Rainer Heuchel - Profile on Academia.edu (original) (raw)

Papers by Rainer Heuchel

Slip-X-Chip: A Sliding Microfluidic Platform with Cross-Flow

2022 IEEE 35th International Conference on Micro Electro Mechanical Systems Conference (MEMS), Jan 9, 2022

In sliding microfluidic platforms (“SlipChip” [1]), two plates with microfluidic wells slide in c... more In sliding microfluidic platforms (“SlipChip” [1]), two plates with microfluidic wells slide in close contact to perform multiplexed reactions in a single operation. In-plane “sliding-flow” liquid transport, however, limits the potential assay operations to 1) sample compartmentalization/metering, 2) reagent addition, 3) mixing, and 4) aliquoting. Here, we introduce a three-plate sliding microfluidics platform, “Slip-X-Chip”, that additionally includes out-of-plane “cross-flow” liquid transport. Slip-X-Chip allows two additional assay operations: 5) sample concentration and 6) liquid exchange/washing, while retaining the simplicity of operation (one-step operation; no precise pipetting required; no external equipment). These additional assay operations extend the range and complexity of applications enabled by sliding microfluidics. We here demonstrate 1) splitting bead solutions in compartments with different concentrations and 2) compartmentalizing human cells from solution, followed by a viability assay. We foresee that Slip-X-Chip could be further adapted to, e.g., cell counting, cell staining, or ELISA.

Developmental Biology, Oct 1, 2013

Members of the bone morphogenetic protein (BMP) superfamily, including transforming growth factor... more Members of the bone morphogenetic protein (BMP) superfamily, including transforming growth factor-betas (TGFβ), regulate multiple aspects of chondrogenesis. Smad7 is an intracellular inhibitor of BMP and TGFβ signaling. Studies in which Smad7 was overexpressed in chondrocytes demonstrated that Smad7 can impact chondrogenesis by inhibiting BMP signaling. However, whether Smad7 is actually required for endochondral ossification in vivo is unclear. Moreover, whether Smad7 regulates TGFβ in addition to BMP signaling in developing cartilage is unknown. In this study, we found that Smad7 is required for both axial and appendicular skeletal development. Loss of Smad7 led to impairment of the cell cycle in chondrocytes and to defects in terminal maturation. This phenotype was attributed to upregulation of both BMP and TGFβ signaling in Smad7 mutant growth plates. Moreover, Smad7-/-mice develop hypocellular cores in the medial growth plates, associated with elevated HIF1α levels, cell death, and intracellular retention of types II and X collagen. Thus, Smad7 may be required to mediate cell stress responses in the growth plate during development.

Scientific Reports, Oct 15, 2020

The pancreatic tumour stroma is composed of phenotypically heterogenous cancer-associated fibrobl... more The pancreatic tumour stroma is composed of phenotypically heterogenous cancer-associated fibroblasts (CAFs) with both pro-and anti-tumorigenic functions. Here, we studied the impact of calcipotriol, a vitamin D 3 analogue, on the activation of human pancreatic CAFs and T cells using 2-and 3-dimensional (2D, 3D) cell culture models. We found that calcipotriol decreased CAF proliferation and migration and reduced the release of the pro-tumorigenic factors prostaglandin E 2 , IL-6, periostin, and leukemia inhibitory factor. However, calcipotriol promoted PD-L1 upregulation, which could influence T cell mediated tumour immune surveillance. Calcipotriol reduced T cell proliferation and production of IFN-γ, granzyme B and IL-17, but increased IL-10 secretion. These effects were even more profound in the presence of CAFs in 2D cultures and in the presence of CAFs and pancreatic tumour cell line (PANC-1) spheroids in 3D cultures. Functional assays on tumour infiltrating lymphocytes also showed a reduction in T cell activation by calcipotriol. This suggests that calcipotriol reduces the tumour supportive activity of CAFs but at the same time reduces T cell effector functions, which could compromise the patients' tumour immune surveillance. Thus, vitamin D 3 analogues appear to have dual functions in the context of pancreatic cancer, which could have important clinical implications.

