative-feedback loop to restrain HPK1 activity. Significance: Our study revealed a direct link of CUL7/Fbxw8 ubiquitin ligase to the MAPK pathway.

Research paper thumbnail of Robust acinar cell transgene expression of CreErT via BAC recombineering

Research paper thumbnail of Human pancreatic acinar cells lack functional responses to cholecystokinin and gastrin

Gastroenterology, 2001

Pancreatic acinar cells from various species express cholecystokinin (CCK) A, CCK-B, or a combina... more Pancreatic acinar cells from various species express cholecystokinin (CCK) A, CCK-B, or a combination of these CCK receptor subtypes. The presence and functional roles of CCK receptors on human acinar cells remain unclear. Acini isolated from human pancreas were treated with CCK receptor agonists, CCK-8 and gastrin, and an agonist for m3 muscarinic acetylcholine receptors (m3 AchR), carbachol. Functional parameters measured included intracellular [Ca(2+)], amylase secretion, and ERK phosphorylation. Binding studies were performed using (125)I-CCK-8. Expression of messenger RNAs (mRNAs) was determined using real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) and localized by in situ hybridization. Human acini did not respond to CCK agonists. In contrast, they responded to carbachol with robust increases in each of the functional parameters. Moreover, the cells responded to CCK agonists after adenoviral-mediated gene transfer of CCK-A or CCK-B receptors. A low level of specific and a high level of nonspecific binding of (125)I-CCK-8 were observed. Quantitative RT-PCR indicated that the message levels for CCK-A receptors were approximately 30-fold lower than those of CCK-B receptors, which were approximately 10-fold lower than those of m3 Ach receptors. In situ hybridization indicated the presence of m3 Ach receptor and insulin mRNA but not CCK-A or CCK-B receptor mRNAs in adult human pancreas. These data indicate that human pancreatic acinar cells do not respond to CCK receptor agonists in terms of expected functional parameters and show that this is due to an insufficient level of receptor expression.

Research paper thumbnail of Acute pancreatitis and systemic inflammatory response induced by NF-κB activation in pancreas

Gastroenterology, 2001

blot arrays of >18,000 murine EST sequences (Mouse GDA, Incyte Genomics). Differential expression... more blot arrays of >18,000 murine EST sequences (Mouse GDA, Incyte Genomics). Differential expression of selected ESTs was confirmed by RNase protection assay using riboprebes prepared from sequenced linearized plasmids from which the ESTs were derived. Relative expression levels of pancreatic transcripts were compared to the pancreatic housekeeping transcript of murine acidic ribosomal protein PO (mARP). RESULTS: A sequence identified as mouse prostatic secretory glycoprotein (MPSG), a serine protease inhibitor, was found to be overexpressed 2.3-fold in the array analysis. The plasmid containing this EST (Incyte Genomics) was sequenced and the insert was established to be a full-length sequence of MPSG, now termed SPINK3. Following a single 6 hr treatment with cerulein, 50/~g/kg/hr, SPINK3 mRNA increased 2 to 2.5-fold at 24-72 hr over the relatively abundant basal transcript levels. The induction of SPINK3 was dose-dependent: no induction followed physiologic or sub-injurious cerulein stimulation (0.1 to 5/~g/kg/hr) whereas 10-50/~g/kg/hr induced a 1.8 to 2.6-fold increase in SPINK3 mRNA levels at 24 hr. CONCLUSION: These experiments identified pancreatic SPINK3 as a gene induced following acute or repetitive pancreatic injury in the mouse. Genetic polymorphisms of a similar serine protease inhibitor, SPINK1, may confer an increased risk of chronic pancreatitis (PfOtzer, Gastro 119:615, 2000). Therefore, pancreatic SPINK3, which we show here to have abundant basal transcript levels in the mouse and to be induced by injury, could be another protease inhibitor important in preventing repetitive acinar cell injury by prematurely activated digestive enzymes.

Research paper thumbnail of Lysophosphatidic acid signaling in the enteric nervous system