The EMBO Journal, Jun 1, 1994

We have described and cloned previously a factor NMTF-1) that binds specifically to heavy metal-r... more We have described and cloned previously a factor NMTF-1) that binds specifically to heavy metal-responsive DNA sequence elements in the enhancer/promoter region of metallothionein genes. MTF-1 is a protein of 72.5 kDa that contains six zinc fingers and multiple domains for transcriptional activation. Here we report the disruption of both alleles of the MTF-1 gene in mouse embryonic stem cells by homologous recombination. The resulting null mutant cell line fails to produce detectable amounts of MTF-1. Moreover, due to the loss of MTF-1, the endogenous metallothionein I and I genes are silent, indicating that MTF-1 is required for both their basal and zinc-induced anscription. In addition to zinc, other heavy metals, including cadmium, copper, nickel and lead, also fail to activate metal-responsive promoters in null mutant cells. However, cotransfection of an MTF-1 expression vector and metal-responsive reporter genes yields strong basal transcription that can be further boosted by zinc treatment of cells. These results demonstrate that MTF-1 is essential for metallothionein gene regulation. Finally, we present evidence that MTF-1 itself is a zinc sensor, which exhibits increased DNA binding activity upon zinc treatment.

Abstract B43: High-throughput drug screening model using 3D cultured human pancreatic ductal adenocarcinoma cells

Background: Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive disease and the fo... more Background: Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive disease and the fourth most common cause of cancer related death. It is lethal in nearly all patients, due to an almost complete chemo-resistance. Most if not all drugs which successfully pass preclinical tests, fail miserably in the clinic. PDAC is characterized by exceptionally fibrotic stroma, which is believed to be important for pancreatic cancer progression and general therapy resistance. Compared to traditional 2D cell culture, cells in a three-dimensional (3D) context demonstrate different behavior and functions as reflected by building up a microenvironment that more closely mimics the one observed in native tissue. This feature is critical for testing drug efficiency, as the cellular response to drugs is profoundly affected by environmental cues. Our lab has previously set up a drug screening system using 3D cell culture containing human PDAC cells. We demonstrated that 3D cultures express more extracellular matrix compared to 2D cell culture and are much more resistant to chemotherapy compared to their 2D counterpart, offering a more predictive approach for drug screening before proceeding to more expensive in vivo models and clinical trials. Methods: To be able to more efficiently screen for compounds, we adjusted our previously developed 3D PDAC 96-well cell culture model to a fully automated high-throughput screening (HTS) system on 384-well format. With this set up we screened a library of over 3000 natural compounds (TimTec_NL3000) on a PDAC cell line (Panc1), at a fixed concentration of 10 µM. As a read out we used a luminescence based cell viability assay (CellTiter Glo, Promega). The compounds found to decrease the cell viability were then validated by performing a dose-response curve in different cell lines. Results: In the initial screen we identified 13 hits which significantly decreased the cell viability of the tested 3D PDAC cell cultures compared to the untreated vehicle control. In the validation phase, 5 out of these 13 compounds, were confirmed to significantly decrease cell viability in in a dose-dependent manner in Panc1 cells. When those 5 compounds have been tested in a different cell line (HPAFII) at least 2 of them showed the ability to dramatically decrease the cell viability. In the future, we are planning to add a level of complexity to the system by inserting a stromal compartment into the 3D cultures to be able to even better resemble the in vivo PDAC situation. Conclusion: We have demonstrated that robotized high-throughput drug screening for pancreatic cancer is feasible even in organotypic 3D culture. Citation Format: Rainer L. Heuchel, Salvatore Nania, Jianping Liu, J. Matthias Löhr.{Authors}. High-throughput drug screening model using 3D cultured human pancreatic ductal adenocarcinoma cells. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B43.

FEBS Letters, Feb 25, 1991

A nuntb¢ r of transcription factors contain so.called zinc finger domains for the inlcr~ction wit... more A nuntb¢ r of transcription factors contain so.called zinc finger domains for the inlcr~ction with their cognate DNA sequence, It tins ~¢n shown that rclnOWd or the zinc io,~s complcxed in these zinc t~n~ers abrol~atcs DNA bindin~ and transcription activation, Therefore we wanted to te~ the hypothesis that the activity of tritnscriptton factors could b~ reguhtted by physolo~,ical chel.ttors of zinc, A pron~inent candid~tv,¢ rot such a chdutor is the Cys-rich protci='b thionein (apometallothionein) that is inducible by heavy metal Io=ids, and by oilier environm~nlal stimuli, Here we show with DNA binding, and in Vitro trtmscription ass;¢ys that thionein indeed can in.ctlvat¢ the zinc flnger.containin~ $pl in != reversible manner. By contrast, transcriptio, fitclor Oct-I, which binds DNA via a borneo.domain, i.~, a helix-turn-helix motif not involving zinc ions, is refntctory to thionein action, We propo~ that modulation ofintracellular thlonein concentration is used for the coordinated reguhttion of =l large subset of genes w!mSe tnm~ription depends on zinc finger proteins.

Embryonic Liver Degeneration and Increased Sensitivity Towards Heavy Metal and H2O2 in Mice Lacking the Metal-Responsive Transcription Factor MTF-1

Springer eBooks, 1999

The expression of metallothioneins, small heavy metal binding proteins, is induced by treatment o... more The expression of metallothioneins, small heavy metal binding proteins, is induced by treatment of cells with heavy metal. This induction depends on the heavy metal-responsive transcriptional activator MTF-1. To investigate the physiological function of MTF-1, we generated null mutant mice by targeted gene disruption. Embryos lacking MTF-1 die in utero at around day 14 of gestation. They show impaired development of hepatocytes and, at later stages, liver decay and generalized oedema. MTF-1-/- embryos fail to transcribe metallothionein I and II genes, and also show diminished transcripts of the gene which encodes the heavy-chain subunit of the gamma glutamylcysteine synthetase, a key enzyme for glutathione biosynthesis. Metallothionein and glutathione are involved in heavy metal homeostasis and detoxification processes, such as scavenging reactive oxygen intermediates. Accordingly, primary mouse embryo fibroblasts lacking MTF-1 show increased susceptibility to the cytotoxic effects of cadmium or hydrogen peroxide. We also note that the MTF-1 null mutant phenotype bears some similarity to those of two other regulators of cellular stress response, namely c-Jun and NF-KB (p65/RelA). Furthermore we find MTF-1 binding sites in the p65 and p50 subuntis of NF-KB, the zinc export pump ZnTl and G6PD. Thus, MTF-1 may help to control metal homeostasis and probably cellular redox state, especially during liver development (Gimes et al, 1998).

Journal of Cellular Biochemistry, 1994

Arteriosclerosis, Thrombosis, and Vascular Biology, Oct 1, 2007

Objective-Knockout studies have demonstrated crucial roles for the platelet-derived growth factor... more Objective-Knockout studies have demonstrated crucial roles for the platelet-derived growth factor-B and its cognate receptor, platelet-derived growth factor receptor-␤ (PDGFR-␤), in blood vessel maturation, that is, the coverage of newly formed vessels with mural cells/pericytes. This study describes the consequences of a constitutively activating mutation of the PDGFR-␤ (Pdgfrb D849V ) introduced into embryonic stem cells with respect to vasculogenesis/ angiogenesis in vitro and in vivo. Methods and Results-Embryonic stem cells were induced to either form teratomas in vivo or embryoid bodies, an in vitro model for mouse embryogenesis. Western blotting studies on embryoid bodies showed that expression of a single allele of the mutant Pdgfrb led to increased levels of PDGFR-␤ tyrosine phosphorylation and augmented downstream signal transduction. This was accompanied by enhanced vascular development, followed by exaggerated angiogenic sprouting with abundant pericyte coating as shown by immunohistochemistry/immunofluorescence. Pdgfrb D849V/ϩ embryoid bodies were characterized by increased expression of vascular endothelial growth factor (VEGF)-A and VEGF receptor-2; neutralizing antibodies against VEGF-A/VEGF receptor-2 blocked vasculogenesis and angiogenesis in mutant embryoid bodies. Moreover, Pdgfrb D849V/ϩ embryonic stem cell-derived teratomas in nude mice were more densely vascularized than wild-type teratomas. receptor-2, acting directly on endothelial cells and resulting in increased vessel formation.