Gastroenterology, 2001

Background. Adenosine 3': 5'-cyclic monophosphate (cAMP) is a key mediator of slow synaptic trans... more Background. Adenosine 3': 5'-cyclic monophosphate (cAMP) is a key mediator of slow synaptic transmission in the ENS. FICRhR / cAMP imaging is providing spatial/temporal information about cAMP content in single gut neurons (Brain Res, 826:253-269,1999), but it does not permit accurate measurement of free cAMP and is less useful for detecting cell-specific changes in cAMP signals that may occur in the ENS. Aim & Methods. A rabbit polyclonal antibody that recognizes acrolein-dedvatizad cAMP was used to quantitate and identify the sub-cellular distribution of free cAMP in cultured myentedc neurons (n = 14 guinea-pigs). A biotinylated goat anti-rabbit lug and FITC-avidin were used to visualize the cAMP by laser confocal imaging. Responsive neurons were identified by PGP9.5 or NeuN staining and visualized by Texas Red-conjugated antibodies. Results. In neurons, forskolin increased cAMP immunofluorescence (IF) dose (0.001-100/~M)-and time-dependently, and its response was augmented by phosphodiesterase inhibitors (PDEI) from 3 to 15 fold higher than PDEI alone (lmM Ro-20-1724/IBMX). About 41% of 1,055 PGP9.5 immunoreactive (IR) neurons showed an increase in cAMP IR in cytoplasm, neudtes, axons, and distant varicose fibers; discrete sub-cellular localization patterns were often evident. Spatial gradients of cAMP IR are revealed over time (0.5-30 min) during uniform exposure to forskolin: The progression of the signal was from cell soma to neudtes, axons and then to distant varicose fibers. Sub-cellular distribution of the cAMP signal is not blocked by 1/~M FIX. The cAMP signal is abolished by absorption of primary antibody by.l-lmM cAMP. The cGMP cocktail (lOOp~U NaNitroprusside, 1raM IBMX, 5raM N-acetylcesteine) increased cAMP like IBMX, in <5% of neurons. Depolarization of neurons was without effect. The slow EPSP-mimetic agents SP, VIP and GRP (5p.M) increased cAMP by 50% of the maximum response to forskolin in <15% of neurons for each agent. Responses were often restricted to cytoplasm, and VIP also elevated cAMP in varicose fibers. VIP (n=41) or forskolin-responsive neurons had Dogiel Type I, filamentous or Dogiel Type II morphology. Conclusions. Cyclic AMP IF provides a suitable method for visualization, quantitation and transmitter analysis of cAMP signals in gut neurons. The spatial profile of the signal is explained by diffusion of free cAMP and differential AC/ cAMP signaling in distinct regions of the neuron (NIH DK44179-07, 3 R01 DK44179-07S1, NIH CA09171 & Nat. Multiple Sclerosis Soc.).

Research paper thumbnail of Genes down-regulated in pancreatic adenocarcinoma affect cell survival: Reintroduction of proapoptotic BNIP3 leads to cancer cell death

Gastroenterology, 2003

with normal physiology and morphology. Results: There was a striking association between carriage... more with normal physiology and morphology. Results: There was a striking association between carriage of the Asp299Gly mutation and presence of premalignant gastric lesions (Table 1). Compared to infected subjects who did not develop ATR/I-IC, the odds ratio for H. pyloriinduced ATR and HC was 11.4 (95% confidence interval 2.2-78.2). Eight of the 13 subjects who were found to have the TLR4 mutation in the ATP,/HC group had the most severe phenotype, i.e. lowest acid outputs and highest atrophy/inflammation scores. Conclusions: Our results indicate that carnage of the TLR4 Asp299Gly mutation predisposes H. pylori infected subjects to a severe phenotype characterised by hypochlorhydna and gastric atrophy, the two most important precursor lesions of GC. This novel finding calls for evaluation of the TLR4 polymorphism as another important host genetic factor in H. pylor/-induced gastric cancer.

Research paper thumbnail of M1961 Galectin-3 Induces Cell Proliferation and Invasion by Binding K-RAS and Activating RAS Signaling in Pancreatic Cancer Cells

Gastroenterology, 2010

strains were less able to activate Gli1 and Snail. Snail expression in Gli1 overexpressed gastric... more strains were less able to activate Gli1 and Snail. Snail expression in Gli1 overexpressed gastric cell was higher than that of Gli1 unexpressed gastric epithelial cell and Gli1 overexpressed gastric cell was 8 fold increased snail promoter activity. In this study, we showed that Hh signaling was clearly upregulated in gastric cancer cell in cagA depedent manner and activated Gli1 upregulate Snail to promote epithelial-to-mesenchymal transition.

Research paper thumbnail of Direct Versus Indirect Action of Cholecystokinin on Human Pancreatic Acinar Cells: Is It Time for a Judgment after a Century of Trial?

Gastroenterology, 2008

sults of a study on radiofrequency ablation for highgrade dysplasia in Barrett's esophagus sugges... more sults of a study on radiofrequency ablation for highgrade dysplasia in Barrett's esophagus suggest that this technique may have similar efficacy to PDT in eliminating dysplasia, but with less patient inconvenience and fewer serious side effects. 12 Nevertheless, far more data are needed before radiofrequency ablation can be recommended for widespread clinical use. In the meantime, Prasad et al have made a valuable contribution, showing how translational research might direct endoscopic therapies for patients with dysplasia in Barrett's esophagus.

Research paper thumbnail of Regulating insulin-receptor-gene expression by differentiation and hormones

Diabetes Care, 1990

Insulin regulates cell function by first binding to the insulin receptor (IR) localized on the ce... more Insulin regulates cell function by first binding to the insulin receptor (IR) localized on the cell surface. With the cloning of IR cDNA and the IR-gene promoter, the regulation of the IR gene during differentiation and by various hormones can be studied. Muscle is a major target tissue for insulin action. BC3H1 cells, a mouse muscle cell line in culture, are a model cell type for studying insulin action. Differentiation in these cells results in a 5- to 10-fold increase in IR binding and a 5- to 10-fold increase in IR content. Studies of IR mRNA by Northern and slot-blot analyses reveal a 10-fold increase in IR mRNA after differentiation. These studies indicate that there is a selective increase in IR-gene expression during muscle differentiation. A similar increase in IRgene expression is observed for the IR during pancreatic acinar cell differentiation. Glucocorticoids increase IR content in several target tissues. Studies in cultured IM-9 lymphocytes indicate that glucocorticoid...

Research paper thumbnail of Abstract 3655: Mast cells promote immunosuppression and pancreatic ductal adenocarcinoma development

Cancer Research, 2011

Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a mostly uniformly lethal disease and is n... more Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a mostly uniformly lethal disease and is notoriously resistant to most therapies. Immunotherapy may be a fourth strategy for treating PDAC, supplementing surgery, chemotherapy, and radiation therapy. Immunosuppression is one of the largest bottlenecks in cancer immunotherapy development. In our preliminary studies, we analyzed a panel of immunocytes in human PDAC tissue samples and found that mast cells infiltrated at the tumor margin. This high infiltration of human mast cells into the tumor microenvironment was associated with poor survival. An early influx of mast cells into the tumor microenvironment combined with the IL-10 and receptor upregulation we found in a transgenic spontaneous PDAC mouse model, was associated with malignant progression. In addition, mast cells were found to be essential to PDAC tumorigenesis in a mast cell-deficient Kitw-sh/w-sh mouse model (Kit-/- mice) compared to wild-type C57BL/6 control mice (WT mice). However, how mast cells regulate immunologic effects in the tumor microenvironment and contribute to PDAC tumorigenesis are unknown. We hypothesized that mast cells mediate the immunosuppression that promotes PDAC development. Methods: We quantified mast cells in 16 normal human pancreas and 22 pancreatitis patients’ tissue samples. Panc-02 PDAC cells were orthotopically implanted into the pancreases of Kit-/-, mast cell-reconstituted, and WT mice. In the mast cell reconstitution model, Kit-/- mice received bone marrow-derived mast cells (BMMCs) from IL-10-/-, PGE2-/-, and WT mice. To determine whether targeting mast cells has a therapeutic effect in PDAC, we treated mice PDAC with cromolyn, a mast cell stabilizer. Saline and gemcitabine were used as controls. Tumor sizes were measured by bioluminescence imaging. Results: Mast cell infiltration to human pancreatic disease was associated with inflammation and malignancy progression in PDAC (P Conclusions: Mast cells promote inflammatory progression and immune suppression in PDAC tumorigenesis. Mast cell-targeted therapy resulted in a higher treatment response rate in mouse PDAC than did classic chemotherapy. Therefore, therapy targeting on mast cells may overcome immune suppression and improve PDAC immunotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3655. doi:10.1158/1538-7445.AM2011-3655

Research paper thumbnail of Abstract 3822: Immunosuppressive role of mast cells is essential for the tumorigenesis of pancreatic ductal adenocarcinoma (PDAC)

Research paper thumbnail of Lysophosphatidic acid stimulates calcium transients in enteric glia

Neuroscience, 2004

The enteric nervous system plays an integral role in the gastrointestinal tract. Within this intr... more The enteric nervous system plays an integral role in the gastrointestinal tract. Within this intricate network, enteric glia are crucial in the maintenance of normal bowel function, yet their signaling mechanisms are poorly understood. Enteric glia, and not enteric neurons, selectively responded to lysophosphatidic acid (LPA), a product of phosphatidylcholine metabolism, with dose-dependent calcium (Ca 2؉) signaling over a range from 100 pM to 10 M. The elicited calcium transients involved both the mobilization of intracellular Ca 2؉ stores and the influx of extracellular Ca 2؉ as LPA signals were obliterated following the depletion of intracellular Ca 2؉ and attenuated by the removal of Ca 2؉ from the perfusion buffer. Pretreatment with pertussis toxin (100 ng/ml) reduced the magnitude of LPA Ca 2؉ transients (95؎20 nM vs 168؎17 nM for controls). Repetitive exposure yielded diminished responsiveness, with a 25% reduction in [Ca 2؉ ] i between first and second exposures. Inhibition of the inositol 1,4,5-trisphosphate (IP 3) receptor with 200 M 2-aminoethoxydiphenylborate (2APB) abolished LPA signals. RT-PCR analysis demonstrated the presence of two LPA-coupled endothelial differentiation gene (EDG) receptor mRNAs (EDG-2 and EDG-7) in myenteric plexus primary cultures. EDG-2 expression in glial cells of the ENS was confirmed immunocytochemically.