Transcriptional Regulation by Heavy Metals, Exemplified at the Metallothionein Genes

Birkhäuser Boston eBooks, 1995

Seventeen of the thirty elements known to be essential for life are metals (Cotton and Wilkinson,... more Seventeen of the thirty elements known to be essential for life are metals (Cotton and Wilkinson, 1980). They can function as structural or catalytic components of bioorganic molecules or even as signal transducers. (Lippard, 1993). The so-called transition metals are found in the groups HIB to IIB of the periodic system. Of these, zinc (Group IIB) is the most widely used in living systems. In 1869 it was discovered that zinc is an essential trace element for higher organisms, and in 1940, it was the first trace element to be recognized as a component of an enzyme, namely carbonic anhydrase (Raulin, 1869; Keilin and Mann, 1940). To date, there are more than 300 enzymes known to require zinc for proper functioning (Vallee and Auld, 1990). Pathological zinc deficiency, due to greatly reduced intestinal zinc uptake as in the recessive, autosomal disorder Acrodermatitis enteropathica, leads to death unless treated by high oral zinc doses (Vallee and Falchuk, 1993). Among the zinc dependent enzymes several are involved in nucleic acid metabolism such as the prokaryotic and eukaryotic RNA polymerases (Vallee and Falchuk, 1993). It has been discovered only recently that zinc is also an integral constituent of proteins that regulate the activity of eukaryotic RNA polymerases.

FEBS Open Bio, Sep 7, 2019

Tamoxifen is very successfully used for the induction of Cre ERT -mediated genomic recombination ... more Tamoxifen is very successfully used for the induction of Cre ERT -mediated genomic recombination in conditional mouse models. Recent studies, however, indicated that tamoxifen might also affect the fibrotic response in several disease models following administration, both in vitro and in vivo. In order to investigate a possible effect of tamoxifen on pancreatic fibrogenesis and to evaluate an optimal treatment scheme in an experimental pancreatitis mouse model, we administered tamoxifen by oral gavage to both male and female C57BL/6J mice, and then waited for different periods of time before inducing chronic pancreatitis by cerulein. We observed a sex-specific and time dependent effect of tamoxifen on the fibrotic response as measured by collagen deposition and the number of myofibroblasts and macrophages. The findings of in vitro studies, in which cerulein was administrated with or without 4-hydroxytamoxifen to stimulate primary murine female and