Research paper thumbnail of Sphingosine-1-phosphate mediates calcium signaling in guinea pig enteroglial cells

Journal of Surgical …, 2004

The enteric nervous system, which regulates multiple aspects of digestive activity, is composed o... more The enteric nervous system, which regulates multiple aspects of digestive activity, is composed of two major cell types, neurons and glial cells. Enteric glia, but not enteric neurons, respond to bioactive lipids with calcium signaling. The sphingomyelin metabolite sphingosine-1-phosphate (S1P) caused dose-dependent calcium (Ca(2+)) signaling using extracellular and intracellular Ca(2+). The signal transduction cascade was pertussis toxin-insensitive and involved an extracellular receptor since repetitive exposure yielded diminished responsiveness. Inhibition of either phospholipase C or the inositol 1,4,5-trisphosphate receptor abolished S1P effects. RT-PCR analysis demonstrated the presence of S1P-coupled endothelial differentiation gene (EDG) receptor mRNAs (EDG-1, EDG-3, and EDG-5) within the enteric nervous system. Immunocytochemical analysis demonstrated strong expression of both EDG-1 and EDG-3 and weak expression of EDG-5 in enteric glial cells. Other sphingomyelin cycle components, including sphingomyelin, sphingomyelinase, and sphingosine caused Ca(2+) transients in enteric glia. Related lipids lysophosphatidic acid and sphingosylphosphorylcholine also induced Ca(2+) signaling in enteric glia, suggesting that multiple lipid-activated signaling mechanisms exist in these cells.

Research paper thumbnail of Reminding Us That We Are "Tough and Competent

American Journal of Critical Care

Reminding Us That We Are "Tough and Competent" Thank you for the inspiring (and for me, timely) e... more Reminding Us That We Are "Tough and Competent" Thank you for the inspiring (and for me, timely) editorial, "Tough and Competent." 1 I grew up in awe of space travel and desperately wanted to be a "nurse in space" on the space shuttle. So I read the editorial that recounted the history of NASA with enjoyment and fascination. In this current health care environment, there is a lack of leaders who will shine the light on "carelessness, incapacity, and neglect." They are not inspiring us to be tough and competent, and to never compromise regarding our responsibilities. But this piece spoke to my head and heart when you referred to NASA's operations and the parallels to critical care. A couple of weeks ago I had lost my inspiration when 5 health care-associated infections were sent to me tagged to the units in my division. This occurred after we had several months of 1 or 2 health care-associated infections. I was feeling defeated and worried that all of the work on reducing them was meaningless. The next day I decided to move forward and embrace the current state of events as a continued challenge. You are right about critical care professionals, we are tough and competent. We just need to be reminded of that.

Research paper thumbnail of Down-regulation of the G-proteins G q α and G 11 α by transfected human M3 muscarinic acetylcholine receptors in Chinese hamster ovary cells is independent of receptor down-regulation

Biochemical Journal, 1995

Chinese hamster ovary cells stably transfected with human M3 muscarinic acetylcholine receptors s... more Chinese hamster ovary cells stably transfected with human M3 muscarinic acetylcholine receptors show a 40-50% reduction in the immunoreactive G-proteins Gq alpha and G11 alpha when stimulated with the cholinergic agonist carbachol. This effect is seen after 9 h, is maximal after 24 h, and occurs over a range of carbachol concentrations that activate phosphoinositide hydrolysis in these cells. The effect is specific for Gq alpha family proteins as Gs alpha was slightly increased after carbachol treatment and G13 alpha was unchanged. Using a urea gel system, we were able to resolve Gq alpha and G11 alpha, both of which were down-regulated by carbachol. An M3 receptor mutant, with C-terminal threonines changed to alanines as described previously, binds ligand and activates phosphoinositide hydrolysis normally but is not down-regulated in response to carbachol. This receptor, however, induces Gq alpha/G11 alpha down-regulation similarly to wild-type M3 receptors, indicating that G-prote...

Research paper thumbnail of Psychosocial and functional outcomes in women after coronary artery bypass surgery

Critical care nurse, 1997

In this longitudinal study of women after CABS, the women were coping admirably with good psychos... more In this longitudinal study of women after CABS, the women were coping admirably with good psychosocial and functional outcomes up to 3 months postoperatively. It is suggested that women be instructed how to find and use their support systems and be told that most women do well and return to normal activities of daily living after CABS.