Journal of Cellular and Molecular Medicine, Feb 6, 2008

Transforming growth factor-␤ (TGF-␤) signalling is induced in liver as a consequence of damage an... more Transforming growth factor-␤ (TGF-␤) signalling is induced in liver as a consequence of damage and contributes to wound healing with transient activation, whereas it mediates fibrogenesis with long-term up-regulation in chronic disease. Smad-dependent TGF-␤ effects are blunted by antagonistic Smad7, which is transcriptionally activated as an immediate early response upon initiation of TGF-␤ signalling in most cell types, thereby providing negative feedback regulation. Smad7 can be induced by other cytokines, e.g. IFN-␥, leading to a crosstalk of these signalling pathways. Here we report on a novel mouse strain, denoted S7⌬E1, with a deletion of exon I from the endogenous smad7 gene. The mice were viable and exhibited normal adult liver architecture. To obtain insight into Smad7-dependent protective effects, chronic liver damage was induced in mice by carbon tetrachloride (CCl 4 ) administration. Subsequent treatment, elevated serum liver enzymes indicated enhanced liver damage in mice lacking functional Smad7. CCl 4 -dependent Smad2 phosphorylation was pronounced in S7⌬E1 mice and accompanied by increased numbers of ␣-smooth muscle actin positive 'activated' HSCs. There was evidence for matrix accumulation, with elevated collagen deposition as assessed morphometrically in Sirius red stained tissue and confirmed with higher levels of hydroxyproline in S7⌬E1 mice. In addition, the number of CD43 positive infiltrating lymphocytes as well as of apoptotic hepatocytes was increased. Studies with primary hepatocytes from S7⌬E1 and wild-type mice indicate that in the absence of functional Smad7 protein, hepatocytes are more sensitive for TGF-␤ effects resulting in enhanced cell death. Furthermore, S7⌬E1 hepatocytes display increased oxidative stress and cell damage in response to CCl 4 , as measured by reactive oxygen species production, glutathione depletion, lactate dehydrogenase release and lipid peroxidation. Using an ALK-5 inhibitor all investigated CCl 4 effects on hepatocytes were blunted, confirming participation of TGF-␤ signalling. We conclude that Smad7 mediates a protective effect from adverse TGF-␤ signalling in damaged liver, re-iterating its negative regulatory loop on signalling.

Hepatology, Oct 1, 2007

Connective tissue growth factor (CTGF) is important for transforming growth factor-␤ (TGF-␤)-indu... more Connective tissue growth factor (CTGF) is important for transforming growth factor-␤ (TGF-␤)-induced liver fibrogenesis. Hepatic stellate cells have been recognized as its major cellular source in the liver. Here we demonstrate the induction of CTGF expression in hepatocytes of damaged livers and identify a molecular mechanism responsible for it. CTGF expression was found by immunohistochemistry in bile duct epithelial cells, hepatic stellate cells, and hepatocytes in fibrotic liver tissue from patients with chronic hepatitis B infection. Similarly, CTGF expression was induced in hepatocytes of carbon tetrachloride-treated mice. CTGF expression and secretion were detected spontaneously in a medium of hepatocytes after 3 days of culture, which was enhanced by stimulation with TGF-␤. TGF-␤induced CTGF expression was mediated through the activin receptor-like kinase 5 (ALK5)/ Smad3 pathway, whereas activin receptor-like kinase 1 activation antagonized this effect. CTGF expression in the liver tissue of TGF-␤ transgenic mice correlated with serum TGF-␤ levels. Smad7 overexpression in cultured hepatocytes abrogated TGF-␤-dependent and intrinsic CTGF expression, indicating that TGF-␤ signaling was required. In line with these data, hepatocyte-specific transgenic Smad7 reduced CTGF expression in carbon tetrachloride-treated animals, whereas in Smad7 knockout mice, it was enhanced. Furthermore, an interferon gamma treatment of patients with chronic hepatitis B virus infection induced Smad7 expression in hepatocytes, leading to decreased CTGF expression and fibrogenesis. Conclusion: Our data provide evidence for the profibrogenic activity of TGF-␤ directed to hepatocytes and mediated via the up-regulation of CTGF. We identify ALK5-dependent Smad3 signaling as the responsible pathway inducing CTGF expression, which can be hindered by an activated activin receptor-like kinase 1 pathway and completely inhibited by TGF-␤ antagonist Smad7. (HEPATOLOGY 2007;46:1257-1270.

Platelet-derived growth factor receptor-β (D849V) activating mutation promotes vascular development and angiogenic sprouting by differentiating embryonic stem cells