Research paper thumbnail of Bisphosphonates Inhibit Stellate Cell Activity and Enhance Antitumor Effects of Nanoparticle Albumin-Bound Paclitaxel in Pancreatic Ductal Adenocarcinoma

Molecular Cancer Therapeutics, 2014

Research paper thumbnail of Yap Part of the Hippo Pathway is Overexpressed in Pancreatic Cancer

Research paper thumbnail of Increased CDC20 expression is associated with pancreatic ductal adenocarcinoma differentiation and progression

Journal of Hematology & Oncology, 2012

Purpose: Cell division cycle 20 (CDC20) homolog is an anaphase-promoting complex activator that i... more Purpose: Cell division cycle 20 (CDC20) homolog is an anaphase-promoting complex activator that is essential for cell division, but whether its expression in pancreatic ductal adenocarcinoma (PDAC) is significant is unknown. In this retrospective study, we determined whether aberrant CDC20 expression can be used as a biomarker in pancreatic ductal adenocarcinoma (PDAC) tumorigenesis and whether its expression reflects clinical progression. Experimental design: We compared CDC20 expression levels in normal, cancerous, and inflamed pancreatic tissues from stage II PDAC patients with clinical outcomes and determined CDC20 levels in seven PDAC cell lines. CDC20 was identified using a cDNA microarray database containing gene expression profiles for PDAC tissues and cell lines and chronic pancreatitis and normal pancreas tissues. Its expression was confirmed by real-time quantitative reverse-transcriptase-polymerase chain reaction (qRT-PCR). An immunohistochemical analysis of tissue microarrays from resected PDAC tumors and paired benign pancreatic tissues was done and CDC20 levels were correlated with clinical outcome. Results: Fifty-six patients were included in this study. A microarray analysis revealed 5-fold higher CDC20 expression in PDAC tissue than in chronic pancreatitis tissue. A qRT-PCR analysis confirmed a mean 20-fold higher CDC20 level in PDAC tissue than in normal pancreas and pancreatitis tissue. RNA and protein CDC20 expression was detected in several PDAC cell lines. An immunohistochemical analysis revealed higher CDC20 protein expression levels in PDAC tissue than in normal pancreas tissue, and high CDC20 expression was associated with poor differentiation (P = 0.020) and a significantly lower 5-year recurrence-free survival rate (P = 0.039); we also found a trend toward a shorter overall survival duration. Conclusions: Aberrant CDC20 expression may play an important role in PDAC tumorigenesis and progression and may thus be useful as a marker of disease progression and prognosis and as a therapeutic target.

Research paper thumbnail of Phosphatidylinositol 3-kinase and trypsin activation in pancreatitis

Journal of Clinical Investigation, 2001

Research paper thumbnail of The CUL7/F-box and WD Repeat Domain Containing 8 (CUL7/Fbxw8) Ubiquitin Ligase Promotes Degradation of Hematopoietic Progenitor Kinase 1

Journal of Biological Chemistry, 2014

Background: HPK1 is lost in Ͼ95% pancreatic cancer through proteasome-mediated degradation. Resul... more Background: HPK1 is lost in Ͼ95% pancreatic cancer through proteasome-mediated degradation. Results: The ubiquitination and degradation of HPK1 mediated by CUL7/Fbxw8 ubiquitin ligase required HPK1 kinase activity and autophosphorylation. Conclusion: Targeted HPK1 degradation by CUL7/Fbxw8 ubiquitin ligase constitutes a negative-feedback loop to restrain HPK1 activity. Significance: Our study revealed a direct link of CUL7/Fbxw8 ubiquitin ligase to the MAPK pathway.

Research paper thumbnail of Robust acinar cell transgene expression of CreErT via BAC recombineering

Research paper thumbnail of Human pancreatic acinar cells lack functional responses to cholecystokinin and gastrin

Gastroenterology, 2001

Pancreatic acinar cells from various species express cholecystokinin (CCK) A, CCK-B, or a combina... more Pancreatic acinar cells from various species express cholecystokinin (CCK) A, CCK-B, or a combination of these CCK receptor subtypes. The presence and functional roles of CCK receptors on human acinar cells remain unclear. Acini isolated from human pancreas were treated with CCK receptor agonists, CCK-8 and gastrin, and an agonist for m3 muscarinic acetylcholine receptors (m3 AchR), carbachol. Functional parameters measured included intracellular [Ca(2+)], amylase secretion, and ERK phosphorylation. Binding studies were performed using (125)I-CCK-8. Expression of messenger RNAs (mRNAs) was determined using real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) and localized by in situ hybridization. Human acini did not respond to CCK agonists. In contrast, they responded to carbachol with robust increases in each of the functional parameters. Moreover, the cells responded to CCK agonists after adenoviral-mediated gene transfer of CCK-A or CCK-B receptors. A low level of specific and a high level of nonspecific binding of (125)I-CCK-8 were observed. Quantitative RT-PCR indicated that the message levels for CCK-A receptors were approximately 30-fold lower than those of CCK-B receptors, which were approximately 10-fold lower than those of m3 Ach receptors. In situ hybridization indicated the presence of m3 Ach receptor and insulin mRNA but not CCK-A or CCK-B receptor mRNAs in adult human pancreas. These data indicate that human pancreatic acinar cells do not respond to CCK receptor agonists in terms of expected functional parameters and show that this is due to an insufficient level of receptor expression.