During development of the mammalian embryo, spatial and temporal expression of fibroblast growth ... more During development of the mammalian embryo, spatial and temporal expression of fibroblast growth factors (FGFs) and their cognate receptors are vital in the regulation of a number of patterning processes. Inappropriate or decreased expression leads to severe malformations and even embryonic death. The objectives of this thesis have been to evaluate the usefulness of differentiating embryonic stem (ES) cells as a model to study FGF and FGF receptors in endothelial and hematopoietic cell function in vitro and in vivo, and the effect of an activating mutation in the platelet-derived growth factor receptor-β (PDGFR-β) on endothelial cells and vessel formation.Aggregates of differentiating ES cells, denoted embryoid bodies, faithfully recapitulate many developmental processes. Embryoid bodies cultured in fetal calf serum spontaneously develop cardiomyocytes and endothelial cells. The endothelial cells organize into lumen-containing vessels carrying erythroblasts. Administration of FGF or vascular endothelial growth factor (VEGF)-A promotes development of specific vascular phenotypes. About 20% of endothelial cells in embryoid bodies and teratomas express FGFR-1, and these FGFR-1-expressing endothelial cells are mitogenically active in the absence of exogenous stimuli and respond to VEGF-A to the same extent as endothelial cells lacking FGFR-1 expression. FGFR-1 deficiency leads to arrest in hematopoietic differentiation, whereas endothelial cell development is enhanced. As a consequence, teratomas derived from ES cells lacking FGFR-1 expression display vessels composed of a double layer of endothelial cells. The hyperactivity of endothelial cells derived from FGFR-1-deficient ES cells is suggested to be due to hyperactivity of VEGF receptor-2, as well as to loss of negative regulators of angiogenesis, such as interleukin-4.Mutation of platelet-derived factor receptor-β (PDGFR-β) to replace D849 in the activating loop in the kinase domain with V leads to ligand-independent kinase activity, increased basal signal transduction, and enhanced expression of VEGF-A as well as VEGFR-2. As a result, endothelial cell sprouts covered with pericyte-like cells are formed in a VEGF-A/VEGFR-2 dependent manner in ES cells expressing the mutated PDGFR-β.In conclusion, embryoid bodies represent a high-quality model for the study of growth factor-regulated vascular development and sprouting angiogenesis.

Experientia, Jun 1, 1995

are normally considered to be sensitive indicators of heavy metal pollution, but it is less clear... more are normally considered to be sensitive indicators of heavy metal pollution, but it is less clear whether the MT gene can actively respond to other environmental stresses. In this study, an MT mRNA molecular sequence of 471 bp (full length) was identified in marine cultured black porgy (Acanthopagrus schlegelii), encoding 60 amino acids containing 20 cysteine residues. The MT sequence was highly homologous to that of other fish belonging to the MT superfamily type 1 family. The three dimensional structure of the deduced MT peptide was composed of two metal-binding domains capable of ligating divalent heavy metals. The MT mRNA transcripts were detected in the 11 tested tissues and the highest quantity was present in the liver. Stresses by two factors, benzo[a]pyrene (B[a]p) exposure and bacterial challenge, were evaluated on MT gene expression. The level of MT gene transcripts in the liver significantly declined 24 h post B[a]p exposure and the quantity was significantly correlated with the exposure time during a 24 h period. In contrast, MT gene expression in the liver was significantly increased 48 h post bacterial infection and the quantity was significantly correlated with the infection time during this period of 48 h. Our results indicated that MT gene expression in black porgy liver was sensitive to environmental stresses other than just the heavy metal pollution reported, suggesting that the development of a reliable biomarker for heavy metal pollution will be more complex than expected.

The EMBO Journal, 1993

Metaliothioneins (MTs) are small cysteine-rich proteins whose structure is conserved from fungi t... more Metaliothioneins (MTs) are small cysteine-rich proteins whose structure is conserved from fungi to man. MTs strongly bind heavy metals, notably zinc, copper and cadmium. Upon exposure of cells to heavy metal and other adverse treatments, MT gene transcription is strongly enhanced. Metal induction is mediated by several copies of a 15 bp consensus sequence (metal-responsive element, MRE) present in the promoter region of MT genes. We and others have demonstrated the presence of an MRE-binding factor in HeLa cell nuclear extracts. We found that this factor, termed MTF-1 (MRE-binding transcription factor) is inactivated/reactivated in vitro by zinc withdrawal/addition. Here we report that the amounts of MTF-1 -DNA complexes are elevated several- fold in zinc-treated cells, as measured by bandshift assay. We have also cloned the cDNA of mouse MTF-1, a 72.5 kDa protein. MTF-1 contains six zinc fingers and separate transcriptional activation domains with high contents of acidic and proline residues. Ectopic expression of MTF-1 in primate or rodent cells strongly enhances transcription of a reporter gene that is driven by four consensus MREd sites, or by the complete mouse MT-I promoter, even at normal zinc levels.

Significance of CEA Antibody Affinity for Recognition of Cancer by Plasma Determinations

Protides of the Biological Fluids, 1985

Abstract It has been stated that high affinity and epitope specificity of CEA antibodies are sign... more Abstract It has been stated that high affinity and epitope specificity of CEA antibodies are significant for discrimination of cancer patients from healthy controls. In order to find out whether this is true, antisera with different affinity and presumptive epitope specificity, conjugated to solid beads, were incubated with plasma of patients with cancer of the lung, intestine or urinary bladder. The test was performed by cross-incubation and sequential incubations. After each incubation step theremnant CEA concentrations were determined. Polyclonal antisera left about 50% CEA in plasma after each incubation, whereas incubation with monoclonal antibodies left no detectable CEA. In addition to these experiments, discrimination of different groups (patients and healthy individuals) by CEA plasma determination was analysed by means of ROC-curves. It turned out that the ROC-curves of the monoclonal and polyclonal tests were practically at the same level. It is, therefore, concluded that the affinity of antisera plays a minor role in cancer discrimination.

Nucleic Acids Research, 1994

Metallothioneins (MTs) are small cysteine-rich proteins that bind heavy metal ions such as zinc, ... more Metallothioneins (MTs) are small cysteine-rich proteins that bind heavy metal ions such as zinc, cadmium and copper with high affinity, and have been functionally implicated in heavy metal detoxification and radical scavenging. Transcription of metallothioneins genes is induced by exposure of cells to heavy metals. This induction is mediated by metal-responsive promoter elements (MREs). We have previously cloned the cDNA of an MRE-binding transcription factor (MTF-1) from the mouse. Here we present the human cDNA equivalent of this metal-regulatory factor. Human MTF-1 is a protein of 753 amino acids with 93% amino acid sequence identity to mouse MTF-1 and has an extension of 78 amino acids at the C-terminus without counterpart in the mouse. The factors of both species have the same overall structure including six zinc fingers in the DNA binding domain. We have physically mapped the human MTF-1 gene to human chromosome 1 where it localizes to the short arm in the region 1p32-34, most likely 1p33. Both human and mouse MTF-1 when produced in transfected mammalian cells strongly bind to a consensus MRE of metallothionein promoters. However, human MTF-1 is more effective than the mouse MTF-1 clone in mediating zinc-induced transcription.

Pancreatology, Jun 1, 2019

database. The meta-analysis of the relation of marginal length on survival was performed with the... more database. The meta-analysis of the relation of marginal length on survival was performed with the inverse variance method in RevMan. Results: There were five studies (902 patients) reporting median survival (MS) for RM 0 mm resection (12.6-23.4), seven studies (1167 patients) reporting MS for RM <1 mm, two studies (259 patients) reporting MS for RM <1.5 mm and three studies (319 patients) reporting MS for RM <2 mm. A total of five studies comprising 1275 patients reported multivariate hazard rations for RM 1 mm vs. RM <1 mm. A RM over 1 mm reduced risk for death compared with patients with RM < 1 mm (HR 1.32 and 95% confidence interval from 1.03 to 1.68, p¼0.03). Three studies reported multivariate HRs for RM 1 mm (929 patients) vs. RM 0 mm (701 patients). The HRs ranged from 0.73-1.31. Two studies reported multivariate HRs for RM >2 mm (63 patients) vs. <2 mm (296 patients). The HRs ranged from 1.2-2.66. Conclusion: RM 1 mm or more leads to longer survival than RM <1 mm. However, not enough evidence exists to analyse benefits of wider RMs.