Research paper thumbnail of Acute pancreatitis and systemic inflammatory response induced by NF-κB activation in pancreas

Gastroenterology, 2001

blot arrays of >18,000 murine EST sequences (Mouse GDA, Incyte Genomics). Differential expression... more blot arrays of >18,000 murine EST sequences (Mouse GDA, Incyte Genomics). Differential expression of selected ESTs was confirmed by RNase protection assay using riboprebes prepared from sequenced linearized plasmids from which the ESTs were derived. Relative expression levels of pancreatic transcripts were compared to the pancreatic housekeeping transcript of murine acidic ribosomal protein PO (mARP). RESULTS: A sequence identified as mouse prostatic secretory glycoprotein (MPSG), a serine protease inhibitor, was found to be overexpressed 2.3-fold in the array analysis. The plasmid containing this EST (Incyte Genomics) was sequenced and the insert was established to be a full-length sequence of MPSG, now termed SPINK3. Following a single 6 hr treatment with cerulein, 50/~g/kg/hr, SPINK3 mRNA increased 2 to 2.5-fold at 24-72 hr over the relatively abundant basal transcript levels. The induction of SPINK3 was dose-dependent: no induction followed physiologic or sub-injurious cerulein stimulation (0.1 to 5/~g/kg/hr) whereas 10-50/~g/kg/hr induced a 1.8 to 2.6-fold increase in SPINK3 mRNA levels at 24 hr. CONCLUSION: These experiments identified pancreatic SPINK3 as a gene induced following acute or repetitive pancreatic injury in the mouse. Genetic polymorphisms of a similar serine protease inhibitor, SPINK1, may confer an increased risk of chronic pancreatitis (PfOtzer, Gastro 119:615, 2000). Therefore, pancreatic SPINK3, which we show here to have abundant basal transcript levels in the mouse and to be induced by injury, could be another protease inhibitor important in preventing repetitive acinar cell injury by prematurely activated digestive enzymes.

Research paper thumbnail of Lysophosphatidic acid signaling in the enteric nervous system

Gastroenterology, 2001

Background. Adenosine 3': 5'-cyclic monophosphate (cAMP) is a key mediator of slow synaptic trans... more Background. Adenosine 3': 5'-cyclic monophosphate (cAMP) is a key mediator of slow synaptic transmission in the ENS. FICRhR / cAMP imaging is providing spatial/temporal information about cAMP content in single gut neurons (Brain Res, 826:253-269,1999), but it does not permit accurate measurement of free cAMP and is less useful for detecting cell-specific changes in cAMP signals that may occur in the ENS. Aim & Methods. A rabbit polyclonal antibody that recognizes acrolein-dedvatizad cAMP was used to quantitate and identify the sub-cellular distribution of free cAMP in cultured myentedc neurons (n = 14 guinea-pigs). A biotinylated goat anti-rabbit lug and FITC-avidin were used to visualize the cAMP by laser confocal imaging. Responsive neurons were identified by PGP9.5 or NeuN staining and visualized by Texas Red-conjugated antibodies. Results. In neurons, forskolin increased cAMP immunofluorescence (IF) dose (0.001-100/~M)-and time-dependently, and its response was augmented by phosphodiesterase inhibitors (PDEI) from 3 to 15 fold higher than PDEI alone (lmM Ro-20-1724/IBMX). About 41% of 1,055 PGP9.5 immunoreactive (IR) neurons showed an increase in cAMP IR in cytoplasm, neudtes, axons, and distant varicose fibers; discrete sub-cellular localization patterns were often evident. Spatial gradients of cAMP IR are revealed over time (0.5-30 min) during uniform exposure to forskolin: The progression of the signal was from cell soma to neudtes, axons and then to distant varicose fibers. Sub-cellular distribution of the cAMP signal is not blocked by 1/~M FIX. The cAMP signal is abolished by absorption of primary antibody by.l-lmM cAMP. The cGMP cocktail (lOOp~U NaNitroprusside, 1raM IBMX, 5raM N-acetylcesteine) increased cAMP like IBMX, in <5% of neurons. Depolarization of neurons was without effect. The slow EPSP-mimetic agents SP, VIP and GRP (5p.M) increased cAMP by 50% of the maximum response to forskolin in <15% of neurons for each agent. Responses were often restricted to cytoplasm, and VIP also elevated cAMP in varicose fibers. VIP (n=41) or forskolin-responsive neurons had Dogiel Type I, filamentous or Dogiel Type II morphology. Conclusions. Cyclic AMP IF provides a suitable method for visualization, quantitation and transmitter analysis of cAMP signals in gut neurons. The spatial profile of the signal is explained by diffusion of free cAMP and differential AC/ cAMP signaling in distinct regions of the neuron (NIH DK44179-07, 3 R01 DK44179-07S1, NIH CA09171 & Nat. Multiple Sclerosis Soc.).

Research paper thumbnail of Genes down-regulated in pancreatic adenocarcinoma affect cell survival: Reintroduction of proapoptotic BNIP3 leads to cancer cell death

Gastroenterology, 2003

with normal physiology and morphology. Results: There was a striking association between carriage... more with normal physiology and morphology. Results: There was a striking association between carriage of the Asp299Gly mutation and presence of premalignant gastric lesions (Table 1). Compared to infected subjects who did not develop ATR/I-IC, the odds ratio for H. pyloriinduced ATR and HC was 11.4 (95% confidence interval 2.2-78.2). Eight of the 13 subjects who were found to have the TLR4 mutation in the ATP,/HC group had the most severe phenotype, i.e. lowest acid outputs and highest atrophy/inflammation scores. Conclusions: Our results indicate that carnage of the TLR4 Asp299Gly mutation predisposes H. pylori infected subjects to a severe phenotype characterised by hypochlorhydna and gastric atrophy, the two most important precursor lesions of GC. This novel finding calls for evaluation of the TLR4 polymorphism as another important host genetic factor in H. pylor/-induced gastric cancer.

Research paper thumbnail of M1961 Galectin-3 Induces Cell Proliferation and Invasion by Binding K-RAS and Activating RAS Signaling in Pancreatic Cancer Cells

Gastroenterology, 2010

strains were less able to activate Gli1 and Snail. Snail expression in Gli1 overexpressed gastric... more strains were less able to activate Gli1 and Snail. Snail expression in Gli1 overexpressed gastric cell was higher than that of Gli1 unexpressed gastric epithelial cell and Gli1 overexpressed gastric cell was 8 fold increased snail promoter activity. In this study, we showed that Hh signaling was clearly upregulated in gastric cancer cell in cagA depedent manner and activated Gli1 upregulate Snail to promote epithelial-to-mesenchymal transition.

Research paper thumbnail of Direct Versus Indirect Action of Cholecystokinin on Human Pancreatic Acinar Cells: Is It Time for a Judgment after a Century of Trial?

Gastroenterology, 2008

sults of a study on radiofrequency ablation for highgrade dysplasia in Barrett's esophagus sugges... more sults of a study on radiofrequency ablation for highgrade dysplasia in Barrett's esophagus suggest that this technique may have similar efficacy to PDT in eliminating dysplasia, but with less patient inconvenience and fewer serious side effects. 12 Nevertheless, far more data are needed before radiofrequency ablation can be recommended for widespread clinical use. In the meantime, Prasad et al have made a valuable contribution, showing how translational research might direct endoscopic therapies for patients with dysplasia in Barrett's esophagus.

Research paper thumbnail of Regulating insulin-receptor-gene expression by differentiation and hormones

Diabetes Care, 1990

Insulin regulates cell function by first binding to the insulin receptor (IR) localized on the ce... more Insulin regulates cell function by first binding to the insulin receptor (IR) localized on the cell surface. With the cloning of IR cDNA and the IR-gene promoter, the regulation of the IR gene during differentiation and by various hormones can be studied. Muscle is a major target tissue for insulin action. BC3H1 cells, a mouse muscle cell line in culture, are a model cell type for studying insulin action. Differentiation in these cells results in a 5- to 10-fold increase in IR binding and a 5- to 10-fold increase in IR content. Studies of IR mRNA by Northern and slot-blot analyses reveal a 10-fold increase in IR mRNA after differentiation. These studies indicate that there is a selective increase in IR-gene expression during muscle differentiation. A similar increase in IRgene expression is observed for the IR during pancreatic acinar cell differentiation. Glucocorticoids increase IR content in several target tissues. Studies in cultured IM-9 lymphocytes indicate that glucocorticoid...

Research paper thumbnail of Abstract 3655: Mast cells promote immunosuppression and pancreatic ductal adenocarcinoma development

Cancer Research, 2011

Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a mostly uniformly lethal disease and is n... more Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a mostly uniformly lethal disease and is notoriously resistant to most therapies. Immunotherapy may be a fourth strategy for treating PDAC, supplementing surgery, chemotherapy, and radiation therapy. Immunosuppression is one of the largest bottlenecks in cancer immunotherapy development. In our preliminary studies, we analyzed a panel of immunocytes in human PDAC tissue samples and found that mast cells infiltrated at the tumor margin. This high infiltration of human mast cells into the tumor microenvironment was associated with poor survival. An early influx of mast cells into the tumor microenvironment combined with the IL-10 and receptor upregulation we found in a transgenic spontaneous PDAC mouse model, was associated with malignant progression. In addition, mast cells were found to be essential to PDAC tumorigenesis in a mast cell-deficient Kitw-sh/w-sh mouse model (Kit-/- mice) compared to wild-type C57BL/6 control mice (WT mice). However, how mast cells regulate immunologic effects in the tumor microenvironment and contribute to PDAC tumorigenesis are unknown. We hypothesized that mast cells mediate the immunosuppression that promotes PDAC development. Methods: We quantified mast cells in 16 normal human pancreas and 22 pancreatitis patients’ tissue samples. Panc-02 PDAC cells were orthotopically implanted into the pancreases of Kit-/-, mast cell-reconstituted, and WT mice. In the mast cell reconstitution model, Kit-/- mice received bone marrow-derived mast cells (BMMCs) from IL-10-/-, PGE2-/-, and WT mice. To determine whether targeting mast cells has a therapeutic effect in PDAC, we treated mice PDAC with cromolyn, a mast cell stabilizer. Saline and gemcitabine were used as controls. Tumor sizes were measured by bioluminescence imaging. Results: Mast cell infiltration to human pancreatic disease was associated with inflammation and malignancy progression in PDAC (P Conclusions: Mast cells promote inflammatory progression and immune suppression in PDAC tumorigenesis. Mast cell-targeted therapy resulted in a higher treatment response rate in mouse PDAC than did classic chemotherapy. Therefore, therapy targeting on mast cells may overcome immune suppression and improve PDAC immunotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3655. doi:10.1158/1538-7445.AM2011-3655

Research paper thumbnail of Abstract 3822: Immunosuppressive role of mast cells is essential for the tumorigenesis of pancreatic ductal adenocarcinoma (PDAC)

Research paper thumbnail of Lysophosphatidic acid stimulates calcium transients in enteric glia

Neuroscience, 2004

The enteric nervous system plays an integral role in the gastrointestinal tract. Within this intr... more The enteric nervous system plays an integral role in the gastrointestinal tract. Within this intricate network, enteric glia are crucial in the maintenance of normal bowel function, yet their signaling mechanisms are poorly understood. Enteric glia, and not enteric neurons, selectively responded to lysophosphatidic acid (LPA), a product of phosphatidylcholine metabolism, with dose-dependent calcium (Ca 2؉) signaling over a range from 100 pM to 10 M. The elicited calcium transients involved both the mobilization of intracellular Ca 2؉ stores and the influx of extracellular Ca 2؉ as LPA signals were obliterated following the depletion of intracellular Ca 2؉ and attenuated by the removal of Ca 2؉ from the perfusion buffer. Pretreatment with pertussis toxin (100 ng/ml) reduced the magnitude of LPA Ca 2؉ transients (95؎20 nM vs 168؎17 nM for controls). Repetitive exposure yielded diminished responsiveness, with a 25% reduction in [Ca 2؉ ] i between first and second exposures. Inhibition of the inositol 1,4,5-trisphosphate (IP 3) receptor with 200 M 2-aminoethoxydiphenylborate (2APB) abolished LPA signals. RT-PCR analysis demonstrated the presence of two LPA-coupled endothelial differentiation gene (EDG) receptor mRNAs (EDG-2 and EDG-7) in myenteric plexus primary cultures. EDG-2 expression in glial cells of the ENS was confirmed immunocytochemically.

Research paper thumbnail of Sphingosine-1-phosphate mediates calcium signaling in guinea pig enteroglial cells

Journal of Surgical …, 2004

The enteric nervous system, which regulates multiple aspects of digestive activity, is composed o... more The enteric nervous system, which regulates multiple aspects of digestive activity, is composed of two major cell types, neurons and glial cells. Enteric glia, but not enteric neurons, respond to bioactive lipids with calcium signaling. The sphingomyelin metabolite sphingosine-1-phosphate (S1P) caused dose-dependent calcium (Ca(2+)) signaling using extracellular and intracellular Ca(2+). The signal transduction cascade was pertussis toxin-insensitive and involved an extracellular receptor since repetitive exposure yielded diminished responsiveness. Inhibition of either phospholipase C or the inositol 1,4,5-trisphosphate receptor abolished S1P effects. RT-PCR analysis demonstrated the presence of S1P-coupled endothelial differentiation gene (EDG) receptor mRNAs (EDG-1, EDG-3, and EDG-5) within the enteric nervous system. Immunocytochemical analysis demonstrated strong expression of both EDG-1 and EDG-3 and weak expression of EDG-5 in enteric glial cells. Other sphingomyelin cycle components, including sphingomyelin, sphingomyelinase, and sphingosine caused Ca(2+) transients in enteric glia. Related lipids lysophosphatidic acid and sphingosylphosphorylcholine also induced Ca(2+) signaling in enteric glia, suggesting that multiple lipid-activated signaling mechanisms exist in these cells.